Taurine reduces high glucose induced leukocyte–endothelial interactions via down-regulation of ICAM-1

Taurine is a semiessential amino acid and naturally occurring antioxidant. One of its main roles is to protect tissues against attack by chlorinated oxidants particularly hypochlorous acid (HOCl). It is found in high concentrations in neutrophils and previous studies showed it possesses potent antimicrobial properties and attenuates high glucose induced endothelial cell apoptosis. In humans taurine has been shown to up-regulate constitutive nitric oxide synthase (cNOS), a known cytoprotector.
No reported studies to date have looked at the possible therapeutic role of taurine in preventing diabetic endothelial dysfunction. We therefore hypothesised that taurine would attenuate the microvascular changes associated with hyperglycaemia in an animal model through alteration of leucocyte–endothelial interactions.
Male Sprague Dawley rats were randomised into control, hyperglycaemia, and taurine + hyperglycaemia groups. Taurine was gavaged (200 mg/kg) for 5 d prior to the experiment. Hyperglycaemia was established by intravenous infusion of 50% glucose. Blood glucose reached a steady state of 3 times baseline at 30 min. Using intravital microscopy leukocyte rolling, adhesion and transendothelial migration was determined in mesenteric postcapillary venules for 3 h. Intracellular adhesion molecule-1 (ICAM-1) was immunohistochemically graded using a scoring system to determine the expression in mesenteric tissue.
Taurine pretreatment significantly attenuates leukocyte-endothelial adhesion and transendothelial migration following acute hyperglycaemia but not leukocyte rolling velocity. The mechanism by which taurine protects against these effects is in part by inhibition of ICAM-1 expression .  

      

2.

Astrocyte–endothelial interactions and blood–brain barrier permeability          下载免费PDF全文

NJ Abbott 《Journal of anatomy》2002,200(5):527

     

3.

Loratadine reduces the expression of ICAM-1   总被引：1，自引：0，他引：1  

G. Ciprandi  A. Catrullo  P. Cerqueti  M. Tosca  N. Fiorino  G. W. Canonica 《Allergy》1998,53(5):545-546

     

4.

Hydroxyethyl starch reduces high stretch ventilation-augmented lung injury via vascular endothelial growth factor     

Li-Fu Li  Chung-Chi Huang  Yung-Yang Liu  Horng-Chyuan Lin  Kuo-Chin Kao  Cheng-Ta Yang  Shuen-Kuei Liao 《Translational Research, The Journal of Laboratory and Clinical Medicine》2011,157(5):293-305

     

5.

HIV-1 Nef通过Erk/Mapk途径调节内皮细胞黏附分子ICAM-1的表达     

杨凡  刘朝奇  覃晓琳  史继静 《细胞与分子免疫学杂志》2010,26(1)

目的:研究HIV-1Nef基因对内皮细胞ECV304细胞ICAM-1表达的影响及其信号途径。方法:选用Nef基因稳定表达细胞株ECV304-Nef和对照细胞株ECV304pcDNA3.1(+),应用Westernblot分析ECV304-Nef细胞ERK的磷酸化水平,利用ERK磷酸化抑制剂通过Westernblot、流式细胞术分析ECV304-Nef细胞ICAM-1的表达水平与信号分子ERK磷酸化的相关性。结果:Westernblot显示ECV304-Nef细胞ICAM-1和p-ERK蛋白的表达水平均高于对照组;流式细胞术检测结果表明ECV304-Nef细胞和对照组ICAM-1阳性细胞百分率分别为(35.3±2.2)%和(12.5±0.8)%(P0.01)。加入p-ERK抑制剂PD98059后,ECV304-Nef细胞p-ERK水平被显著抑制,ICAM-1降至对照组水平,ECV304-Nef细胞和对照组细胞ICAM-1阳性细胞百分率分别为(11.4±1.1)%和(10.4±1.5)%(P0.05)。结论:HIV-1Nef基因上调血管内皮细胞细胞黏附分子ICAM-1的表达与ERK信号分子磷酸化有关,为HIV-1感染的致病机制及临床治疗提供实验基础。     

6.

Hyaluronan inhibits cytokine production by lipopolysaccharide-stimulated U937 macrophages through down-regulation of NF-κB via ICAM-1     

T. Yasuda 《Inflammation research》2007,56(6):246-253

Objective: This study investigated the inhibitory mechanism of hyaluronan (HA) on lipopolysaccharide (LPS)-stimulated production of proinflammatory cytokines in U937 macrophages. Methods: HA was added to U937 macrophage cultures in the presence of LPS, with or without pretreatment with anti-intercellular adhesion molecule-1 (ICAM-1) antibody. Secreted levels of tumor necrosis factor α (TNFα), interleukin (IL)-1β, and IL-6 were determined by enzyme-linked immunosorbent assay. The phosphorylation of nuclear factor (NF)-κB, IκBα, and mitogen-activated protein kinases (MAPKs) was analyzed by immunoblotting. Results: LPS stimulated production of TNFα, IL-1β, and IL-6. In contrast to 800 kDa HA, 2700 kDa HA at 1 mg/ml inhibited LPS-induced cytokine production. Anti-ICAM-1 antibody blocked the effects of HA on the LPS actions on U937 cells. LPS activated NF-κB and MAPK pathways, whereas HA down-regulated p65 NF-κB and IκBα phosphorylation by LPS without affecting MAPKs. Inhibition studies revealed the requirement of NF-κB for LPS-stimulated cytokine production. Anti-ICAM-1 antibody reversed the inhibitory effects of HA on phosphorylation of p65 NF-κB and IκBα. Conclusion: HA of intrinsic molecular weight suppresses LPS-stimulated production of proinflammatory cytokines via ICAM-1 through down-regulation of NF-κB and IκB. Exogenous HA injected into arthritic joints could act as an anti-NF-κB agent by the mechanism demonstrated in the present study. Received 23 September 2006; returned for revision 12 October 2006; accepted J. Di Battista 18 December 2006     

7.

Vaspin alleviates dysfunction of endothelial progenitor cells induced by high glucose via PI3K/Akt/eNOS pathway     

Ning Sun  Hui Wang  Lin Wang 《International journal of clinical and experimental pathology》2015,8(1):482-489

Improving the dysfunction of endothelial progenitor cell (EPCs) in patients with diabetes mellitus is important for preventing vascular complication. Vaspin, an adipocytokine, has the anti-atherogenic properties rely on its positive effect on nitric oxide (NO) bioavailability. We hypothesis that vaspin may ameliorate high glucose induced dysfunction of EPCs. In rat bone morrow derived EPCs, glucose treatment results in a decrease in the proliferation and migration capacity in a dose dependent manner. These detrimental effects can be alleviated by vaspin. Furthermore, vaspin increased the production of NO and the effect of vaspin on EPCs can be diminished partly by the eNOS inhibitor (_L-NAME). We assessed total eNOS protein expression and Ser¹¹⁷⁷-phospho-eNOS expression and found that vaspin not only induced eNOS protein expression but also up regulate the eNOS activation. Subsequently, we investigated protein kinase B (Akt) activation in the presence and absence of phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor (LY-2940002). Vaspin increased total Akt and Ser⁴⁷³-phospho-Akt expression and these effects can be blocked by LY-2940002. The results of our study indicate a novel effect of vaspin to regulate eNOS expression and function in EPCs via a PI3K/Akt/eNOS pathway; vaspin may have a protective effect in patients with diabetes to prevent the occurrence of vascular complication.     

8.

Production of soluble ICAM-1 from human endothelial cells induced by IL-1β and TNF-α     

Michi Hashimoto  Masao Shingu  Ichiko Ezaki  Masashi Nobunaga  Makoto Minamihara  Keiji Kato  Hisao Sumioki 《Inflammation》1994,18(2):163-173

The present study was designed to establish the effects of cytokines on soluble ICAM-1 (sICAM-1) production by human endothelial cells (EC) and ICAM-1 expression on these cells and the effects of purified sICAM-1 on lympho-cyte-EC adhesion. Expression of ICAM-1 and production of sICAM-1 were measured by a specific ELISA method. ICAM-1 expression was enhanced by IL-1β, TNF-α, and most effectively by IFN-γ. IL-4, IL-6, M-CSF, or GM-CSF showed no effects on ICAM-1 expression. IL-4 (100 units/ml) or IL-6(100 units/ml) abolished the enhancing effect of IL-1β, while TNF-α (1, 10, 100 units/ml) synergized with IL-1β to promote ICAM-1 expression in EC. In contrast with the transient increase of cell-associated ICAM-1 expression after activiation by IL-1β, which peaked 40 h poststimulation and declined thereafter, sICAM-1 continued to accumulate in culture supernatants even after 48 h poststimulation in IL-1β-stimulated EC. IL-1β treatment resulted in an increase in adhesion. sICAM-1, purified from cell-free supernatants obtained after a 48-h culture of EC in IL-1β by affinity chromatography using monoclonal ICAM-1 antibody coupled to Sepharose beads, significantly inhibited lymphocyte EC adhesion. Preincubation of lymphocytes with conditioned medium of EC cultured with 100 units/ml IL-1β for 48 h, which contained a considerable amount of sICAM-1, resulted in a significant inhibition of lymphocyte adhesion to IL-1β-stimulated EC. These results suggest that there is a cumulative increase in sICAM-1 concentration in the vicinity of cytokine-stimulated EC and that this sICAM-1 modulates ICAM-1-mediated cell to cell interaction.     

9.

Hepatopoietin Cn reduces ethanol‐induced hepatoxicity via sphingosine kinase 1 and sphingosine 1‐phosphate receptors     

Yang Liu  Saiyan Saiyan  Tong‐Yi Men  Hui‐Ying Gao  Chuan Wen  Yong Liu  Xu Zhou  Chu‐Tse Wu  Li‐Sheng Wang  Chun‐Ping Cui 《The Journal of pathology》2013,230(4):365-376

The hepatic growth factor hepatopoietin Cn (HPPCn) prevents liver injury induced by carbon tetrachloride in rats. Sphingosine 1‐phosphate (S1P) is a bioactive sphingolipid produced by sphingosine kinase (SphK). S1P and S1P receptors (S1PRs) are involved in liver fibrogenesis and oxidative injury. This work sought to understand the mechanism by which SphK/S1P/S1PRs are involved in the protective effects of HPPCn on ethanol‐induced liver injury and fibrosis. Transgenic mice with liver‐specific overexpression of HPPCn (HPPCn^liver+/+) were generated. Two ethanol feeding protocols were used to assess the protective effect of HPPCn on acute and chronic liver injury in mice. Specific inhibitors of S1PR1, S1PR2 and S1PR3 and siRNA were used to examine the roles of S1PRs in hepatic stellate cell (HSC) activation and hepatocyte apoptosis. Increased HPPCn expression in transgenic mice attenuated fibrosis induced by ethanol and carbon tetrachloride (CCl₄). Treatment with recombinant human HPPCn prevented human hepatocyte apoptosis and HSC activation. JTE‐013 or S1PR2‐siRNA attenuated the effect of HPPCn on HSC activation induced by tumour necrosis factor‐α (TNF‐α). Consistent with the effect of N,N‐dimethylsphingosine (DMS), suramin or S1PR3‐siRNA treatment blocked HPPCn‐induced Erk1/2 phosphorylation in human hepatocytes. This study demonstrated that HPPCn attenuated oxidative injury and fibrosis induced by ethanol feeding and that the SphK1/S1P/S1PRs signalling pathway contributes to the protective effect of HPPCn on hepatocyte apoptosis and HSC activation. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

10.

辛伐他汀抑制高糖诱导人脐静脉内皮细胞的凋亡     

徐茂凤  李永杰  王婉丽 《中国组织工程研究》2011,15(41):7726-7729

背景：他汀类药物对血管内皮细胞的凋亡是否有影响目前尚不明确。 目的：探讨辛伐他汀对高糖诱导的人脐静脉内皮细胞凋亡的影响。 方法：用DMEM细胞培养液培养人脐静脉内皮细胞,将细胞分成空白对照组、高糖组和高糖+辛伐他汀组,用四甲基偶氮唑蓝比色法测定人脐静脉内皮细胞的存活率,流式细胞仪和Western blot分别检测细胞早期凋亡率及P53蛋白表达。 结果与结论：高糖组及高糖+辛伐他汀组细胞增殖率较空白对照组明显降低(P < 0.01),而高糖组细胞增殖率较高糖+辛伐他汀组亦降低(P < 0.01);高糖组P53蛋白表达量及凋亡率较空白对照组及高糖+辛伐他汀组明显增加(P < 0.01),高糖+辛伐他汀组P53蛋白表达及凋亡率亦明显高于空白对照组(P< 0.01)。表明高糖可通过促进促凋亡蛋白P53的表达进而促进人脐静脉内皮细胞的凋亡,而辛伐他汀可抑制此作用。     

11.

NicE-C efficiently reveals open chromatin–associated chromosome interactions at high resolution     

Zhengyu Luo  Ran Zhang  Tengfei Hu  Yuting Zhu  Yueming Wu  Wenfei Li  Zhi Zhang  Xuebiao Yao  Haiyi Liang  Xiaoyuan Song 《Genome research》2022,32(3):534

Enhancer–promoter communication is known to regulate spatiotemporal dynamics of gene expression. Several methods are available to capture enhancer–promoter interactions, but they either require large amounts of starting materials and are costly, or provide a relative low resolution in chromatin contact maps. Here, we present nicking enzyme-assisted open chromatin interaction capture (NicE-C), a method that leverages nicking enzyme–mediated open chromatin profiling and chromosome conformation capture to enable robust and cost-effective detection of open chromatin interactions at high resolution, especially enhancer–promoter interactions. Using TNF stimulation and mouse kidney aging as models, we applied NicE-C to reveal characteristics of dynamic enhancer–promoter interactions.

Enhancers and promoters are key cis-regulatory elements located at open chromatin regions, which can be detected by open chromatin profiling methods, such as FAIRE-seq (Giresi and Lieb 2009), DNase-seq (Song and Crawford 2010), ATAC-seq (Buenrostro et al. 2013), and nicking enzyme-assisted sequencing (NicE-seq) (Ponnaluri et al. 2017). Long-range chromatin interactions between enhancers and promoters can control cell type– and condition-specific gene expression, which play important roles in diverse biological processes, including neural development (Bonev et al. 2017), cell differentiation (Isoda et al. 2017), and etiopathology of diseases (Hua et al. 2018). The development of chromosome conformation capture (3C)-based methods, including Hi-C (Lieberman-Aiden et al. 2009; Rao et al. 2014) and Micro-C (Hsieh et al. 2020; Krietenstein et al. 2020), has greatly promoted our understanding of high-order chromatin organization and enhancer–promoter interactions. However, these genome-wide methods need extremely deep sequencing to provide sufficient spatial resolution to identify enhancer–promoter interactions.To enrich cis-regulatory elements–associated chromatin interactions, especially between enhancers and promoters, methods such as Capture Hi-C (Mifsud et al. 2015), OCENA-C (Li et al. 2018), and Trac-looping (Lai et al. 2018) have been developed. Although Capture Hi-C can identify genome-wide chromatin interactions associated with both active and inactive promoters, it requires a costly, species-specific predesigned biotinylated RNA bait library to target known promoters. OCEAN-C and Trac-looping are probe-free methods that use open chromatin features. OCEAN-C combines Hi-C with FAIRE-seq to select open chromatin interactions, whereas Trac-looping uses transposase enzyme and a bivalent ME linker. Although potentially very promising, Trac-looping requires about 100 million (M) cells per experiment, yet identifies only a relatively low fraction of long-range (>20 kb) cis chromatin interactions. Limitation of OCEAN-C falls in its relative lower resolution and lower enrichment of open chromatin regions. Accordingly, an improved method for capturing open chromatin interactions would be highly desirable. To address these challenges, we developed a new probe-free method named nicking enzyme-assisted open chromatin interaction capture (NicE-C) for capturing open chromatin interactions.     

12.

LY333531对高糖所致心肌微血管内皮细胞通透性上调的拮抗作用   总被引：3，自引：0，他引：3  

尹志勇  魏丽萍  郝媛媛  张荣庆  李聪叶  王海昌 《细胞与分子免疫学杂志》2009,25(4)

目的:观察LY333531对高糖所致心肌微血管内皮细胞通透性上调的拮抗作用,并探讨其机制.方法:将分离培养并鉴定后的大鼠心肌微血管内皮细胞分为正常组,高糖组(葡萄糖浓度25 mmol/L),高糖(葡萄糖浓度25 mmol/L)+LY333531(10 μmol/L)组,高糖(葡萄糖浓度25 mmol/L)+生理盐水组,通过离体血管通透性检测试剂盒检测单层细胞通透性,TUNEL法检测细胞凋亡,免疫荧光和Western blot法检测PKCβⅡ的表达.结果:与正常组比较,高糖组单层细胞通透性明显上调(400.0±20.00 vs 223.3±25.17;P<0.01),凋亡率升高(55.00%±5.000% vs 2.333%±1.155%;P<0.01),PKCβⅡ的表达增加(0.4767±0.07506 vs 0.1733±0.02082;P<0.01);LY333531可明显降低PKCβⅡ的表达(0.2800±0.070 vs 0.4767±0.07506;P<0.01)并拮抗上述病理性通透功能上调(360±17.32 vs 400.0±20.00;P<0.05)和凋亡率升高(25.00%±5.000% vs 55.00%±5.000%;P<0.01),而生理盐水对细胞通透性、凋亡率和PKCβⅡ表达的影响不明显.结论:高糖损害心肌微血管内皮细胞的通透功能,LY333531可拮抗此损伤作用.     

13.

Soluble ICAM-1 and VCAM-1 as markers of endothelial activation   总被引：1，自引：0，他引：1  

Videm V  Albrigtsen M 《Scandinavian journal of immunology》2008,67(5):523-531

Activated endothelium releases the soluble adhesion molecules vascular cell adhesion molecule-1 (sVCAM-1) and intercellular adhesion molecule-1 (sICAM-1). Measurement of fluid-phase adhesion molecules is therefore used to quantify endothelial activation, but it is unclear which is the better marker. The aims of the study were to compare the relationships between mRNA, surface and total expression and released VCAM-1 and ICAM-1 in endothelial cell cultures during activation, and to compare human umbilical vein endothelial cells (HUVEC) with the microvascular cell line HMEC-1. sVCAM-1 better represented mRNA and surface expression changes in HUVEC undergoing endotoxin stimulation than did sICAM-1. Very little VCAM-1 was released from endotoxin-stimulated HMEC-1, and sICAM-1 seemed a better activation marker for these cells. During incubation of HUVEC in media with glucose concentrations of 5.6, 10.6 or 20.6 m m , VCAM-1 was released to the media in a dose-dependent way without changes in surface expression. ICAM-1 was not influenced by the glucose concentration. There are situations when VCAM-1 concentrations in the media do not mirror the surface expression on HUVEC in culture, indicating that measurements of soluble adhesion molecules may not necessarily be representative of the conditions on the cell surface. Endothelium from different locations showed varying responses with respect to VCAM-1 and ICAM-1 liberation upon endotoxin stimulation. Thus, both sVCAM-1 and sICAM-1 should be quantified in clinical studies of endothelial activation until their characteristics are better clarified.     

14.

Nrf2 activation via Keap1 deletion or sulforaphane treatment reduces Ova‐induced sinonasal inflammation     

Nyall R. London  Anuj Tharakan  Michelle Mendiola  Mengfei Chen  Alex Dobzanski  Thomas E. Sussan  Michael Zaykaner  Andrew H. Han  Andrew P. Lane  Venkataramana Sidhaye  Shyam Biswal  Murugappan Ramanathan 《Allergy》2019,74(9):1780-1783

     

15.

肥胖抑制素减轻高糖致大鼠INS1细胞的凋亡     

徐敏  王威  姜方旭  乔虹  张玲 《基础医学与临床》2018,(1):7-12

目的探讨肥胖抑制素(OB)减轻高糖致大鼠胰岛细胞系INS1细胞的凋亡作用。方法 INS1细胞在不同糖浓度环境下培养24 h,用MTT法检测INS1细胞存活率和增殖;Hoechst33258细胞核染色法检测细胞核形态学;酶标法检测caspase-3活性及OB的保护作用是否涉及了PI3K;real-time PCR检测FOXO1、SREBP1c、Bax和PDX-1mRNA的表达。结果在高糖条件下,OB促进INS1细胞的增殖,在100 nmol/L浓度时能最大程度促进INS1细胞增殖(与对照组、高糖组相比,P<0.01)。OB能减轻高糖诱导的凋亡(P<0.01);在高糖组,FOXO1、SREBP1c和Bax基因表达较对照组增多,PDX-1表达减少;反之在OB干预的高糖组;FOXO1、SREBP1c和Bax表达减少,PDX-1表达增多。结论 OB能够减轻高糖致大鼠INS1细胞的损伤作用。     

16.

MEK1/2在脂多糖诱导人脐静脉内皮细胞ICAM-1表达中的作用          下载免费PDF全文

黄起壬  陈和平  刘丹  曾国华  何明 《中国病理生理杂志》2005,21(12):2320-2323

目的：探讨MEK1/2在脂多糖诱导人脐静脉内皮细胞（HUVECs）ICAM-1表达中的作用。 方法： 用不同浓度LPS或LPS加MEK1/2特异性抑制剂PD98059和HUVECs孵育不同时间后，分别采用RT-PCR和Western blotting检测ICAM-1 mRNA和蛋白的表达。 结果： LPS呈时间-浓度依赖性地上调HUVECs ICAM-1 mRNA和蛋白的表达。LPS预处理后2 h，HUVECs ICAM-1 mRNA和蛋白的表达即开始升高，LPS(100 μg·L-1)作用后6 h,ICAM-1 mRNA和蛋白的表达基本达到高峰；PD98059(10 μg·L-1)可显著抑制LPS(100 μg·L-1)诱导6 h的ICAM-1 mRNA和蛋白的表达，抑制率分别为54.4%和44.9%（P<0.01 vs LPS）。 结论： 调控MEK1/2通路可能为内毒素休克诱导血管内皮损伤的防治提供新的策略。     

17.

Selective eosinophil transendothelial migration triggered by eotaxin via modulation of Mac-1/ICAM-1 and VLA-4/VCAM-1 interactions   总被引：7，自引：1，他引：6  

Jia GQ  Gonzalo JA  Hidalgo A  Wagner D  Cybulsky M  Gutierrez-Ramos JC 《International immunology》1999,11(1):1-10

We have recently cloned eotaxin, a highly efficacious eosinophilicchemokine involved in the development of lung eosinophilia duringallergic inflammatory reactions. To understand more preciselyhow eotaxin facilitates the specific migration of eosinophils,we have studied which adhesion receptors are essential for eotaxinaction both in vivo and in vitro. Experiments using mice geneticallydeficient in adhesion receptors demonstrated that moleculespreviously reported to be involved in both leukocyte tethering/rolling(P-selectin and E-selectin) and in sticking/transmigration (ICAM-1and VCAM-1) are required for eotaxin action in vivo. To furtherelucidate the mechanism(s) involved in this process, we haveused an in vitro transendothelial chemotaxis model. mAb neutralizationstudies performed in this system suggest that the integrinsMac-1 (CD11b/18), VLA-4 (₄ß₁) and LFA-1 (CD11a/18) areinvolved in the transendothelial chemotaxis of eosinophils toeotaxin. Accordingly, the expression of these integrins on eosinophilsis elevated by direct action of this chemokine in a concentration-dependentmanner. Taken together, our results suggest that eotaxin-inducedeosinophil transendothelial migration in vivo and in vitro relieson Mac-1/ICAM-1 and VLA-4/VCAM-1 interactions, the latter onesbecoming more relevant at later time points of the eotaxin-inducedrecruitment process.     

18.

Corrigendum: NicE-C efficiently reveals open chromatin–associated chromosome interactions at high resolution     

Zhengyu Luo  Ran Zhang  Tengfei Hu  Yuting Zhu  Yueming Wu  Wenfei Li  Zhi Zhang  Xuebiao Yao  Haiyi Liang  Xiaoyuan Song 《Genome research》2022,32(6):12292

     

19.

红景天苷上调HIF-1α减轻高糖诱导的大鼠肾小球内皮细胞损伤          下载免费PDF全文

谢瑞燕  方雪玲  Hamze I. RAGE  崔彤霞  朱伟平 《中国病理生理杂志》2019,35(2):237-242

目的:观察体外不同浓度葡萄糖对大鼠肾小球内皮细胞(rRGECs)表达低氧诱导因子1α(HIF-1α)、血管内皮生长因子A(VEGFA)及血管内皮钙黏素(VE-cadherin)的影响,探讨红景天苷减轻高糖诱导rRGECs损伤的作用及可能相关机制。方法:体外培养rRGECs,分为正常糖组、高糖(20、30和50 mmol/L)组、高渗组及红景天苷+高糖组。采用MTT法检测rRGECs的活力;RT-qPCR法检测rRGECs内HIF-1α、VEGFA及VE-cadherin的mRNA表达;Western blot法检测rRGECs内HIF-1α蛋白的表达。结果:与正常糖组相比,培养24 h后,高糖(20mmol/L)组rRGECs HIF-1α的mRNA及蛋白表达均上调(P 0.05);培养120 h后,高糖组HIF-1αmRNA表达下调(P 0.05)。与正常糖组相比,培养24 h和120 h后,高糖组rRGECs内VE-cadherin的mRNA表达下调(P 0.05)。与正常糖组相比,培养24 h后,高糖组rRGECs内VEGFA的mRNA表达上调(P 0.05);培养120 h后,高糖组rRGECs内VEGFA的mRNA表达下调(P 0.05)。与正常糖组相比,培养24 h和120 h后,高渗组rRGECs内HIF-1α、VE-cadherin及VEGFA的mRNA表达无变化。与高糖组相比,培养24 h后,红景天苷(50μmol/L)组rRGECs的活力增加(P 0.01),且细胞内HIF-1α和VE-cadherin的mRNA及HIF-1α蛋白表达均上调(P 0.05)。结论:体外高糖培养能影响rRGECs表达HIF-1α,可能与细胞活力、葡萄糖的浓度、作用时间及HIF/VEGF通路有关。红景天苷能减轻高糖诱导的rRGECs损伤,其机制可能与增加rRGECs内HIF-1α的表达有关。     

20.

Snail1 siRNA 对高糖诱导肾小管上皮细胞表型转变的影响          下载免费PDF全文

方开云  石明隽  肖瑛  桂华珍  郭兵  张国忠 《中国病理生理杂志》2009,25(12):2424-2429

目的： 观察Snail1 siRNA对高糖诱导的肾小管上皮细胞向间充质细胞转变（TEMT）的影响。方法: 原代培养肾小管上皮细胞分为5组：（1）对照组（含糖5.5 mmol/L）;（2）高糖组（含糖25 mmol/L）;（3）Snail1 siRNA处理组,转染Snail1 siRNA,6 h后更换为高糖（含糖25 mmol/L）培养;（4）control siRNA处理组,转染control siRNA作为siRNA阴性对照,6 h后换为高糖（含糖25 mmol/L）培养;（5）高渗组（含D-manntio19.5 mmol/L）;72 h后收集细胞,用Western blotting和半定量RT-PCR检测Snail1、TGF-β1、α-平滑肌肌动蛋白（α-SMA）、vimentin和E-cadherin蛋白和mRNA表达。结果: 与高糖组比较,肾小管上皮细胞转染Snail1 siRNA后,Snail1 mRNA和蛋白表达水平分别下降62%和68%（P<0.01）。同时,Snail1 siRNA处理组α-SMA和vimentin蛋白和mRNA表达显著下调（P<0.01）,而E-cadherin蛋白和mRNA表达显著上调（P<0.01）。结论: Snail1参与了高糖诱导TEMT的调节。     

HIV-1 Nef通过Erk/Mapk途径调节内皮细胞黏附分子ICAM-1的表达

目的:研究HIV-1Nef基因对内皮细胞ECV304细胞ICAM-1表达的影响及其信号途径。方法:选用Nef基因稳定表达细胞株ECV304-Nef和对照细胞株ECV304pcDNA3.1(+),应用Westernblot分析ECV304-Nef细胞ERK的磷酸化水平,利用ERK磷酸化抑制剂通过Westernblot、流式细胞术分析ECV304-Nef细胞ICAM-1的表达水平与信号分子ERK磷酸化的相关性。结果:Westernblot显示ECV304-Nef细胞ICAM-1和p-ERK蛋白的表达水平均高于对照组;流式细胞术检测结果表明ECV304-Nef细胞和对照组ICAM-1阳性细胞百分率分别为(35.3±2.2)%和(12.5±0.8)%(P0.01)。加入p-ERK抑制剂PD98059后,ECV304-Nef细胞p-ERK水平被显著抑制,ICAM-1降至对照组水平,ECV304-Nef细胞和对照组细胞ICAM-1阳性细胞百分率分别为(11.4±1.1)%和(10.4±1.5)%(P0.05)。结论:HIV-1Nef基因上调血管内皮细胞细胞黏附分子ICAM-1的表达与ERK信号分子磷酸化有关,为HIV-1感染的致病机制及临床治疗提供实验基础。     

Hyaluronan inhibits cytokine production by lipopolysaccharide-stimulated U937 macrophages through down-regulation of NF-κB via ICAM-1

Objective: This study investigated the inhibitory mechanism of hyaluronan (HA) on lipopolysaccharide (LPS)-stimulated production of proinflammatory cytokines in U937 macrophages. Methods: HA was added to U937 macrophage cultures in the presence of LPS, with or without pretreatment with anti-intercellular adhesion molecule-1 (ICAM-1) antibody. Secreted levels of tumor necrosis factor α (TNFα), interleukin (IL)-1β, and IL-6 were determined by enzyme-linked immunosorbent assay. The phosphorylation of nuclear factor (NF)-κB, IκBα, and mitogen-activated protein kinases (MAPKs) was analyzed by immunoblotting. Results: LPS stimulated production of TNFα, IL-1β, and IL-6. In contrast to 800 kDa HA, 2700 kDa HA at 1 mg/ml inhibited LPS-induced cytokine production. Anti-ICAM-1 antibody blocked the effects of HA on the LPS actions on U937 cells. LPS activated NF-κB and MAPK pathways, whereas HA down-regulated p65 NF-κB and IκBα phosphorylation by LPS without affecting MAPKs. Inhibition studies revealed the requirement of NF-κB for LPS-stimulated cytokine production. Anti-ICAM-1 antibody reversed the inhibitory effects of HA on phosphorylation of p65 NF-κB and IκBα. Conclusion: HA of intrinsic molecular weight suppresses LPS-stimulated production of proinflammatory cytokines via ICAM-1 through down-regulation of NF-κB and IκB. Exogenous HA injected into arthritic joints could act as an anti-NF-κB agent by the mechanism demonstrated in the present study. Received 23 September 2006; returned for revision 12 October 2006; accepted J. Di Battista 18 December 2006     

Vaspin alleviates dysfunction of endothelial progenitor cells induced by high glucose via PI3K/Akt/eNOS pathway

Improving the dysfunction of endothelial progenitor cell (EPCs) in patients with diabetes mellitus is important for preventing vascular complication. Vaspin, an adipocytokine, has the anti-atherogenic properties rely on its positive effect on nitric oxide (NO) bioavailability. We hypothesis that vaspin may ameliorate high glucose induced dysfunction of EPCs. In rat bone morrow derived EPCs, glucose treatment results in a decrease in the proliferation and migration capacity in a dose dependent manner. These detrimental effects can be alleviated by vaspin. Furthermore, vaspin increased the production of NO and the effect of vaspin on EPCs can be diminished partly by the eNOS inhibitor (_L-NAME). We assessed total eNOS protein expression and Ser¹¹⁷⁷-phospho-eNOS expression and found that vaspin not only induced eNOS protein expression but also up regulate the eNOS activation. Subsequently, we investigated protein kinase B (Akt) activation in the presence and absence of phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor (LY-2940002). Vaspin increased total Akt and Ser⁴⁷³-phospho-Akt expression and these effects can be blocked by LY-2940002. The results of our study indicate a novel effect of vaspin to regulate eNOS expression and function in EPCs via a PI3K/Akt/eNOS pathway; vaspin may have a protective effect in patients with diabetes to prevent the occurrence of vascular complication.     

Production of soluble ICAM-1 from human endothelial cells induced by IL-1β and TNF-α

The present study was designed to establish the effects of cytokines on soluble ICAM-1 (sICAM-1) production by human endothelial cells (EC) and ICAM-1 expression on these cells and the effects of purified sICAM-1 on lympho-cyte-EC adhesion. Expression of ICAM-1 and production of sICAM-1 were measured by a specific ELISA method. ICAM-1 expression was enhanced by IL-1β, TNF-α, and most effectively by IFN-γ. IL-4, IL-6, M-CSF, or GM-CSF showed no effects on ICAM-1 expression. IL-4 (100 units/ml) or IL-6(100 units/ml) abolished the enhancing effect of IL-1β, while TNF-α (1, 10, 100 units/ml) synergized with IL-1β to promote ICAM-1 expression in EC. In contrast with the transient increase of cell-associated ICAM-1 expression after activiation by IL-1β, which peaked 40 h poststimulation and declined thereafter, sICAM-1 continued to accumulate in culture supernatants even after 48 h poststimulation in IL-1β-stimulated EC. IL-1β treatment resulted in an increase in adhesion. sICAM-1, purified from cell-free supernatants obtained after a 48-h culture of EC in IL-1β by affinity chromatography using monoclonal ICAM-1 antibody coupled to Sepharose beads, significantly inhibited lymphocyte EC adhesion. Preincubation of lymphocytes with conditioned medium of EC cultured with 100 units/ml IL-1β for 48 h, which contained a considerable amount of sICAM-1, resulted in a significant inhibition of lymphocyte adhesion to IL-1β-stimulated EC. These results suggest that there is a cumulative increase in sICAM-1 concentration in the vicinity of cytokine-stimulated EC and that this sICAM-1 modulates ICAM-1-mediated cell to cell interaction.     

Hepatopoietin Cn reduces ethanol‐induced hepatoxicity via sphingosine kinase 1 and sphingosine 1‐phosphate receptors

The hepatic growth factor hepatopoietin Cn (HPPCn) prevents liver injury induced by carbon tetrachloride in rats. Sphingosine 1‐phosphate (S1P) is a bioactive sphingolipid produced by sphingosine kinase (SphK). S1P and S1P receptors (S1PRs) are involved in liver fibrogenesis and oxidative injury. This work sought to understand the mechanism by which SphK/S1P/S1PRs are involved in the protective effects of HPPCn on ethanol‐induced liver injury and fibrosis. Transgenic mice with liver‐specific overexpression of HPPCn (HPPCn^liver+/+) were generated. Two ethanol feeding protocols were used to assess the protective effect of HPPCn on acute and chronic liver injury in mice. Specific inhibitors of S1PR1, S1PR2 and S1PR3 and siRNA were used to examine the roles of S1PRs in hepatic stellate cell (HSC) activation and hepatocyte apoptosis. Increased HPPCn expression in transgenic mice attenuated fibrosis induced by ethanol and carbon tetrachloride (CCl₄). Treatment with recombinant human HPPCn prevented human hepatocyte apoptosis and HSC activation. JTE‐013 or S1PR2‐siRNA attenuated the effect of HPPCn on HSC activation induced by tumour necrosis factor‐α (TNF‐α). Consistent with the effect of N,N‐dimethylsphingosine (DMS), suramin or S1PR3‐siRNA treatment blocked HPPCn‐induced Erk1/2 phosphorylation in human hepatocytes. This study demonstrated that HPPCn attenuated oxidative injury and fibrosis induced by ethanol feeding and that the SphK1/S1P/S1PRs signalling pathway contributes to the protective effect of HPPCn on hepatocyte apoptosis and HSC activation. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

辛伐他汀抑制高糖诱导人脐静脉内皮细胞的凋亡

背景：他汀类药物对血管内皮细胞的凋亡是否有影响目前尚不明确。 目的：探讨辛伐他汀对高糖诱导的人脐静脉内皮细胞凋亡的影响。 方法：用DMEM细胞培养液培养人脐静脉内皮细胞,将细胞分成空白对照组、高糖组和高糖+辛伐他汀组,用四甲基偶氮唑蓝比色法测定人脐静脉内皮细胞的存活率,流式细胞仪和Western blot分别检测细胞早期凋亡率及P53蛋白表达。 结果与结论：高糖组及高糖+辛伐他汀组细胞增殖率较空白对照组明显降低(P < 0.01),而高糖组细胞增殖率较高糖+辛伐他汀组亦降低(P < 0.01);高糖组P53蛋白表达量及凋亡率较空白对照组及高糖+辛伐他汀组明显增加(P < 0.01),高糖+辛伐他汀组P53蛋白表达及凋亡率亦明显高于空白对照组(P< 0.01)。表明高糖可通过促进促凋亡蛋白P53的表达进而促进人脐静脉内皮细胞的凋亡,而辛伐他汀可抑制此作用。     

NicE-C efficiently reveals open chromatin–associated chromosome interactions at high resolution

Enhancer–promoter communication is known to regulate spatiotemporal dynamics of gene expression. Several methods are available to capture enhancer–promoter interactions, but they either require large amounts of starting materials and are costly, or provide a relative low resolution in chromatin contact maps. Here, we present nicking enzyme-assisted open chromatin interaction capture (NicE-C), a method that leverages nicking enzyme–mediated open chromatin profiling and chromosome conformation capture to enable robust and cost-effective detection of open chromatin interactions at high resolution, especially enhancer–promoter interactions. Using TNF stimulation and mouse kidney aging as models, we applied NicE-C to reveal characteristics of dynamic enhancer–promoter interactions.

Enhancers and promoters are key cis-regulatory elements located at open chromatin regions, which can be detected by open chromatin profiling methods, such as FAIRE-seq (Giresi and Lieb 2009), DNase-seq (Song and Crawford 2010), ATAC-seq (Buenrostro et al. 2013), and nicking enzyme-assisted sequencing (NicE-seq) (Ponnaluri et al. 2017). Long-range chromatin interactions between enhancers and promoters can control cell type– and condition-specific gene expression, which play important roles in diverse biological processes, including neural development (Bonev et al. 2017), cell differentiation (Isoda et al. 2017), and etiopathology of diseases (Hua et al. 2018). The development of chromosome conformation capture (3C)-based methods, including Hi-C (Lieberman-Aiden et al. 2009; Rao et al. 2014) and Micro-C (Hsieh et al. 2020; Krietenstein et al. 2020), has greatly promoted our understanding of high-order chromatin organization and enhancer–promoter interactions. However, these genome-wide methods need extremely deep sequencing to provide sufficient spatial resolution to identify enhancer–promoter interactions.To enrich cis-regulatory elements–associated chromatin interactions, especially between enhancers and promoters, methods such as Capture Hi-C (Mifsud et al. 2015), OCENA-C (Li et al. 2018), and Trac-looping (Lai et al. 2018) have been developed. Although Capture Hi-C can identify genome-wide chromatin interactions associated with both active and inactive promoters, it requires a costly, species-specific predesigned biotinylated RNA bait library to target known promoters. OCEAN-C and Trac-looping are probe-free methods that use open chromatin features. OCEAN-C combines Hi-C with FAIRE-seq to select open chromatin interactions, whereas Trac-looping uses transposase enzyme and a bivalent ME linker. Although potentially very promising, Trac-looping requires about 100 million (M) cells per experiment, yet identifies only a relatively low fraction of long-range (>20 kb) cis chromatin interactions. Limitation of OCEAN-C falls in its relative lower resolution and lower enrichment of open chromatin regions. Accordingly, an improved method for capturing open chromatin interactions would be highly desirable. To address these challenges, we developed a new probe-free method named nicking enzyme-assisted open chromatin interaction capture (NicE-C) for capturing open chromatin interactions.     

LY333531对高糖所致心肌微血管内皮细胞通透性上调的拮抗作用

目的:观察LY333531对高糖所致心肌微血管内皮细胞通透性上调的拮抗作用,并探讨其机制.方法:将分离培养并鉴定后的大鼠心肌微血管内皮细胞分为正常组,高糖组(葡萄糖浓度25 mmol/L),高糖(葡萄糖浓度25 mmol/L)+LY333531(10 μmol/L)组,高糖(葡萄糖浓度25 mmol/L)+生理盐水组,通过离体血管通透性检测试剂盒检测单层细胞通透性,TUNEL法检测细胞凋亡,免疫荧光和Western blot法检测PKCβⅡ的表达.结果:与正常组比较,高糖组单层细胞通透性明显上调(400.0±20.00 vs 223.3±25.17;P<0.01),凋亡率升高(55.00%±5.000% vs 2.333%±1.155%;P<0.01),PKCβⅡ的表达增加(0.4767±0.07506 vs 0.1733±0.02082;P<0.01);LY333531可明显降低PKCβⅡ的表达(0.2800±0.070 vs 0.4767±0.07506;P<0.01)并拮抗上述病理性通透功能上调(360±17.32 vs 400.0±20.00;P<0.05)和凋亡率升高(25.00%±5.000% vs 55.00%±5.000%;P<0.01),而生理盐水对细胞通透性、凋亡率和PKCβⅡ表达的影响不明显.结论:高糖损害心肌微血管内皮细胞的通透功能,LY333531可拮抗此损伤作用.     

Soluble ICAM-1 and VCAM-1 as markers of endothelial activation

Activated endothelium releases the soluble adhesion molecules vascular cell adhesion molecule-1 (sVCAM-1) and intercellular adhesion molecule-1 (sICAM-1). Measurement of fluid-phase adhesion molecules is therefore used to quantify endothelial activation, but it is unclear which is the better marker. The aims of the study were to compare the relationships between mRNA, surface and total expression and released VCAM-1 and ICAM-1 in endothelial cell cultures during activation, and to compare human umbilical vein endothelial cells (HUVEC) with the microvascular cell line HMEC-1. sVCAM-1 better represented mRNA and surface expression changes in HUVEC undergoing endotoxin stimulation than did sICAM-1. Very little VCAM-1 was released from endotoxin-stimulated HMEC-1, and sICAM-1 seemed a better activation marker for these cells. During incubation of HUVEC in media with glucose concentrations of 5.6, 10.6 or 20.6 m m , VCAM-1 was released to the media in a dose-dependent way without changes in surface expression. ICAM-1 was not influenced by the glucose concentration. There are situations when VCAM-1 concentrations in the media do not mirror the surface expression on HUVEC in culture, indicating that measurements of soluble adhesion molecules may not necessarily be representative of the conditions on the cell surface. Endothelium from different locations showed varying responses with respect to VCAM-1 and ICAM-1 liberation upon endotoxin stimulation. Thus, both sVCAM-1 and sICAM-1 should be quantified in clinical studies of endothelial activation until their characteristics are better clarified.     

肥胖抑制素减轻高糖致大鼠INS1细胞的凋亡

目的探讨肥胖抑制素(OB)减轻高糖致大鼠胰岛细胞系INS1细胞的凋亡作用。方法 INS1细胞在不同糖浓度环境下培养24 h,用MTT法检测INS1细胞存活率和增殖;Hoechst33258细胞核染色法检测细胞核形态学;酶标法检测caspase-3活性及OB的保护作用是否涉及了PI3K;real-time PCR检测FOXO1、SREBP1c、Bax和PDX-1mRNA的表达。结果在高糖条件下,OB促进INS1细胞的增殖,在100 nmol/L浓度时能最大程度促进INS1细胞增殖(与对照组、高糖组相比,P<0.01)。OB能减轻高糖诱导的凋亡(P<0.01);在高糖组,FOXO1、SREBP1c和Bax基因表达较对照组增多,PDX-1表达减少;反之在OB干预的高糖组;FOXO1、SREBP1c和Bax表达减少,PDX-1表达增多。结论 OB能够减轻高糖致大鼠INS1细胞的损伤作用。     

MEK1/2在脂多糖诱导人脐静脉内皮细胞ICAM-1表达中的作用

目的：探讨MEK1/2在脂多糖诱导人脐静脉内皮细胞（HUVECs）ICAM-1表达中的作用。 方法： 用不同浓度LPS或LPS加MEK1/2特异性抑制剂PD98059和HUVECs孵育不同时间后，分别采用RT-PCR和Western blotting检测ICAM-1 mRNA和蛋白的表达。 结果： LPS呈时间-浓度依赖性地上调HUVECs ICAM-1 mRNA和蛋白的表达。LPS预处理后2 h，HUVECs ICAM-1 mRNA和蛋白的表达即开始升高，LPS(100 μg·L-1)作用后6 h,ICAM-1 mRNA和蛋白的表达基本达到高峰；PD98059(10 μg·L-1)可显著抑制LPS(100 μg·L-1)诱导6 h的ICAM-1 mRNA和蛋白的表达，抑制率分别为54.4%和44.9%（P<0.01 vs LPS）。 结论： 调控MEK1/2通路可能为内毒素休克诱导血管内皮损伤的防治提供新的策略。     

Selective eosinophil transendothelial migration triggered by eotaxin via modulation of Mac-1/ICAM-1 and VLA-4/VCAM-1 interactions

We have recently cloned eotaxin, a highly efficacious eosinophilicchemokine involved in the development of lung eosinophilia duringallergic inflammatory reactions. To understand more preciselyhow eotaxin facilitates the specific migration of eosinophils,we have studied which adhesion receptors are essential for eotaxinaction both in vivo and in vitro. Experiments using mice geneticallydeficient in adhesion receptors demonstrated that moleculespreviously reported to be involved in both leukocyte tethering/rolling(P-selectin and E-selectin) and in sticking/transmigration (ICAM-1and VCAM-1) are required for eotaxin action in vivo. To furtherelucidate the mechanism(s) involved in this process, we haveused an in vitro transendothelial chemotaxis model. mAb neutralizationstudies performed in this system suggest that the integrinsMac-1 (CD11b/18), VLA-4 (₄ß₁) and LFA-1 (CD11a/18) areinvolved in the transendothelial chemotaxis of eosinophils toeotaxin. Accordingly, the expression of these integrins on eosinophilsis elevated by direct action of this chemokine in a concentration-dependentmanner. Taken together, our results suggest that eotaxin-inducedeosinophil transendothelial migration in vivo and in vitro relieson Mac-1/ICAM-1 and VLA-4/VCAM-1 interactions, the latter onesbecoming more relevant at later time points of the eotaxin-inducedrecruitment process.     

红景天苷上调HIF-1α减轻高糖诱导的大鼠肾小球内皮细胞损伤

目的:观察体外不同浓度葡萄糖对大鼠肾小球内皮细胞(rRGECs)表达低氧诱导因子1α(HIF-1α)、血管内皮生长因子A(VEGFA)及血管内皮钙黏素(VE-cadherin)的影响,探讨红景天苷减轻高糖诱导rRGECs损伤的作用及可能相关机制。方法:体外培养rRGECs,分为正常糖组、高糖(20、30和50 mmol/L)组、高渗组及红景天苷+高糖组。采用MTT法检测rRGECs的活力;RT-qPCR法检测rRGECs内HIF-1α、VEGFA及VE-cadherin的mRNA表达;Western blot法检测rRGECs内HIF-1α蛋白的表达。结果:与正常糖组相比,培养24 h后,高糖(20mmol/L)组rRGECs HIF-1α的mRNA及蛋白表达均上调(P 0.05);培养120 h后,高糖组HIF-1αmRNA表达下调(P 0.05)。与正常糖组相比,培养24 h和120 h后,高糖组rRGECs内VE-cadherin的mRNA表达下调(P 0.05)。与正常糖组相比,培养24 h后,高糖组rRGECs内VEGFA的mRNA表达上调(P 0.05);培养120 h后,高糖组rRGECs内VEGFA的mRNA表达下调(P 0.05)。与正常糖组相比,培养24 h和120 h后,高渗组rRGECs内HIF-1α、VE-cadherin及VEGFA的mRNA表达无变化。与高糖组相比,培养24 h后,红景天苷(50μmol/L)组rRGECs的活力增加(P 0.01),且细胞内HIF-1α和VE-cadherin的mRNA及HIF-1α蛋白表达均上调(P 0.05)。结论:体外高糖培养能影响rRGECs表达HIF-1α,可能与细胞活力、葡萄糖的浓度、作用时间及HIF/VEGF通路有关。红景天苷能减轻高糖诱导的rRGECs损伤,其机制可能与增加rRGECs内HIF-1α的表达有关。     

Snail1 siRNA 对高糖诱导肾小管上皮细胞表型转变的影响

目的： 观察Snail1 siRNA对高糖诱导的肾小管上皮细胞向间充质细胞转变（TEMT）的影响。方法: 原代培养肾小管上皮细胞分为5组：（1）对照组（含糖5.5 mmol/L）;（2）高糖组（含糖25 mmol/L）;（3）Snail1 siRNA处理组,转染Snail1 siRNA,6 h后更换为高糖（含糖25 mmol/L）培养;（4）control siRNA处理组,转染control siRNA作为siRNA阴性对照,6 h后换为高糖（含糖25 mmol/L）培养;（5）高渗组（含D-manntio19.5 mmol/L）;72 h后收集细胞,用Western blotting和半定量RT-PCR检测Snail1、TGF-β1、α-平滑肌肌动蛋白（α-SMA）、vimentin和E-cadherin蛋白和mRNA表达。结果: 与高糖组比较,肾小管上皮细胞转染Snail1 siRNA后,Snail1 mRNA和蛋白表达水平分别下降62%和68%（P<0.01）。同时,Snail1 siRNA处理组α-SMA和vimentin蛋白和mRNA表达显著下调（P<0.01）,而E-cadherin蛋白和mRNA表达显著上调（P<0.01）。结论: Snail1参与了高糖诱导TEMT的调节。