脂肪源性血管基质成分对皮肤光老化的影响

 目的:观察人脂肪源性血管基质成分 (stromal vascular fractions, SVF) 对中波紫外线(ultraviolet B, UVB) 诱导的裸大鼠皮肤光老化的影响。方法:6周龄裸大鼠随机分为正常对照组及实验组,实验组持续UVB照射8周建立皮肤光老化模型并随机分为模型组,以及SVF低剂量 (low-dose, LD)、中剂量(middle-dose, MD) 和高剂量 (high-dose, HD) (100 μL悬液中的细胞数分别为10⁴、10⁵和10⁶) 治疗组;采用酶消化法提取健康女性脂肪组织中的SVF,分别取低、中、高剂量的SVF悬液注射于裸大鼠背部皮下;模型组注射相同剂量的生理盐水,正常对照组不做任何处理;7 d后取材,观察注射部位表皮厚度及结构的改变;28 d后取材观察注射部位真皮厚度及结构的改变。结果:SVF治疗7 d后,中、高剂量治疗组裸大鼠皮肤较实验对照组表皮层厚度减小、角质层比例下降、基底层处于异常增殖状态的细胞核减少(P<0.05)。治疗28 d后,中剂量及高剂量治疗组I型胶原蛋白mRNA相对表达量增加,III型胶原蛋白及基质金属蛋白酶-3 (matrix metalloproteinases-3, MMP-3) mRNA相对表达量下降(P<0.05),高剂量治疗组皮肤真皮层厚度增加(P<0.05)。结论:SVF具有抗皮肤光老化的潜能,有潜在的临床应用价值。     

基质金属蛋白酶及其组织抑制因子在皮肤增生性瘢痕中的作用

目的探讨基质金属蛋白酶（MMPs）MMP2、MMP9及其抑制物（TIMPs）TIMP-1在皮肤增生性瘢痕中的作用。方法取3-6个月、6-12个月皮肤增生性瘢痕，以正常皮肤组织为对照，ELISA方法测定其MMP2、MMP9以及TIMP-1的含量，采用SPSS统计软件，以成组设计的单因素方差分析，分析它们在不同时段瘢痕组织中变化的意义。结果（1）3-6月瘢痕组织和6-12月瘢痕组织的MMP2，均较正常皮肤显著增高（110.70±5.23ng／ml VS 54.59±3.01，P〈0.01；77.23±7.10ng／ml VS 54.59±3.01，P〈0.01），而3-6月瘢痕组织的MMP2较6-12月瘢痕组织的MMP2亦有显著增高（110.70±5.23 VS 77．23±7．10，P〈0．01）；3-6月瘢痕组织和6-12月瘢痕组织的MMP9之间（3.85±0.88 VS 3.61±0.43，P〉0.05）以及它们与正常皮肤组织的MMP9相比（3.85±0.88 VS 4.13±0.33，P〉0.05；3.61±0.43 VS 4.13±0.33，P〉0.05）均无显著性差异；（2）3-6月瘢痕组织和6-12月瘢痕组织的TIMP-1与正常皮肤组织相比有显著增高（4．74±0．35 VS 3．01±0．11，P〈0．01；5．12±0．34 VS 3.01±0．11，P〈0．01），6-12月瘢痕组织较3-6月瘢痕组织的TIMP-1有显著增高（5.12±0.34 VS 4.74±0．35，P〈0．05）；（3）3-6月瘢痕组织和6-12月瘢痕组织的MMP2与TIMP-1的比值与正常皮肤组织相比均有显著增高（23．38±1．01 VS 18．15±0．58，P〈0.01；15.10±1.45 VS 18．15±0．58，P〈0．01），3-6月瘢痕组织较6-12月瘢痕组织的MMP2与TIMP-1的比值有显著降低（23.38±1.01 VS 15.10±1.45，P〈0.01）。结论MMPs以及TIMPs参与了皮肤瘢痕的增生过程，MMP2、TIMP-1含量及其比值可作为瘢痕生长和预后的指标。     

紫外线诱导人皮肤成纤维细胞形态改变及对基质金属蛋白酶表达的影响

目的研究紫外线(UVB)诱导人皮肤成纤维细胞中端粒长度及基质金属蛋白酶(MMPs)的变化,及其在UVB诱导光老化中的作用。方法原代培养人成纤维细胞,用第5代细胞进行UVB照射处理。观察细胞形态,实时定量PCR检测COL1a1和h TERT的mRNA表达细胞端粒长度,Western blot检测MMP-3和MMP-1蛋白。结果 30 m J/cm2UVB照射人成纤维细胞24 h后,细胞逐渐变圆、皱缩和排列紊乱;COL1a1和TERT的mRNA水平表达显著升高,基质金属蛋白酶MMP-3和MMP-1蛋白水平也显著升高,端粒长度显著缩短。结论 UVB可能诱导人皮肤成纤维细胞形态改变,及MMP-3和MMP-1的表达明显升高,启动了光老化的早期进程。     

BMSCs-exo对紫外线诱导成纤维细胞光老化的保护作用

目的探讨小鼠骨髓间充质干细胞来源的外泌体(bone marrow mesenchymal stem cell-derived exosomes,BMSCs-exo)对紫外线辐射后的鼠真皮成纤维细胞(Fibroblasts,FBs)的保护作用。方法用流式细胞术鉴定小鼠骨髓间充质干细胞(BMSCs),提取并鉴定其上清液来源的外泌体(exosome)。用紫外线照射FBs建立细胞光老化模型,采用不同浓度的BMSCs-exo处理照射后的FBs。采用β-半乳糖苷酶染色法进行FBs衰老检测。通过Western blot检测处理后FBs的I型胶原蛋白,基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)和弹性蛋白的蛋白质水平。结果β-半乳糖苷酶染色法显示BMSCs-exo浓度越高细胞衰老数目越少。Western blot检测显示BMSCs-exo浓度越高MMP-1表达越低,I型胶原蛋白和弹性蛋白表达越高。结论 BMSCsexo可以缓解紫外线诱导的鼠真皮成纤维细胞光老化进展。     

MMP-3和TIMP-3在卵巢癌中的表达及其意义

目的探讨MMP-3和TIMP-3在卵巢癌的表达及其意义。方法本文采用光镜、透射电镜和免疫组化方法对27例卵巢癌组织中的MMP-3及TIMP-3表达进行检测。结果结果表明,临床分期为晚期的卵巢癌组织中MMP-3阳性细胞的数密度和面密度明显高于早期,而TIMP-3则低于早期。电镜下,早期淋巴细胞和树突状细胞浸润较多,癌细胞没有穿过基底膜;晚期淋巴细胞和树突状细胞浸润较少,可见癌细胞穿基底膜。结论MMP-3和TIMP-3的表达程度及MMP-3/TIMP-3的比值,可作为判断卵巢癌病程的指标。     

丹酚酸B对小鼠皮肤光老化的保护效应

背景：中药及其提取物在改善皮肤组织结构、提高抗氧化酶活性、抑制基质金属蛋白酶高表达、增强皮肤免疫防御功能等多方面对皮肤光老化起着防治作用。 目的：验证丹酚酸B对皮肤光老化的保护作用。 方法：同时建立小鼠皮肤体外细胞培养紫外线损伤模型及小鼠紫外照射致皮肤衰老模型,给予不同剂量的丹酚酸B,通过MTT法, 单细胞凝胶电泳等观察各组皮肤细胞损伤程度以及丹酚酸B对细胞损伤的保护作用,小鼠灌胃口服不同剂量丹酚酸B,通过对皮肤组织羟脯氨酸、超氧化物歧化酶等含量的检测,评价丹酚酸B是否具有抑制皮肤光老化的作用。 结果与结论：紫外照射可使皮肤丙二醛含量明显上升,羟脯氨酸含量,超氧化物歧化酶和谷胱甘肽氧化酶活性下降。丹酚酸B能有效地对紫外线引起细胞致死性损伤起到保护作用,能显著降低皮肤组织丙二醛含量,提高羟脯氨酸含量,超氧化物歧化酶和谷胱甘肽氧化酶活性而起到抑制皮肤光老化作用。结果证实丹酚酸B具有保护皮肤光老化作用。     

二甲双胍抑制UVA诱导的人皮肤成纤维细胞的光老化效应

目的研究二甲双胍(MF)对长波紫外线(UVA)诱导的人皮肤成纤维细胞(HSF)光老化效应的抑制作用及相关机制。方法体外培养人皮肤成纤维细胞,分为对照组(control)、UVA组、UVA+MF组。以CCK-8法检测细胞增殖;对各组细胞进行细胞衰老β-半乳糖苷酶染色;以荧光探针DCF-DA染色流式细胞仪检测细胞内ROS水平;real-time PCR检测衰老相关基因MMP1、MMP3的mRNA相对表达量;Western blot检测蛋白MMP1、MMP3、SOD1和SOD2的表达。结果 0.01 mmol/L二甲双胍对HSF增殖无显著影响,0.1和1 mmol/L二甲双胍则显著抑制HSF的细胞增殖(P0.05)。与对照组相比,UVA组β-半乳糖苷酶染色阳性率显著增高(P0.01);ROS水平显著升高(P0.05);MMP1和MMP3 mRNA相对表达量显著增加(P0.01);MMP1和MMP3蛋白表达量显著增多(P0.01);SOD1、SOD2蛋白表达显著减少(P0.01)。与UVA组相比,UVA+MF组β-半乳糖苷酶染色阳性率显著降低(P0.05);ROS水平显著下降(P0.05);MMP1和MMP3的mRNA相对表达量显著降低(P0.05);蛋白MMP1和MMP3表达量显著降低(P0.05);蛋白SOD1表达显著增加(P0.01);SOD2表达量增加。结论二甲双胍可通过抑制细胞内ROS水平和相关基质金属蛋白酶的表达,提高细胞抗氧化能力,从而抑制UVA诱导的皮肤成纤维细胞的光老化效应。     

ω-3多不饱和脂肪酸对急性肺损伤炎症反应及免疫功能影响的机制研究进展

烧(创)伤、吸入性损伤、严重感染以及休克等疾病过程中,机体内多种炎性介质的释放,引发炎性细胞向肺组织迁移,激活呈级联放大的炎症反应,常常可造成以肺毛细血管内皮细胞和肺泡上皮细胞损伤为主的急性肺损伤(ALI),严重者短时间内可迅速发展为急性呼吸窘迫综合征(ARDS),病死率极高。ω-3多不饱和脂肪酸(ω-3PUFA)作为药理性免疫营养素的重要组成部分,目前已经超越了以往单纯提供能量、恢复正氮平衡的范畴,发挥着调控机体炎症反应、免疫功能的全面作用,并逐渐演变为现代危重性肺损伤治疗的重要组成部分。因此,本文就ω-3PUFA对ALI炎症反应及免疫功能影响的机制作一综述,旨在为临床治疗ALI提供理论依据。     

基质金属蛋白酶-2、3及其组织抑制剂TIMP-1在子宫内膜异位症中的表达

目的探讨基质金属蛋白酶(MMP-2、MMP-3)及其抑制剂(TIMP-1)在子宫内膜异位症发生及发展中的作用.方法采用免疫组化SP法分别测定MMP-2、MMP-3 、TIMP-1在卵巢子宫内膜异位症异位内膜60例(A组),子宫腺肌症异位内膜40例(B组),子宫肌瘤子宫内膜30例(对照组C)的表达强度.结果 A、B组中MMP-2、MMP-3的表达强度明显高于对照组(P<0.05)而TIMP-1的表达明显低于对照组(P<0.05);A、B组间MMP-2、MMP-3 、TIMP-1 的表达无明显差异(P>0.05).结论在子宫内膜异位症中MMP-2、MMP-3的过度表达及TIMP-1的低表达可能与内异症的发生与发展有关.     

Dietary n-3 polyunsaturated fatty acids modulate purified murine T-cell subset activation

Studies in humans and murine disease models have clearly shown dietary fish oil to possess anti-inflammatory properties, apparently mediated by the n-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). To determine the mechanisms by which dietary EPA and DHA modulate mouse T-cell activation, female C57BL/6 mice were fed diets containing either 2% safflower oil (SAF), 2% fish oil (FO), or a 2% purified EPA/DHA ethyl ester mixture for 14 days. Splenic CD4 T cells ( approximately 90% purity) or CD8 T cells ( approximately 85% purity) were incubated with agonists which act at the plasma membrane receptor level [anti(alpha)-CD3/anti(alpha)-CD28], the intracellular level (PMA/Ionomycin), or at both the receptor and intracellular levels (alphaCD3/PMA). CD4 T cells stimulated with alphaCD3/alphaCD28 or PMA/Ionomycin proliferated and produced principally IL-2 (i.e. a Th1 phenotype), whereas the proliferation of CD4 T cells stimulated with alphaCD3/PMA was apparently driven principally by IL-4 (i.e. a Th2 phenotype). The IL-4 driven proliferation of putative Th2 CD4 cells was enhanced by dietary n-3 fatty acids (P = 0.02). Conversely, IL-2 production by alphaCD3/alpha CD28-stimulated CD4 T cells was reduced in FO-fed animals (P < 0.0001). The alphaCD3/alphaCD28-stimulated CD8 cells cultured from FO-fed animals exhibited a significant decrease (P < 0.05) in proliferation. There were no dietary effects seen in alphaCD3/PMA-stimulated CD8 cells, which produced both IL-2 and IL-4, or in PMA/Ionomycin-stimulated CD8 cells, which produced principally IL-2. These data suggest that dietary n-3 fatty acids down-regulated IL-2 driven CD4 and CD8 activation, while up-regulating the activation of the Th2 CD4 T-cell subset. Thus, the anti-inflammatory effects of n-3 fatty acids may result in both the direct suppression of IL-2-induced Th1 cell activation and the indirect suppression of Th1 cells by the enhanced cross-regulatory function of Th2 cells.     

饱和及多不饱和脂肪酸对金黄地鼠肝LDL受体影响的研究

雄性金黄地鼠６０只随机分成４组：Ｃ１组（对照组），Ｃ２组（胆固醇组），Ｓ组（胆固醇加饱和脂肪酸组）及Ｐ组（胆固醇加多不饱和脂肪酸组）。选用玉米油为多不饱和脂肪酸和猪油为饱和脂肪酸的来源，５周后进行血脂测定和肝ＬＤＬ受体分析。结果表明：在等量胆固醇条件下，和Ｃ２组比较，多不饱和脂肪酸可显著降低血清甘油三酯，总胆固醇，低密度脂蛋白胆固醇和极低密度脂蛋白胆固醇，而高密度脂蛋白胆固醇则升高，饱和脂肪酸使这     

Effect of n-3 fatty acids on free tryptophan and exercise fatigue

Free tryptophan (Trp), which is augmented by liberated free fatty acids (FFA) from adipose tissue, can induce mental fatigue via serotonin during exercise. Since an attenuation in FFA has been observed with omega-3 fatty acid (n-3fa) use, our purpose was to examine the effect of n-3fa supplementation on free Trp availability and exercise fatigue. Ten recreationally trained men (n=5) and women (n=5), with maximal oxygen consumption (O_2max)of 51.6 (3.0) and 44.3 (1.4) ml kg^–1 min^–1, respectively, were studied on two occasions following an overnight fast, before and after n-3fa supplementation (4 g day^–1 for 4 weeks). The exercise trials consisted of a 75-min treadmill run at 60% O_2max followed immediately by a high-intensity incremental bout to fatigue. Measurements included exercise monitors, plasma volume (PV), triglycerides (TG), FFA, glycerol, lactate, and glucose. Free Trp and branched-chain amino acids (BCAA) were measured and correlated with time to fatigue; all blood variables were corrected for PV. Free Trp, lactate, glucose, FFA, and glycerol were not significantly different between trials, but TG (P<0.001) and the free Trp/BCAA ratio were significantly lower after n-3fa use [1.76 (0.18)×10^-2 g ml^–1] versus before supplementation [2.17 (0.22), P=0.033]. There was a non-significant increase in time to fatigue after supplementation [10.2 (0.3) min] versus before n-3fa use [9.7 (0.2), P=0.068], and a tendency for higher BCAA levels after supplementation, P=0.068. However, neither free Trp nor the free Trp/BCAA ratio significantly predicted time to fatigue. In conclusion, n-3fa supplementation did not diminish free Trp concentrations or significantly improve endurance performance during a maximal bout of exercise.     

不饱和脂肪酸影响HepG-2细胞PAI-1表达的机制初探

目的：探讨不饱和脂肪酸对HepG-2细胞纤溶酶原激活物抑制剂-1(PAI-1)表达的影响及其机制。方法： 以发色底物法检测PAI-1活性，RT-PCR法检测PAI-1 mRNA水平；构建两个含不同片段缺失的PAI-1启动子序列控制表达的氯霉素转移乙酰酶（CAT）报告基因质粒，转染HepG-2细胞，ELISA法检测CAT表达量。结果： 油酸、亚油酸诱导下HepG-2细胞PAI-1mRNA表达及蛋白活性显著高于对照组；共转染过氧化体增殖物激活型受体表达质粒（PPARα-pSG5）PAI-1转录活性显著增加；转染NF-κB样蛋白结合序列缺失的重组质粒，亚油酸诱导下PAI-1转录活性显著增加，而转染VLDL/脂肪酸反应元件缺失的重组质粒则无显著变化。结论： 不饱和脂肪酸增强HepG-2细胞PAI-1 mRNA表达及活性；PPARα可能是其上调PAI-1表达所涉及的转录因子之一，且VLDL/脂肪酸反应元件在该调控中具有重要作用，但可能并不涉及NF-κB信号转导途径。     

结肠癌干细胞样细胞的体外培养、鉴定及n-3多不饱和脂肪酸对结肠癌干细胞样细胞的抗增殖作用

 目的:采取无血清培养法培养出SW620细胞球,并对细胞球细胞进行干细胞鉴定;在细胞水平研究n-3多不饱和脂肪酸（n-3 PUFAs）对结肠癌干细胞样细胞的作用。 方法: 正常培养人结肠癌细胞株SW620并使其逐步适应无血清培养条件,经无血清培养1周后收集SW620细胞球。用免疫荧光法检测胚胎干细胞标志物SSEA-1和TRA-1-81;采用real-time PCR的方法检测干细胞相关基因Sox-2和Oct-4的表达情况;对比SW620贴壁细胞和干细胞样细胞（CSCLC）在软琼脂上的克隆形成能力;采用裸鼠移植瘤模型比较2种细胞的成瘤能力;用MTS法对比2种细胞在递增浓度的5-氟尿嘧啶（5-FU）或mitomycin C处理下的生长抑制情况;用MTS法、Annexin V/PI染色和台盼蓝染色分别观察递增浓度二十二碳六烯酸（DHA）和二十碳五烯酸（EPA）作用于SW620 CSCLC后细胞生长抑制情况、凋亡情况和死亡情况;MTS法检测5-FU或mitomycin C联合n-3 PUFAs对结肠癌CSCLC增殖的影响。 结果: 无血清培养法成功从SW620中培养出细胞球。细胞球细胞高表达SSEA-1和TRA-1-81并一过性表达Sox-2和Oct-4基因;对5-FU及mitomycin C相对抵抗;在软琼脂上克隆形成率及在裸鼠皮下的成瘤率均显著高于贴壁细胞,表明这些细胞具有干细胞特性,即为来自于SW620的CSCLC。DHA和（或）EPA作用于SW620 CSCLC能抑制细胞生长、诱导细胞凋亡,并能增强5-FU及mitomycin C对其抑制作用。 结论: 无血清培养法能够从SW620细胞中培养出具有干细胞特性的细胞,它们具有高克隆形成能力及高致瘤性,对化疗药物相对抗拒;DHA和EPA能够诱导SW620来源CSCLC发生凋亡并增强化疗药物的抗肿瘤活性。     

Dietary n-3 polyunsaturated fatty acids enhance hormone ablation therapy in androgen-dependent prostate cancer

Hormone ablation therapy typically causes regression of prostate cancer and represents an important means of treating this disease, particularly after metastasis. However, hormone therapy inevitably loses its effectiveness as tumors become androgen-independent, and this conversion often leads to death because of subsequent poor responses to other forms of treatment. Because environmental factors, such as diet, have been strongly linked to prostate cancer, we examined the affects of dietary polyunsaturated fatty acids (PUFAs; at 1.5 wt%) on growth of androgen-dependent (CWR22) and androgen-independent (CWR22R) human prostatic cancer xenografts, the acute response of CWR22 tumors to ablation therapy, and their progression to androgen independence. Significant diet-induced changes in tumor n-3 or n-6 PUFA content had no affect on CWR22 or CWR22R tumors growing with or without androgen support, respectively. However, dietary changes that increased tumor eicosapentaenoic acid and linoleic acid content enhanced responses to ablation therapy, measured by cancer cell apoptosis and mitosis. In addition, relapse to androgen-independent growth (measured by renewed increases in tumor volume and serum prostate-specific antigen after ablation) positively correlated with tumor arachidonic acid content. There was no correlation between expression of 15-lipoxygenase isozymes or their products and tumor growth or responses to ablation. In conclusion, dietary n-3 PUFA may enhance the response of prostate cancer to ablation therapy and retard progression to androgen-independent growth by altering tumor PUFA content.     

Role of dietary n-3 polyunsaturated fatty acids in type 2 diabetes: A review of epidemiological and clinical studies

The worldwide increasing prevalence of type 2 diabetes mellitus (T2DM) poses an immense public health hazard leading to a variety of complications such as cardiovascular diseases, nephropathy and neuropathy. Diet, as a key component of a healthy human lifestyle, plays an important role in the prevention and management of T2DM and its complications. The dietary n-3 polyunsaturated fatty acids (PUFAs) have been associated with various favourable functions such as anti-inflammatory effects, improving endothelial function, controlling the blood pressure, and reducing hypertriglyceridemia and insulin insensitivity. According to some epidemiological studies, a lower prevalence of T2DM was found in populations consuming large amounts of seafood products, which are rich in n-3 PUFAs. However, the evidence on the relation between fish intake, dietary n-3 PUFAs, and risk of T2DM is controversial. Therefore, this paper aimed to review the epidemiological and clinical studies on the role of dietary n-3 PUFAs in T2DM. Also, the limitations of these studies and the need for potential further research on the subject are discussed.