蝉花菌丝对单侧输尿管结扎大鼠肾间质TGF-β_1和CTGF蛋白及mRNA表达的影响

目的：探讨蝉花菌丝对肾间质纤维化大鼠肾间质TGF-β1、CTGF蛋白及mRNA表达的影响。方法：采用单侧输尿管结扎的方法制备单侧输尿管梗阻（UUO）肾间质纤维化大鼠模型。实验分组为正常组、模型组、假手术组、冬虫夏草组、蝉花高剂量组、蝉花中剂量组和蝉花低剂量组,实验周期为25d。采用免疫组织化学和原位杂交方法,分别检测肾组织TGF-β1、CTGF蛋白及其mRNA表达,使用图像分析系统对结果进行处理,采用SPSS11.5统计软件统计分析。结果：UUO模型组大鼠肾间质中TGF-β1、CTGF蛋白及mRNA的表达量均明显高于正常组和假手术组（P〈0.05～0.01）;不同剂量的蝉花菌丝组TGF-β1、CTGF蛋白及mRNA的表达量明显低于模型组（P〈0.05～0.01）,呈明显的量效依赖关系;蝉花菌丝大剂量组的TGF-β1、CTGF蛋白及mRNA的表达量较虫草组为低（P〈0.05～0.01）。结论：蝉花菌丝可以通过下调TGF-β1、CTGF蛋白及其mRNA的表达,对UUO肾间质纤维化大鼠肾脏起一定的保护作用。     

肾纤康抗大鼠肾间质纤维化的实验研究

目的:探讨肾纤康防治肾间质纤维化的作用机制.方法:将大鼠随机分为4组,即假手术组、模型组、肾纤康组、苯那普利组,采用左侧输尿管梗阻方法造模,用免疫组织化学方法作肾间质转化生长因子-β1(TGF-β1)、Ⅲ型胶原(ColⅢ)、纤维连结蛋白(FN)检测,并观察肾脏病理学变化.结果:模型组和各治疗组肾间质TGF-β1、ColⅢ、FN面积比明显增加,与假手术组相比统计学差异(P＜0.05).与模型组相比,肾纤康治疗组、苯那普利治疗组肾间质TGF-β1、ColⅢ、FN面积比减少,有统计学差异(P＜0.05).肾纤康组与苯那普利组肾间质TGF-β1、ColⅢ、FN面积比无统计学差异.结论:肾纤康通过下调TGF-β1在肾间质的表达,抑制ECM的增生、积聚,防止肾间质纤维化,对大鼠单侧输尿管梗阻所致肾损害有明显保护作用,其作用与苯那普利相当.     

活性维生素D3抑制UUO模型大鼠肾间质纤维化作用的实验研究

目的:研究活性维生素D3[1,25(OH)2D3]在肾间质纤维化(RIF)中的保护作用,探讨1,25(OH)2D3的抗肾间质纤维化作用机制。方法:将40只SD大鼠随机分为UUO空白对照组、1,25(OH)2D3治疗组、贝那普利组、假手术组和假手术+1,25(OH)2D3组。采用单侧输尿管梗阻(UUO)致大鼠RIF模型。于术后第9周处死动物,取血标本做血钙浓度和肾功能检测。取肾组织行苏木素-伊红(HE)染色观察肾间质病理变化。免疫组化法检测HGF、TGF-β1和α-SMA蛋白表达,RT-PCR法检测肾组织HGF和TGF-β1mRNA的表达水平。结果:1,25(OH)2D3治疗组、贝那普利组血肌酐、尿素氮均较UUO空白对照组显著降低(P<0.05);各组间血钙浓度差异无统计学意义(P>0.05)。光镜下观察1,25(OH)2D3治疗组和贝那普利组RIF程度轻于UUO空白对照组。1,25(OH)2D3治疗组HGF和TGF-β1蛋白和mRNA表达水平与UUO空白对照组相比,分别显著上调(P<0.05)和下调(P<0.05)。结论:1,25(OH)2D3抑制RIF的作用与其抑制TGF-β1表达和肌成纤维细胞转分化,同时上调HGF表达有关。     

厄贝沙坦对糖尿病大鼠肾脏骨桥蛋白表达及肾纤维化的影响

目的 探讨不同剂量厄贝沙坦对糖尿病大鼠肾组织骨桥蛋白( OPN)表达及小管间质纤维化的影响.方法 将63只8周龄雄性Wistar大鼠按随机数字表法分为正常对照组(Ctrl组,n=7)、糖尿病组(DM组,n=14)、30 mg/kg肼屈嗪干预组(DM+ Hyd组,n=12)、25mg·kg-1· d-1厄贝沙坦干预组(DM+Irb25组,n=10)、50 mg·kg-1·d-1厄贝沙坦干预组(DM+Irb50组,n=9)和200 mg · kg-1·d-1厄贝沙坦干预组(DM+Irb200组,n=11).糖尿病模型造模成功后4周,灌胃法给予相应剂量的肼屈嗪和厄贝沙坦.第12周末,观察各组大鼠24h尿白蛋白排泄率(UAER)、内生肌酐清除率(Ccr);PAS及Masson染色观察各组大鼠肾脏病理形态和胶原纤维沉积;ELISA法测定各组大鼠肾组织AngⅡ含量;实时定量PCR检测大鼠肾组织转化生长因子(TGF)β1、OPN mRNA的表达.结果 药物干预各组大鼠UAER、Ccr较DM组显著减少(均P＜ 0.05).DM组大鼠肾小球肥大,系膜基质增生,肾小球及小管间质有大量胶原纤维沉积;药物干预后,肾小球及肾小管上述病变减轻.DM组大鼠肾组织AngⅡ水平及TGF-β1、OPN mRNA表达均显著升高(均P＜0.05),给予肼屈嗪及厄贝沙坦干预后,AngⅡ、TGF-β1、OPN mRNA表达显著降低(均P＜0.05),且随着厄贝沙坦剂量的递增而递减.相关分析结果显示,DM组大鼠肾组织AngⅡ水平与OPN mRNA的表达呈正相关(r=0.74,P＜0.01).结论 厄贝沙坦通过减少肾脏局部AngⅡ水平,进而减少肾脏TGF-β1、OPN mRNA的表达,最终减轻小管间质纤维化,发挥肾脏保护作用,且这种保护作用具有剂量依赖性.     

肾炎防衰液对糖尿病肾病大鼠的防治作用及对于MCP-1表达的影响

目的:研究肾炎防衰液对糖尿病肾病大鼠的防治作用。方法:建立大鼠糖尿病肾病模型,随机分为模型组、贝那普利组和肾炎防衰液组,另设假手术组;给予贝那普利组和肾炎防衰液组分别灌胃以贝那普利混悬液(1.0 mg·kg-1·d-1)和肾炎防衰液(11.4 g·kg-1·d-1);模型组和假手术组灌以等量生理盐水。实验进入第8周后取材,检测血清肌酐、尿素氮,采用HE和Masson染色检测肾脏病理、免疫组化法检测肾组织MCP-1的表达。结果:各组大鼠血清尿素氮水平差异无统计学意义(P>0.05);模型组血清肌酐水平明显高于假手术组(P<0.05),而肾炎防衰液组较模型组为低(P<0.05);模型组肾小球硬化指数、肾小管损伤指数、肾小管间质纤维化指数以及肾组织中MCP-1表达显著高于假手术组(P<0.05);肾炎防衰液组肾小球硬化指数、肾小管损伤指数、肾小管间质纤维化指数以及MCP-1表达均较模型组为低(P<0.05)。结论:肾炎防衰液可以降低血清肌酐水平,减轻糖尿病肾病大鼠肾小球硬化及肾间质纤维化的程度,下调糖尿病肾病大鼠肾组织MCP-1的表达,延缓糖尿病肾病进展。     

γ-干扰素对梗阻性积水肾间质纤维化的治疗作用

目的 探讨γ干扰素(IFN-γ)对梗阻性肾积水肾间质纤维化的抑制作用及其可能的机制.方法 将65只雄性SD大鼠随机分成4组:治疗组(20只)、模型组(20只)、药物对照组(20只)、假手术组(5只).在建模和给药后的第3、7、14、21、28天,每组各随机处死动物4只(假手术组1只).采用苏木素-伊红(HE)和Masson染色观察病变肾脏间质纤维化的情况;采用聚合酶链反应(RT-PCR)技术检测肾组织中转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原(Col-Ⅰ)的mRNA表达情况;免疫组织化学法观察以上三种蛋白的变化.结果 模型组大鼠术后肾间质逐渐出现纤维化改变,并随梗阻时间的延长逐渐加重,TGF-β1、α-SMA和Col-Ⅰ的mRNA和蛋白表达亦逐渐升高.第14天,模型组TGF-β1和α-SMA的表达量达到高峰,28 d时各指标的表达均达到最高,TGF-β1、α-SMA和Col-Ⅰ分别为51.84%、72.59%和68.73%.治疗组各指标在不同时间点均低于模型组,第28天时,三项指标依次为33.84%、32.59%和48.73%,与模型组比较P＜0.05.Banff评分显示治疗组间质纤维化程度明显减轻(P＜0.01).其余两组未见肾间质纤维化改变.结论 IFN-γ具有减轻积水后肾间质纤维化程度的作用,该作用与其下调TGF-β1的表达、抑制肌成纤维细胞(MyoF)激活和减少Col-Ⅰ生成有关.     

单侧输尿管梗阻大鼠肾脏转化生长因子β受体亚型表达的变化

目的 探讨转化生长因子β（TGF-β）Ⅰ型受体（RⅠ）、Ⅱ型（RⅡ）受体以及下游Smad蛋白在单侧输尿管梗阻（UUO）大鼠模型肾脏中表达及意义。 方法 90只雌性Wistar大鼠随机分为正常对照组（CON组）、假手术组（SOR组）和单侧输尿管梗阻组（UUO组）,分别于术后1、3、7、14、21 d处死,检测各组大鼠肾功能;PAS与Masson染色观察大鼠肾间质病理形态改变;实时定量PCR基因芯片分析正常大鼠和肾间质纤维化大鼠肾组织TGF-βⅠ、Ⅱ、Ⅲ型受体及Smad蛋白家族表达。筛选出差异表达的受体亚型,进一步应用实时荧光定量PCR、蛋白免疫印迹法、免疫荧光法检测和验证筛选出的差异受体亚型在不同分期肾间质纤维化大鼠肾组织的分布和表达。 结果 与CON组相比,UUO组大鼠的Scr及BUN于术后3 d开始升高（P < 0.05）,第21天达峰值（P < 0.01）;UUO组术后3 d肾间质可见明显炎性细胞浸润;14 d后出现明显肾小管萎缩;21 d可见明显肾间质纤维化。UUO组肾组织TGF-βⅠ型受体ALK-5、ALK-7和TGF-βRⅡ的mRNA表达于术后3 d上升并随梗阻时间延长逐渐增加（P < 0.05）,于14 d达到峰值（均P < 0.01）;ALK-6的mRNA表达于术后3 d下降（P < 0.05）并随梗阻时间延长逐渐减少,于14 d达谷值（P < 0.01）。ALK-5、ALK-6、ALK-7和TGF-βRⅡ蛋白表达与基因表达一致。Smad2/3及磷酸化（p）-Smad2/3的蛋白表达于术后3 d上升（均P < 0.05）并随梗阻时间延长逐渐增加,于14 d达到峰值（均P < 0.01）。 结论 在肾间质纤维化进展中不同TGF-β受体亚型存在不同的变化规律并与肾间质纤维化进展密切关联。     

黏膜方对IgA肾病大鼠异常糖基化IgA1及肾间质纤维化的干预及机制研究

目的:研究血清和肾组织中的异常糖基化IgA1水平与IgA肾病的间质纤维化程度的相关性;阐明黏膜方治疗IgA肾病的作用机制。方法:采用口服牛血清白蛋白、四氯化碳皮下注射和尾静脉注射脂多糖免疫复合法建立IgA肾病大鼠模型,随机分为正常组、模型组、中药组(黏膜方)及西药组(福辛普利钠)、中西医结合组(黏膜方+福辛普利),观察治疗后各组血清和肾组织异常糖基化的IgA1水平、血清IgA-纤维连接蛋白(IgA-FN)、肾组织转化生长因子-β1(TGF-β1)和α-平滑肌肌动蛋白(α-SMA)及其mRNA的表达量。结果:血清IgA-FN,中药组及中西医结合组低于模型组(P<0.001);血清异常糖基化IgA1量,中药组及西药组均低于模型组(P<0.05);肾组织中的异常糖基化IgA1水平,模型组高于正常组(P<0.05),各治疗组低于模型组(P<0.05);肾组织中TGF-β1mRNA及蛋白表达量模型组均显著高于正常组(P<0.01),各治疗组TGF-β1mRNA及蛋白表达均低于模型组(P<0.05);模型组α-SMAmRNA高于正常组(P<0.05),各治疗组低于模型组(P<0.001),模型组α-SMA蛋白表达阳性面积高于正常组(P<0.001),中西医结合组和西药组阳性面积低于模型组(P<0.05);肾组织α-SMAmRNA及蛋白表达与血清及肾组织中异常糖基化IgA1呈正相关(P<0.005);肾组织TGF-β1mRNA与血清及肾组织异常糖基化IgA1呈正相关(P<0.005)。结论:血清中异常糖基化IgA1与肾间质纤维化程度密切相关,黏膜方能减轻实验性IgAN大鼠血清及肾脏异常糖基化的IgA1水平,改善肾间质纤维化。     

和解聚散方对单侧输尿管梗阻大鼠肾间质纤维化的防治作用

目的：研究和解聚散方对单侧输尿管梗阻（unilateral ureteral obstruction,UUO）大鼠肾间质纤维化的防治作用。方法：采用UUO建立大鼠肾间质纤维化模型,随机分为模型组、贝那普利组和和解聚散方组,另设假手术组;术后7d、14d、21d分批取材,检测血清肌酐、尿素氮,采用HE和Masson染色检测肾脏病理、免疫组化法检测肾组织TGF-β1的表达。结果：与模型组相比,和解聚散方组血清尿素氮、血清肌酐水平均降低（P〈0.05）,肾小管损害百分比和肾间质损害分值较低（P〈0.05）,肾组织TGF-β1表达减少（P〈0.05）。结论：和解聚散方可以降低尿素氮、血清肌酐水平,减轻大鼠肾间质纤维化的程度,下调UUO大鼠肾组织TGF-β1的表达,验证了和解聚散法的正确性。     

坎地沙坦对单侧输尿管梗阻大鼠肾间质纤维化及肾脏骨桥蛋白表达的影响

目的 观察坎地沙坦(CAN)对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响,并观察骨桥蛋白(OPN)与肾间质纤维化的关系及CAN干预对肾脏OPN表达的影响,探讨其在纤维化中的作用机制.方法 (1)36只成年雄性SD大鼠随机分为3组,每组12只:假手术组(Sham)、UUO模型组(UUO)、坎地沙坦治疗组(CAN).UUO模型组和CAN组大鼠行左侧输尿管结扎术,Sham组只游离左侧输尿管但不结扎.CAN组于术前1 d开始用CAN治疗[10 mg/(kg·d)]灌胃.术后第7、14天分别处死大鼠,左侧肾脏组织行Masson染色,免疫组织化学方法检测肾组织中OPN的表达,逆转录-聚合酶链反应(RT-PCR)检测OPN mRNA表达水平.结果 Masson染色结果显示术后7 d和14 d UUO组大鼠肾纤维化阳性面积分别为15.2%和24.8%,CAN组为1O.1%和18.5%.CAN组与UUO组大鼠术后7 d和14 d大鼠肾纤维化阳性面积差异有统计学意义(P＜0.05).UUO模型大鼠OPN蛋白及mRNA水平均较Sham组明显升高(P＜0.01),CAN组OPN蛋白及mRNA水平较UUO组明显降低(P＜0.05),但较Sham组高(P＜0.01).结论 CAN能有效地延缓UUO大鼠肾间质纤维化的进展,其延缓肾间质纤维化作用可能与下调OPN蛋白和mRNA有关.     

Lycium barbarum polysaccharides alleviate kidney injury and oxidative stress in unilateral ureteral obstruction rats

Objective To investigate the effect of lycium barbarum polysaccharides (LBP) on oxidative stress in renal tissue of rats with renal interstitial fibrosis (RIF). Methods The RIF rat model was established by unilateral ureteral obstruction (UUO). A total of 108 specified pathogen free (SPF) class healthy adult male Sprague-Dawley (SD) rats were randomly divided into sham operation group, UUO model group and treatment group. The treatment group was further divided into low, medium and high dose of LBP groups and benazapril group. From the next day of the operation, the rats were given continuous intragastric administration for 3 weeks. The LBP low, medium and high dose groups were given 400, 600, 800 mg?kg-1?d-1 LBP, respectively. The benazapril group was administered with 1.05 mg?kg-1?d-1 benazepril hydrochloride. The sham operation group and UUO model group were daily fed normal saline solution by gavage. Six rats were sacrificed randomly at 7, 14 and 21 days after operation. Their blood samples were collected to detect the serum creatinine (Scr) and the kidney organ index was calculated. The pathological changes on the surgical side were observed by both HE staining and Masson staining. Meanwhile, the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the renal tissue were detected by colorimetry detection. The expression of transforming growth factor-β1 (TGF-β1) protein was detected by immunohistochemical staining and the expression of TGF-β1 mRNA was detected by real time PCR. Results (1) Compared with the sham group, the Scr and kidney organ index of the UUO model group and treatment groups increased at each time point (all P＜0.05). Compared with the UUO model group, the kidney organ index of LBP low dose group in the 7th days, the LBP medium and high dose group in the 21st days as well as benazapril group in the 7th and 21st days were significantly lower (all P＜0.05). (2) Renal pathological change: compared with the sham operation group, both the renal tubular interstitial injury index and collagen positive area of the else groups were higher at each time point (all P＜0.05). Compared with the UUO group, the tubulointerstitial injury index and collagen staining positive area of LBP dose groups and benazapril group significantly decreased at different time points (all P＜0.05). (3) Compared with the sham group, in renal tissue of the other groups the level of MDA increased, SOD level decreased, while the expressions of TGF-1 mRNA and protein increased (all P＜0.05). Compared with the UUO model group, LBP low, medium and high dose group as well as benazapril group had lower MDA level, higher SOD level as well as lower expressions of TGF-1 mRNA and protein at each time point (all P＜0.05). Conclusions The pathological injury in UUO rats can be improved by the LBP. The LBP can alleviate the oxidative stress status of the kidney tissue by decreasing MDA and increasing SOD. The further study on the LBP delaying the progression of RIF is to be conducted.     

Histone methyltransferase MLL1 accelerates the development of renal fibrosis via promoting the epithelial-mesenchymal transition of HK-2 cells

Objective To investigate the possible effects of histone methyltransferase MLL1 on renal interstitial fibrosis and epithelial-mesenchymal transition (EMT). Methods Forty-two male SD rats were randomly divided into normal group, sham operation group and unilateral ureteral occlusion (UUO) group, and then UUO group was further divided into group 1 d, 1 week, 2 week, 3 week and 4 week after operation. The expression of MLL1, E-cadherin, α-SMA, Vimentin and Col3α1 in UUO rat kidney tissue as well as TGF-β1 stimulated HK-2 cells were detected by real-time PCR and Western blotting. siRNA-MLL1 plasmids was used to inhibit the expression of MLL1 and the protein levels of MLL1, α-SMA, Vimentin, E-cadherin, Col3α1 and H3K4me3 induced by TGF-β1 stimulation were detected by Western blotting. The level of H3K4me3 in promoter region of EMT related genes was observed by chromatin immunoprecipitation (CHIP). Results Compared with normal and sham operated groups, the loss of renal function in UUO group was more obvious with the obstruction time (P＜0.05). The renal fibrosis was most obvious 1 week and 2 weeks after the rats underwent the UUO operation (all P＜0.05), with the highest protein expressions of MLL1, E-cadherin, α-SMA, Vimentin and Col3α1 (all P＜0.05). Compared with the control group, 3 ng/ml TGF-β1 induced the highest expression of MLL1 and the most obvious EMT in HK-2 cells (all P＜0.05). Moreover, the EMT and the high level of H3K4me3 in HK-2 triggered by TGF-β1 were all inhibited by siRNA-MLL1 plasmids transfection (all P＜0.05). Conclusions MLL1 can enhance the occurrence of EMT induced by TGF-β1 in HK-2 cells by increasing the level of H3K4me3 in the promoter region of α-SMA and Vimentin.     

缺氧诱导因子-1α在肾间质纤维化中的表达及意义

目的：探讨缺氧诱导因子-1α（HIF-1α）致肾间质纤维化的作用。方法：60只雄性SD大鼠随机分为假手术组（SOR）和单侧输尿管梗阻（UUO）模型组。术后第3天,第7天,第14天各处死大鼠10只,肾组织行HE染色并观察病理变化。采用免疫组织化学和荧光定量逆转录聚合酶链反应（Q-RT-PCR）法检测肾脏组织中HIF-1α、CTGF、TGF-β1蛋白和mRNA表达。结果：与假手术组相比,模型组大鼠肾脏病理损害进行性加重;HIF-1α、CTGF、TGF-β1蛋白表达随梗阻时间的延长逐渐增加;与假手术组相比HIF-lα、CTGF、TGF-β1mRNA表达明显增加（P〈0.05）;相关分析显示,模型组第3天HIF-lαmRNA表达与CTGFmRNA,TGF-β1mRNA表达分别呈正相关（r=0.748,0.659,P〈0.05）,模型组第7天组HIF-1αmRNA分别和CTGFmRNA、TGF-β1mRNA呈正相关（分别为r=0.663,0.645,P〈0.05）,模型组第14天组HIF-1αmRNA分别和CTGFmRNA、TGF-β1mRNA呈正相关（分别为r=0.515,0.752,P〈0.05）。结论：HIF-1α可能通过调节CTGF、TGF-β1的表达参与了肾间质纤维化发生和发展的过程。     

Inhibition of Src kinase can ameliorate renal interstitial fibrosis in unilateral ureteral obstruction mice

Objective To investigate the effect and mechanism of Src kinase in renal interstitial fibrosis of unilateral ureteral obstruction (UUO) mice. Methods Male C57BL/6J mice were randomly divided into 4 groups, including sham operation group (n=8), sham operation+PP2 group (n=8), UUO operation group (n=8) and UUO operation+PP2 group (n=8). The mice were injected 2 mg/kg PP2 by intraperitoneal everyday after surgery in sham+PP2 group and UUO+PP2 group. PP2 dissolved in 1% DMSO (formulated with normal saline). Sham and UUO group were given equal 1% DMSO. The mice were sacrificed at 7th day. Renal collagen was observed with Sirius red stain. The activities of Src, protein kinase B (PKB, AKT), p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK) and the protein expressions of α-smooth muscle actin (α-SMA) and fibronectin (FN) were detected by Western blotting. The expression of collagen I (COLⅠ) was detected by immunohistochemistry and the expressions of matrix metalloprotein 9 (MMP-9), tissue inhibitor of metalloproteinase 1 (TIMP-1), transforming growth factor-β1 (TGF-β1), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6) were measured by ELISA. Results Compared with sham mice, UUO mice on 7th day displayed obvious renal fibrosis. Meanwhile, UUO mice had increased expressions of COLⅠ and FN, and activities of AKT, ERK and p38 MAPK (all P＜0.05). Their renal expressions of α-SMA, TGF-β1, MMP-9, TIMP-1, MCP-1 and IL-6 were also raised (all P＜0.05). Compared with those in UUO group, in UUO+PP2 group the activities of Src, AKT, p38 MAPK and ERK, and expressions of TGF-β1, MCP-1 and IL-6 decreased (all P＜0.05). Additionally, expressions of COLⅠ, FN and α-SMA, collagen deposition and renal fibrosis receded in UUO+PP2 group (all P＜0.05). However, the expressions of MMP-9 and TIMP-1 were not influenced by PP2 treatment. Conclusions Src kinase promotes myofibroblasts accumulation and inflammatory reaction through activating its downstream signaling pathway in the progressing of renal interstitial fibrosis.     

肾淋巴循环障碍致肾小管间质纤维化及对转化生长因子β1和Smad2/3表达的影响

目的 探讨肾淋巴循环障碍对大鼠肾小管间质纤维化的作用及其与TGF-β1、Smad2/3表达变化的关系。 方法 雄性Wistar大鼠48只，随机分为淋巴循环障碍模型组和假手术对照组各24只。分别在术后1、2、4、8周每组各处死6只。测定尿蛋白量（24 h）和Scr。PAS和Masson染色观察肾组织病理改变。用实时PCR检测TGF-β1、Smad2/3、I型胶原（ColⅠ）mRNA的表达量。用免疫组化和（或）Western 印迹方法检测肾组织ColⅠ、TGF-β1、Smad2/3和磷酸化Smad2/3（p-Smad2/3）的蛋白表达量及主要表达部位。 结果 模型组大鼠尿蛋白显著增加，随着时间推移肾功能逐渐减退，并出现明显的组织病理改变，小管间质损伤指数明显高于对照组（P < 0.05或P < 0.01），并随时间延长而逐渐加重。肾组织中ColⅠ、TGF-β1、Smad2/3、p-Smad2/3的蛋白和（或）基因表达水平也明显增高（P < 0.01），且主要表达在肾小管上皮细胞及肾间质。 结论 肾淋巴循环障碍可导致大鼠肾脏功能及小管间质的损害，并随着时间的延长而加重。其作用机制可能与激活TGF-β-Smad途径，导致肾小管间质纤维化有关。     

和解聚散方对单侧输尿管梗阻肾间质纤维化大鼠细胞外基质表达的影响

目的:研究和解聚散方对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾间质纤维化(renal interstitial fibrosis,RIF)、细胞外基质(extracellular matrix,ECM)、纤维连接蛋白(FN)和Ⅳ型胶原(Col Ⅳ)表达的影响.方法:...     

三七总皂苷对单侧输尿管梗阻大鼠肾组织BMP-7及TGF-β1/samds信号传导通路的影响

目的:观察三七总皂苷(PNS)对单侧输尿管梗阻(UUO)大鼠肾组织骨形成蛋白-7(BMP-7)及TGF-β1/Smads信号传导通路的影响。方法:将50只雄性Wistar大鼠随机分为5组:假手术组、模型组、肾康注射液组(对照组)、PNS低剂量组(低剂量组)、PNS高剂量组(高剂量组),每组10只,除假手术组,其余各组用UUO法建立肾间质纤维化动物模型,术前1d起各组给予相应浓度和剂量的药物,术后第14天处死所有大鼠,检测大鼠血肌酐(Scr)和尿素氮(BUN)变化,梗阻侧肾组织行HE染色、PAS染色和Masson染色,观察肾组织病理变化并半定量计算肾小管损伤指数(TII),免疫组化法检测BMP-7、TGF-β1、Smad2、Smad7在肾组织的表达。结果:与模型组相比,PNS和肾康注射液能明显降低UUO大鼠术后14dScr、BUN(P<0.05);对梗阻侧肾组织HE染色、PAS染色进行TII评分发现,2个试验组大鼠肾小管损伤指数显著低于模型组(P<0.01);Masson染色观察发现,2个实验组大鼠肾小管间质病变明显轻于模型组;免疫组化法染色并对其灰度值测定发现,与模型组相比,2个试验组大鼠TGF-β1、Smad2在肾组织的表达下调(P<0.01),BMP-7、Smad7在肾脏组织的表达上调(P<0.01)。结论:PNS可能通过干预BMP-7/Smads/TGF-β1信号转导通路抑制了TGF-β1信号的细胞内转导,而起到抗肾间质纤维化的作用。