缬沙坦抑制腹膜纤维化作用机制的进一步研究

发布时间：2018-05-01 04:13

本文选题：缬沙坦 + 腹膜透析　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:观察缬沙坦对腹膜纤维化大鼠腹膜结构和超滤功能的影响,以及对大鼠腹膜组织TGF-β1、p-Smad2/3、Smad7信号蛋白表达的影响,进一步探讨缬沙坦对腹膜透析大鼠腹膜纤维化的抑制作用及作用机制,以协助发现治疗腹膜纤维化作用的新靶点。方法:采用浓度为0.1%的葡萄糖氯己定1.0ml/(100g·d)腹腔注射以建立大鼠腹膜纤维化的模型。实验共包括三组,30只雄性SD大鼠(清洁级)随机分为对照组、模型组(腹膜纤维化组)、实验组(模型+缬沙坦组),每组大鼠各10只。三组大鼠均予自由进食水,每12小时昼夜节律交替照明。适应性喂养大鼠1周后开始进行实验。给予对照组大鼠生理盐水腹腔注射1.0ml/(100g·d);给予模型组大鼠0.1%葡萄糖氯己定腹腔注射1.0ml/(100g·d);给予实验组大鼠腹腔注射0.1%葡萄糖氯己定1.0ml/(100g·d)建立大鼠腹膜透析腹膜纤维化模型的同时,将缬沙坦2.0mg/(kg·d)溶于0.5m L 0.9%的生理盐水中每日予腹腔注射;注射部位均选择大鼠右下腹部,以上操作共进行14天。第14天后,分别测定三组大鼠的腹膜超滤量,以评估各组大鼠的腹膜超滤功能。各组大鼠的腹膜超滤量测量完毕后,处死大鼠,避开腹腔注射进针部位留取各组大鼠的壁层及脏层腹膜组织。采取HE染色的方法观察并对比三组大鼠壁层腹膜组织的病理学变改变,采用免疫组化检测各组大鼠的壁层腹膜组织转化生长因子β1(TGFβ1)的表达情况,采用Western-blot检测各组大鼠脏层腹膜组织p-Smad2/3及Smad7信号蛋白的表达情况。全部实验数据采用SPSS 9.0统计软件进行统计学处理,计量资料用均数±标准差表示,组间比较采用单因素方差分析。P0.05认为差异有统计学意义。结果:通过给予大鼠0.1%葡萄糖氯己定腹腔注射,成功复制了伴有腹膜纤维化的腹膜透析大鼠模型。模型组和实验组大鼠较对照组大鼠腹膜超滤量降低,腹膜增厚且伴有腹膜结构紊乱等腹膜纤维化表现,且腹膜组织对TGF-β1、p-Smad2/3、Smad7信号蛋白的表达显著增加。而实验组与模型组相比较,实验组大鼠腹膜超滤情况较模型组大鼠明显改善,腹膜变薄,腹膜结构有一定程度的改善,且实验组大鼠腹膜对TGF-β1、p-Smad2/3蛋白的表达受抑制,作为能够抑制腹膜TGF-β1表达的负反馈因子Smad7蛋白的表达与能够上调TGF-β1的Smad2/3蛋白一样受到抑制。结论:(1)缬沙坦能够一定程度地改善大鼠腹膜结构及超滤功能,抑制大鼠腹膜纤维化进程。其抑制作用是过下调了TGF-β/Smad通路的表达实现的。(2)予缬沙坦对腹膜纤维化的发展进行干预后,腹膜组织对TGF-β1及pSmad2/3表达减少,而对腹膜纤维化具有抑制作用的信号蛋白Smad7蛋白的表达同样受抑制。
[Abstract]:Aim: to observe the effects of valsartan on peritoneal structure and ultrafiltration function in rats with peritoneal fibrosis, and on the expression of TGF- 尾 1 p-Smad2 / 3 Smad7 signal protein in peritoneal tissue of rats. To further investigate the inhibitory effect and mechanism of valsartan on peritoneal fibrosis in peritoneal dialysis rats in order to find a new target for the treatment of peritoneal fibrosis. Methods: the peritoneal fibrosis model of rats was established by intraperitoneal injection of 0.1% glucose chlorhexidine (1.0ml/(100g d). Three groups of 30 male SD rats (clean grade) were randomly divided into two groups: model group (peritoneal fibrosis group) and experimental group (model valsartan group, 10 rats in each group). All the rats in the three groups were fed freely, and the circadian rhythm was alternately illuminated every 12 hours. The experiment began after 1 week of adaptive feeding. Rats in the control group were given normal saline intraperitoneal injection of 1.0ml/(100g dai; the model group rats were given 0.1% glucoclohexidine intraperitoneal injection of 1.0ml/(100g dai; and the experimental group rats were injected intraperitoneally with 0.1% glucoclohexidine 1.0ml/(100g d) to establish peritoneal fibrosis model of peritoneal dialysis in rats. The valsartan 2.0mg/(kg d was dissolved in 0.5 mL 0.9% normal saline daily and injected intraperitoneally into the right lower abdomen of rats for 14 days. After 14 days, peritoneal ultrafiltration was measured in three groups to evaluate the peritoneal ultrafiltration function of each group. After the peritoneal ultrafiltration measurement was completed, the rats were killed, and the wall and visceral peritoneal tissues of each group were removed from the intraperitoneal injection. The pathological changes of parietal peritoneum in three groups were observed and compared by HE staining. The expression of transforming growth factor 尾 1(TGF 尾 1 (TGF 尾 1) in parietal peritoneum of rats in each group was detected by immunohistochemistry. Western-blot was used to detect the expression of p-Smad2/3 and Smad7 signal proteins in the visceral peritoneum of rats. All the experimental data were statistically processed by SPSS 9.0 statistical software. The measurement data were expressed as mean 卤standard deviation, and the differences were statistically significant by single factor ANOVA (P0.05). Results: the peritoneal dialysis rat model with peritoneal fibrosis was successfully established by intraperitoneal injection of 0.1% glucose chlorhexidine. Compared with the control group, the peritoneal ultrafiltration (UF) of the model group and the experimental group decreased, the peritoneal thickening accompanied with peritoneal structural disorder and other peritoneal fibrosis manifestations, and the expression of TGF- 尾 1 p-Smad2 / 3 Smad7 signal protein in the peritoneal tissue was significantly increased. Compared with the model group, the peritoneal ultrafiltration in the experimental group was obviously improved, the peritoneum thinned, the peritoneal structure was improved to some extent, and the expression of TGF- 尾 1p-Smad2 / 3 protein was inhibited in the experimental group. The expression of Smad7 protein, a negative feedback factor that can inhibit the expression of TGF- 尾 1 in peritoneum, was inhibited as well as that of Smad2/3 protein which could up-regulate TGF- 尾 1. Conclusion Valsartan can improve the peritoneal structure and ultrafiltration function to some extent and inhibit the process of peritoneal fibrosis in rats. The inhibitory effect of valsartan on the development of peritoneal fibrosis was that the expression of TGF- 尾 1 and pSmad2/3 in peritoneal tissue decreased after the downregulation of the expression of TGF- 尾 / Smad pathway. The expression of signal protein Smad7 was also inhibited in peritoneal fibrosis.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692.5

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