论文部分内容阅读
目的:探讨糖尿病肾病(DN)肾小管上皮细胞间充质转分化(EMT)进程中整合素连接激酶(ILK)/锌指转录因子(Snail)信号通路的表达情况及大黄酸对其的影响。方法:将8周龄的健康雄性Wistar大鼠，随机分为正常组、糖尿病肾病组、大黄酸组及缬沙坦组，每组各12只。使用链脲佐菌素(STZ)诱导糖尿病肾病模型，大黄酸组及缬沙坦组分别给予大黄酸100 mg/(kg·d)、缬沙坦30 mg/(kg·d)灌胃。于第8、16周末，以上四组大鼠各处死6只，原位灌洗肾脏，取出大鼠的肾脏组织后固定于蜡块并切片，使用HE及Masson染色分别对肾小管间质损伤指数、间质胶原相对面积进行评价；免疫组织化学方法测定E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、ILK、Snail及基质金属蛋白酶-2(MMP-2)的表达并作半定量分析。结果:与正常组比较，糖尿病肾病组大鼠肾小管间质损伤指数升高、肾间质胶原相对面积增加，肾小管上皮细胞E-cadherin表达下调、α-SMA表达上调(n P<0.05)。与糖尿病肾病组比较，大黄酸组、缬沙坦组肾小管间质损伤指数下降、肾间质胶原相对面积减少，肾小管上皮细胞E-cadherin表达上调、α-SMA表达下调(n P0.05)。与正常组比较，糖尿病肾病大鼠肾小管上皮细胞ILK、Snail及MMP-2的表达均随病情发生进行性升高(n P<0.05)。与糖尿病肾病组比较，大黄酸组、缬沙坦组ILK、Snail及MMP-2的表达均有所下降(n P0.05)。n 结论:大黄酸可通过下调DN大鼠肾小管上皮细胞ILK/Snail信号通路的表达阻抑EMT的进展。“,”Objective:To investigate the expression of integrin-linked kinase (ILK)/zinc finger transcription factor (Snail) signaling pathway in renal tubular epithelial-mesenchymal transition (EMT) in diabetic nephropathy (DN) and the effect of rhein.Methods:Healthy male Wistar rats of 8 weeks old were randomly divided into normal group, diabetic nephropathy group, rhein intervention group and valsartan intervention group, with 12 rats in each group. Streptozotocin (STZ) was used to induce the diabetic nephropathy model, then rhein intervention group and valsartan intervention group were given rhein 100 mg/(kg·d) and valsartan 30 mg/(kg·d), respectively. At the end of the 8th and 16th week, six rats of each group were killed, in situ lavage kidney, take out the kidney tissue of rats after fixed in wax block and slices. Renal tubular interstitial damage index and the relative area of interstitial collagen evaluated by hematoxylin-eosin (HE) and Masson staining respectively. The protein expression of E-cadherin, α-smooth muscle actin (α-SMA), ILK, Snail and matrix metalloproteinase-9 (MMP-2) in renal tubular epithelial cells were detected by immunohistochemistry.Results:Comparing to normal group, the renal tubular interstitial damage index and relative area of renal interstitial collagen of diabetic nephropathy rats were both increased. The expression of E-cadherin in renal tubular epithelial cells decreased and the expression of α-SMA significantly increased ( n P<0.05). Comparing with diabetic nephropathy group, in rhein and valsartan intervention groups, the expression of E-cadherin in renal tubular epithelial cells increased, while the expression of α-SMA significantly decreased (n P0.05). Compared to normal group, the expressions of ILK, Snail, MMP-2 increased progressively with the disease (n P<0.05). Compared with diabetic nephropathy group, rhein and valsartan intervention groups showed significant decrease in expression of ILK, Snail, MMP-2 (n P0.05).n Conclusions:Rhein could inhibit EMT progression by down-regulating the expression of ILK/Snail signaling pathway in renal tubular epithelial cells of DN rats.