摘要
目的:探讨细胞自噬与上皮间质转化(epithelial-to-mesenchymal transition,EMT)之间的关系,以及自噬是否可以通过调节EMT进程影响梗阻性肾病肾纤维化进展。方法:构建大鼠单侧输尿管梗阻(unilateral ureteral obstruction,UUO)肾纤维化模型,并将实验大鼠随机分为假手术组、UUO组、UUO+低剂量雷帕霉素组、UUO+高剂量雷帕霉素组。采用HE染色观察肾结构,Masson染色观察肾间质胶原沉积;采用蛋白质印迹法检测钙黏附蛋白E(E-cadherin)、α平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)、锌指转录因子1(Snail 1)表达以反映细胞EMT,检测微管相关蛋白1轻链3II(microtubule-associated protein-1 light chain 3II,LC3II)表达以反映细胞自噬。构建转化生长因子-β1(transforming growth factor-β1,TGF-β1)处理的大鼠肾小管上皮细胞NRK52E模型,并将其分为对照组、TGF-β1组、TGF-β1+Snail 1 siRNA组;为探讨自噬对EMT的影响,再将细胞分为对照组、雷帕霉素组、Beclin 1 siRNA组。采用蛋白质印迹法检测E-cadherin、α-SMA、Snail 1、LC3II、I型胶原(collagen I)和纤维黏连蛋白(fibronectin)的表达。结果:HE染色结果显示:与假手术组比较,UUO组的肾损伤明显加重;Masson染色结果显示:与假手术组比较,UUO组的肾组织中胶原沉积明显增加;与UUO组比较,UUO+高剂量雷帕霉素组和UUO+低剂量雷帕霉素组胶原沉积明显减轻。蛋白质印迹法结果显示:与假手术组比较,UUO组肾组织中E-cadherin水平明显降低,α-SMA及LC3II表达水平明显升高(均P<0.05);与UUO组比较,UUO+高剂量雷帕霉素组和UUO+低剂量雷帕霉素组Ecadherin和LC3II表达水平均明显升高(分别P<0.01和P<0.05),α-SMA表达水平明显降低(分别P<0.01和P<0.05)。TGF-β1组中Snail 1、α-SMA、collagen I和fibronectin表达水平均显著高于对照组,E-cadherin水平明显低于对照组(均P<0.05)。与TGF-β1组比较,TGF-β1+Snail 1 siRNA组E-cadherin的表达显著增加,α-SMA、 collage
Objective: To explore the relationship between autophagy and epithelial-to-mesenchymal transition(EMT), and to evaluate whether autophagy can affect the progression of renal fibrosis in obstructive nephropathy by regulating the EMT process.Methods: Unilateral ureteral obstruction(UUO) renal fibrosis model of rat was constructed, and the animals were divided into a sham group, an UUO group, an UUO+low-dose rapamycin group, and an UUO+high-dose rapamycin group. HE staining was used to observe the structure of the kidney, and Masson staining was used to observe renal interstitial collagen deposition. The expressions of E-cadherin, alpha-smooth muscle actin(α-SMA), Snail 1, and microtubule-associated protein-1 light chain 3 II(LC3 II) were detected by Western blotting, reflecting cellular EMT and autophagy. Transforming growth factor β1(TGF-β1) induced-NRK52 E cells model was constructed, and the cells were divided into a control group, a TGF-β1 group, and a TGF-β1+ Snail 1 siRNA group. To explore the effect of autophagy on EMT, the cells were also divided into a control group, a rapamycin group, and a Beclin 1 siRNA group. Western blotting was used to detect the expressions of E-cadherin, α-SMA, Snail 1, LC3II, collagen I, and fibronectin.Results: Compared with the sham group, the kidney damage in the UUO group was significantly worse;compared with the sham group, the collagen deposition in the kidney tissues in the UUO group was significantly increased, which were significantly reduced in the UUO+high-dose rapamycin group and the UUO+low-dose rapamycin group compared with the UUO group;compared with the sham group, the E-cadherin level in the kidney of the UUO group was significantly reduced, and the expression levels of α-SMA and LC3 II were significantly increased(all P<0.05). Compared with the UUO group, the expression levels of E-cadherin and LC3II in the UUO+high-dose rapamycin group and the UUO+lowdose rapamycin group were significantly increased(P<0.01 and P<0.05, respectively), and the expression lev
作者
张波
汝峰
陈湘
陈志
ZHANG Bo;RU Feng;CHEN Xiang;CHEN Zhi(Department of Urology,Xiangya Hospital,Central South University,Changsha 410008,China)
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2021年第6期601-608,共8页
Journal of Central South University ：Medical Science
基金
湖南省自然科学基金(2020JJ4870)。
关键词
自噬
上皮间质转化
肾纤维化
autophagy
epithelial-to-mesenchymal transition
renal fibrosis
