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剪切型X盒结合蛋白1(spliced X-box binding protein 1,XBP1S)是内质网应激关键信号分子。有研究表明XBP1S在肾脏系膜细胞具有抗氧化应激和抗凋亡作用,而氧化应激是肾纤维化发生发展重要因素。缬沙坦(valsartan)具有改善肾纤维化作用。本研究旨在探讨valsartan能否通过XBP1S进而影响肾间质纤维化发展。C57BL/6J小鼠行左侧输尿管结扎(unilateral ureteral obstruction,UUO)制作肾纤维化模型,valsartan组于造模前1 d起每天给予valsartan(20 mg/kg)灌胃,UUO组和对照组给予等容积生理盐水灌胃,于手术后第7天处死动物,取左侧肾。HE、Masson和天狼星红(Sirius red)染色观察肾间质纤维化;免疫组化染色观察XBP1S在肾间质表达;Western blot检测XBP1S、纤维连接蛋白(fibronectin)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、BAX和BCL2蛋白水平;实时荧光定量PCR检测NADPH氧化酶亚基p47-phox与p67-phox m RNA水平。结果显示,与对照组相比,UUO组XBP1S蛋白水平明显降低,valsartan明显升高UUO小鼠XBP1S蛋白水平;HE、Masson和Sirius red染色结果显示valsartan明显改善UUO小鼠肾间质纤维化;Western blot结果显示valsartan明显降低UUO小鼠fibronectin、BAX/BCL2、α-SMA蛋白水平。实时荧光定量PCR结果显示:UUO组p47-phox与p67-phox m RNA水平明显较对照组升高,相比较于UUO组,valsartan明显降低UUO小鼠p47-phox与p67-phox m RNA水平。以上结果提示,XBP1S蛋白水平下调与UUO模型的肾间质纤维化有关,其机制可能与XBP1S抗氧化应激相关。
The spliced X-box binding protein 1 (XBP1S) is a key signaling molecule of endoplasmic reticulum stress. Some studies have shown that XBP1S in renal mesangial cells with anti-oxidative stress and anti-apoptotic effect, and oxidative stress is an important factor in the development of renal fibrosis. Valsartan improves renal fibrosis. This study aimed to investigate whether valsartan can affect the development of renal interstitial fibrosis through XBP1S. C57BL / 6J mice were subjected to left unilateral ureteral obstruction (UUO) to establish a model of renal fibrosis. The valsartan group was orally administered with valsartan (20 mg / kg) once a day prior to model establishment. The UUO group and the control group Equal volume of normal saline gavage, the animals were sacrificed on the 7th day after surgery, the left kidney. The expression of XBP1S in renal interstitium was observed by immunohistochemical staining. The expressions of XBP1S, fibronectin, α-smooth muscle actin (α -smooth muscle actin, α-SMA), BAX and BCL2 protein levels; real-time fluorescent quantitative PCR detection of NADPH oxidase subunits p47-phox and p67-phox m RNA levels. The results showed that compared with the control group, the level of XBP1S protein in UUO group was significantly decreased and the valsartan significantly increased the level of XBP1S protein in UUO mice. The results of HE, Masson and Sirius red staining showed that valsartan significantly improved renal interstitial fibrosis in UUO mice. blot results showed that valsartan significantly reduced the fibronectin, BAX / BCL2 and α-SMA protein levels in UUO mice. Real-time PCR results showed that the levels of p47-phox and p67-phox m RNA in UUO group were significantly higher than those in control group. Compared with UUO group, valsartan significantly reduced the levels of p47-phox and p67-phox m RNA in UUO mice. The above results suggest that the down-regulation of XBP1S protein is associated with renal interstitial fibrosis in UUO model, which may be related to the anti-oxidative stress of XBP1S.