Fan Yan-Jun, Fang Jing-Ai, Li Su-Fen, Liu Ting, Liu Wen-Yuan, Hu Ya-Ling, Wang Rui-Hua, Li Hui, Sun Da-Lin, Zhang Guang, Zhang Zi-Yuan
Department of Nephrology, the First Hospital of Shanxi Medical University, Taiyuan, 030001, China.
Department of Pathology, the First Hospital of Shanxi Medical University, Taiyuan, 030001, China.
Chin J Integr Med. 2025 May 26. doi: 10.1007/s11655-025-3829-6.
To explore the mechanism of colon dialysis with Yishen Decoction (YS) in improving the autophagy disorder of intestinal epithelial cells in chronic renal failure (CRF) in vivo and in vitro.
Thirty male SD rats were randomly divided into normal, CRF, and colonic dialysis with YS groups by a random number table method (n=10). The CRF model was established by orally gavage of adenine 200 mg/(kg•d) for 4 weeks. CRF rats in the YS group were treated with colonic dialysis using YS 20 g/(kg•d) for 14 consecutive days. The serum creatinine (SCr) and urea nitrogen (BUN) levels were detected by enzyme-linked immunosorbent assay. Pathological changes of kidney and colon tissues were observed by hematoxylin and eosin staining. Autophagosome changes in colonic epithelial cells was observed with electron microscopy. In vitro experiments, human colon cancer epithelial cells (T84) were cultured and divided into normal, urea model (74U), YS colon dialysis, autophagy activator rapamycin (Ra), autophagy inhibitor 3-methyladenine (3-MA), and SIRT1 activator resveratrol (Re) groups. RT-PCR and Western blot were used to detect the mRNA and protein expressions of zonula occludens-1 (ZO-1), Claudin-1, silent information regulator sirtuin 1 (SIRT1), LC3, and Beclin-1 both in vitro and in vivo.
Colonic dialysis with YS decreased SCr and BUN levels in CRF rats (P<0.05), and alleviated the pathological changes of renal and colon tissues. Expressions of SIRT1, ZO-1, Claudin-1, Beclin-1, and LC3II/I were increased in the YS group compared with the CRF group in vivo (P<0.05). In in vitro study, compared with normal group, the expressions of SIRT1, ZO-1, and Claudin-1 were decreased, and expressions of Beclin-1, and LC3II/I were increased in the 74U group (P<0.05). Compared with the 74U group, expressions of SIRT1, ZO-1, and Claudin-1 were increased, whereas Beclin-1, and LC3II/I were decreased in the YS group (P<0.05). The treatment of 3-MA and rapamycin regulated autophagy and the expression of SIRT1. SIRT1 activator intervention up-regulated autophagy as well as the expressions of ZO-1 and Claudin-1 compared with the 74U group (P<0.05).
Colonic dialysis with YS could improve autophagy disorder and repair CRF intestinal mucosal barrier injury by regulating SIRT1 expression in intestinal epithelial cells.
探讨益肾汤结肠透析改善慢性肾衰竭（CRF）大鼠体内及体外肠上皮细胞自噬紊乱的机制。
将30只雄性SD大鼠按随机数字表法随机分为正常组、CRF组和益肾汤结肠透析组（n = 10）。采用腺嘌呤200 mg/(kg•d)灌胃4周建立CRF模型。益肾汤组CRF大鼠采用20 g/(kg•d)益肾汤进行结肠透析，连续14天。采用酶联免疫吸附测定法检测血清肌酐（SCr）和尿素氮（BUN）水平。苏木精-伊红染色观察肾和结肠组织的病理变化。电子显微镜观察结肠上皮细胞自噬体变化。体外实验，培养人结肠癌上皮细胞（T84），分为正常组、尿素模型组（74U）、益肾汤结肠透析组、自噬激活剂雷帕霉素（Ra）组、自噬抑制剂3-甲基腺嘌呤（3-MA）组和SIRT1激活剂白藜芦醇（Re）组。采用RT-PCR和蛋白质免疫印迹法检测体内外紧密连接蛋白1（ZO-1）、闭合蛋白1（Claudin-1）、沉默信息调节因子sirtuin 1（SIRT1）、微管相关蛋白1轻链3（LC3）和Beclin-1的mRNA和蛋白表达。
益肾汤结肠透析可降低CRF大鼠的SCr和BUN水平（P<0.05），减轻肾和结肠组织的病理变化。与CRF组相比，益肾汤组体内SIRT1、ZO-1、Claudin-1、Beclin-1和LC3II/I的表达增加（P<0.05）。体外研究中，与正常组相比，74U组SIRT1、ZO-1和Claudin-1的表达降低，Beclin-1和LC3II/I的表达增加（P<0.05）。与74U组相比，益肾汤组SIRT1、ZO-1和Claudin-1的表达增加，而Beclin-1和LC3II/I的表达降低（P<0.05）。3-MA和雷帕霉素处理可调节自噬及SIRT1的表达。与74U组相比，SIRT1激活剂干预上调了自噬以及ZO-1和Claudin-1的表达（P<0.05）。
益肾汤结肠透析可通过调节肠上皮细胞中SIRT1的表达改善自噬紊乱，修复CRF肠黏膜屏障损伤。
