Jia Qi, Zhang Xiaoyu, Hao Gaimei, Zhao Yun, Lowe Scott, Han Lin, Qin Jianguo
Department of Nephropathy, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing, China.
Institute of Basic Theory for Chinese Medicine, China Academy of Chinese Medical Sciences, Beijing, China.
Front Pharmacol. 2022 Jul 6;13:936853. doi: 10.3389/fphar.2022.936853. eCollection 2022.
In this study, we investigated the mechanism of Tongluo Yishen (TLYS) decoction in more detail, from the perspective of pyroptosis in the unilateral ureteral ligation (UUO) model and the hypoxia-induced renal tubular epithelial (NRK-52E) cell. The UUO model was used, and after 14 days of TLYS intervention, rats were tested for blood creatinine and urea nitrogen, HE staining was used to observe the pathological changes in the kidney, Masson staining was used to assess the degree of interstitial fibrosis, western blot was used to detect the changes of α-smooth muscle actin (α-SMA) protein expression level, immunohistochemistry and western blot detected the changes in protein expression levels of NOD-like receptor protein 3 inflammasome (NLRP3), gasdermin D (GSDMD), cysteinyl aspartate specific proteinase (caspase-1), interleukin 18 (IL-18) and interleukin 1β (I L-1β). A hypoxia model was created using NRK-52E cell, and after different concentrations of TLYS decoction intervention, the changes in the expression levels of pyroptosis were used with immunofluorescence and western blot methods. TLYS decoction improved renal function, delayed the advancement of renal interstitial fibrosis, and inhibited pyroptosis in UUO rats. Furthermore, we observed that TLYS can mitigate hypoxia-induced NRK-52E cell damage via the suppression of the NLRP3-mediated pyroptosis. TLYS decoction exert renoprotective effects by inhibiting NLRP3-mediated pyroptosis.
在本研究中，我们从单侧输尿管结扎（UUO）模型中的细胞焦亡以及缺氧诱导的肾小管上皮（NRK-52E）细胞的角度，更详细地研究了通络益肾（TLYS）汤的作用机制。采用UUO模型，在TLYS干预14天后，检测大鼠的血肌酐和尿素氮，用苏木精-伊红（HE）染色观察肾脏的病理变化，用Masson染色评估间质纤维化程度，用蛋白质印迹法检测α平滑肌肌动蛋白（α-SMA）蛋白表达水平的变化，用免疫组织化学和蛋白质印迹法检测NOD样受体蛋白3炎性小体（NLRP3）、gasdermin D（GSDMD）、半胱天冬酶-1（caspase-1）、白细胞介素18（IL-18）和白细胞介素1β（IL-1β）蛋白表达水平的变化。利用NRK-52E细胞建立缺氧模型，在不同浓度的TLYS汤干预后，用免疫荧光和蛋白质印迹法检测细胞焦亡表达水平的变化。TLYS汤改善了肾功能，延缓了肾间质纤维化的进展，并抑制了UUO大鼠的细胞焦亡。此外，我们观察到TLYS可以通过抑制NLRP3介导的细胞焦亡减轻缺氧诱导的NRK-52E细胞损伤。TLYS汤通过抑制NLRP3介导的细胞焦亡发挥肾脏保护作用。
