南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (6): 924-934.doi: 10.12122/j.issn.1673-4254.2023.06.07

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肾病III号方治疗慢性肾脏病大鼠肾脏纤维化的作用机制:基于网络药理学和分子对接技术

罗冠峰，刘华熙，谢 钡，邓伊健，谢鹏辉，赵晓山，孙晓敏   

  1. 南方医科大学珠江医院中医科，广东 广州 510280；南方医科大学中医药学院，广东 广州 510515
  • 出版日期:2023-06-20 发布日期:2023-07-06

Therapeutic mechanism of Shenbing Decoction III for renal fibrosis in chronic kidney disease: a study with network pharmacology, molecular docking and validation in rats

LUO Guanfeng, LIU Huaxi, XIE Bei, DENG Yijian, XIE Penghui, ZHAO Xiaoshan, SUN Xiaomin   

  1. Department of Traditional Chinese Medicine, Zhujiang Hospital Affiliated to Southern Medical University, Guangzhou 510280, China; School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China
  • Online:2023-06-20 Published:2023-07-06

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摘要： 目的 观察肾病III号方对5/6肾切除大鼠肾功能、肾脏病理的改善作用及联合网络药理学和分子对接技术分析肾病III号方抗慢性肾脏病肾脏纤维化的作用机制。方法 SPF级雄性SD大鼠40只，从中随机选取10只作为假手术组，30只制作5/6肾切除模型，造模2周后测血清肌酐，并将造模组随机分为肾病III号方组、氯沙坦组、模型组。肾病III号方组予肾病III号方灌胃，氯沙坦组予氯沙坦钾灌胃，模型组与假手术组予生理盐水灌胃，给药16周后测定血清肌酐、尿素氮，HE染色及免疫印迹法观察肾脏病理学改变和纤维化相关因子。网络药理学联合分子对接技术探索肾病III号方抗慢性肾脏病肾脏纤维化的作用机制，免疫印迹法验证核心靶点表达。结果 与模型组比较，肾病III号方治疗后，5/6肾切除大鼠的血清肌酐、尿素氮降低（P<0.05），肾组织的病理损害减轻，且肾组织中上皮间充质转化相关蛋白经治疗后也得到相应改善。进一步通过网络药理学分析，发现肾病III号方主要活性成分有金合欢素、芹菜素、异泽兰黄素、槲皮素、山奈酚、木樨草素等，关键靶点为STAT3、SRC、CTNNB1、PIK3R1、AKT1等。分子对接结果显示，肾病III号方的活性成分与STAT3、SRC、CTNNB1、PIK3R1、AKT1靶点都有良好的结合活性。免疫印迹法验证发现肾病III号方能恢复5/6肾切除大鼠肾组织中STAT3、PI3K、AKT的蛋白表达（P<0.05）。结论 肾病III号方能减轻5/6肾切除大鼠所致的肾脏损伤，其主要的药效成分可能通过调控STAT3、PIK3R1、AKT1等多靶点发挥抗慢性肾脏病肾脏纤维化的作用。

关键词: 肾病III号方；5/6肾切除；肾脏纤维化；网络药理学；分子对接技术

Abstract: Objective To observe the effect of Shenbing Decoction III for improving renal function and pathology in rats with 5/6 nephrectomy and analyze its therapeutic mechanism for renal fibrosis in chronic kidney disease using network pharmacology combined with molecular docking. Methods Forty male SD rats were randomized into two groups to receive two-staged 5/6 nephrectomy (n=30) or sham operation (n=10), and 2 weeks after the final operation, serum creatinine level of the rats was measured. The rats with nephrectomy were further randomized into Shenbing Decoction III group, losartan group and model group for daily treatment with the corresponding drugs via gavage starting at 1 week after 5/6 nephrectomy. After 16 weeks of treatment, serum creatinine and urea nitrogen levels of the rats were measured, and HE staining and Western blotting were used to examine the changes in renal pathology and fibrosis-related factors. Network pharmacology combined with molecular docking study was performed to explore the therapeutic mechanism Shenbing Decoction III against renal fibrosis in chronic kidney disease, and Western blotting was used to verify the expressions of the core targets. Results Compared with those in the model group, the rats receiving 5/6 nephrectomy and Shenbing Decoction III treatment showed significantly reduced serum creatinine and urea nitrogen levels, lessened renal pathologies, and improvement of the changes in epithelial mesenchymal transition-related proteins. Network pharmacological analysis showed that the main active ingredients of Shenbing Decoction III were acacetin, apigenin, eupatilin, quercetin, kaempferol and luteolin, and the key targets included STAT3, SRC, CTNNB1, PIK3R1 and AKT1. Molecular docking study revealed that the active ingredients of Shenbing Decoction III had good binding activity to the key targets. Western blotting showed that in rats with 5/6 nephrectomy, treatment with Shenbing Decoction III obviously restored the protein expression of STAT3, PI3K, and AKT in renal tissue. Conclusion Shenbing Decoction III can reduce renal injury induced by 5/6 nephrectomy in rats, and its therapeutic effects are mediated possibly by its main pharmacologically active ingredients that alleviate renal fibrosis via modulating multiple targets including STAT3, PIK3R1, and AKT1.

Key words: Shenbing Decoction III; 5/6 nephrectomy; renal fibrosis; network pharmacology; molecular docking techniques