Zhao Tingting, Li Minyi, Xiang Qian, Lie Beifeng, Chen Deqi, Wang Weiming, Li Xuling, Xu Tiancheng, Zhang Xi, Li Yuntong, Dong Ruixue, Du Xinwen, Wang Yilin, Yang Junzheng, He Bao, Zhu Quan, Duan Tingting, Li Zhenghai, Xu Youhua
State Key Laboratory of Quality Research in Chinese Medicines, Faculty of Chinese Medicine, Macau University of Science and Technology, Macao, China.
Institute of Consun Co., for Chinese Medicine in Kidney Diseases, Guangdong Consun Pharmaceutical Group, Guangzhou, China.
Front Pharmacol. 2022 Jul 22;13:872940. doi: 10.3389/fphar.2022.872940. eCollection 2022.
Diabetic nephropathy (DN) is one of the most common complications of diabetes and the primary cause of end-stage renal disease. At present, renin-angiotensin-aldosterone system (RAAS) blockers have been applied as first-class drugs to restrain development of DN; however, its long-term effect is limited. Recent evidence has shown definite effects of Chinese medicine on DN. Yishen Huashi (YSHS) granule is a traditional Chinese Medicine prescription that has been used in the clinic to treat DN, but its mechanism is not understood. In the present study, both and studies were carried out. The DN model was induced by STZ in Wistar rats, and GEnC and HPC cell lines were applied in the study. Quality of YSHS was evaluated by LC-MS/MS. A metabolomic study of urine was carried out by LC-MS; influence of YSHS on composition of DN was analyzed by network pharmacology. Mechanism of the YSHS on DN was analyzed by Q-PCR, Western Blot, and multi-immunological methods. We found YSHS administration significantly reduced levels of HbA1c and mALB. Histopathological analysis found that YSHS preserved integrity of glomerular filtration barrier by preserving viability of glomerular endothelial cells and podocytes, inhibiting glomerular fibrosis, reducing oxidative stress damage, and enhancing cross-talk among glomerular endothelial cells and podocytes. Network pharmacology, differential metabolite analysis, as well as intracellular pathway experimental study demonstrated that the PI3K/AKT/mTOR signaling pathway played a pivotal role in it. Our present findings supplied new understanding toward the mechanism of YSHS on inhibiting DN.
糖尿病肾病（DN）是糖尿病最常见的并发症之一，也是终末期肾病的主要病因。目前，肾素 - 血管紧张素 - 醛固酮系统（RAAS）阻滞剂已被用作抑制DN进展的一线药物；然而，其长期效果有限。最近的证据表明中药对DN有确切疗效。益肾化湿（YSHS）颗粒是一种已在临床上用于治疗DN的中药方剂，但其作用机制尚不清楚。在本研究中，我们进行了体内和体外研究。通过链脲佐菌素（STZ）诱导Wistar大鼠建立DN模型，并在体外研究中应用肾小球内皮细胞（GEnC）和人足细胞（HPC）细胞系。采用液相色谱 - 串联质谱法（LC-MS/MS）对YSHS进行质量评价。通过液相色谱 - 质谱法（LC-MS）对尿液进行代谢组学研究；采用网络药理学分析YSHS对DN成分的影响。通过实时定量聚合酶链反应（Q-PCR）、蛋白质免疫印迹法（Western Blot）和多种免疫学方法分析YSHS对DN的作用机制。我们发现给予YSHS可显著降低糖化血红蛋白（HbA1c）和微量白蛋白（mALB）水平。组织病理学分析发现，YSHS通过维持肾小球内皮细胞和足细胞的活力、抑制肾小球纤维化、减少氧化应激损伤以及增强肾小球内皮细胞和足细胞之间的相互作用，保持肾小球滤过屏障的完整性。网络药理学、差异代谢物分析以及细胞内信号通路实验研究表明，磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路在其中起关键作用。我们目前的研究结果为YSHS抑制DN的机制提供了新的认识。
