Jian-Pi-Yi-Shen formula alleviates renal fibrosis by restoring NAD+ biosynthesis in vivo and in vitro.
Jian-Pi-Yi-Shen formula alleviates renal fibrosis by restoring NAD+ biosynthesis in vivo and in vitro.
复制标题
健脾益肾方通过恢复体内、外NAD生物合成，减轻肾纤维化。
DOI：
10.18632/aging.205352
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发表时间：
2023-12-28
期刊：
影响因子：
5.2
通讯作者：
Liu, Xinhui
中科院分区：
文献类型：
--
作者：
Gao, Liwen;Huang, Xi;Deng, Ruyu;Wu, Shanshan;Peng, Yu;Xiong, Guoliang;Lu, Jiandong;Liu, Xinhui
背景：慢性肾脏病（CKD）患者缺乏有效的治疗。健脾益肾方治疗慢性肾脏病已有数十年的临床疗效。然而，其肾脏保护机制尚未完全阐明。本研究旨在确定JPYSF是否可以通过恢复烟酰胺腺嘌呤二核苷酸（NAD+）的生物合成来延缓CKD中的肾纤维化进展。方法：采用腺嘌呤饲料喂养C57 BL/6小鼠，建立慢性肾脏病模型。JPYSF口服给药4周。用转化生长因子-β1（TGF-β1）刺激人近端肾小管上皮细胞（HK-2），同时加入或不加入JPYSF。以血清肌酐和血尿素氮水平评价小鼠肾功能。采用过碘酸-希夫和Masson三色染色法评价肾组织病理学变化。使用细胞计数试剂盒-8测定法评估细胞活力。通过NAD+/NADH测定试剂盒检测NAD+浓度。采用Western blotting、免疫组织化学和免疫荧光法检测CKD肾脏和TGF-β1诱导的HK-2细胞中纤维化相关蛋白和关键NAD+生物合成酶的表达。结果：健脾益肾方能明显改善CKD小鼠肾功能，减轻病理损伤，抑制肾纤维化。此外，JPYSF逆转TGF-β1诱导的HK-2细胞的纤维化反应。此外，JPYSF通过恢复NAD+生物合成中关键酶的表达，包括喹啉酸磷酸核糖基转移酶、烟酰胺单核苷酸腺苷酰转移酶1和烟酰胺核苷激酶1，挽救CKD小鼠和TGF-β1诱导的HK-2细胞中NAD+含量降低。结论：JPYSF可减轻CKD小鼠肾纤维化，逆转TGF-β1诱导的HK-2细胞纤维化反应，可能与恢复NAD+生物合成有关。
Background: Patients with chronic kidney disease (CKD) lack efficacious treatment. Jian-Pi-Yi-Shen formula (JPYSF) has demonstrated significant clinical efficacy in treating CKD for decades. However, its renoprotective mechanism has not been fully elucidated. This study aimed to determine whether JPYSF could delay renal fibrosis progression in CKD by restoring nicotinamide adenine dinucleotide (NAD+) biosynthesis. Methods: Adenine-diet feeding was used to model CKD in C57BL/6 mice. JPYSF was orally administered for 4 weeks. Human proximal tubular epithelial cells (HK-2) cells were stimulated with transforming growth factor-β1 (TGF-β1) with or without JPYSF treatment. Renal function of mice was assessed by serum creatinine and blood urea nitrogen levels. Renal histopathological changes were assessed using Periodic acid-Schiff and Masson’s trichrome staining. Cell viability was assessed using a cell counting kit-8 assay. NAD+ concentrations were detected by a NAD+/NADH assay kit. Western blotting, immunohistochemistry, and immunofluorescence were employed to examine fibrosis-related proteins and key NAD+ biosynthesis enzymes expression in the CKD kidney and TGF-β1-induced HK-2 cells. Results: JPYSF treatment could not only improve renal function and pathological injury but also inhibit renal fibrosis in CKD mice. Additionally, JPYSF reversed fibrotic response in TGF-β1-induced HK-2 cells. Moreover, JPYSF rescued the decreased NAD+ content in CKD mice and TGF-β1-induced HK-2 cells through restoring expression of key enzymes in NAD+ biosynthesis, including quinolinate phosphoribosyltransferase, nicotinamide mononucleotide adenylyltransferase 1, and nicotinamide riboside kinase 1. Conclusions: JPYSF alleviated renal fibrosis in CKD mice and reversed fibrotic response in TGF-β1-induced HK-2 cells, which may be related to the restoration of NAD+ biosynthesis.