Research Paper Volume 16, Issue 1 pp 106—128
Jian-Pi-Yi-Shen formula alleviates renal fibrosis by restoring NAD+ biosynthesis in vivo and in vitro
- 1 Department of Nephrology, Shenzhen Traditional Chinese Medicine Hospital, Guangzhou University of Chinese Medicine, Shenzhen, Guangdong 518033, China
- 2 The Fourth Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, Guangdong 518033, China
- 3 Shenzhen Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Shenzhen, Guangdong 518033, China
Received: May 30, 2023 Accepted: November 15, 2023 Published: December 28, 2023
https://doi.org/10.18632/aging.205352How to Cite
Copyright: © 2024 Gao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
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.
Abbreviations
α-SMA: α-smooth muscle actin; AKI: Acute kidney injury; ATP: Adenosine triphosphate; BUN: Blood urea nitrogen; CKD: Chronic kidney disease; Col-IV: Type IV collagen; FAO: Fatty acid oxidation; FN: Fibronectin; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; HK-2: The human renal proximal tubular epithelial cell; JPYSF: Jian-Pi-Yi-Shen Formula; NA: Nicotinic acid; NAD+: Nicotinamide adenine dinucleotide; NAM: Nicotinamide; NAMPT: Nicotinamide phosphoribosyl transferase; NAPRT1: Nicotinic acid phosphoribosyl transferase 1; NMN: Nicotinamide mononucleotide; NMNAT1: Nicotinamide mononucleotide adenylyltransferase 1; NR: Nicotinamide riboside; NRK1: Nicotinamide riboside kinase 1; QPRT: Quinolinate phosphoribosyl transferase; ROS: Reactive oxygen species; Scr: Serum creatinine; TGF-β1: Transforming growth factor-beta 1.