Abstract
The formula of Compound Centella mainly contains 3 traditional Chinese herbs: Centella asiatica (L.) Urb. (JiXueCao), Astragalus Membranaceus Fish. (HuangQi), and Tripterygium wilfordii Hook. f. (LeiGongTeng). Though this formula is effective for treating diabetic kidney disease (DKD) in clinic, the exact mechanism is still unclear. This study aims to investigate the effect and antioxidant mechanism of Compound Centella on DKD rats. High-performance liquid chromatography (HPLC) was applied to analyse 3 herbs in Compound Centella. Sprague-Dawley rats were divided into the normal group (NG), DKD group (DKDG), Compound Centella group (CCG), and losartan group (LG), with 8 rats in each group. On the first day of the experiment, rats in the NG were fed with ordinary –feed, while the other groups were fed with high-fat and high-sugar feed. On the 29th day, except the NG, the other 3 groups received a single intraperitoneal injection of streptozocin (STZ, 35 mg/kg). Fasting blood glucose (FBG) was measured on the 1st day, 32nd day, 46th day, 56th day, 84th day, and 112th day. Total protein/creatinine ratio of urine was determined by the phenol red assay on the 1st day and 112th day. Serum creatinine (Scr) was determined by an automatic biochemical analyser on the 112th day. Kidneys were collected on the 112th day for analysis and evaluation. Periodic acid-Schiff (PAS) staining, hematoxylin-eosin (HE) staining, and transmission electron microscopy were used to observe kidney pathological changes. The mRNA and protein expressions of Kelch-like ECH-associated protein 1 (Keap1) and nuclear factor-erythroid 2-related factor 2 (Nrf2) in renal tissues were detected by RT-qPCR, Western blot, and immunohistochemistry. Oxidative stress was evaluated by detecting the levels of malondialdehyde (MDA) and heme oxidase-1 (HO-1). The results showed that the content of asiaticoside, astragaloside, and triptolide in the herb was 5960, 809, and 2.42 μg/g and in the Compound Centella was 340, 64, and 0.1403 μg/mL by HPLC. Compound Centella reduced the urine protein/creatinine ratio and improved renal pathology in the kidneys of DKD rats. In addition, the mRNA and protein expressions of Keap1 and Nrf2 in kidneys were upregulated by Compound Centella. The expressions of MDA were downregulated but HO-1 were upregulated by Compound Centella. Therefore, the protective effect of Compound Centella in the kidney of DKD rats may be related to regulating the Keap1-Nrf2-ARE pathway under oxidative stress, suggesting Compound Centella as a promising treatment against DKD.
Highlights
Diabetic kidney disease (DKD) is one of the most serious chronic complications of diabetes
USA, CAS no. 18883-66-4; Kelch-like ECH-associated protein 1 (Keap1) antibody was bought from Proteintech Group, Wuhan, China, no. 13161-1-ap; nuclear factor-erythroid 2-related factor 2 (Nrf2) antibody was purchased from Proteintech Group, Wuhan, China, no. 16396-1-ap; heme oxidase-1 (HO-1) antibody was purchased from Proteintech Group, Wuhan, China, no. 107011-ap; the malondialdehyde (MDA) test box was purchased
The incidence of dialysis has stabilized both in the general population and in diabetics, but in whom it remains far higher by comparison [7]. erefore, it is important to clarify the pathogenesis of DKD as early as possible and to find effective methods to delay the progress of DKD
Summary
Diabetic kidney disease (DKD) is one of the most serious chronic complications of diabetes. With the development of medicine, there are more and more ways to treat DKD, including lifestyle adjustment, glycemic control, blood pressure control, renin-angiotensin system (RAS), sodium-dependent glucose transporter-2 (SLGT-2) inhibitors, and glucagonlike peptide-1 (GLP-1) agonists [2]. In China, traditional Chinese medicine (TCM) has been widely used in treating diabetes and its complications for many years. TCM gets more attention from other countries and is becoming a promising new therapeutic agent for DKD [3]. One of such medicines is the Compound Centella. One of such medicines is the Compound Centella. is formula is founded by Professor Yongjun Wang, a famous contemporary doctor of TCM
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