Yan Xiaotong, Li Ping, Liu Chang, Yin Fengting, Han Jinwei, Sun Hui, Zheng Ying, Chen Xiangmei, Guan Shihan, Wang Xijun
State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine, National Chinmedomics Research Center, National TCM Key Laboratory of Serum Pharmacochemistry, Metabolomics Laboratory, Department of Pharmaceutical Analysis, Heilongjiang University of Chinese Medicine, Heping Road 24, Harbin, 150040, China.
Department of Nephrology, First Medical Center of Chinese PLA General Hospital, Nephrology Institute of the Chinese People's Liberation Army, National Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Disease Research, Beijing, 100853, China.
J Ethnopharmacol. 2025 Jan 31;340:119223. doi: 10.1016/j.jep.2024.119223. Epub 2024 Dec 18.
Huangkui capsule (HKC), a patent traditional Chinese medicine, has shown significant efficacy in managing chronic kidney disease (CKD), particularly diabetic nephropathy (DN). Previous studies have shown that HKC can alleviate kidney damage in DN. However, the exact mechanisms through which it exerts its effects remain unclear.
This study aimed to elucidate the potential molecular mechanisms of HKC in treating kidney injury in type 1 diabetic nephropathy (T1DN) models through serum metabolomics, Chinmedomics, and molecular docking techniques.
T1DN mouse models were induced by intraperitoneal injection of streptozotocin (STZ), resulting in the ACR value ten times that of the control group. The efficacy of HKC on T1DN was comprehensively evaluated in general conditions, renal coefficient, histopathology, and related biochemical indicators. UPLC-Q-TOF-MS/MS based serum metabolomics was employed to identify biomarkers of T1DN and evaluate the effects of HKC. Relevant pathways were analyzed, and followed by Protein-Protein Interaction network analysis to screen for key enzymes. By integrating the Chinmedomics strategy and molecular docking the relationship between these targets and active components was elucitaed.
HKC resulted in a significant reduction in renal inflammation and fibrosis, as evidenced by the decreased levels of urinary ACR, blood TG, T-CHO, BUN, and renal TNF-α and VEGF-A, along with a reduction in the positive area of COL-1. Palmitic acid, stearic acid, arachidonic acid, pantothenic acid, and sphingosine-1-phosphate serve as key serum metabolite biomarkers for T1DN, involved in the biosynthesis of unsaturated fatty acids, arachidonic acid metabolism, pantothenate and CoA biosynthesis, and sphingolipid metabolism. FASN, Cyp2e1, and Cyp4a32 are the key enzymes in the treatment of T1DN with HKC. Additionally, 8 key active components were identified in the serum of HKC-H, including quercetin, myricetin, isoquercitrin, hyperoside, hibifolin, gentisic acid 5-O-β-glucoside, floramanoside F, and quercetin-4'-O-glucoside, which are believed to interact with key enzymes.
The active components of HKC influence Fasn, Cyp2e1, and Cy4a32, improving renal injury in T1DN. These findings provide new molecular insights for the future clinical application and research of HKC in treating T1DN.
黄葵胶囊（HKC）是一种专利中药，在治疗慢性肾脏病（CKD），尤其是糖尿病肾病（DN）方面已显示出显著疗效。先前的研究表明，HKC可减轻DN中的肾损伤。然而，其发挥作用的确切机制仍不清楚。
本研究旨在通过血清代谢组学、中药组学和分子对接技术阐明HKC治疗1型糖尿病肾病（T1DN）模型肾损伤的潜在分子机制。
通过腹腔注射链脲佐菌素（STZ）诱导T1DN小鼠模型，使其尿白蛋白肌酐比值（ACR）值为对照组的10倍。从一般状况、肾系数、组织病理学和相关生化指标等方面综合评价HKC对T1DN的疗效。采用基于超高效液相色谱-四级杆-飞行时间串联质谱（UPLC-Q-TOF-MS/MS）的血清代谢组学方法鉴定T1DN的生物标志物并评估HKC的作用效果。分析相关通路，随后进行蛋白质-蛋白质相互作用网络分析以筛选关键酶。通过整合中药组学策略和分子对接阐明这些靶点与活性成分之间的关系。
HKC可显著减轻肾脏炎症和纤维化，表现为尿ACR、血甘油三酯（TG）、总胆固醇（T-CHO）、尿素氮（BUN）水平以及肾组织肿瘤坏死因子-α（TNF-α）和血管内皮生长因子-A（VEGF-A）水平降低，同时Ⅰ型胶原蛋白（COL-1）阳性面积减小。棕榈酸、硬脂酸、花生四烯酸、泛酸和鞘氨醇-1-磷酸是T1DN的关键血清代谢物生物标志物，参与不饱和脂肪酸生物合成、花生四烯酸代谢、泛酸和辅酶A生物合成以及鞘脂代谢。脂肪酸合酶（FASN）、细胞色素P450 2E1（Cyp2e1）和细胞色素P450 4A32（Cyp4a32）是HKC治疗T1DN的关键酶。此外，在HKC-H血清中鉴定出8种关键活性成分，包括槲皮素、杨梅素、异槲皮苷、金丝桃苷、山奈酚-3-O-葡萄糖苷、龙胆酸5-O-β-葡萄糖苷、花旗松素F和槲皮素-4'-O-葡萄糖苷，它们被认为与关键酶相互作用。
HKC的活性成分影响Fasn、Cyp2e1和Cy4a32，改善T1DN中的肾损伤。这些发现为HKC未来治疗T1DN的临床应用和研究提供了新的分子见解。
