Cai Hong-Die, Su Shu-Lan, Qian Da-Wei, Guo Sheng, Tao Wei-Wei, Cong Xu Dong, Tang Renmao, Duan Jin-Ao
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China; Center for Translational Systems Biology and Neuroscience, Laboratory of Integrative Biomedicine of Brain Diseases, School of Basic Biomedical Science, Nanjing University of Chinese Medicine, Nanjing 210023, China.
J Ethnopharmacol. 2017 Jul 12;206:152-159. doi: 10.1016/j.jep.2017.02.046. Epub 2017 Apr 10.
The flower of Abelmoschus manihot (Linn.) Medicus (A. manihot), as a traditional Chinese Herbal medicine, was used widely in China with efficacy of inducing diuresis for treating strangurtia, and subdhing swelling and detoxicating. It has been reported that Huangkui capsule, prepared by the extract of the flower of A. manihot, can reduce the content of urinary protein, serum creatinine and serum urea nitrogen in nephropathy rats and processes renoprotective activity, while the action mechanism need to illuminate deeply.
In this study, we investigated the protection effect of Huangkui capsule on tubulointerstitial fibrosis in chronic renal failure (CRF) rats and its mechanism against high glucose-induced epithelial to mesenchymal transition (EMT) in renal tubular epithelial cells (HK-2) of its bioactive components.
The animals were divided into normal group, CRF model group and Huangkui capsule-treated group. Hematoxylin eosin (HE) staining and Masson staining were applied to observe pathological changes in renal tissue of different groups. Biochemical indicators including serum urea nitrogen (BUN), urine protein (UP) and serum creatinine (Scr) were measured according to the manufacturer's instructions of kits. HK-2 cell damaged model was established to access the protection effect and action mechanism of five main flavonoids from Huangkui capsule. The experimental cells were divided into eight groups: control group, model group, positive drug group and five main flavonoids treated groups. The dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used to determine the reactive oxygen species (ROS) in different groups. Western blot was applied to analyze the expression of pathogenesis-related proteins in different groups.
The results stated that Huangkui capsule significantly inhibited the elevation of Scr, BUN, UP, the expression of α-smooth muscle actin (α-SMA), phosphorylation-extracellular signal-regulated kinase (p-ERK1/2), NADPH Oxidase 1, NADPH Oxidase 2 and NADPH Oxidase 4 in adenine-induced CRF rats. The main bioactive components of quercetin (QT), hyperoside (HY), isoquercitrin (IQT), gossypetin-8-O-β-D-glucuronide (GG) and quercetin-3'-O-glucoside (QG) at the dosage of 100µM, like NADPH oxidase inhibitor diphenyleneiodonium, exhibited a significant effect on inhibiting the expression of α-SMA, p-ERK1/2, NADPH Oxidase 1, NADPH Oxidase 2 and NADPH Oxidase 4 in high glucose-induced HK-2 cells, especially GG.
These results demonstrated that Huangkui capsule and the flavonoids components prevent tubulointerstitial fibrosis in CRF rat involvement in the action mechanism of inhibiting NADPH oxidase/ROS/ERK pathway.
苘麻花作为一种传统中药，在中国被广泛应用，具有利尿通淋、消肿解毒的功效。据报道，由苘麻花提取物制备的黄葵胶囊可降低肾病大鼠尿蛋白、血清肌酐和血清尿素氮的含量，并具有肾脏保护活性，但其作用机制仍需深入阐明。
本研究旨在探讨黄葵胶囊对慢性肾衰竭（CRF）大鼠肾小管间质纤维化的保护作用及其活性成分对高糖诱导的肾小管上皮细胞（HK-2）上皮-间质转化（EMT）的作用机制。
将动物分为正常组、CRF模型组和黄葵胶囊治疗组。采用苏木精-伊红（HE）染色和Masson染色观察不同组肾组织的病理变化。根据试剂盒说明书检测血清尿素氮（BUN）、尿蛋白（UP）和血清肌酐（Scr）等生化指标。建立HK-2细胞损伤模型，以研究黄葵胶囊中五种主要黄酮类化合物的保护作用及作用机制。将实验细胞分为八组：对照组、模型组、阳性药物组和五种主要黄酮类化合物处理组。采用二氯二氢荧光素二乙酸酯（DCFH-DA）法检测不同组细胞内活性氧（ROS）水平。应用蛋白质免疫印迹法分析不同组中与发病机制相关蛋白的表达。
结果表明，黄葵胶囊能显著抑制腺嘌呤诱导的CRF大鼠血清Scr、BUN、UP水平升高，以及α-平滑肌肌动蛋白（α-SMA）、磷酸化细胞外信号调节激酶（p-ERK1/2）、NADPH氧化酶1、NADPH氧化酶2和NADPH氧化酶4的表达。槲皮素（QT）、金丝桃苷（HY）、异槲皮苷（IQT）、棉黄素-8-O-β-D-葡萄糖醛酸苷（GG）和槲皮素-3'-O-葡萄糖苷（QG）这五种主要活性成分在100µM剂量下，与NADPH氧化酶抑制剂二苯基碘鎓一样，对高糖诱导的HK-2细胞中α-SMA、p-ERK1/2、NADPH氧化酶1、NADPH氧化酶2和NADPH氧化酶4的表达具有显著抑制作用，尤其是GG。
这些结果表明，黄葵胶囊及其黄酮类成分可通过抑制NADPH氧化酶/ROS/ERK途径参与预防CRF大鼠肾小管间质纤维化。
