Zhu Xiao-Ting, Chen Zhi-Qiang
Hebei University of Chinese Medicine.
Tangshan People's Hospital, Tangshan 063000, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Jan 28;36(1):67-72. doi: 10.12047/j.cjap.5928.2020.015.
Effects of Yiqi Huashi Tongluo Formula on oxidative stress and renal fibrosis of residual kidney were investigated in five/sixth nephrectomy rats.
The rat model of chronic renal failure after nephrectomy was established by Platt method. Two weeks after the operation, the rats were randomly divided into model group, Yiqi Huashi Tongluo Formula (YHT) group, benazepril online (BH) group and sham group, with 8 rats in each group. Treatment was initiated once a day for 12 weeks after successful modeling. Animals were treated once a day with intragastric administration for 12 weeks. (Aqueous solution of free decoction granules in YHT group was 0.276 g/100 g·d. BH group benazepril hydrochloride tablet aqueous solution 0.09 mg/100 g·d gavage; sham group and model group were gavage with 1 ml/100 g normal saline). Urine was collected with a metabolic cage at the end of the 12th week, and urine protein content was detected for 24 hours. The rats were then anesthetized to extract blood from the abdominal aorta and the kidneys. The pathological changes of left kidney were observed by HE staining and Masson staining. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in kidney homogenate were determined by colorimetry. Western blot assay was used to detect the expressions of nuclear factor-erythroid 2-related factor 2 (Nrf2), kelch-like ech-associated protein-1 (Keap1), NADPH oxidase 4 (Nox4), transforming growth factor-binding 1(TGF-β1), type I collagen (Collagen1) and Nrf2 in the nucleus in renal tissue.
Compared with sham group, model group rats had severe glomerular injury and obvious fibrosis. Levels of Scr, BUN, MDA and 24-hour urine protein excretion, protein expressions of Keap1, Nox4, TGF-β1 and Collagen1 were significantly increased (P＜0.01), while SOD activity and Nrf2 expression were significantly decreased (P＜0.01).Compared with the model group, the degree of glomerular lesion was reduced and fibrosis was less after YHT or BH intervention, and the levels of Scr, BUN, MDA, 24-hour urine protein excretion, protein expressions of Keap1, Nox4, TGF-β1 and Collagen1 were significantly decreased (P＜0.01), while SOD activity and Nrf2 expression were significantly increased (P＜0.01).
Through affecting the Nrf2/Keap1 signaling pathway and down-regulating the expression of TGF-β1 protein, Yiqi Huashi Tongluo Formula improved the oxidative stress damage and fibrosis degree of residual kidney in the model rats with renal failure.
在五/六肾切除大鼠中研究益气化湿通络方对残肾氧化应激及肾纤维化的影响。
采用普拉特法建立肾切除术后慢性肾衰竭大鼠模型。术后两周，将大鼠随机分为模型组、益气化湿通络方（YHT）组、苯那普利（BH）组和假手术组，每组8只。造模成功后每天给药1次，共12周。动物每天灌胃给药12周。（YHT组为免煎颗粒剂水溶液0.276 g/100 g·d。BH组为盐酸苯那普利片水溶液0.09 mg/100 g·d灌胃；假手术组和模型组用1 ml/100 g生理盐水灌胃）。第12周结束时用代谢笼收集尿液，检测24小时尿蛋白含量。然后将大鼠麻醉，经腹主动脉取血并摘取肾脏。通过苏木精-伊红（HE）染色和Masson染色观察左肾病理变化。采用比色法测定肾匀浆中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。采用蛋白质免疫印迹法检测肾组织中核因子E2相关因子2（Nrf2）、kelch样ECH相关蛋白1（Keap1）、NADPH氧化酶4（Nox4）、转化生长因子β1（TGF-β1）、Ⅰ型胶原（Collagen1）以及肾组织细胞核中Nrf2的表达。
与假手术组相比，模型组大鼠肾小球损伤严重且纤维化明显。血清肌酐（Scr）、尿素氮（BUN）、MDA水平及24小时尿蛋白排泄量、Keap1、Nox4、TGF-β1和Collagen1蛋白表达均显著升高（P＜0.01），而SOD活性和Nrf2表达显著降低（P＜0.01）。与模型组相比，YHT或BH干预后肾小球病变程度减轻，纤维化程度减轻，Scr、BUN、MDA水平、24小时尿蛋白排泄量、Keap1、Nox4、TGF-β1和Collagen1蛋白表达均显著降低（P＜0.01），而SOD活性和Nrf2表达显著升高（P＜0.01）。
益气化湿通络方通过影响Nrf2/Keap1信号通路，下调TGF-β1蛋白表达，改善了肾衰竭模型大鼠残肾的氧化应激损伤及纤维化程度。
