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泽泻乙醇提取物对大鼠的肾毒性及其分子机制研究

[Study on nephrotoxicity of ethanol extract from Alismatis Rhizoma in rats and its molecular mechanism].

作者信息

Wang Chun-Fei, Ma Liang, Hou Xue-Feng, Chen Hai-Feng, Feng Liang, Jia Xiao-Bin

机构信息

Key Laboratory of New Drug Delivery System of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing 210028, China.

College of Pharmacy, Anhui University of Chinese Medicine, Hefei 230038, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Sep;41(18):3432-3438. doi: 10.4268/cjcmm20161819.

Abstract

Female rats were fed with ethanol extraction of Alismatis Rhizoma for 6 months to study its nephrotoxicity and molecular mechanism. HPLC was used to determine the components in ethanol extraction of Alismatis Rhizoma. An assessment of renal pathology was determined by HE staining. Meanwhile Western blot, immunohistochemical assay and q-PCR were used to assess the protein expression and mRNA level of Kim-1, clusterin, LCN2, osteopontin, ceruloplasmin and TIMP1 in rat kidney. Eight components were identified in ethanol extraction of Alismatis Rhizoma. Tubule-interstitial inflammation, renal tubular epithelial cell exfoliation and morphological changes of cell were observed in rat kidney. Comparing with control blank group, the protein expression of clusterin, Kim-1, LCN2, osteopontin and TIMP1 in rat kidney was significantly increased while the protein expression of ceruloplasmin was significantly decreased. The mRNA level of Kim-1, TIMP1, osteopontin, clusterin and LCN2 was significantly increased while the mRNA level of ceruloplasmin was significantly decreased. In this study, it was inferred that there is chronic toxicity in kidney by using high dosage of ethanol extraction of Alismatis Rhizoma for a long time. And the underlying molecular mechanism was related to regulate the protein expression of ceruloplasmin, clusterin, Kim-1, LCN2, osteopontin and TIMP1.

摘要

给雌性大鼠喂食泽泻乙醇提取物6个月,以研究其肾毒性及分子机制。采用高效液相色谱法测定泽泻乙醇提取物中的成分。通过苏木精-伊红染色对肾脏病理进行评估。同时,采用蛋白质免疫印迹法、免疫组织化学法和定量聚合酶链反应法评估大鼠肾脏中Kim-1、簇集素、脂质运载蛋白2、骨桥蛋白、铜蓝蛋白和金属蛋白酶组织抑制因子1的蛋白表达及mRNA水平。在泽泻乙醇提取物中鉴定出8种成分。观察到大鼠肾脏出现肾小管间质炎症、肾小管上皮细胞脱落及细胞形态改变。与空白对照组相比,大鼠肾脏中簇集素、Kim-1、脂质运载蛋白2、骨桥蛋白和金属蛋白酶组织抑制因子1的蛋白表达显著增加,而铜蓝蛋白的蛋白表达显著降低。Kim-1、金属蛋白酶组织抑制因子1、骨桥蛋白、簇集素和脂质运载蛋白2的mRNA水平显著增加,而铜蓝蛋白的mRNA水平显著降低。本研究推断,长期高剂量使用泽泻乙醇提取物对肾脏存在慢性毒性。其潜在的分子机制与调节铜蓝蛋白、簇集素、Kim-1、脂质运载蛋白2、骨桥蛋白和金属蛋白酶组织抑制因子1的蛋白表达有关。

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