丹红注射液通过降低氧化应激损伤发挥抗肾纤维化的作用机制研究  

Protective effect of Danhong injection against renal fibrosis by reducing oxidative stress damage

作　　者：王松[1] 陈娟娟[1] 赵院霞[1] 鲁春云 薛莹 于沛 潘豹 杜书章[1] WANG Song;CHEN Juan-juan;ZHAO Yuan-xia;LU Chun-yun;XUE Ying;YU Pei;PAN Bao;DU Shu-zhang(Department of Pharmacy,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052

出　　处：《中南药学》2024年第10期2602-2608,共7页Central South Pharmacy

基　　金：河南省医学科技攻关计划项目(No.2018020135)。

摘　　要：目的探索丹红注射液对慢性肾衰竭大鼠肾损伤及肾纤维化的保护作用及其机制。方法采用腺嘌呤法建立大鼠慢性肾衰竭体内模型,采用叔丁基过氧化氢诱导NRK-52E细胞,建立肾损伤体外模型,HE染色评估丹红注射液对慢性肾衰竭大鼠的肾脏病理学损伤,免疫组化法检测肾损伤及纤维化的关键蛋白表达,免疫荧光法检测NRK-52E细胞GRP78蛋白表达,Western blot法进行组织和细胞中的关键蛋白定量,活性氧(ROS)荧光探针检测NRK-52E细胞中的ROS水平,JC-1检测试剂盒检测NRK-52E细胞中线粒体膜电位(MMP)水平。结果丹红注射液可以显著缓解慢性肾衰竭的肾间质纤维化状态,减轻病理损伤;抑制KIM-1、GRP78和CHOP表达(P<0.01),从而缓解内质网过度应激状态;降低纤维化胶原CollagenⅠ沉积(P<0.01),减少α-平滑肌肌动蛋白(α-SMA)(P<0.01),从而抑制肾间质纤维化形成;降低NRK-52E细胞内ROS的水平(P<0.01),减轻NRK-52E线粒体氧化损伤,稳定MMP水平。结论丹红注射液能够通过降低氧化损伤和缓解内质网过度应激状态,从而发挥抗肾纤维化的作用。Objective To determine the protective effect of Danhong injection against renal injury and renal fibrosis caused by chronic renal failure and related mechanism.Methods Adenine method was used to establish the rat model of chronic renal failure in vivo.NRK-52E cells were induced by TBHP to establish the model of renal injury in vitro.HE staining was used to evaluate the renal pathological injury of Danhong injection on chronic renal failure rats,and immunohistochemical method was used to detect the expression of key proteins in the renal injury and fibrosis.The expression of GRP78 protein in NRK-52E cells was detected by immunofluorescence method,key proteins in the tissues and cells were quantified by Western blot.ROS levels in NRK-52E cells were detected by ROS fluorescence probe,and the membrane potential levels in NRK-52E cells by JC-1 detection kit.Results Danhong injection greatly relieved the renal interstitial fibrosis in chronic renal failure and alleviated the pathological injury.It inhibited the expression of KIM-1,GRP78 and CHOP,thereby inhibiting the state of endoplasmic reticulum overstress.It inhibited the deposition of fibrotic CollagenⅠ,thus inhibiting the formation of renal interstitial fibrosis.Inhibition of myofibroblast proliferation(α-SMA).It also inhibited the excessive stress of the endoplasmic reticulum of NRK-52E cells,alleviated the oxidative damage of NRK-52E mitochondria,and stabilized the level of membrane potential.Conclusion Danhong injection plays an anti-renal fibrosis role by reducing the oxidative damage and alleviating endoplasmic reticulum overstress.

关 键 词：丹红注射液 慢性肾衰竭 肾间质纤维化 内质网过度应激状态 

分 类 号：R286[医药卫生—中药学]