摘要
目的探讨茵陈蒿汤对阻塞性黄疸大鼠肾氧化应激损伤的作用与调节核因子E2相关因子2(Nrf2)表达及核异位的关系。方法32只雄性SD大鼠,随机分为假手术组(S组)、模型组(O组)、低剂量茵陈蒿汤组(LY组)和高剂量茵陈蒿汤组(HY组),每组8只,S组大鼠仅游离上段胆总管但不予结扎,剩余各组大鼠胆总管中上1/3行双重结扎建立阻塞性黄疸模型,7天后LY组、HY组分别予茵陈蒿汤6.3 mL/kg和18.9 mL/kg灌胃,S组、O组每日给予等体积蒸馏水灌胃,连续7天,于第14天处理大鼠。采用ELISA法测定大鼠血清总胆红素(TBil)、直接胆红素(DBil)、丙氨酸转氨酶(ALT)、谷氨酰转移酶(GGT)、尿素氮(BUN)和血肌酐(Cr)水平;分光光度法检测大鼠肾组织氧化应激因子超氧化物歧化酶(SOD)和丙二醛(MDA)活性;实时荧光定量PCR和蛋白免疫印迹法分别检测肾组织中Nrf2、Kelch样环氧氯丙烷相关蛋白1(Keap1)、醌氧化还原酶1(NQO1)mRNA和蛋白表达水平;免疫组化检测肾组织中Nrf2蛋白的核异位情况。计量资料多组间比较采用单因素方差分析,组内进一步两两比较采用LSD-t检验。结果与S组比较,O组大鼠TBil、DBil、ALT、GGT、BUN、Cr水平升高,SOD活性减弱,MDA水平升高,差异均有统计学意义(P值均<0.05);与O组比较,LY组和HY组肝、肾功能指标水平均下降,SOD活性升高,MDA水平降低,差异均有统计学意义(P值均<0.05)。与S组比较,O组大鼠肾组织中Nrf2、NQO1 mRNA和蛋白表达水平均明显下降(P值均<0.05);与O组比较,LY组和HY组Nrf2、NQO1 mRNA和蛋白表达水平均明显升高(P值均<0.05);各组大鼠肾组织中Keap1蛋白表达水平无明显差异(P>0.05)。与S组比较,O组大鼠肾组织中Nrf2细胞核内阳性率显著降低(P<0.05);与O组比较,LY组和HY组Nrf2细胞核内阳性率均显著升高(P值均<0.05)。结论茵陈蒿汤可以有效减轻阻塞性黄疸引起的肾损伤,其作用机制可能是通过上调大鼠肾组织中Nrf2�
Objective To investigate the effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its association with the regulation of the expression of nuclear factor erythroid 2-related factor 2(Nrf2)and nuclear translocation.Methods A total of 32 male Sprague-Dawley rats were randomly divided into sham-operation group(S group),model group(O group),low-dose Yinchenhao decoction group(LY group),and high-dose Yinchenhao decoction group(HY group),with 8 rats in each group.For the rats in the S group,the upper common bile duct was isolated without ligation,and for those in the other groups,double ligation of the middle and upper 1/3 of the common bile duct was performed to establish a model of obstructive jaundice.After 7 days,the rats in the LY group and the HY group were given Yinchenhao decoction by gavage at a dose of 6.3 and 18.9 mL/kg,respectively,while those in the S and O groups were given an equal volume of distilled water by gavage every day for 7 consecutive days,and the rats were treated on day 14.ELISA was usedto measure the serum levels of total bilirubin(TBil),direct bilirubin(DBil),alanine aminotransferase(ALT),gamma-glutamyl transpeptidase(GGT),blood urea nitrogen(BUN),and creatinine(Cr);spectrophotometry was used to measure the activity of the oxidative stress factors superoxide dismutase(SOD)and malondialdehyde(MDA)in renal tissue;quantitative real-[JP3]time PCR and Western blotting were used to measure the mRNA and protein expression levels of Nrf2,Kelch-like ECH-associated protein 1(Keap1),and NAD(P)H quinone dehydrogenase 1(NQO1)in renal tissue;immunohistochemistry was used to measure observe the nuclear translocation of Nrf2 protein in renal tissue.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further pairwise comparison within groups.Results Compared with the S group,the O group had significant increases in the levels of TBil,DBil,ALT、GGT,BUN,and C
作者
刘军舰
陈帅
袁红霞
许艳
张西波
李忠廉
LIU Junjian;CHEN Shuai;YUAN Hongxia;XU Yan;ZHANG Xibo;LI Zhonglian(Second Department of Hepatopancreatobiliary Surgery,Tianjin Hospital of Integrated Traditional Chinese and Western Medicine,Tianjin 300100,China;Department of Thoracic Surgery,Xuzhou Hospital of Traditional Chinese Medicine,Xuzhou,Jiangsu 221003,China;School of Management,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Graduate School of Tianjin Medical University,Tianjin 300070,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2023年第5期1126-1133,共8页
Journal of Clinical Hepatology
基金
国家自然科学基金面上项目(81273952)
天津市应用基础研究多元投入基金(21JCZDJC01150)
天津市卫生健康委员会中医药重点领域科研项目(2019003)
天津市卫生健康委员会中医中西医结合科研课题(2021042)
2022中央财政转移支付地方项目。
