竹节香附素A对肝缺血再灌注大鼠的保护作用机制  

Mechanism of Protective Effect of Raddeanin A on Hepatic Ischemia-reperfusion in Rats

作　　者：李洪波 黄锐[2] 代将 钟振东[4] LI Hongbo;HUANG Rui;DAI Jiang;ZHONG Zhendong(Department of Second Surgeny,Guanghan People's Hospital,Guanghan 618300,China;Department of Hepatobiliary&Pancreatic Center,Sichuan Academy of Medical Science and Provincial People's Hospital,Chengdu 610007,China;Department of First-aid Center,Sichuan Academy of Medical Science and Provincial People's Hospital,Chengdu 610007,China;Center of Animal Research,Sichuan Academy of Medical Science and Provincial People's Hospital,Chengdu 610007,China)

机构地区：[1]四川省广汉市人民医院外二科,广汉618300 [2]四川省医学科学院·四川省人民医院肝胆外科,成都610007 [3]四川省医学科学院·四川省人民医院急救中心,成都610007 [4]四川省医学科学院·四川省人民医院动物研究中心,成都610007

出　　处：《实验动物与比较医学》2021年第1期55-60,65,共7页Laboratory Animal and Comparative Medicine

基　　金：国家自然科学基金项目(81302161);四川省卫生厅资助项目(17PJ587)。

摘　　要：目的探讨竹节香附素A(raddeani n A,RA)对大鼠肝缺血再灌注损伤的保护作用机制。方法将30只S D大鼠随机分为假手术组、模型组和RA组,每组10只。RA组大鼠于造模前24 h和1 h分别按体质量尾静脉注射RA(10 mg/kg),假手术组与模型组分别注射等体积的0.9%NaCl溶液(生理盐水)。大鼠麻醉后夹闭入肝血管60 min致肝脏70%缺血,恢复血流6 h后检测血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(L DH)的含量,E LI SA法检测肝脏中超氧化物歧化酶(SOD)、丙二醛(MDA)、白细胞介素-1β(IL-1β)、白细胞介素-6(I L-6)和肿瘤坏死因子-α(TNF-α)的含量,HE与TUNEL染色观察肝脏病理损伤与肝细胞凋亡情况,蛋白质印迹法检测肝脏中B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2蛋白(Bcl-2)和半胱氨酸蛋白酶-3(Caspase-3)的表达水平。结果与模型组比较,RA组血清中ALT、AST和LDH含量减少(P<0.05),肝组织中MDA、I L-1 b、IL-6和TNF-a含量减少(P<0.05),SOD含量增加(P<0.05);RA组肝脏组织结构完好,凋亡细胞数量减少,凋亡率降低(P<0.01),Bax和Caspase-3水平下降,Bcl-2水平升高(P<0.05)。结论缺血再灌注前注射RA能提升缺血再灌注后大鼠肝功能,减少氧化与炎性因子的释放,通过下调Bax和Caspase-3蛋白表达减少肝细胞凋亡,从而发挥保护肝脏的作用。Objective To explore the protective mechanism of raddeanin A(RA)on hepatic ischemiareperfusion injury in rats.Methods Thirty rats were randomly assigned into three groups including sham,model and RA groups(h=10).The rats in the RA group were injected with RA(10 mg/kg)in the tail vein 24 h and 1 h before model construction.The rats in the sham and model groups were injected with normal saline of equal volume.After the rats were anesthetized,the blood vessels entering the liver were blocked for 60 min,causing 70%of the liver to be ischemia.The contents of aspartate aminotransferase(AST),alanine aminotransferase(ALT)and lactate dehydrogenase(LDH)in serum were detected 6 h after liver blood flow was restored,the superoxide dismutase(SOD),malondialdehyde(MDA),interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels in liver were detected by ELISA.Liver specimens were stained with hematoxylin-eosin(HE)and TUNEL for observation of pathologic and apoptotic changes.Bcl2-associated X protein(Bax),Bcl-2 and Caspase-3 protein levels in liver were detected by Western blotting.Results The levels of ALT,AST and LDH in the RA groups were lower than those in the model group(P<0.01).The levels of MDA,IL-1 b,IL-6,TNF-α,Bax and Caspase-3 in the RA groups were lower than those in the model group(P<0.05),the levels of SOD and Bcl-2 were higher(P<0.05).The structure of hepatic lobules was complete in the RA group.The apoptotic cells and the apoptosis rate in the RA group were lower than those in the model group(P<0.01).Conclusion Preoperative injection of RA can protect the liver by improving the liver function of rats after ischemia-reperfusion,reducing the release of oxidative and inflammatory factors,and reducing the apoptosis of hepatocytes via down-regulating the expressions of Bax and Caspase-3 proteins.

关 键 词：竹节香附素A 肝缺血再灌注 细胞凋亡 氧化应激 体内活性 中药有效成分 大鼠 

分 类 号：Q95-33[生物学—动物学] R965.2[医药卫生—药理学]