水飞蓟宾对肢体缺血再灌注大鼠肝损伤及线粒体凋亡途径的影响  

Effects of silybin on liver injury and mitochondrial apoptosis pathway in limb ischemia-reperfusion rats

作　　者：杨小春 吕建[2] 孙景毅 汪文月 王媛媛[5] YANG Xiaochun;LYU Jian;SUN Jingyi;WANG Wenyue;WANG Yuanyuan(Digestive Department,Hebei Port Group Co.,Ltd.Qinhuangdao Integrated Traditional Chinese and Western Medicine Hospital,Qinhuangdao,Hebei 066000,China;Digestive Endoscopy Center,Hebei Port Group Co.,Ltd.Qinhuangdao Integrated Traditional Chinese and Western Medicine Hospital,Qinhuangdao,Hebei 066000,China;Department of Cardiology,Hebei Port Group Co.,Ltd.Qinhuangdao Integrated Traditional Chinese and Western Medicine Hospital,Qinhuangdao,Hebei 066000,China;Nephrology and Blood Purification Department,Cangzhou Central Hospital,Cangzhou,Hebei 061000,China;Department of Gastroenterology,Peking University Third Hospital,Qinhuangdao Hospital,Qinhuangdao,Hebei 066000,China)

机构地区：[1]河北港口集团有限公司秦皇岛中西医结合医院消化科,河北秦皇岛066000 [2]河北港口集团有限公司秦皇岛中西医结合医院消化内镜中心,河北秦皇岛066000 [3]河北港口集团有限公司秦皇岛中西医结合医院心内科,河北秦皇岛066000 [4]沧州市中心医院肾病血液净化科,河北沧州061000 [5]北京大学第三医院秦皇岛医院消化科,河北秦皇岛066000

出　　处：《安徽医药》2025年第3期494-501,共8页Anhui Medical and Pharmaceutical Journal

基　　金：秦皇岛市科学技术研究与发展计划项目(202101A104)。

摘　　要：目的探讨水飞蓟宾对肢体缺血再灌注大鼠肝损伤的保护作用及其具体机制。方法2022年1月至2023年1月将50只大鼠按照随机数字表法分为正常组、模型组、水飞蓟宾低浓度组、水飞蓟宾中浓度组和水飞蓟宾高浓度组,每组10只。水飞蓟宾低、中和高浓度组大鼠分别给予100、200和400 mg·kg^(-1)·d^(-1)水飞蓟宾灌胃,连续7 d。于末次给药2 h后,双后肢根部环扎橡皮筋阻断血流4 h,再灌注4 h,制备肢体缺血再灌注肝损伤模型。计算肝指数和脾指数,检测肝脏损伤指标[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)以及乳酸脱氢酶(LDH)]、氧化应激指标[超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSHPx)和丙二醛]以及血清中炎症反应指标[白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)],线粒体通透性转运孔(mPTP)孔开放程度,肝组织进行苏木精-伊红(HE)染色和脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)染色,逆转录实时荧光定量聚合酶链式反应(RT-qPCR)法检测B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)mRNA表达水平,蛋白质印迹法检测肝组织中活化胱天蛋白酶(cleaved caspase)-3和cleaved caspase-9蛋白表达水平以及Bax/Bcl-2,并分别检测肝细胞胞质和线粒体中细胞色素C(Cyt-c)蛋白表达水平。结果模型组肝细胞排列紊乱,肝细胞肿胀,部分出现空泡变性,大量炎症细胞浸润;水飞蓟宾低、中和高浓度组大鼠肝组织病理损伤减轻,出现空泡化、坏死的细胞数减少,炎症细胞浸润减少。与正常组比较,模型组肝指数、脾指数、ALT、AST和LDH、TNF-α和IL-1β以及丙二醛含量、细胞凋亡率均升高,SOD和GSH-Px活性降低,mPTP孔开放程度增强,BaxmRNA表达水平以及cleaved caspase-3、cleaved caspase-9蛋白表达水平和Bax/Bcl-2升高,胞质中Cyt-c蛋白表达水平升高,Bcl-2mRNA表达水平以及线粒体中Cyt-c蛋白表达水平降低(P<0.05);与模型组比较,�Objective To investigate the protective effect and mechanism of silybin on liver injury in limb ischemia-reperfusion rats.Methods From January 2022 to January 2023,50 rats were randomly divided into normal group,model group,silybin low concentration group,silybin medium concentration group and silybin high concentration group according to the random number table method,with 10 rats in each group.Rats in low,medium,and high concentration groups were given 100,200 and 400 mg·kg^(-1)·d^(-1) silybin intragastric administration,for consecutive 7 days,respectively.2 h after the last administration,the blood flow was blocked for 4 h by tying rubber bands around the root of the double hind limbs,and the limb ischemia-reperfusion liver injury model was prepared for 4 h.The liver index and spleen index were calculated,liver injury indexes[alanine aminotransferase(ALT),aspartate transaminase(AST),and lactate dehydrogenase(LDH)],oxidative stress indexes[superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)],serum inflammatory response indexes[interleukin-1β(IL-1β)and tumor Necrosis factorα(TNF-α)],mitochondrial permeability transport pore(mPTP)openness were detected,liver tissue was stained by hematoxylin-eosin(HE)and Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL),and the mRNA expression levels of B lymphoblastoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)were detected by Reverse Transcription quantitative PCR(RT-qPCR).The expression levels of spliced cysteine aspartate proteolytic enzyme 3(cleaved caspase 3)and cleaved caspase 9 and Bax/Bcl-2 were detected by Western blotting,and the cytochrome C(Cyt-c)protein expression levels were detected in the cytoplasm and mitochondria of hepatocytes,respectively.Results In the model group,the arrangement of hepatocytes was disordered,the hepatocytes were swollen,some of them showed vacuolar degeneration,and a large number of inflammatory cells infiltrated.The pathological changes of liver tissue in the low,medium and high concent

关 键 词：肝损伤 再灌注损伤 线粒体凋亡 水飞蓟宾 丙氨酸转氨酶 超氧化物歧化酶 胱天蛋白酶-3 胱天蛋白酶-9 

分 类 号：R285.5[医药卫生—中药学]