紫草素在大鼠肝移植缺血再灌注损伤中的作用  

Role of shikonin in ischemia-reperfusion injury in rat liver transplantation

作　　者：李明虎 陆兵 何真 朱丽容 文宁 李海滨 吴基华[1] 蓝柳根[1] 董建辉 孙煦勇 Li Minghu;Lu Bing;He Zhen;Zhu Lirong;Wen Ning;Li Haibin;Wu Jihua;Lan Liugen;Dong Jianhui;Sun Xuyong(Transplant Medical Center of The Second Affiliated Hospital of Guangxi Medical University Guangxi Clinical Research Center for Organ Transplantation Guangxi Key Laboratory of Organ Donation and Transplantation,Nanning 530007,China)

机构地区：[1]广西医科大学第二附属医院移植医学中心,广西器官移植临床医学研究中心,广西器官捐献与移植研究重点实验室,南宁530007

出　　处：《中华实验外科杂志》2024年第6期1205-1208,共4页Chinese Journal of Experimental Surgery

基　　金：广西医学高层次骨干人才培养"139"计划培养项目(G202002016);广西自然科学基金区域高发疾病研究联合专项资助(2023GXNSFAA026142)。

摘　　要：目的探讨紫草素在大鼠肝移植缺血再灌注损伤的作用机制。方法采用随机数字表法将36只健康雄性SD大鼠随机分为4组:对照组(CTRL)、假手术组(SH)、肝移植组(LT)、紫草素组(SKLT)。CTRL和SH每组6只大鼠,LT和SKLT供、受体各6只大鼠。采用二袖套法建立大鼠原位肝移植模型,SKLT术后给予紫草素15 mg/kg灌胃。于术后24 h获取大鼠血清和肝脏组织。免疫组织化学法检测肝脏组织中B淋巴细胞瘤-2基因(bcl-2)、bcl-2相关X蛋白(bax)和半胱氨酰天冬氨酸特异性蛋白酶-3(Caspase-3)的蛋白表达;蛋白质印迹法(Western blot)检测肝脏组织中大肿瘤抑制因子1(LATS1)、丝氨酸苏氨酸激酶3(MST1)和Yes相关蛋白(YAP)的蛋白表达。组间比较采用独立样本t检验。结果免疫组织化学显示,SKLT的bax和Caspase-3阳性面积比例显著低于LT组(bax:0.061±0.002比0.084±0.005,t=11.470,P<0.05;Caspase-3:0.148±0.008比0.210±0.029,t=7.328,P<0.05),而bcl-2阳性面积比例显著高于LT组(0.127±0.036比0.069±0.014,t=3.752,P<0.05)。Western blot结果显示,SKLT的LATS1和MST1蛋白的表达显著高于LT组(LATS1:1.56±0.14比1.11±0.30,t=5.715,P<0.05;MST1:1.80±0.38比1.32±0.31,t=4.190,P<0.05),而YAP蛋白的表达显著低于LT组(1.42±0.10比1.80±0.07,t=11.580,P<0.05)。结论紫草素可能通过激活Hippo信号通路来减少大鼠肝移植缺血再灌注损伤中的肝细胞凋亡。Objective To investigate the mechanism of shikonin in ischemia-reperfusion injury(IRI)in rat liver transplantation.Methods Through the stochastic indicator method,36 healthy male SD rats were randomly divided into 4 groups:control(CTRL)and sham operated control(SH),liver transplantation(LT),shikonin group(SKLT).There were 6 rats in each group of CTRL and SH,and 6 rats served as donors and 6 rats as recipients for LT and SKLT.A rat orthotopic liver transplantation model was established by the two-sleeve method,and the SKLT group was given 15 mg/kg gavage after surgery.Serum and liver tissue were obtained 24 h after operation.Immunohistochemistry was used to detect the protein expression of B lymphocytoma-2 gene(bcl-2),bcl-2 associated X protein(bax)and caspase-3 in liver tissues.Western blotting was used to detect the protein expression of large tumor suppressor factor 1(LATS1),serine threonine kinase 3(MST1)and Yes-related protein(YAP)in liver tissues.Comparison of continuous data between two groups was performed by independent samples t-test.Results Immunohistochemistry results showed that the proportion of bax and Caspase-3 positive area was significantly reduced in SKLT group as compared with LT group(bax:0.061±0.002 vs.0.084±0.005,t=11.47,P<0.05;Caspase-3:0.148±0.008 vs.0.210±0.029,t=7.328,P<0.05),while the proportion of bcl-2 positive area was significantly increased(0.127±0.036 vs.0.069±0.014,t=3.752,P<0.05).The Western blotting results showed that the expression of LATS1 and MST1 proteins in SKLT group was significantly increased as compared with LT group(LATS1:1.56±0.141 vs.1.11±0.30,t=5.715,P<0.05;MST1:1.80±0.38 vs.1.32±0.31,t=4.190,P<0.05),while the expression of YAP protein was significantly decreased(1.42±0.10 vs.1.80±0.07,t=11.580,P<0.05).Conclusion Shikonin may reduce hepatocyte apoptosis of IRI in rat liver transplantation by activating the Hippo signaling pathway.

关 键 词：紫草素 肝移植 肝脏缺血再灌注损伤 凋亡 Hippo信号通路 

分 类 号：R-332[医药卫生] R657.3