京尼平对大鼠肾移植缺血再灌注损伤中的作用  

The role of genipin in ischemia-reperfusion injury in rat kidney transplantation

作　　者：陆兵 李明虎 何贵柠 张秋雯 文宁 李海滨 吴基华[1] 蓝柳根[1] 董建辉 孙煦勇 Lu Bing;Li Minghu;He Guining;Zhang Qiuwen;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9期2041-2044,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 探讨京尼平对大鼠肾移植缺血再灌注损伤的作用机制.方法 将36只健康成年雄性SD大鼠按照随机数字表法分为4组:对照组(Control)、假手术组(Sham)、肾移植组(KT)、京尼平干预组(GP),Control组和Sham组,每组6只,KT组和GP组供、受体每组各6只.采用大鼠原位肾移植模型,GP组术前1 h予50 mg/kg京尼平灌胃.于术后24 h获取大鼠血清和肾脏组织.采用苏木精-伊红(HE)染色观察肾组织病理结果;酶联免疫吸附法(ELISA)检测大鼠血清血肌酐(Scr)和尿素氮(BUN)水平,大鼠肾组织超氧化物歧化酶(SOD)和丙二醛(MDA)水平;用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法(TUNEL)检测肾组织细胞凋亡情况;蛋白免疫印迹法(Western blot)检测肾组织中腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子-1(SIRT1)和解耦联蛋白2(UCP2)的蛋白表达.组间比较采用独立样本t检验.结果 HE结果显示,KT组大鼠肾脏有炎性细胞浸润、肾小管萎缩变性、间质细胞水肿、皮质出血,GP组肾组织结构损害较轻.ELISA结果显示,GP组血清 Scr、BUN 和肾组织 MDA 低于 KT组(42.06±4.03 比 53.18±3.59、28.82±2.31 比42.49±5.05、4.01±0.39 比 5.13±0.31,i=5.04、6.03、5.42,P<0.05),而 SOD 水平高于 KT 组(8.58±0.75比6.88±0.45,t=4.77,P<0.05).TUNEL检测结果显示,GP组大鼠肾组织细胞凋亡率低于 KT组(3.53±1.795 比 17.53±6.91,t=4.81,P<0.05).Western blot 结果显示,GP 组AMPK、SIRT1 蛋白表达水平高于 KT组(1.07±0.17 比 0.52±0.11、0.84±0.15 比 0.48±0.23,t=6.49、3.29,P<0.05),而UCP2 蛋白表达水平低于 KT组(0.85±0.20 比 1.18±0.07,t=3.89,P<0.05).结论 京尼平可能通过激活AMPK/SIRT1通路减轻移植肾缺血再灌注损伤.Objective To investigate the action mechanism of genipin on ischemia-reperfusion in-jury in rat kidney transplantation.Methods Totally,36 healthy adult male SD rats were divided into four groups according to the randomized numerical table method:the control group(Control),the sham opera-tion group(Sham),the kidney transplantation group(KT),and the genipin intervention group(GP),with 6 rats in each of the Control and Sham groups,and 6 rats in each of the donor and recipient groups of the KT and GP groups.The rat in situ kidney transplantation model was used,and saline gavage was given to the Sham and KT groups 1 h before operation,and 50 mg/kg of genipin was given to the GP group 1 h be-fore operation.After 24 h of modeling,hematoxylin-eosin(HE)staining was used to observe the pathologi-cal results of renal tissue.The enzyme-linked immunosorbent assay(ELISA)was used to detect serum cre-atinine(Scr)and blood urea nitrogen(BUN)levels,superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the rat kidney tissue.The notched-end-labeling method of terminal deoxynucleotidyl transferase dUTP transferase(TUNEL)was used to detect apoptosis.The protein expression of adenylate-activated protein kinase(AMPK),silencing information regulator-1(SIRT1),and uncoupling protein 2(UCP2)was detected in renal tissues by protein immunoblotting(Western blotting).The t-test was used for comparison between groups.Comparison of continuous data between two groups was performed by inde-pendent samples t-test.Results HE results showed that the kidneys of rats in the KT group had inflamma-tory cell infiltration,tubular atrophy and degeneration,interstitial cell edema,and cortical hemorrhage,while the GP group had less structural damage to the renal tissues.ELISA results showed that serum Scr,BUN,and MDA of renal tissues in the GP group were lower than those of the KT group(42.06±4.03 vs.53.18±3.59,28.82±2.31 vs.42.49±5.05,4.01±0.39 vs.5.13±0.31,t=5.04,6.03,5.42,P<0.05),while SOD level was higher than that in the KT group(8.58±0.75 vs.

关 键 词：京尼平 肾移植 缺血再灌注损伤 腺苷酸活化蛋白激酶/沉默信息调节因子-1/解耦联蛋白2 多炎性因子 

分 类 号：R699.2[医药卫生—泌尿科学]