CAJ | 학술논문

目的观察intermedin(IMD)对大鼠肾脏缺血再灌注损伤(IRI)的保护作用并探讨其机制.方法健康雄性Wistar大鼠24只随机分为对照组、IRI组、转空质粒组、转IMD质粒组.动物右肾切除后,用超声微泡技术将质粒转染入肾脏,1周后制作肾脏IRI模型.PAS染色观察肾脏病理损伤,比色法检测肾组织超氧化物岐化酶(SOD)、髓过氧化物酶(MPO)和天冬氨酸半胱氨酸蛋白酶3(caspase-3),以及脂质过氧化物丙二醛(MDA)含量.免疫组织化学方法检测细胞问黏附分子1(ICAM-1)、P选择素及内皮素1(ET-1)表达.TUNEL染色检测肾组织细胞凋亡.结果 PAS染色结果显示,IRI组肾小管及间质病理损伤显著重于对照组(P<0.01);转IMD组肾组织病理损伤则显著轻于IRI组(P<0.01).IRI组肾组织SOD活性显著低于对照组(P<0.05),MPO活性、活性caspase-3、MDA含量及ICAM-1、P选择素和ET-1表达均显著高于对照组(均P<0.01);转IMD组SOD活性显著高于IRI组(P<0.05),MPO活性、活性caspase-3、MDA含量及ICAM-1、P选择素和ET-1表达均显著低于IRI组(均P<0.01).TUNEL染色显示,IRI组肾组织凋亡细胞数显著高于对照组(34.83%±8.75%比3.33%±0.47%,P<0.01);转IMD组肾组织凋亡细胞数(20.67%±7.71%)则较IRI组显著减轻(P<0.01).转空质粒组和IRI组以上指标差异均无统计学意义.结论 IMD能减轻肾脏IRI,其机制至少部分与抑制氧自由基生成、炎细胞浸润及炎性因子ICAM-1、P选择素生成、ET-1生成、细胞凋亡有关,从而减轻肾组织局部氧化应激反应产生的活性氧.
목적 관찰intermedin(IMD)대대서신장결혈재관주손상(IRI)적보호작용병탐토기궤제.방법 건강웅성Wistar대서24지수궤분위대조조、IRI조、전공질립조、전IMD질립조.동물우신절제후,용초성미포기술장질립전염입신장,1주후제작신장IRI모형.PAS염색관찰신장병리손상,비색법검측신조직초양화물기화매(SOD)、수과양화물매(MPO)화천동안산반광안산단백매3(caspase-3),이급지질과양화물병이철(MDA)함량.면역조직화학방법검측세포문점부분자1(ICAM-1)、P선택소급내피소1(ET-1)표체.TUNEL염색검측신조직세포조망.결과 PAS염색결과현시,IRI조신소관급간질병리손상현저중우대조조(P<0.01);전IMD조신조직병리손상칙현저경우IRI조(P<0.01).IRI조신조직SOD활성현저저우대조조(P<0.05),MPO활성、활성caspase-3、MDA함량급ICAM-1、P선택소화ET-1표체균현저고우대조조(균P<0.01);전IMD조SOD활성현저고우IRI조(P<0.05),MPO활성、활성caspase-3、MDA함량급ICAM-1、P선택소화ET-1표체균현저저우IRI조(균P<0.01).TUNEL염색현시,IRI조신조직조망세포수현저고우대조조(34.83%±8.75%비3.33%±0.47%,P<0.01);전IMD조신조직조망세포수(20.67%±7.71%)칙교IRI조현저감경(P<0.01).전공질립조화IRI조이상지표차이균무통계학의의.결론 IMD능감경신장IRI,기궤제지소부분여억제양자유기생성、염세포침윤급염성인자ICAM-1、P선택소생성、ET-1생성、세포조망유관,종이감경신조직국부양화응격반응산생적활성양.
Objective To investigate the protective effect of intermedin(IMD)on renal ischemia reperfusion injury(IRI)and its mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups: control group, IRI group, empty plasmid group and IMD group. After remove of right kidney, plasmid was transfected into the kidney by ultrasonic microbubbles technology, and IRI model was made after 1 week. Renal pathology was observed by PAS staining. Renal tissue superoxide dismutase(SOD), myeloperoxidase(MPO), caspase-3 activity, and malondialdehyde(MDA)content were detected by colorimetric method. The intercellular cell adhesion molecule-1(ICAM-1), endothelin 1(ET-1)and P-selection expression of renal tissue were detected by immunohistochemical method. Apoptosis of renal tubular cell was detected by TUNEL.Results Compared with control group, tubulointerstitial pathological injury was significant aggravated in IRI group(P<0.01);compared with IRI group, IMD pretreatment significantly alleviated the degree of renal injury(P<0.01). Compared with control group, in IRI group, SOD activity was significantly decreased(P<0.05), MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 were increased significantly(all P< 0.01). Compared with IRI group, IMD pretreatment significantly increased SOD activity(P <0.05), decreased the MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 (all P<0.01). The apoptosis rate of renal tubular epithelial cells in IRI group was significantly higher than that in control group(34.83%±8.75% vs 3.33%±0.47%, P<0.01), while the apoptosis rate of IMD group(20.67%±7.71%)was significantly lower than that of IRI group. There was no difference of above indexes between empty plasmid group and IRI group. Conclusions IMD pretreatment protects against renal IRI. The mechanism may be at least partly related to the clearance of oxygen free radicals, the improvement of lipid peroxidation, inflammatory cell infiltration and cell apoptosis, leading to the decrease of the production of reactive oxygen species caused by oxidative stress.