CAJ | 학술논문

目的观察和分析A20基因转染对大鼠肾脏缺血再灌注后诱发的肾小管损伤及凋亡的影响.方法通过构建人源的A20基因表达载体pcDNA3.1-A20及空质粒载体pcDNA3.1,并将其表达载体及pcDNA3.1空质粒载体转化到感受态大肠杆菌.对该大肠杆菌进行培养,并提取质粒DNA进行10％琼脂糖凝胶电泳分析,以确定其是目的基因.雄性SD大鼠24只,通过手术方法制成大鼠肾脏缺血再灌注模型,随机分为3组:生理盐水组、A20组、空质粒组,每组8只大鼠.术前48h,分别注射生理盐水、脂质体pcD-NA3.1-A20+生理盐水至250uL(15uL脂质体加10ug构建的质粒DNA混匀后加入生理盐水至250uL)、脂质体-pcDNA3.1+生理盐水至250uL(方法同上),混匀室温静置30min,分别通过大鼠尾静脉注射.术后24h收获大鼠,采用全自动生化分析仪检测肾功能,肾组织HE染色行病理学检测,TUNEL法检测肾组织细胞凋亡的情况.结果 A20组Scr、BUN水平明显低于空质粒组和生理盐水组(P＜0.05),差异有统计学意义.各组肾小管都有不同程度的损伤,A20组与生理盐水组及空质粒组比较,肾小管扩张,肾小管上皮细胞刷状缘脱落、坏死,蛋白管型等肾脏组织病理损伤明显减轻(P＜0.05),差异有统计学意义；空质粒组与生理盐水组比较,差异无统计学意义(P＞0.05).肾小管上皮细胞凋亡多见于远端小管,A20组与生理盐水组和空质粒组比较,凋亡细胞数明显减少,差异有统计学意义(P＜0.05)；生理盐水组和空质粒组比较无显著性差异(P＞0.05).结论 A20基因转染能明显减轻肾脏缺血再灌注损伤,其机制可能与抑制肾小管上皮细胞凋亡有关.
목적 관찰화분석A20기인전염대대서신장결혈재관주후유발적신소관손상급조망적영향.방법 통과구건인원적A20기인표체재체pcDNA3.1-A20급공질립재체pcDNA3.1,병장기표체재체급pcDNA3.1공질립재체전화도감수태대장간균.대해대장간균진행배양,병제취질립DNA진행10％경지당응효전영분석,이학정기시목적기인.웅성SD대서24지,통과수술방법제성대서신장결혈재관주모형,수궤분위3조:생리염수조、A20조、공질립조,매조8지대서.술전48h,분별주사생리염수、지질체pcD-NA3.1-A20+생리염수지250uL(15uL지질체가10ug구건적질립DNA혼균후가입생리염수지250uL)、지질체-pcDNA3.1+생리염수지250uL(방법동상),혼균실온정치30min,분별통과대서미정맥주사.술후24h수획대서,채용전자동생화분석의검측신공능,신조직HE염색행병이학검측,TUNEL법검측신조직세포조망적정황.결과 A20조Scr、BUN수평명현저우공질립조화생리염수조(P＜0.05),차이유통계학의의.각조신소관도유불동정도적손상,A20조여생리염수조급공질립조비교,신소관확장,신소관상피세포쇄상연탈락、배사,단백관형등신장조직병리손상명현감경(P＜0.05),차이유통계학의의；공질립조여생리염수조비교,차이무통계학의의(P＞0.05).신소관상피세포조망다견우원단소관,A20조여생리염수조화공질립조비교,조망세포수명현감소,차이유통계학의의(P＜0.05)；생리염수조화공질립조비교무현저성차이(P＞0.05).결론 A20기인전염능명현감경신장결혈재관주손상,기궤제가능여억제신소관상피세포조망유관.
Objectives Observe and analyze the effect of A20 transfection on damage and apoptosis of renal tubular in rats induced by renal ischemia-reperfusion.Methods Constructing anthropogenic A20 gene expression vector pcDNA3.1-A20 and empty plasmid vector pcDNA3.1,wrapped with liposome.Twenty-four male SD rats were randomly divided into three groups (n =8):saline group,A20 group,empty plasmid group.Forty-eight hours before operation every group was injected with Saline 250ul,liposome pcDNA3.1-A20 + saline to 250ul,empty plastid-PcDNA3.1 + saline to 250ul,respectively.then renal ischemia reperfusion injury model was done by clamping left renal artery and vein for 45 min,and cut the right renal before reperfusion of left renal vascular.Twenty-four hours after operation harvested the rats,then the renal function was detected by automatic biochemical analyzer,renal pathological examination was analyzed by HE staining,renal tubular cell apoptosis was detected by TUNEL assay.Results Serum Scr and BUN level of A20 group was lower than the other two groups,renal pathological damage was alleviated significantly,renal tubular epithelial cell apoptosis was decreased.Conclusions A20 transfection can significantly reduce the renal IRI,which may be related to the inhibition of apoptosis of renal tubular epithelial cells.