CAJ | 학술논문

目的评价盐酸戊乙奎醚预先给药对内毒素诱发大鼠急性肺损伤(ALI)时一氧化氮合酶(NOS)表达和细胞凋亡的影响.方法成年雄性SD大鼠40只,体重230 ～ 270 g,采用随机数字表法,将其分为5组(n=8):对照组(C组)、内毒素性ALI组(ALI组)、盐酸戊乙奎醚0.03、0.10、0.30mg/kg组(P1-,组).尾静脉注射LPS 5 mg/kg建立ALI模型,P1-3组于LPS注射前1h分别腹腔注射盐酸戊乙奎醚0.03、0.10、0.30 mg/kg,C组和ALI组给予等容量生理盐水.模型制备成功后4h处死大鼠,取肺组织标本,采用TUNEL法确定细胞凋亡指数(AI),RT-PCR法测定NOS mRNA表达,Western blot法检测Bcl-2和Bax蛋白表达,并计算Bcl-2/Bax蛋白表达比值,化学比色法检测NOS活性,硝酸还原酶法检测NO含量,光镜和电镜下观察病理学结果.结果与C组比较,ALI组、P1-3组肺组织NOS mRNA和Bax蛋白表达上调,NOS活性、NO含量和AI升高,Bcl-2蛋白表达下调,Bcl-2/Bax蛋白表达比值降低(P＜0.01),肺组织病理学损伤程度加重;与ALI组和P1组比较,P2-3组肺组织NOS mRNA和Bax蛋白表达下调,NOS活性、NO含量和AI降低,Bcl-2蛋白表达上调,Bcl-2/Bax蛋白表达比值升高(P＜0.01),肺组织病理学损伤程度减轻;ALI组与P1组,P2组与P3组比较,上述指标差异无统计学意义(P＞0.05),肺组织病理学结果差异不明显.结论盐酸戊乙奎醚预先给药可能通过抑制肺组织NOS表达和细胞凋亡,减轻大鼠内毒素性ALI.
목적 평개염산무을규미예선급약대내독소유발대서급성폐손상(ALI)시일양화담합매(NOS)표체화세포조망적영향.방법 성년웅성SD대서40지,체중230 ～ 270 g,채용수궤수자표법,장기분위5조(n=8):대조조(C조)、내독소성ALI조(ALI조)、염산무을규미0.03、0.10、0.30mg/kg조(P1-,조).미정맥주사LPS 5 mg/kg건립ALI모형,P1-3조우LPS주사전1h분별복강주사염산무을규미0.03、0.10、0.30 mg/kg,C조화ALI조급여등용량생리염수.모형제비성공후4h처사대서,취폐조직표본,채용TUNEL법학정세포조망지수(AI),RT-PCR법측정NOS mRNA표체,Western blot법검측Bcl-2화Bax단백표체,병계산Bcl-2/Bax단백표체비치,화학비색법검측NOS활성,초산환원매법검측NO함량,광경화전경하관찰병이학결과.결과 여C조비교,ALI조、P1-3조폐조직NOS mRNA화Bax단백표체상조,NOS활성、NO함량화AI승고,Bcl-2단백표체하조,Bcl-2/Bax단백표체비치강저(P＜0.01),폐조직병이학손상정도가중;여ALI조화P1조비교,P2-3조폐조직NOS mRNA화Bax단백표체하조,NOS활성、NO함량화AI강저,Bcl-2단백표체상조,Bcl-2/Bax단백표체비치승고(P＜0.01),폐조직병이학손상정도감경;ALI조여P1조,P2조여P3조비교,상술지표차이무통계학의의(P＞0.05),폐조직병이학결과차이불명현.결론 염산무을규미예선급약가능통과억제폐조직NOS표체화세포조망,감경대서내독소성ALI.
Objective To evaluate the effects of penehyclidine hydrochloride pretreatment on the expression of nitric oxide synthase (NOS) and cell apoptosis in lung tissues in rats with endotoxin-induced acute lung injury (ALI).Methods Forty adult male Sprague-Dawley rats,weighing 220-270 g,were randomly divided into 5 groups (n =8 each) using a random number table:control group (group C),endotoxin-induced ALI group (ALI group),and penehyclidine hydrochloride 0.03,0.10,0.30 mg/kg groups (Pi-3 groups).ALI was induced with lipopolysaccharide (LPS) 5 mg/kg which was injected via the caudal vein.In P1-3 groups,penehyclidine hydmchloride 0.03,0.10 and 0.30 mg/kg were injected intraperitoneally,respectively,at 1 h before LPS injection,while the equal volume of normal saline was administered in C and ALI groups.The animals were sacrificed at 4 h after ALI models were successfully established and pulmonary specimens were obtained for determination of the cell apoptosis (by TUNEL),expression of NOS mRNA (using PT-PCR),and Bcl-2 and Bax protein (by Western blot),NOS activity (using chemical colorimetry) and NO content (by using nitrate reductase method) and for examination of pathological changes (by light and electron microscopes).Apoptotic index (AI) and the ratio of Bcl-2/Bax was calculated.Results Compared with group C,NOS mRNA and Bax protein expression was significantly up-regulated,NOS activity,NO content and AI were increased,Bcl-2 protein expression was down-regulated,the ratio of Bcl-2/Bax was decreased (P ＜ 0.01),and the degree of pathological changes of the lung was aggravated in ALI and P1-3 groups.Compared with ALI and P1 groups,NOS mRNA and Bax protein expression was significantly down-regulated,NOS activity,content of NO and AI were decreased,Bcl-2 protein expression was up-regulated,the ratio of Bcl-2/Bax was increased (P ＜ 0.01),and the degree of pathological changes of the lung was alleviated in P2-3 groups.There was no significant difference in the indexes mentioned above and results of pathological changes of the lung between group ALI and group P1,and between group P2 and group P3 (P ＞ 0.05).Conclusion Penehyclidine hydrochloride pretreatment ameliorates endotoxin-induced ALI by inhibiting NOS expression and cell apoptosis in rat lung tissues.