摘要
目的:观察氯气致大鼠肺损伤过程中,线粒体的结构与功能变化及红景天苷的干预作用,探讨红景天苷可能的保护机制。方法:36只雄性SD大鼠随机分为6组,分别为未染毒组(给予空气),以及氯气染毒后0.5、1.5、3、6和9 h组,给予1200mg/m^3氯气暴露处理5 min。另24只雄性SD大鼠随机分为4组,分别为氯气暴露组、红景天苷干预组、红景天苷对照组和阴性对照组,氯气染毒条件同上,红景天苷干预组和红景天苷对照组在氯气染毒前30 min和染毒后15 min各给予1次300 mg/kg红景天苷灌胃,阴性对照组给予生理盐水灌胃。比色法检测各组大鼠肺泡灌洗液(BALF)和血清中乳酸脱氢酶(LDH)的活力;透射电镜下观察氯气暴露后3 h肺组织血管内皮细胞线粒体超微结构改变;定磷法检测肺组织Na^+,K-ATP酶和Ca^(2+),Mg^(2+)-ATP酶的活力,低温差速离心法分离并提取线粒体蛋白;Western blot法检测肺组织或线粒体中Tom 20、PINK1以及Parkin蛋白表达水平。结果:与未染毒组相比,氯气暴露0.5 h后,大鼠BALF中LDH活力显著升高(P<0.05);肺组织PINK1、Parkin、Tom20蛋白表达水平均显著升高(P<0.05),且Tom20蛋白表达在染毒后3 h最高(P<0.05);故选取染毒后3 h为干预时间点。透射电镜结果显示,氯气暴露后3 h,肺血管内皮细胞中线粒体明显肿胀,嵴数量减少或消失。与阴性对照组相比,氯气染毒后大鼠肺组织中PINK1、Parkin以及线粒体PINK1蛋白表达显著升高(P<0.05),线粒体Parkin蛋白表达显著下降(P<0.05);肺组织Na^+,K-ATP酶和Ca^(2+),Mg^(2+)-ATP酶以及血清LDH的活力均显著升高(P<0.05)。红景天苷干预可显著逆转因氯气暴露引起的上述检测指标的异常表达(P<0.05)。结论:氯气致大鼠急性肺损伤过程中,血管内皮细胞线粒体结构与功能受损,呈时间效应关系。红景天苷可能通过保护线粒体,改善氯气暴露引起的急性肺损伤。
OBJECTIVE：To observe the effects of chlorine on structure and function of mitochondria of lungs in rats and explore the protective effects of salidroside on acute lung injury induced by chlorine. METHODS：36 healthy male Sprague-Dawley （SD） rats were divided into 6 groups：1 control and 5 exposure. Rats in the exposure groups were 3exposed to an initial concentration of 1 200 mg/m chlorine for 5 min in a devised cabinet or room air. Rats were sacrificed at 0.5,1.5,3,6 and 9 h,respectively,after exposure. In addition,24 rats were randomly divided into 4 groups：control,chlorine,chlorine＋salidroside and salidroside. Chlorine and chlorine＋salidroside groups were exposed to chlorine as mentioned above. Chlorine＋salidroside and salidroside groups were treated with salidroside （300 mg/kg） at 30 min before chlorine exposure and at 15 min after chlorine exposure. Then the rats were sacrificed at 3 h after chlorine exposure,and the lung tissues were harvested. Lung mitochondria ultra structures were observed with electron microscope. ＋ ＋ 2＋ 2＋The Na ,K -ATPase,Ca ,Mg -ATPase in lung tissues and lactate dehydrogenase （LDH） in serum and bronchoalveolar lavage fluid （BALF） were measured using kits,respectively. The mitochondria in lung tissues were isolated with differential centrifugation. The protein levels of Tom 20,PINK1 and Parkin in lung tissues or mitochondria were detectedby western blotting. RESULTS：Compared with control group,LDH of BALF in exposure groups were significantly increased （P〈0.05）. Tom 20,PINK1 and Parkin pro
出处
《癌变．畸变．突变》
CAS
CSCD
2018年第1期31-36,41,共7页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
军事医学创新工程专项(16CXZ021)
陕西省自然科学基金重点项目(2014J2009)
关键词
氯气
线粒体
急性肺损伤
红景天苷
chlorine
mitochondria
acute lung injury
salidroside
