慢性低氧性肺动脉高压大鼠肺组织内质网应激介导的凋亡的变化  

Changes of endoplasmic reticulum stress-induced apoptosis in pulmonary tissue of rats with hypoxic pulmonary hypertension

作　　者：范小芳[1] 李文娟[1] 陈兆琴[1] 王雪瑞[1] 孔晓霞[1] 毛孙忠[1] 胡良冈[1] 龚永生[1] 

机构地区：[1]温州医学院低氧医学研究所肺心病研究室机能实验教学中心,浙江温州325035

出　　处：《中国应用生理学杂志》2011年第3期270-274,385,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(30871031/C140305);浙江省自然科学基金资助项目(Y2091033);高原医学教育部重点实验室(第三军医大学)资助项目(2009GK04);温州市科技计划项目(Y20070083)

摘　　要：目的:探讨内质网应激介导的凋亡在低氧性肺动脉高压大鼠肺组织中的变化及意义。方法:清洁级雄性SD大鼠22只随机被均分成对照组和低氧组(n=11)。采用常压低氧法复制慢性低氧高二氧化碳性肺动脉高压模型,4周后,测定肺动脉平均压(mPAP)、右心室游离壁(RV)和左心室加室间隔(LV+S)重量比、肺细小动脉管壁面积/管总面积(WA/TA)、管腔面积/管总面积(CA/TA)及肺细小动脉中膜厚度(PAMT)作为模型指标。Hoechst染色检测肺组织凋亡细胞核。RT-PCR检测肺组织中胱冬酶12(caspase12)和葡萄糖调节蛋白78与94(GRP78、GRP94)mR-NA的表达水平。免疫组化法检测肺组织GRP78、GRP94蛋白的表达。结果:①与对照组相比,低氧组mPAP高50.5%,RV/(LV+S)高37.3%,WA/TA高72.5%,PAMT高137%,而CA/TA低41.9%(均P<0.01)。②Hoechst染色结果显示低氧组肺泡上皮细胞及肺血管内皮细胞可见较多呈高蓝光细胞核,肺血管中膜平滑肌细胞见低蓝光细胞核。③半定量RT-PCR结果显示,与对照组相比,低氧组肺组织caspase12 mRNA高144%,GRP78 mRNA高137%,GRP94 mRNA高80.7%(均P<0.05)。④免疫组化半定量结果显示,低氧组GRP78、GRP94蛋白明显高于对照组(均P<0.01),棕黄色颗粒表达主要位于肺泡上皮细胞及肺血管内皮细胞,而中层平滑肌层表达不明显。结论:内质网应激介导的凋亡与大鼠慢性低氧性肺动脉高压、肺血管改建的病理过程有关。Objective： To investigate the changes of endoplasmie reticultun stress-induced apoptosis in pulmonary tissue of rats with hypaxic pulmonary hypertension. Mehods： Twenty two male SD rats were randomly divided into control group and 4-week hypoxia-hypercapnia group（ n = 11）. The mean pulmonary arterial pressure （mPAP） and the mean carotid arterial pressure （reCAP） were monitored, and the weight ratio of right ventricle （RV） to left ventricle plus septum （LV ＋ S） were measured. The rattish pathological model were assessed by mPAP, reCAP, RV/（LV ＋ S）, vessel wall area/total area （WA/TA）, vessel cavity area/total area （CA/TA） and media thickness of pulmonary arteriole （PAMT）. The pulmonary apoptotic cells were detected by Hoechst staining. RT-PCR was used to study the genetic expression of caspasel2, glucose regulated protein 78（GRP78） and GRP94 in pulmonary tissue. The expression of GRP94 and GRP78 proteins in pulmonmy tissue were determined by using immunohistochemistry. Results： （1）The mPAP, RV/（LV ＋ S）, WA/TA and PAMT were respectively higher by 50.5%, 37.3%, 72.5% and 137% in hypoxic group than those in control group, while CA/TA was lower by 41.9 % （all P 〈 0.01）. There was not significant difference of reCAP between the two groups. （2）Heechst staining showed that the pulmonary apoptotic ceils in hypoxic group outnumbered markedly than those in control group, and the apaptotic cells were mainly in pulmonary tissue, while they were rare in pulmonary vascular smooth muscle cell. （3）Compared with control group, the expression of pulmonary caspase12, GRF78 and GRP94 mRNA in hypoxic group were higher by 144 %, 137 % and 80.7 % （all P 〈 0.05）, respectively. （4）The expression of pulmonary GRP78 and GRP94 proteins were up-regnlated in hypoxic group, and these proteins mainly localized in pulmonary vascular endothelial cell. Conclusion： The endoplasmic reticulum stress-induced apoptosis may be one of the mechanism of hypoxic pulmonary h

关 键 词：内质网应激 凋亡 高血压 肺性 低氧 

分 类 号：R363.2[医药卫生—病理学]