Calpain-1通过诱导内质网应激加速肺动脉高压的内皮细胞凋亡  

Calpain-1 accelerates endothelial cell apoptosis in pulmonary hypertension by inducing endoplasmic reticulum stress

作　　者：孙鹤宁 鲁美丽 田小雪 邓海艳 刘欢 王洪新 SUN He-ning;LU Mei-li;TIAN Xiao-xue;DENG Hai-yan;LIU Huan;WANG Hong-xin(Key Lab of Cardiovascular and Cerebrovascular Drugs,Jinzhou Medical University,Jinzhou Liaoning 121001,China)

机构地区：[1]锦州医科大学,辽宁省心脑血管药物重点实验室,辽宁锦州121001

出　　处：《中国药理学通报》2023年第4期723-730,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81973553)。

摘　　要：目的 探究calpain-1通过内质网应激促进低氧诱导肺动脉高压肺动脉内皮细胞凋亡的作用机制。方法 C57BL/6野生型(WT)和calpain-1基因敲除小鼠(KO)低氧仓中(10%O2)饲养4周诱导肺动脉高压模型,分为WT常氧对照(WT Con)、WT低氧模型(WT Hypoxia)、KO常氧对照(KO Con)和KO低氧(KO Hypoxia)4组。HPAECs在低氧(3%O2)培养箱中培养24 h模拟体内肺高压微环境,分为对照(Con)、模型(Hypoxia)、低氧+MDL28170(Hypoxia+MDL)和低氧+4-PBA(Hypoxia+4-PBA)4组。应用HE染色、TUNEL、免疫组化、免疫荧光、流式、Western blot检测组织形态、细胞凋亡,calpain-1,pPERK,CHOP,GRP78,caspase-12,caspase-3,Bax, Bcl-2表达水平。结果 与WT Con组相比,WT Hypoxia组小鼠血流动力学指数、肺组织中血管壁厚度比率和血管壁面积比率明显升高,calpain-1上调、内质网应激加重(pPERK,CHOP,GRP78升高)、内皮细胞凋亡增多,caspase-12、caspase-3、Bax增加,Bcl-2降低,而KO Hypoxia组逆转上述指标异常表达,凋亡和内质网应激减轻。与HPAECs Con组相比,Hypoxia组calpain-1上调,细胞凋亡、内质网应激明显,MDL28170处理后,上述效应逆转。4-PBA处理Hypoxia组细胞,calpain-1水平变化无意义但凋亡相关蛋白受到抑制。结论 Calpain-1通过促进内质网应激加速低氧诱导肺动脉高压肺动脉内皮细胞的凋亡。Aim To explore the mechanism by which calpain-1 promotes hypoxia-induced pulmonary hypertension pulmonary artery endothelial cell apoptosis through endoplasmic reticulum stress.Methods C57BL/6 wild-type(WT)and calpain-1 gene knockout mice(KO)were reared in a hypoxic chamber(10%O 2)for four weeks to induce pulmonary hypertension models,then they were divided into WT Con group,WT Hypoxia group,KO Con group and KO Hypoxia group.HPAECs were cultured in a hypoxic(3%O 2)incubator for 24 hours to simulate the microenvironment of pulmonary hypertension in vivo,then divided into Con group,Hypoxia group,Hypoxia+MDL group and Hypoxia+4-PBA group.HE staining,TUNEL,immunohistochemistry,immunofluorescence,flow cytometry,Western blot were applied to detect tissue morphology,cell apoptosis,calpain-1,pPERK,CHOP,GRP78,caspase-12,caspase-3,Bax,Bcl-2 expression level.Results Compared with the WT Con group,the hemodynamic index,the ratio of blood vessel wall thickness and the ratio of blood vessel wall area in the lungs of the WT Hypoxia group increased significantly,calpain-1 was up-regulated,endoplasmic reticulum stress increased(pPERK,CHOP,GRP78 increased),endothelial cell apoptosis increased,caspase-12,caspase-3,Bax increased,and Bcl-2 decreased,while the KO Hypoxia group reversed the abnormal expression of the above indicators,and decreased apoptosis and endoplasmic reticulum stress.Compared with the HPAECs Con group,calpain-1 was up-regulated in the Hypoxia group,and apoptosis and endoplasmic reticulum stress were significant.After MDL28170 treatment,the above effects were reversed.In Hypoxia group cells treated with 4-PBA,the level of calpain-1 was meaningless but apoptosis-related proteins were inhibited.Conclusion Calpain-1 accelerates hypoxia-induced pulmonary hypertension pulmonary artery endothelial cell apoptosis by promoting endoplasmic reticulum stress.

关 键 词：calpain-1 低氧 肺动脉高压 肺动脉内皮细胞 内质网应激 凋亡 

分 类 号：R-332[医药卫生] R329.25R544R845.22R977.3