摘要
目的从细胞自噬的角度,探讨外源性Apelin对肺动脉高压(PAH)的作用及相关机制。方法 26只雄性SD大鼠随机分为对照组(n=6)、模型组(n=10)和干预组(n=10),模型组和干预组采用左肺切除术联合野百合碱注射法(PE+MCT)制备大鼠PAH模型,对照组仅打开胸腔+注射等量生理盐水。从术后第2周开始,干预组每日按大鼠体质量腹腔注射10 nmol/kg Apelin-13,连续3周,对照组与模型组均注射等量生理盐水。3组大鼠均于术后第5周测量平均肺动脉压力(mPAP),计算右心室肥厚指数(RVHI),行HE染色观察肺组织及肺血管形态结构,肺组织免疫荧光染色检测自噬相关蛋白LC3的表达,实时荧光定量(qRT)-PCR测量肺组织自噬相关蛋白P62、Beclin-1的mRNA表达量,Western blot测量肺组织LC3、LC3-Ⅱ/LC3-Ⅰ、P62、Beclin-1的蛋白表达量,肺组织免疫荧光染色检测自噬相关蛋白LC3的定位及表达。结果与对照组相比,模型组mPAP、RVHI升高(P<0.05),肺组织结构紊乱、肺血管壁增厚,肺组织LC3蛋白表达量升高,LC3-Ⅱ/LC3-Ⅰ升高,Beclin-1的mRNA和蛋白表达量升高,P62的mRNA和蛋白表达量下降(P<0.05);Apelin-13干预后,上述指标均得到改善(P<0.05,与模型组比较)。结论外源性Apelin对PAH的形成具有一定的防治作用,其作用机制可能与Apelin抑制自噬有关。
Objective To investigate the effect of exogenous Apelin on pulmonary artery hypertension(PAH and its related mechanism. Methods 26 male SD rats were randomly divided into Control group(n=6), Model group(n=10) and Intervention group(n=10). The rat model of PAH was established by left pneumonectomy combined with monocrotaline injection(PE+MCT) in the Model group and the Intervention group, while the Control group rats were opened chest cavity and injected the same amount of normal saline. From the 2 nd week after operation, the Intervention group was intraperitoneally injected with 10 nmol/(kg·d) Apelin-13 for 3 weeks, while the Control group and Model group were injected the same volume of normal saline. The mean pulmonary arterial pressure(mPAP) was measured and the right ventricular hypertrophy index(RVHI) was calculated in all three groups of rats at the 5 th week after operation.The pulmonary tissue HE staining was performed to observe the pulmonary tissue and pulmonary vascular morphology.Protein LC3 was detected by immunofluorescence staining of lung tissues, the mRNA expression level of P62 and Beclin-1 in lung tissues was measured by RT-PCR, and the protein expressions of LC3, LC3-Ⅱ/LC3-Ⅰ, P62 and Beclin-1 in lung tissues were measured by Western blot. Results Compared with the Control group, the Model group showed increased mPAP and RVHI(P<0.05), disordered pulmonary tissue structure and thicker pulmonary vascular wall. In Model group rats, expression of LC3 protein and LC3-Ⅱ/LC3-Ⅰ increased in lung tissues, and the expression of Beclin-1 mRNA and the Beclin-1 protein also increased in lung tissues, while the level of P62 mRNA and the expression of P62 protein decreased(P<0.05). After Apelin-13 intervention, the above indexes were all improved(P<0.05, compared with the Model group). Conclusion Exogenous Apelin has a certain preventive and therapeutic effect on the formation of PAH, and the mechanism may be related to its inhibition effect on autophagy.
作者
苟永英
刘东
李刚
胡娅南
贾鹏
刘瀚旻
刘斌
GOU Yong-ying;LIU Dong;LI Gang;HU Ya-nan;JIA Peng;LIU Han-min;LIU Biu(Department of Pediatrics,the First Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Pediatrics,West China Second University Hospital,Sichuan University,Chengdu 610041,China)
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2020年第2期193-199,共7页
Journal of Sichuan University(Medical Sciences)
基金
四川省科技厅-泸州市科技局课题(No.Z1432)资助。
作者简介
通信作者:刘斌,E-mail:lblyfy@126.com。
