胰高血糖素样肽1受体激动剂...素诱导大鼠心肌纤维化的作用_程晓洪.pdf

安 徽 医 药 Anhui Medical and Pharmaceutical Journal 2023 Feb，27（2）胰高血糖素样肽1受体激动剂对阿霉素诱导大鼠心肌纤维化的作用程晓洪，李家富，宋昕雨，陈俞瑾，王欢作者单位：西南医科大学附属医院心血管内科，四川 泸州646000通信作者：李家富，男，主任医师，硕士生导师，研究方向为心脏起搏器植入、心内电生理检查，Email：基金项目：国家自然科学基金项目（31300946）；四川省心血管疾病防治协同创新中心基金项目（xtcx2019-14）；四川省泸州市-川医大联合课题 2015LZCYD-S03（7/7）摘要：目的 研究胰高血糖素样肽1受体（GLP-1R）激动剂对阿霉素诱导的心肌纤维化大鼠的影响及机制。方法 将40只SD大鼠随机数字表法分为正常组（N组）、阿霉素组（DOX组）、阿霉素+度拉糖肽组（GLP-1RA组）、阿霉素+度拉糖肽+Exendin（9-39）组（EX9-39组）。除N组外，利用阿霉素（3 mg/kg，每周3次）建立心肌纤维化大鼠模型，GLP-1RA组予以度拉糖肽1 mg kg-1 w-1 皮下注射，EX-9-39组予以度拉糖肽 1 mg kg-1 w-1皮下注射及Ex9-39 200 g kg-1 d-1腹腔注射。4周后完善心脏超声测量左室收缩末期内径（LVDs），左室舒张末期内径（LVDd）和左室射血分数（LVEF）后处死大鼠，HE染色观察心脏组织形态学变化；Masson染色检测胶原含量；蛋白质印迹法（Western blotting）测GLP-1R、肌质网钙泵（SERCA2a）、受磷蛋白（PLB）、磷酸化受磷蛋白（p-PLB）、蛋白激酶G（PKG）、钙调蛋白激酶（CaMK）、磷酸化兰尼碱受体（p-RyR）；酶联免疫吸附试验（ELISA）测N端脑钠肽前体（NT-proBNP）、环单磷酸鸟苷（cGMP）、胰高血糖素样肽-1（GLP-1）、血糖（GLU）、胰岛素（INS）。结果 各组间血糖及胰岛素的浓度均差异无统计学意义（P0.05）；HE染色提示DOX组纤维化程度较N组重，度拉糖肽可改善心肌纤维化，Exendin9-39 可部分抑制度拉糖肽作用；Masson 染色结果显示，与 N 组相比，DOX 组 CVF 明显升高 （5.880.13）%比（0.30.05）%；给予度拉糖肽治疗后，GLP-1RA组 （0.830.14）%CVF较DOX组下降；Exendin9-39削弱了度拉糖肽的保护作用（均P0.05）；蛋白质印迹法（Western blotting）结果显示DOX组与N组相比，GLP-1R、PKG、SERCA2a、p-PLB水平降低，PLB、p-RYR、CaMK水平升高，给予度拉糖肽治疗后可使GLP-1R、PKG、SERCA2a、p-PLB水平升高，PLB、p-RYR、CaMK水平降低，Exendin9-39可部分削弱度拉糖肽作用（均P0.05）；ELISA结果显示与N组相比，DOX组cGMP、GLP-1有所下降，GLP-1RA组则可使cGMP、GLP-1升高，Exendin9-39可使cGMP、GLP-1部分下降（均P0.05）；心脏超声与NT-proBNP结果显示，与N组相比，DOX组LVDs（4.000.34）mm、LVDd（6.000.52）mm值明显上升，LVEF（68.331.53）%降低，给予度拉糖肽治疗后，LVDs（2.990.06）mm、LVDd（4.600.21）mm值降低，LVEF（81.003.61）%升高，Exendin9-39可拮抗度拉糖肽的保护作用（均P0.05）。与N组相比，DOX组NT-proBNP明显升高，予以度拉糖肽治疗后NT-proBNP有所下降，而予以Exendin9-39可抵消部分度拉糖肽保护作用（均P0.05）。结论 GLP-1RA可改善阿霉素诱导的心肌纤维化而逆转心肌重构；GLP-1RA逆转心肌重构可能与cGMP-PKG通路相关。关键词：胰高血糖素样肽-1受体；心肌纤维化；心室重构；cGMP-PKG信号通路Effect of GLP-1R agonist on doxorubicin-induced cardiac fibrillation in ratsCHENG Xiaohong,LI Jiafu,SONG Xinyu,CHEN Yujin,WANG HuanAuthor Affiliation:Department of Cardiology,the Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000,ChinaAbstract：Objective Effect of GLP-1R agonist on doxorubicin-induced cardiac fibrillation in rats.Methods 40 SD rats were divided into four groups randomly:normal group(N group),doxorubicin group(DOX group),doxorubicin+dulapeptide group(GLP-1RA group),doxorubicin+dulapeptide+Exendin(9-39)group(EX9-39 group).doxorubicin(3 mg/kg tiw)was used to establish the rat model of myocardial fibrosis,except N group.GLP-1RA rats received subcutaneous injections of dulapeptide at a dose of 1 mg kg-1 w-1,and EX9-39 rats received subcutaneous injections of dulapeptide at a dose of 1 mg kg-1 w-1 and Ex9-39 at a dose of 200 g kg-1 d-1.Four weeks later,after ultrasonic measurements of LVDd,LVDs,LVEF and FS,the rats were sacrificed.HE staining was used to observe morphological changes in heart tissue,Masson staining was used to detect collagen content,Western blotting was used to detect GLP-1R,SERCA2a,PLB,p-PLB,PKG,CaMK,and p-RyR.NT-proBNP,cGMP,GLP-1,blood glucose and insulin were measured by Elisa.Results Blood glucose and insulin levels were not significantly different between the three groups.HE staining showed that the 药学研究引用本文：程晓洪，李家富，宋昕雨，等.胰高血糖素样肽1受体激动剂对阿霉素诱导大鼠心肌纤维化的作用 J.安徽医药，2023，27（2）：236-240.DOI：10.3969/j.issn.1009-6469.2023.02.006.236安 徽 医 药 Anhui Medical and Pharmaceutical Journal 2023 Feb，27（2）degree of fibrosis in DOX group was more severe than that in N group,and Exendin9-39 could partially inhibit myocardial fibrosis.Masson staining showed that CVF in DOX group(5.88 0.13)%was significantly higher than that in N group.After treatment with Dura glycopeptide,CVF in GLP-1RA group was lower than that in DOX group,and exendin9-39 antagonized the protective effect of Dura glycopeptide(all P 0 05).Western blotting results showed that the levels of GLP-1R,PKG,SERCA2a and p-PLB in DOX group were lower than those in N group,while the levels of PLB,p-RYR and CaMK were higher in Dulapeptide group.After treatment with Dulapeptide,the levels of GLP-1R,PKG,SERCA2a and p-PLB were increased,and the levels of PLB,p-RYR and CaMK were decreased.Exendin9-39 antagonized part of the effect of Dulapeptide.The results of Elisa showed that compared with N group,cGMP and GLP-1 decreased in DOX group,cGMP and GLP-1 increased in GLP-1RA group,and cGMP and GLP-1 decreased partly in exendin9-39 group.The results of echocardiography and NT-proBNP showed that the values of LVDs(4.000.34)mm and LVDD(6.000.52)mm in DOX group were significantly higher than those in N group,while the values of LVEF(68.331.53)%in Dulapeptide group were lower than those in N group.After treatment with Dulapeptide,LVDs(2.990.06)mm and LVDd(4.600.21)mm decreased,while LVEF(4.600.21)mm increased.Exendin9-39 inhibited the protective effect of Dulapeptide.Compared with N group,NT-proBNP in DOX group increased significantly,NT-proBNP decreased after treatment with dulapeptide,but exendin9-39 partially counteracted the protective effect of dulapeptide.Conclusions Doxorubicin-induced myocardial fibrosis can be reversed with GLP-1RA.cGMP-PKG signal pathway may b