摘要
目的探讨司美格鲁肽对高糖诱导的心肌成纤维细胞(CFs)增殖中的作用,及其可能的机制。方法取原代小鼠CFs,通过CFs的特异性抗体波形蛋白Vimentin鉴定后进行下一步实验。以25 mmol·L^(-1)葡萄糖刺激细胞增殖,5~20 nmol·L^(-1)司美格鲁肽进行干预,并筛选浓度。分别设置正常组:含5.5 mmol·L^(-1)葡萄糖DMEM;高糖组:25 mmol·L^(-1)葡萄糖;司美格鲁肽组:25 mmol·L^(-1)葡萄糖+10 nmol·L^(-1)司美格鲁肽。应用Cell Cycle Staining Kit检测细胞周期;RT-qPCR方法检测α-平滑肌肌动蛋白(α-SMA)、TGF-β1、Smad3的mRNA表达变化;用ELISA检测Ⅰ型胶原(CoLⅠ)和Ⅲ型胶原(CoLⅢ)。结果与对照组相比,高糖诱导的CFs增殖明显增强(P<0.05),细胞周期中S期细胞比例增加;与高糖组相比,司美格鲁肽抑制了CFs的增殖(P<0.05),减少CFs细胞周期中S期细胞比例。与对照组相比,高糖组α-SMA、CoLⅠ、CoLⅢ显著增加(P<0.01);与高糖组相比,司美格鲁肽组α-SMA、CoLⅠ、CoLⅢ显著减少(P<0.01)。与对照组相比,高糖组TGF-β1、Smad3显著增加(P<0.01);与高糖组相比,司美格鲁肽组TGF-β1、Smad3显著减少(P<0.01)。结论司美格鲁肽可以抑制高糖诱导的CFs增殖和胶原蛋白的合成,进而发挥抗心肌纤维化的作用,其机制可能与下调TGFβ/Smad3信号通路有关。
Objective To investigate the effect of semaglutide on high glucose-induced proliferation of cardiac fibroblasts(CFs)and explore its possible mechanism.Methods Primary mouse CFs,identified by detecting vimentin expression,were stimulated with 25 mmol/L and treated with 5-20 nmol/L semaglutide,and the cell proliferation was examined with CKK-8 assay for concentration screening.Cultured CFs exposed to high glucose(25 mmol/L)were treated with 5 nmol/L semaglutide,and the changes in cell cycle were detected using Cell Cycle Staining Kit;The mRNA expressions ofα-smooth muscle actin(α-SMA),transforming growth factor-β1(TGF-β1)and Smad3 were detected using RT-qPCR,and the levels of type I collagen(CoLⅠ)and type III collagen(CoLIII)in the cell cultures were determined with ELISA.Results Compared with the control cells,the CFs cultured in high glucose exhibited significantly enhanced proliferative activity(P<0.05)with increased percentage of S-phase cells.Semagutide treatment obviously inhibited high glucose-induced proliferation of the CFs(P<0.05)and reduced the percentage of S-phase cells.High glucose stimulation significantly increased the mRNA expressions ofα-SMA,CoLⅠand CoLIII in the cells(P<0.01),which were effectively lowered by semaglutide treatment(P<0.01).The expressions of TGF-β1 and Smad3 were significantly increased in high glucose-stimulated CFs(P<0.01)and were lowered by semaglutide treatment(P<0.01).Conclusion Semaglutide can inhibit high glucose-induced proliferation and collagen synthesis in mouse CFs possibly by downregulating the TGFβ/Smad3 signaling pathway.
作者
李京娟
李凡凡
牛璐
安斌斌
成晓琼
刘媛媛
王金羊
LI Jingjuan;LI Fanfan;NIU Lu;AN Binbin;CHENG Xiaoqiong;LIU Yuanyuan;WANG Jinyang(First Clinical Medical School,Gansu University of Chinese Medicine,Lanzhou 730000,China;Department of Endocrinology,Gansu Provincial People's Hospital,Lanzhou 730000,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2023年第11期1935-1940,共6页
Journal of Southern Medical University
基金
国家自然科学基金(81560143,82360163)
甘肃省自然科学基金(21JR1RA013)
甘肃省兰州市城关区科学技术局项目(2020JSCX0039)。
