左、右归丸干预PGC-1α/PPARγ对骨髓间充质干细胞线粒体生物发生的影响  

Effect of Zuogui Wan and Yougui Wan on Mitochondrial Biogenesis in BMSCs Through PGC-1α/PPARγ

作　　者：杨莹 冯秀芝[1] 陈怡然[1] 王智民[2] 郭晛 任艳玲[1] YANG Ying;FENG Xiuzhi;CHEN Yiran;WANG Zhimin;GUO Xian;REN Yanling(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110033,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学附属医院,沈阳110033

出　　处：《中国实验方剂学杂志》2025年第9期28-36,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174260);辽宁省科学技术厅联合基金博士科研启动项目(2023-BSBA-224);辽宁省科技厅2024年度优秀研究生项目(2024-JYTCB-039)。

摘　　要：目的:基于“阳化气,阴成形”理论,探讨左、右归丸通过过氧化物酶体增殖激活受体γ共激活因子-1α(PGC-1α)及过氧化物酶体增殖物激活受体γ(PPARγ)影响大鼠骨髓间充质干细胞成脂分化过程中线粒体生物发生的分子机制,为左、右归丸防治绝经后骨质疏松症(PMOP)奠定更多理论依据。方法:将骨髓间充质干细胞(BMSCs)分为成脂诱导组(模型组)、左归丸组、右归丸组、补佳乐组,采用流式细胞仪进行细胞鉴定;噻唑蓝(MTT)比色法绘制BMSCs的生长曲线,并检测左、右归丸对BMSCs增殖的影响;油红O染色法检测脂滴的形成;蛋白免疫印迹法(Western blot)检测成脂相关因子PPARγ、CCAAT增强子结合蛋白(C/EBP)α、C/EBPβ、脂蛋白脂肪酶(LPL)蛋白,棕色脂肪相关蛋白PGC-1α、解偶联蛋白1(UCP1)、PR结构域蛋白16(PRDM16)蛋白,线粒体生物发生相关PGC-1α、核呼吸因子1(Nrf1)、核转录因子红系2相关因子2(Nrf2)、线粒体转录因子A(TFAM)蛋白的表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测成脂相关因子PPARγ、C/EBPα、C/EBPβ、LPL mRNA及细胞色素B(CytoB)基因拷贝数的表达;透射电镜观察线粒体超微结构。结果:与模型组比较,各给药组BMSCs增殖能力随干预时间增强而增强,药物干预后脂滴明显减少,成脂相关PPARγ、C/EBPα、C/EBPβ、LPL mRNA及蛋白表达水平显著下调(P<0.01);棕色脂肪相关PGC-1α、UCP1、PRDM16蛋白表达水平显著上调(P<0.01);线粒体数量增多,肿胀减少,双层膜和嵴结构清晰,同时内嵴断裂减少;CytoB基因拷贝数显著上调(P<0.01);线粒体生物发生相关PGC-1α、Nrf1、Nrf2、TFAM蛋白表达水平显著上调(P<0.01)。结论:左、右归丸均能通过PGC-1α/PPARγ影响BMSCs线粒体生物发生以防治PMOP。Objective:Based on the TCM theory of"Yang transforms materials to Qi while Yin constitutes material form",this paper explored the effects of Zuogui Wan and Yougui Wan on the molecular mechanism of mitochondrial biogenesis during the adipogenic differentiation process of rat bone marrow mesenchymal stem cells(BMSCs)by mediating peroxisome proliferatoractivated receptorγcoactivator-1α(PGC-1α)and peroxisome proliferators-activated receptorγ(PPARγ),providing theoretical support for the prevention and treatment of postmenopausal osteoporosis(PMOP)using Zuogui Wan and Yougui Wan.Methods:BMSCs were divided into a blank group,Zuogui Wan(ZGW)group,Yougui Wan(YGW)group,and Progynova group.Cell identification was performed using flow cytometry.The growth curves of BMSCs were plotted using the methylthiazolyldiphenyltetrazolium bromide(MTT)method,and the effects of Zuogui Wan and Yougui Wan on the proliferation of BMSCs were detected.The Oil red O staining method was used to detect lipid droplet formation.The Western blot method was used to detect the expression of adipogenesis-related factors PPARγ,CCAAT/enharcer-binding protein(C/EBP)α,C/EBPβ,lipoprotein lipase(LPL)protein,brown adipose tissue-related(BAT)proteins PGC-1α,uncoupcing protein 1(UCP1),PR domdin-containing protein 16(PRDM16),mitochondrial biogenesis-related PGC-1α,nuclear respiratory factor 1(Nrf1),nuclear factor E_(2)-related factor 2(Nrf2),and mitochondrial transcription factor A(TFAM).The expression of adipogenesis-related factors PPARγ,C/EBPα,C/EBPβ,LPL genes,and the copy number of cytochrome B(CytoB mtDNA)gene was detected using real-time polymerase chain reaction(Real-time PCR).Mitochondrial ultrastructure was detected using transmission electron microscopy.Results:Compared with that in the blank group,the proliferation ability of BMSCs in each treatment group increased continuously as the intervention progressed,and lipid droplets significantly decreased after the drug intervention.The mRNA and protein expression levels of adipogenesis-relat

关 键 词：左归丸 右归丸 绝经后骨质疏松症 过氧化物酶体增殖激活受体γ共激活因子-1α(PGC-1α) 线粒体生物发生 

分 类 号：R681[医药卫生—骨科学] R285[医药卫生—外科学] R289[医药卫生—临床医学]