摘要
目的:探讨丹参来源纳米囊泡(SDNV)通过氧化物酶体增殖物激活受体γ(PPAR-γ)/肝X受体α(LXR-α)/三磷酸腺苷结合盒转运体A1(ABCA1)信号通路抑制氧化低密度脂蛋白(ox-LDL)诱导的平滑肌细胞源性泡沫细胞形成的作用机制。方法:采用透射电镜、纳米颗粒跟踪分析对超高速差速离心法提取的SDNV进行鉴定;选择MOVAS小鼠主动脉血管平滑肌细胞株和ox-LDL诱导建立泡沫细胞模型;通过激光共聚焦观察MOVAS细胞对SDNV的摄取情况;采用CCK8法检测SDNV对MOVAS细胞存活率的影响;将细胞分为对照组、模型组、SDNV组及辛伐他汀组,微板法检测各组细胞内总胆固醇(TC)、游离胆固醇(FC)、胆固醇酯(CE)含量,并计算CE/TC比值;油红O染色观察各组细胞内脂质蓄积的情况;Western Blot分析各组细胞中PPAR-γ、LXR-α、ABCA1蛋白表达水平。结果:SDNV通过透射电镜、纳米颗粒跟踪分析鉴定SDNV符合纳米囊泡的表征及粒径要求;激光共聚焦观察到SDNV可以被MOVAS细胞摄取;CCK8实验结果显示SDNV对MOVAS细胞无明显毒性;油红O染色及CE/TC值的改变表明SDNV能够抑制MOVAS细胞的脂质蓄积,抑制泡沫细胞的形成;与模型组比较,SDNV组PPAR-γ、LXR-α、ABCA1蛋白表达水平显著升高(P<0.01)。结论:SDNV可以通过PPAR-γ/LXR-α/ABCA1信号通路抑制平滑肌源性泡沫细胞的形成,进而防治动脉粥样硬化。
Objective:To investigate the mechanism by which Salvia miltiorrhiza-derived nanometer vesicles(SDNV)inhibit the formation of smooth muscle cell-derived foam cells induced by oxidized low density lipoprotein(ox-LDL)via the PPAR-γ/LXR-α/ABCAI signal pathway.Methods:Transmission electron microscopy and nanoparticle tracking analysis were used to identify SDNV extracted by ultra-high speed differential centrifugation method.The foam cell model was established by using MOVAS mouse aortic smooth muscle cell line and oxidized low density lipoprotein.The uptake of SDNV by MOVAS cells was observed by confocal laser.The effect of SDNV on the survival rate of MOVAS cells was detected by CCK8 method.The cells were divided into control group,model group,SDNV group and simvastatin group.The contents of total cholesterol(TC),free cholesterol(FC)and cholesteryl ester(CE)in each group were detected by microplate method,and the CE/TC ratio was calculated.The intracellular lipid accumulation in each group was observed by oil red O staining.The protein expression levels of PPAR-γ,LXR-αand ABCAI in cells of each group were analyzed by Western Blot.Results:SDNV was identified by transmission electron microscopy and nanoparticle tracking analysis to meet the requirements of characterization and particle size of nanovesicles.The laser confocal observation showed that SDNV could be taken up by MOVAS cells.The results of CCK8 experiment showed that SDNV had no obvious toxicity to MOVAS cells.Oil red O staining and CE/TC values showed that SDNV could inhibit the accumulation of lipids and the formation of foam cells in MOVAS cells.Compared with the model group,the protein expression levels of PPAR-γ,LXR-αand ABCA1 in SDNV group significantly increased(P<0.01).Conclusion:SDNV can inhibit the formation of smooth muscle derived foam cells through PPAR-γ/LXR-α/ABCA1 signal pathway,and then prevent atherosclerosis.
作者
张林琦
武若愚
徐红俊
陈鸿旭
安胜军
ZHANG Linqi;WU Ruoyu;XU Hongjun;CHEN Hongxu;AN Shengjun(Hebei University of Chinese Medicine,Shijiazhuang 050200,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2024年第7期3685-3689,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.82204658)
河北省中医药管理局科研计划项目(No.2019073)
河北省重点研发计划项目(No.213777100D)
河北省自然科学基金中医药联合基金重点项目(No.H2022423319)。
