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目的探讨中药活性成分黄芪甲苷(Astragaloside IV,ASIV)、羟基红花黄色素A(hydroxysafflor yellow A,HSYA)和芍药苷(paeoniflorin,PAE)对小鼠主动脉平滑肌细胞(ASMCs)泡沫化过程的影响,并探讨其可能的分子机制。方法体外培养小鼠ASMCs,并利用免疫荧光染色对其进行鉴定。利用氧化型低密度脂蛋白(oxLDL)和肿瘤坏死因α(TNFα)诱导ASMCs泡沫化,并应用ASIV、HSYA和PAE进行干预,观察其对泡沫细胞形成的影响,应用油红O染色检测细胞形态学改变、酶促比色法测定细胞内胆固醇含量变化;应用RT-PCR和Western blot方法检测凝集素样氧化型低密度脂蛋白1(LOX-1)表达的变化。结果 oxLDL(80μg/m L)和TNFα(30 ng/mL)联用72 h可诱导ASMCs泡沫化,细胞内脂质颗粒明显增多,细胞内胆固醇含量增加,LOX-1表达上调;ASIV、HSYA和PAE显著抑制上述各过程。结论 ASIV、HSYA和PAE能够抑制ASMCs泡沫化,抑制oxLDL诱导的LOX-1表达可能是其作用机制之一。
Objective To investigate the effects of astragaloside IV (ASIV), hydroxysafflor yellow A (HSYA) and paeoniflorin (PAE) on the foaming process of mouse aortic smooth muscle cells (ASMCs) Influence, and explore its possible molecular mechanism. Methods Mouse ASMCs were cultured in vitro and identified by immunofluorescence staining. ASMCs were induced to foam by using oxidized low density lipoprotein (oxLDL) and tumor necrosis factor α (TNFα), and ASIV, HSYA and PAE were used to interfere the formation of foam cells. Oil red O staining was used to detect cell morphology The change of intracellular cholesterol content was determined by enzymatic colorimetry. The expression of LOX-1 was detected by RT-PCR and Western blot. Results The combination of oxLDL (80μg / mL) and TNFα (30 ng / mL) for 72 h could induce the foaming of ASMCs, the intracellular lipid granules increased obviously, the intracellular cholesterol increased and the expression of LOX-1 increased. ASIV, HSYA and PAE significantly inhibited the above processes. Conclusions ASIV, HSYA and PAE can inhibit the foaming of ASMCs and the inhibition of oxLDL-induced LOX-1 expression may be one of its mechanisms.