基于NF-κB/NLRP3炎症小体途径探讨丹参通脉方抗动脉粥样硬化机制

Exploration on the anti-atherosclerotic mechanism of Danshen Tongmai Formula based on the NF-κB/NLRP3 inflammasome pathway

目的:探讨丹参通脉方含药血清通过NF-κB/NLRP3信号通路抗动脉粥样硬化(AS)的作用机制。方法:通过免疫荧光技术标记人主动脉血管平滑肌细胞(HA-VSMC)细胞标志蛋白α-平滑肌肌动蛋白(α-SMA)的表达进行细胞鉴定;采用油红“O”染色验证实验造模成功;制备含药血清,MTT法检测HA-VSMC增殖活性;应用免疫荧光法检测NF-κB P65、NLRP3炎症小体的荧光表达水平;应用蛋白质免疫印迹实验检测NF-κB P65、NLRP3炎症小体蛋白表达水平。结果:与模型组比较,丹参通脉方组可抑制HA-VSMC细胞过度增殖(P<0.05),抑制NF-κB P65活化磷酸化入核及NLRP3炎症小体的荧光表达,下调NF-κB P65、NLRP3蛋白水平(P<0.05)。结论:丹参通脉方可减少NF-κB通路诱导的NLRP3炎症小体表达,具有一定的抗炎、抗AS作用。

Objective:To investigate the anti-atherosclerotic(AS)mechanism of Danshen Tongmai Formula medicated serum through NF-κB/NLRP3 signaling pathway.Methods:The expression ofα-smooth muscle actin(α-SMA)was labeled by immunofluorescence technique for HA-VSMC;oil red‘O’staining was used to verify whether the experiment was successful;the drug containing serum was prepared,and the proliferative activity of HA-VSMC was screened by MTT method;NF-κB P65 and NLRP3 inflammatory bodies was detected by immunofluorescence method and Western Blot was used to detect the protein expression of NF-κB P65 and NLRP3.Results:Compared with model group,Danshen Tongmai Formula group could inhibit the excessive proliferation of HA-VSMC cells(P<0.05),inhibit the activation and phosphorylation of NF-κB P65 into nucleus and the fluorescence expression of NLRP3 inflammatory bodies,and down regulate the protein levels of NF-κB P65 and NLRP3(P<0.05).Conclusion:Danshen Tongmai Formula can reduce the expression of NLRP3 inflammatory bodies induced by NF-κB pathway,and has certain anti-inflammatory and anti-AS effects.