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目的研究丹参通脉方对人主动脉血管内皮细胞(human vascular endothelial cell,HVEC)同型半胱氨酸(HCY)诱导模型内质网应激(ERS)偶联凋亡及炎症调节作用的影响,探讨丹参通脉方抗动脉粥样硬化的详细机制。方法应用HCY制作HVEC凋亡模型,丹参通脉方干预24h后,采取Annexin V-FITC/PI法检测细胞凋亡,免疫荧光检测ERS通路中BIP蛋白,实时荧光定量PCR法检测ERS通路中BIP、Chop的m RNA表达,同时使用ELISA法检测HVEC培养上清液中炎症因子表达。结果丹参通脉方抑制HVEC凋亡,与空白对照组及HCY模型组相比有统计学意义(P<0.05);丹参通脉方同时可抑制BIP、Chop细胞内m RNA的表达,通脉方组与空白组及模型组相比有统计学意义(P<0.05);丹参通脉方可抑制IL-6,IL-8及MCP-1炎症因子的表达(P<0.01)。结论丹参通脉方可抑制HCY诱导的HVEC凋亡,减轻HVEC炎症反应,其具体机制可能与抑制ERS有关。
Objective To investigate the effect of Danshen Tongmai Fang on endoplasmic reticulum stress (ERS) -coupling apoptosis and inflammation in human homocysteine (HCY) -induced model of human vascular endothelial cell (HVEC) To explore the detailed mechanism of Danshen Tongmai against artherosclerosis. Methods HVEC apoptosis model was made by using HCY. The cells were stained with Annexin V-FITC / PI for 24 hours, then BIP protein was detected by immunofluorescence and BIP in ERS pathway by real-time fluorescence quantitative PCR. Chop m RNA expression, while using ELISA method to detect the expression of inflammatory cytokines in HVEC culture supernatant. Results Danshen Tongmai Fang inhibited the apoptosis of HVEC and had statistical significance compared with the blank control group and the HCY model group (P <0.05). The Danshen Tongmai prescription could inhibit the expression of m RNA in BIP and Chop cells simultaneously. (P <0.05). Compared with the blank group and model group, Danshen Tongmai could inhibit the expression of IL-6, IL-8 and MCP-1 inflammatory factors (P <0.01). Conclusion Danshen Tongmai can inhibit the apoptosis of HVEC induced by HCY and reduce the inflammatory reaction of HVEC. The specific mechanism may be related to the inhibition of ERS.