CAJ | 학술논문

目的探讨天麻素对血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)诱导血管平滑肌细胞(vascular smooth muscle cell,VSMC)迁移的影响及其可能机制.方法酶消化法获取大鼠主动脉VSMC并传代纯化.免疫荧光染色法对VSMC标记蛋白进行鉴定.建立PDGF-BB诱导细胞迁移模型.Transwell小室法评价天麻素对PDGF-BB诱导VSMC迁移的影响.蛋白质印迹法检测c-Jun N末端激酶(c-jun N-terminal kinase,JNK)磷酸化水平.结果原代培养的VSMC纯度达99％以上.PDGF-BB组VSMC迁移数量为(85.2 ±3.486)个/视野,显著多于对照组的(42.5±1.927)个/视野(=9.981,P＜0.001),而天麻素能使PDGF-BB诱导VSMC迁移数量显著减少为(71.3 ±1.783)个/视野(t=3.550,P=0.002).蛋白质印迹分析显示,天麻素能抑制PDGF-BB诱导的JNK磷酸化(0.190±0.015对0.190±0.015;t=14.548,P=0.000).结论天麻素可抑制PDGF-BB诱导VSMC迁移,其机制可能与抑制JNK信号通路激活有关.
목적 탐토천마소대혈소판연생생장인자-BB(platelet-derived growth factor-BB,PDGF-BB)유도혈관평활기세포(vascular smooth muscle cell,VSMC)천이적영향급기가능궤제.방법 매소화법획취대서주동맥VSMC병전대순화.면역형광염색법대VSMC표기단백진행감정.건립PDGF-BB유도세포천이모형.Transwell소실법평개천마소대PDGF-BB유도VSMC천이적영향.단백질인적법검측c-Jun N말단격매(c-jun N-terminal kinase,JNK)린산화수평.결과 원대배양적VSMC순도체99％이상.PDGF-BB조VSMC천이수량위(85.2 ±3.486)개/시야,현저다우대조조적(42.5±1.927)개/시야(=9.981,P＜0.001),이천마소능사PDGF-BB유도VSMC천이수량현저감소위(71.3 ±1.783)개/시야(t=3.550,P=0.002).단백질인적분석현시,천마소능억제PDGF-BB유도적JNK린산화(0.190±0.015대0.190±0.015;t=14.548,P=0.000).결론 천마소가억제PDGF-BB유도VSMC천이,기궤제가능여억제JNK신호통로격활유관.
Objective To investigate the effect of gastrodin on rat vascular smooth muscle cell (VSMC) migration induced by platelet-derived growth factor-BB (PDGF-BB) and its possible mechanisms.Methods Enzyme digestion method was used to obtain rat aortic VSMCs and be purified by passage.Immunofluorescence staining was used to identify VSMC marker proteins.A PDGF-BB induced cell migration model was established.Transwell chamber assay was used to evaluate the effect of gastrodin on PDGF-BB induced VSMC migration.Western blots were performed to detect the phosphohorylation levels of c-jun N-terminal kinase (JNK).Results The purity of primary cultured VSMC was more than 99％.The VSMC migrated number in the PDGF-BB group was 85.2 ± 3.486 per field.It was significantly more than 42.5 ± 1.927 per field in the control group (t =9.981,P＜0.001),and gastrodin was enable to make PDGF-BB induced the number of VSMC migration significantly reduce to 71.3 ± 1.783 per filed (t=3.550,P =0.002).Western blots analysis showed that gastrodin inhibited PDGF-BB induced JNK phosphorylation (0.190 ± 0.015 vs.0.190 ± 0.015; t =14.548,P =0.000).Conclusions Gastrodin inhibits PDGF-BB induced VSMC migration,its mechanisms may be associated with the inhibition of the JNK signaling pathway activation.