替普瑞酮对香烟提取物引起的气道平滑肌细胞凋亡中p-JNK表达的影响

Effect of Geranylgeranylacetone on the p-JNK Expression During the Apoptotic Process of Airway Smooth Muscle Cells Induced by Cigarette Smoke Extract

目的:探讨替普瑞酮(GGA)诱导的热休克蛋白70(HSP70)在香烟提取物(CSE)引起的气道平滑肌细胞(ASMCs)凋亡中对C-Jun N-terminal激酶(JNKs)的影响。方法:在体外培养Wistar大鼠的气道平滑肌细胞中,给予GGA处理前后,给予CSE刺激3 h。用流式细胞仪(FCM)检测细胞凋亡情况,Western blot的方法检测HSP70和p-JNK的表达。结果:CSE诱导ASMCs凋亡随CSE浓度的增加而逐渐增加,两者相比有显著性差异(P<0.05);GGA预处理后再加入不同浓度的CSE处理的气道平滑肌细胞的凋亡较单纯CSE组相比有显著性差异(P<0.05)。在给予CSE刺激后,各组中的HSP70的表达较对照组有明显的降低,而且随CSE浓度越高降低越明显(P<0.05);p-JNK的表达随CSE浓度的增高而增多(P<0.05)。GGA预处理后的各组细胞的HSP70的表达与对照组比较有明显的增多(P<0.05)。在给予GGA预处理后,再加入CSE处理的气道平滑肌细胞中,与对照组比较HSP70的表达有所降低但无显著差异,p-JNK的表达在GGA+CSE组中的表达较单纯CSE组中有所下降(P<0.05)。结论:较高浓度的CSE能引起气道平滑肌细胞的凋亡并且伴随p-JNK的表达的增多。GGA处理气道平滑肌细胞后引起HSP70表达的增加,并且抑制较高浓度CSE所致的平滑肌细胞的凋亡,这可能与HSP70下调p-JNK的活性有关。

To study the effect of HSP70 induced by geranylgeranylacetone （GGA） on p-JNK expression in the apoptosis of airway smooth muscle ceils （ASMCs） induced by cigarette smoke extract （CSE）. Methods： The ASMCs in Wistar rats were cultured in vitro and stimulated with different concentrations of CSE for three hours before and after treated by GGA. The apoptosis was determined by flow cytometry （FCM）, and the expression of HSPT0 and p-JNK were deter-mined by Western blot. Results： The apoptosis of ASMCs increased in CSE groups and it was associated with the rising CSE concentration （P〈0. 05, n= 4）; the apoptosis of ASMCs in CSE groups with previously treated by GGA were significantly decreased compared with those of the CSE groups （P%0.05）. After stimulating by CSE, the expression of HSPT0 was lower than that of the control group, and it was associated with the increased CSE concentration （P〈0.05） ; the expression of p-JNK was higher than that of control group, and it was also associated with the increased CSE concentration （P〈0.05）. The expression of HSPT0 in GGA pretreatment groups was significantly and respectively increased compared with that of the control （all P〈0.05）. The expression of HSPT0 was mildly decreased in CSE groups with GGA pretreatment but showed no significant difference, while the expression of p-JNK in CSE groups previously treated by GGA was significantly lower than that of in CSE groups respectively （all P〈0.05）. Conclusion： High- er concentration of CSE may induce the apoptosis of ASMCs and increase the expression of p-JNK. The expression of HSPT0 was increased and the apoptosis of ASMCs induced by higher concentration CSE was suppressed after the treatment of GGA, and it suggests that HSPT0 could down-regulate the expression of p-JNK during the process.