补肾益气方对血管内皮细胞p16基因表达的影响

Impact of Bushen Yiqi prescription on p16 gene expression in vascular endothelial cells

目的研究补肾益气方对大鼠血管内皮细胞p16基因表达的的影响。方法 24只健康雄性SD大鼠,随机分为中药组(补肾益气方灌胃)、黄芪组(黄芪水煎液灌胃)、生理盐水组(生理盐水灌胃),各8只。提取含药血清,培养人脐静脉内皮细胞株,采用β-半乳糖苷酶(β-gal)染色鉴定细胞衰老;流式细胞术分析细胞周期变化,用反转录聚合酶链式反应(RT-PCR)方法检测p16基因的表达。结果中药组、黄芪组G0/G1期细胞比例(63.18±1.07)、(71.73±1.32)%小于生理盐水组(79.56±2.88)%,中药组G0/G1期细胞比例小于黄芪组,差异均具有统计学意义(P<0.05)。中药组、黄芪组m RNA灰度值/β-肌动蛋白(β-actin)灰度值比值(0.1231±0.0064)、(0.1835±0.0057)小于生理盐水组(0.5126±0.0110),中药组m RNA灰度值/β-actin灰度值比值小于黄芪组,差异均具有统计学意义(P<0.05)。结论补肾益气方具有延缓血管内皮细胞凋亡的作用,其分子机制可能通过调控p16的表达使细胞周期停滞于G1期发挥作用,具有一定参考价值。

Objective To study the impact of Bushen Yiqi prescription on pl6 gene expression in vascular endothelial cells of rats. Methods A total of 24 healthy male SD rats were randomly divided into traditional Chinese medicine group （garage of Bushen Yiqi prescription）, astragalus group （garage of astragalus water decoction） and normal saline group （gavage of normal saline）, with 8 rats in each group. Drug-containing serum was extracted to cultivate human umbilical vein endothelial cell strain, and ±- galactosidase （β-gal） dyeing was used to identify cell aging. Cell cycle changes was analyzed by flow cytometry, and pl6 gene expression was detected by reverse transcriptase polymerase chain reaction （RT-PCR）. Results The traditional Chinese group and astragalus group had G0/G1 phase cell proportion as （63.18 ± 1.07） and （71.73 ± 1.32）%, which were less than （79.56 ± 2.88）% in normal saline group, and the traditional Chinese group had less G0/G1 phase cell proportion than astragalus group. Their difference had statistical significance （P〈0.05）. The traditional Chinese group and astragalus group had mRNA gray value/13-actin gray value ratio as （0.1231 ± 0.0064） and （0.1835 ± 0.0057）, which were less than （0.5126 ± 0.0110） in normal saline group, and the traditional Chinese group had less mRNA gray value/β -actin gray value ratio than astragalus group. Their difference had statistical significance （P〈0.05）. Conclusion Bushen Yiqi prescription plays a role in delaying vascular endothelial apoptosis, and its molecular mechanism may regulate the expression of p16 to stagnate the cell cycle at GI phase, along with certain reference value.