高血压病气虚血瘀证患者血清对CRL-1730凋亡相关基因、miRNA表达的影响及补阳还五汤的干预作用

Effects of Serum of a Patient with Hypertensive Syndrome of Qi Deficiency and Blood Stasis on CRL-1730 Apoptosis-related Genes,miRNA Expression and Buyanghuanwu Decoction’s Intervening Action

为探讨高血压病气虚血瘀证患者血清对人脐静脉内皮细胞株(CRL-1730)凋亡的影响与其作用机制。应用实时定量PCR(polymerase chain reaction)芯片技术检测与miRNA芯片技术分别检测与人细胞凋亡相关的89个关键基因和384个miRNAs的表达。高血压病气虚血瘀证组与非气虚血瘀证组比较,有1个上调及3个下调miRNA,与健康对照组比较,89个基因有22个上调基因和1个下调基因,有8个上调及2个下调miRNA,有统计学差异(P<0.05);高血压病非气虚血瘀证组与健康对照组比较,89个基因有19个上调基因和1个下调基因,有12个上调及2个下调miRNA,有统计学差异(P<0.05)。补阳还五汤高剂量组与高血压病气虚血瘀证组比较,89个基因中有5个上调基因和1个下调基因,有6个上调miRNA,有统计学差异(P<0.05);中剂量组与高血压病气虚血瘀证组比较,89个基因有7个上调基因和3个下调基因,有3个下调miRNA,有统计学差异(P<0.05);低剂量组与高血压病气虚血瘀证组比较,89个基因中有5个上调基因和3个下调基因,有3个上调及1个下调miRNA,有统计学差异(P<0.05)。高血压病气虚血瘀证患者血清诱导内皮细胞凋亡的机制可能与凋亡相关的mRNA和miRNA表达失调有关,其中,TNFSF10具有关键作用,补阳还五汤对上述部分差异表达的凋亡相关分子表达具有调节作用。

In order to investigate the apoptosis of human umbilical vein endothelial cell line(CRL-1730)induced by the serum of patients with hypertension with Qi deficiency and blood stasis syndrome and observe its mechanism.The expression of 89 key genes and 384 miRNAs related to human cell apoptosis were detected by real-time quantitative PCR(polymerase chain reaction)chip technology and the miRNA chip technology respectively.Compared with the non-qi-deficiency and blood-stasis syndrome group,there were 1 up-regulated miRNA and 3 down-regulated miRNAs had beed detected in Qi-deficiency and blood-stasis syndrome group.Compared with the healthy control group,there were 22 up-regulated genes and 1 down-regulated gene in 89 genes,and there were 8 up-regulated and 2 down-regulated miRNAs had beed detected in Qi-deficiency and blood-stasis syndrome group,the difference was statistically significant(P<0.05).Compared with the healthy control group,there were 19 up-regulated genes and 1 down-regulated gene in 89 genes,and there were 12 up-regulated and 2 down-regulated miRNAs had beed detected in hypertension non-Qi deficiency blood stasis syndrome group,the difference was statistically significant(P<0.05).Compared with the hypertension Qi deficiency blood stasis syndrome group,there were 5 up-regulated genes and 1 down-regulated gene in 89 genes,and there were 6 up-regulated miRNAs had beed detected in the high-dose group of Buyang Huanwu decoction,the difference was statistically significant(P<0.05).Compared with the hypertension Qi deficiency blood stasis syndrome group,there were 7 up-regulated genes and 3 down-regulated gene in 89 genes,and there were 3 down-regulated miRNAs had beed detected in the middle-dose group of Buyang Huanwu Decoction,the difference was statistically significant(P<0.05).Compared with the hypertension Qi deficiency blood stasis syndrome group,there were 5 up-regulated genes and 3 down-regulated gene in 89 genes,and there were 3 up-regulated and 1 down-regulated miRNAs had beed detected in the low-dose group of Bu Yang Huan Wu Decoction,the difference was statistically significant(P<0.05).The mechanism of endothelial cell apoptosis induced by serum of hypertensive patients with qi deficiency and blood stasis syndrome may be related to the imbalance of apoptosis-related expression of mRNA and miRNA,in which TNFSF10 plays a key role,and Buyanghuanwu decoction can regulate the expression of apoptosis-related molecules mentioned above.