加味芍药甘草汤调控P53-273H对子宫腺肌细胞增殖的影响

Influence of Jiawei Shaoyao Gancao Tang on proliferation of adenomyosis cells through regulating P53-273H

目的研究加味芍药甘草汤通过调控肿瘤抑制因子的突变型P53-273H对子宫腺肌细胞增殖的影响。方法取人子宫腺肌细胞进行细胞培养,随机分为加味芍药甘草汤组、米非司酮组和空白组3组,分别给予对应的药物处理24 h和48 h后,通过流式细胞术(FCM)检测加味芍药甘草汤对人子宫腺肌病病灶细胞凋亡的影响;反转录实时定量PCR(RT-q PCR)技术检测子宫腺肌细胞P53-273H m RNA、白血病抑制因子受体(LIFR)m RNA表达的影响;蛋白质印迹法(Western blot)检测药物对细胞中P53-273H、白血病抑制因子(LIF)、信号转导与转录激活子3(STAT3)、磷酸化的信号转导与转录激活子3(p-STAT3)蛋白表达的影响。结果 FCM检测显示,加味芍药甘草汤组细胞的凋亡率明显高于空白组(P<0.01),其差异具有统计学意义。RT-q PCR检测显示,加味芍药甘草汤组细胞的P53-273H m RNA、LIFR m RNA的相对表达量较空白组低,与空白组比较P<0.05,其差异有统计学意义。Western blot检测显示,加味芍药甘草汤处理24 h和48 h后,子宫腺肌细胞中的P53-273H、LIF、STAT3、p-STAT3蛋白的表达量均低于米非司酮组和空白组,与米非司酮组比较P<0.05,与空白组比较P<0.05,差异均具有统计学意义。结论加味芍药甘草汤能在一定程度上抑制子宫腺肌细胞的增殖、并促进其凋亡,其作用机制可能是加味芍药甘草汤通过调控P53/LIF/STAT3信号通路而实现的。

Objective To study the influence of Jiawei Shaoyao Gancao Tang （ Modified Debark Peony Root and Liquorice Root Decoction, JSGT） on proliferation of adenomyosis（AM） cells through regulating P53-273H. Methods The human AM cells were collected and cultured, and then divided randomly into JSGT group, mifepristone group and blank group. After treatment with corresponding medicinals for 24 h and 48 h, the influence of JSGT on apoptosis of AM focus cells was detected by using flow cytometry （FCM）. The expressions of P53-273H mRNA and LIFR mRNA of AM cells were detected by using real- time reverse transcription quantitative polymerase chain reaction （RT-PCR）. The protein expressions of P53-273H, LIF, STAT3 and p-STAT3 were detected by using Western blotting assay. Results The results of FCM showed that the apoptosis rate was significantly higher in JSGT group than that in blank group （P 〈 0.01 ）. The results of real-time RT-PCR showed that the relative expressions of P53-273H mRNA and LIFR mRNA were lower in JSGT group than those in blank group （P 〈 0.05）. The results of Western blotting assay showed that, after intervention with JSGT for 24 h and 48 h, the protein expres- sions of P53-273H, LIF, STAT3 and p-STAT3 were lower in JSGT group than those in mifepristone group （P 〈 0.05） and blank group （P 〈 0.05 ）. Conclusion JSGT can inhibit the proliferation and improve the apoptosis of AM cells, and the mechanism may be related to that JSGT can regulate P53/LIF/STAT3 signal pathway.