摘要
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 P53-273 H. 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 P53-273 H m RNA and LIFR m RNA of AM cells were detected by using realtime reverse transcription quantitative polymerase chain reaction(RT-PCR). The protein expressions of P53-273 H,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 P53-273 H m RNA and LIFR m RNA 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 expressions of P53-273 H,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.
引文
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