补肾解毒法对HBV感染免疫耐受期产妇脐带血浆细胞样树突状细胞中PI3K、mTOR、p70S6K、IFN-α表达的影响

The effects of Bushen Jiedu decoction on expression of PI3K,mTOR,p70S6K and IFN-α in umbilical cord blood pDCs of HBV infected puerpera at immune tolerance stage

目的:研究补肾解毒法对HBV感染免疫耐受期产妇脐带血浆细胞样树突状细胞(pDCs)中磷脂酰肌醇3-激酶(PI3K)、哺乳动物雷帕霉素靶蛋白(mTOR)、p70核糖体S6蛋白激酶(p70S6k)、干扰素-α(IFN-α)表达的影响。方法:收集HBV感染免疫耐受期产妇脐带血10例,分离培养pDCs。同时制备补肾方、解毒方、补肾解毒方含药血浆及无药血浆。将培养至第7天的pDCs随机分为5组,即补肾组、解毒组、补肾解毒组、无药血浆组及空白组,干预48 h后,分别收集细胞及上清液,实时PCR检测pDCs PI3K、mTOR、p70S6K mRNA的表达,Western Blot技术检测pDCs PI3K、mTOR、p70S6K蛋白表达,ELISA检测pDCs培养上清IFN-α水平。结果:补肾方和补肾解毒方能提高HBV感染免疫耐受期产妇脐带血pDCs中PI3K、mTOR、p70S6K mRNA、蛋白及IFN-α的表达,以补肾解毒方更明显,而解毒方不能明显提高HBV感染免疫耐受期产妇脐带血pDCs中PI3K、mTOR、p70S6K mRNA、蛋白及IFN-α的表达。结论:补肾解毒方能明显提高HBV感染免疫耐受期产妇脐带血pDCs中PI3K、mTOR、p70S6K mRNA、蛋白的表达,使IFN-α分泌增加,改善HBV感染患者pDCs的功能。

Objective: To study the effects of Bushen Jiedu decotion on expression of PI3 K,mTOR,p70 S6 K and IFN-α in umbilical cord blood pDCs of HBV infected puerpera at immune tolerance stage. Methods: 10 cases of HBV infected puerpera at immune tolerance stage were selected,and pDCs of umbilical cord blood were isolated and cultured.At the same time,the plasma containing drugs of Bushen decotion,Jiedu decotion and Bushen Jiedu decotion were prepared.On the 7 th day,the pDCs were randomly divided into 5 groups,Bushen drug plasma group,Jiedu drug plasma group,Bushen Jiedu drug plasma group,blank plasma group and blank group.After cultured with plasma for 48 hours,pDCs and supernatant were collected,PI3 K,mTOR,p70 S6 K mRNA were detected by real time PCR,PI3 K,mTOR,p70 S6 K protein were detected by Western blot,IFN-α in supernatant were measured by ELISA. Results :The plasma containing drugs of Bushen Decoction and Bushen Jiedu Decoction can improve the expression of PI3 K,mTOR,p70 S6 K mRNA,protein,and IFN-α in umbilical cord blood pDCs of HBV infected puerpera at immune tolerance stage,especially in the Bushen Jiedu group.Jiedu decotion can’t improve the expression of PI3 K,mTOR,p70 S6 K mRNA,protein,and IFN-α in umbilical cord blood pDCs of HBV infected puerpera at immune tolerance stage statistically. Conclusion:Bushen Jiedu Decoction can significantly improve the expression of PI3 K,mTOR,p70 S6 K mRNA and protein in umbilical cord blood pDCs of HBV infected puerpera at immune tolerance stage,increase the secretion of IFN-α and improve the function of pDCs in patients infected with HBV.