健脾补肾方对再生障碍性贫血患者CD+34和Fas表达的影响

Effects of Jianpibushen prescription on the expression levels of CD+34 and Fas in patients wit aplastic anemia

目的观察健脾补肾方对再生障碍性贫血(AA)患者骨髓单个核细胞(BMNC)CD+34细胞占比、Fas表达的影响,并从自噬、凋亡角度探讨其作用机制。方法将2016年1月至2018年6月收治的90例AA患者纳入研究范围,按照随机数字表法随机分为对照组和治疗组,每组45例。对照组给予环孢素A治疗,同时根据病情给予抗感染、补血及止血药物。治疗组给予健脾补肾方治疗,1个月为1个疗程,共治疗2个疗程。流式细胞术测定2组治疗前后BMNC CD+34细胞占比、Fas表达及凋亡率。Real time-PCR和Western blot测定2组治疗前后BMNC自噬、凋亡相关基因mRNA和蛋白表达。结果治疗前2组骨髓CD+34细胞占比、Fas表达、凋亡率及LC3-Ⅱ、Bcl-2、caspase-3、PARP mRNA和蛋白表达比较差异无统计学意义(P>0.05)。治疗后2组骨髓CD+34细胞占比、LC3-Ⅱ、Bcl-2 mRNA和蛋白表达均明显高于治疗前,Fas表达、凋亡率及caspase-3、PARP mRNA和蛋白表达均明显低于治疗前,组间比较差异有统计学意义(P<0.05),且治疗组CD+34细胞占比、LC3-Ⅱ、Bcl-2 mRNA和蛋白表达高于对照组,Fas表达、凋亡率及caspase-3、PARP mRNA和蛋白表达低于对照组(P<0.05)。结论健脾补肾方治疗AA疗效确切,能够明显上调CD+34细胞占比,下调Fas表达,并抑制BMNC凋亡,其机制与调节自噬、凋亡相关基因表达有关。

Objective To investigate the effects of Jianpibushen prescription on the expression levels of CD+34 and Fas in bone marrow mononuclear cells(BMNC)of patients with aplastic anemia(AA),and to explore its action mechanism from the aspects of autophagy and apoptosis.Methods A total of 90 patients with AA were enrolled in the study,who were divided into observation group and control group,with 45 cases in each group.The patients in control group were treated by cyclosporine A and anti-infection,nourishing blood and hemostatic drugs,however,the patients in observation group were treated by Jianpibushen prescription,with a treatment course of two months for both geoups.The levels of CD+34 cells proportion,Fas expression and apoptosis rate before and after treatment were detected by flow cytometry.The autophagy and the expression levels of apoptosis related genes mRNA and protein before and after treatment were measured by Real time-PCR and Western Blot,respectively.Results Before treatment,there were no significant differences in CD+34 cells proportion,Fas expression levels,apoptosis rate and the expression levels of LC3-Ⅱ,Bcl-2,caspase-3 and PARP mRNA and proteins in BMNC between the two groups(P>0.05).After treatment,the CD+34 cells proportion and the expression levels of LC3-Ⅱand Bcl-2 mRNA and proteins in BMNC were significantly increased in both groups,however,the Fas expression levels,apoptosis rate and the expression levels of caspase-3 and PARP mRNA and proteins in BMNC were significantly decreased in both groups(P<0.05).Moreover the improvement degree of the indexes mentioned above in observation group was more significant than that in control group(P<0.05).Conclusion Jianpibushen prescription has definite clinical efficacy in treatment of AA,which can up-regulate CD+34 cells proportion,down-regulate Fas expression and apoptosis rate,inhibit the apoptosis of BMNC,and its action mechanism may be associated with regulating the expression of autophagy and apoptosis related genes.