活血祛瘀法对真性红细胞增多症细胞凋亡及相关基因表达的影响

Influence of Blood- activating and Blood- stasis- removing Therapy on Cell Apoptosis and Expression of Associated Gene in Polycythemia Vera

目的探讨活血祛瘀法治疗真性红细胞增多症的作用机理。方法选取真性红细胞增多症初治或未缓解的住院及门诊患者10例,以活血祛瘀药治疗12周,并设立正常对照组。在治疗前,治疗1周、4周、8周、12周采用TUNEL法检测患者骨髓单个核细胞的凋亡率,以免疫组化法检测治疗前后Bcl-2、p53基因,用原位杂交法检测上述基因mRNA表达。结果治疗后1周、4周、8周,患者骨髓单个核细胞凋亡率较前一个观察时点逐步增高(P值均小于0.05);但治疗后12周与治疗后8周比较,无显著性差异(P>0.05)。治疗前患者Bcl-2基因及其mR-NA、p53基因及其mRNA水平均高于正常对照组(P<0.05);治疗后4周Bcl-2mRNA、治疗后8周Bcl-2基因及其mRNA、治疗后4周和8周p53基因及其mRNA表达均低于治疗前(P<0.05)。结论活血祛瘀法治疗真性红细胞增多症的起效时间可能在治疗后第8周达到高峰,此后若继续用药,其疗效可能趋于平稳。其机理之一可能是通过调节Bcl-2、P53基因及其mRNA水平表达,从而诱导真性红细胞增多症骨髓单个核细胞凋亡。

Objective To explore the therapeutic mechanism of blood- activating and blood- stasis- removing (BABAR) therapy for polycythemia vera (PV).Methods Ten first- visit or un- relieved outpatients and inpatients were treated with BABAR therapy for 12 weeks. Healthy volunteers served as the normal control. Before treatment and 1, 4, 8 and 12 weeks after treatment, the apoptotic rates of bone marrow mononuclear cells (BMMC) was analyzed with the TUNEL method. The expression of bcl- 2 and p53 was observed by the method of immunohistochemistry and their mRNA expression by the method of hybridization in site.Results One, four and eight weeks after treatment, the apoptotic rates of BMMC was increased as compared with those before treatment (P < 0.05); the apoptotic rate 12 weeks after treatment did not differ from that 8 weeks after treatment (P > 0.05).The gene expression level of Bcl- 2 and p53 and their RNA expression level in the patients before treatment were higher than those in the normal control(P < 0.05). Expression level of p53 gene and mRNA level of bcl- 2 and p53 4 weeks after treatment were lower than those before treatment (P < 0.05). So did the expression level of bcl- 2 gene and p53 gene and their mRNA levels 8 weeks after treatment.Conclusion The effect of BABAR in treating PV arrived the peak 8 weeks after treatment and then the effect will be mild. Its therapeutic mechanism may be related to the regulation of expression of Bcl- 2 and p53 and their mRNA expression, which can induce the apoptosis of BMMC in PV patients. 