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目的:观察穿龙薯蓣皂苷治疗再生障碍性贫血小鼠的疗效及骨髓单个核细胞Caspase-3、Caspase-8蛋白的表达。方法:选取健康ICR小鼠105只,取15只设为正常对照组,其余90只参照孙氏[1]等报道方法建立再障模型,造模成功后将小鼠随机分为6组(①模型组②低剂量组③中剂量组④高剂量组⑤环孢菌素A组⑥雷公藤多苷组)。每组15只。于造模成功后第1～14天,各组分别以相应药物灌胃,每日1次。于第15天,检测小鼠血常规,验证小鼠血象,并提取小鼠骨髓单个核细胞,进行细胞培养,培养24小时后,应用Western Blot方法测定Caspase-3、Caspase-8蛋白的表达水平。结果:模型组caspase-3表达百分率明显高于其余各组(P<0.05);与正常组相比,低、中、高剂量组阳性率稍有增加;雷公藤组高于低、中、高剂量组和CsA组(P<0.05),薯蓣皂苷3个剂量组与CsA组之间未见差异。模型组caspase-8表达百分率与其余各组相比显著增加(P<0.05),与正常组相比,各组均显著高于正常组(P<0.05),薯蓣皂苷3个剂量组之间差异不明显,其中除高剂量组外,其余两组与CsA组、雷公藤组之间差异不明显(P>0.05)。结论:穿龙薯蓣皂苷可以通过下调Caspase-3、Caspase-8蛋白的表达,抑制Fas/FasL细胞凋亡信号传导,减少由T淋巴细胞异常活化导致的骨髓细胞凋亡,发挥免疫调节,从而起到对再障的治疗作用。
Objective: To observe the therapeutic effect of Diosmin in Aplastic Anemia mice and the expression of Caspase-3 and Caspase-8 in bone marrow mononuclear cells. Methods: A total of 105 healthy ICR mice were selected and 15 were selected as the normal control group. The remaining 90 mice were randomly divided into 6 groups according to the method of Sun et al. [1] Model group ② low dose group ③ medium dose group ④ high dose group ⑤ cyclosporin A group ⑥ tripterygium glycosides group). 15 in each group On the 1st to 14th days after the model was successfully established, the rats in each group were gavaged with the corresponding drugs once daily. On the 15th day, the blood routine of the mice was tested, the mice blood samples were verified, and the mouse bone marrow mononuclear cells were extracted for cell culture. After cultured for 24 hours, the expression levels of Caspase-3 and Caspase-8 protein were determined by Western Blot . Results: The expression of caspase-3 in the model group was significantly higher than that in the other groups (P <0.05). Compared with the normal group, the positive rate of caspase-3 in the model group increased slightly; Dose group and CsA group (P <0.05). There was no difference between three dosages of diosgenin and CsA group. Compared with normal group, the expression of caspase-8 in model group was significantly higher than that in normal group (P <0.05), and the difference was statistically significant The difference was not significant between the other two groups and CsA group and Tripterygium wilfordii group (P> 0.05). Conclusion: Diosmin can decrease the signal transduction of Fas / FasL cells and down-regulate the expressions of Caspase-3 and Caspase-8 proteins, decrease the apoptosis of bone marrow cells induced by the abnormal activation of T lymphocytes and play an immunomodulatory role To the treatment of aplastic anemia.