中药何首乌饮对大鼠卵巢颗粒细胞凋亡相关蛋白bcl-2,bax表达的影响

目的通过测定不同剂量的中药何首乌饮含药血清作用后的大鼠卵巢颗粒细胞的凋亡相关基因bcl-2、bax的蛋白表达及caspase-3活性,探讨何首乌饮对卵巢自身功能的影响。方法制备排卵前卵巢模型,收集颗粒细胞分别与雌性SD幼鼠空白血清及含不同剂量何首乌饮的药物血清在体外共同培养,Western blotting检测GC内凋亡相关基因bcl-2、bax的蛋白表达情况;分光光度法检测颗粒细胞内caspase-3活性。结果何首乌饮含药血清可上调颗粒细胞bcl-2蛋白表达,下调bax蛋白表达,中、高剂量组何首乌饮含药血清与对照组比较,可明显上调bcl-2蛋白表达(P均<0.01),中、高剂量组何首乌饮含药血清与对照组比较,明显下调bax蛋白表达(P均<0.01)。何首乌饮含药血清可下调颗粒细胞caspase-3活性,中、高剂量何首乌饮含药血清组与对照组比较有显著性差异(P均<0.01)。结论何首乌饮增加GC内凋亡抑制蛋白bcl-2表达及减少凋亡促进蛋白bax表达的作用,可能是其抑制GC凋亡,防止卵泡闭锁的作用途径之一;同时何首乌饮可能是在凋亡的较早阶段,通过阻断caspase-3的级联裂解即可开始起效,而发挥凋亡抑制作用。

Effects of serum containing Hswy on apoptosis ralated protein expression of rat's granulosa cells of ovary

Objective It is to explore the effect of Heshouwuyin on ovarian function by determining the expression of the bcl-2 and bax gene protein and activity of caspase-3 in cultured rat ovarian granulosa cells(GCs) after different dosage of this decoction’s treatment.Methods Femal S.D.rats were used as a model and ovarian granulosa cells were prepared.Two groups were set up: study group with rat serum contained Heshouwuyin decoction in different dosage and control group with normal serum.The bcl-2 and bax gene protein expression of granulosa cells were assessed by Western blotting;the activity of cellular caspase-3 were measured by colorimetric method.Results The serum with medicine could increase the expression of bcl-2 protein and decrease the expression of bax protein of GCs,compared with blank serum group,medium and high dosage serum groups could obviously increase bcl-2 protein and decrease bax protein expression of GCs respectively(P<0.01).The activity of caspase-3 of granulosa cells in study groups may be lower than the blank serum group,compared with blank serum group,medium and high dosage serum groups could decrease the activity of caspase-3 obviously(P<0.01).Conclusion Heshouwuyin decoction could improve ovarian function.One of possible mechanism is the decoction can inhibit the rate of apoptosis by increasing the protein expression of bcl-2 and suppressing bax expression.The activity of casepase-3 could decreased under the stimulation of serum with medicine.It presumed that the decoction may regulate follicle development and follicle atresia.