不同浓度的二仙汤及其拆方对大鼠卵泡颗粒细胞凋亡因子影响的比较研究

Comparative Study of Erxian Decoction and Its Demolition Parties in Different Concentrations on Rats Follicular Granulosa Cells Apoptosis Factors

目的:观察不同浓度的二仙汤及其拆方温补肾阳组、滋阴泻火组、补血组对原代培养的雌性大鼠卵巢颗粒细胞凋亡因子Bcl-2、Bax蛋白表达及Caspase-3蛋白活性的影响。方法:采用氯化镉复制的大鼠卵泡颗粒细胞凋亡模型,加入不同浓度二仙汤及其拆方,作用24 h后,酶联免疫吸附法检测卵泡颗粒细胞内Bcl-2、Bax蛋白表达及Caspase-3活性。结果:1二仙汤及其拆方均可抑制卵泡颗粒细胞的凋亡;2二仙汤及其拆方抑制卵泡颗粒细胞凋亡的作用靶点与其浓度相关:高浓度(1 mg/m L)二仙汤主要降低卵泡颗粒细胞中Bax蛋白表达;高浓度(1 mg/m L)二仙汤、低浓度(0.01 mg/m L)补肾阳组主要上调卵泡颗粒细胞中Bcl-2蛋白表达,二者比较无差异;低浓度(0.01 mg/m L)补血组主要抑制卵泡颗粒细胞内Caspase-3活性。结论:二仙汤主要是通过补肾阳组上调卵泡颗粒细胞Bcl-2蛋白表达而发挥君药的"扶正"作用,通过补血组抑制Caspase-3蛋白活性发挥使药的调和作用,初步证明了二仙汤配伍的合理性。

Objective： To investigate the effect of Erxian Decoction and its demolition parties in different concentrations,including the warmly invigorating kidney Yang group,the nourishing Yin and purging fire group and the replenishing blood group,on the originally cultured rats follicle granule cell apoptosis factors Bcl-2 and Bax expressions and Caspase-3 activity.Methods： By the model of the rats follicular granulosa cell apoptosis made by the cadmium chloride,mix Erxian Decoction and its demolition parties in different concentrations.After 24 h,detect the Bcl-2 and Bax protein expressions and the Caspase-3 activity of follicular granulosa cells by enzyme-linked immunosorbent assay（ELISA）.Results： 1Erxian Decoction and its demolition parties all can inhibit the apoptosis of follicular granulosa cells.2The effect of Erxian Decoction and its demolition parties on inhibiting the follicular granulosa cells apoptosis was related to their concentrations：the high concentration（1 mg/m L） Erxian Decoction group significantly degraded Bax protein expression in follicular granulosa cells and the high concentration（1 mg/m L） Erxian Decoction group and the low concentration（0.01 mg/m L） warmly invigorating kidney Yang group significantly upgraded the Bcl-2 protein expression in follicular granulosa cells,with no difference between them in statistics.The low concentration（0.01 mg/m L） replenishing blood group significantly suppressed Caspase-3 activity in the follicular granulosa cells.Conclusion： Erxian Decoction plays the Sovereign Herb role of Fuzheng by the warmly invigorating kidney Yang group raising the Bcl-2 protein expression of follicular granulosa cells and gentleman medicine should play the Conductant Herb role of Tiaohe by the replenishing blood group restraining the activity of Caspase 3 protein,thereby preliminarily proving the rationality of the concerted application of Erxian Decoction.