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目的研究在体外条件下葛根素对顺铂引发的大鼠卵巢颗粒细胞损伤的影响。方法分离获取21~25日龄Wistar雌性大鼠的卵巢颗粒细胞,建立稳定的生长和传代体系,鉴定获取颗粒细胞纯度,以MTT法绘制细胞生长曲线。取对数生长期细胞,以浓度分别为1、2.5、5、10、20μg/ml的顺铂作用于细胞,继续培养48 h后行MTT法检测,并由吸光度计算出细胞相对活力。描述顺铂药物浓度和颗粒细胞相对活力之间的关系。选定5、10、20μg/ml三组浓度的顺铂致颗粒细胞损伤,各组均加入0.1、0.5、1 mg/ml不同浓度的葛根素稀释液培养,MTT法检测。判断葛根素对颗粒细胞相对活力的回复能力,以及浓度与效应之间的关系。结果顺铂可以抑制颗粒细胞的生长,并且抑制效应具有剂量依赖性。当顺铂浓度为1μg/ml时,可抑制颗粒细胞的生长,与对照组相比较差异有统计学意义(P<0.05);当顺铂浓度≥2.5μg/ml时,明显抑制颗粒细胞的生长,与对照组比较差异有统计学意义(P<0.01),抑制效果呈剂量依赖性。当顺铂浓度达到20μg/ml,颗粒细胞生长的相对活力较对照组下降近30%。选用5、10、20μg/ml三种浓度顺铂引发卵巢颗粒细胞损伤,在三种条件下分别加入0.1、0.5及1 mg/ml葛根素三种药物浓度共同作用。实验结果显示,葛根素对于3种浓度顺铂引发的卵巢颗粒细胞损伤均有一定程度的抑制作用,且抑制效用具有剂量依赖性。当顺铂浓度升高而导致细胞活力明显下降时,葛根素的保护效应并不发生明显改变。结论三种浓度的葛根素均能够在体外条件下对顺铂引发的大鼠卵巢颗粒细胞损伤起到抑制作用,作用效应具有剂量依赖性;当顺铂浓度升高导致卵巢颗粒细胞损伤增加明显时,葛根素的保护作用未被减弱。
Objective To study the effect of puerarin on cisplatin-induced ovarian granulosa cell injury in vitro. Methods Ovarian granulosa cells from 21 to 25 days old Wistar female rats were isolated and cultured. The stable growth and passage system was established. The purity of granulosa cells was identified and the cell growth curve was drawn by MTT method. The cells in logarithmic growth phase were treated with cisplatin at concentrations of 1, 2.5, 5, 10 and 20μg / ml, respectively. The cells were cultured for 48 hours and detected by MTT assay. The relative cell viability was calculated by absorbance. Describe the relationship between cisplatin drug concentration and the relative viability of granulosa cells. Granulocyte damage was induced by cisplatin in concentrations of 5, 10 and 20μg / ml in three groups. All groups were incubated with different concentrations of Puerarin (0.1,0.5,1 mg / ml) and MTT assay. Determine the ability of Puerarin to restore the relative viability of granulosa cells and the relationship between concentration and effect. Results Cisplatin could inhibit the growth of granulosa cells, and the inhibitory effect was dose-dependent. When the concentration of cisplatin was 1μg / ml, the growth of granulosa cells was inhibited, which was significantly different from the control group (P <0.05). When the concentration of cisplatin was more than 2.5μg / ml, the growth of granulosa cells was obviously inhibited , The difference was statistically significant (P <0.01) compared with the control group, the inhibitory effect was dose-dependent. When the cisplatin concentration reached 20μg / ml, the relative activity of granulosa cells decreased by nearly 30% compared with the control group. Ovarian granulosa cells were induced by three concentrations of cisplatin (5, 10, 20μg / ml). The three drug concentrations of Puerarin at concentrations of 0.1,0.5 and 1 mg / ml were added respectively under the three conditions. The experimental results showed that puerarin inhibited the ovarian granulosa cell damage induced by cisplatin at three concentrations in a dose-dependent manner. When the concentration of cisplatin caused cell viability decreased significantly, the protective effect of puerarin did not change significantly. CONCLUSION: Puerarin at all three concentrations can inhibit the ovarian granulosa cells induced by cisplatin in vitro in a dose-dependent manner. When the concentration of cisplatin is increased, the damage of ovarian granulosa cells is significantly increased , The protective effect of puerarin has not been weakened.