何首乌饮含药血清对大鼠卵巢颗粒细胞端粒酶活性的影响

Effects of serum containing Hswy on telomerase activity of rat's granulosa cells of ovary

目的观察不同剂量的中药何首乌饮对大鼠卵巢颗粒细胞端粒酶活性的影响,探讨其在调节卵泡微环境过程中所起的作用。方法制备大鼠排卵前卵巢模型,收集颗粒细胞(GC)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。用TRAP-ELISA方法分析GC的端粒酶活性;运用流式细胞术分析GC凋亡、坏死情况。结果各剂量组何首乌饮含药血清可明显上调GC端粒酶活性,与对照组比较,中、高剂量何首乌饮含药血清能够显著提高GC端粒酶活性(P均<0.05)。随着中药血清药物剂量的增加,颗粒细胞凋亡率逐渐下降,与对照组比较,中、高剂量组何首乌饮含药血清均能够显著降低离体GC的凋亡率(P均<0.01),各含药血清组细胞坏死率虽较对照组高,但与各组比较差别无统计学意义。结论何首乌饮通过上调离体培养的大鼠卵巢GC端粒酶活性、抑制凋亡从而对卵泡局部微环境的调节,可能是其调节下丘脑-垂体-卵巢轴的重要靶点之一。

Objective It is to observe different dosage of Heshouwu Yin decoction on teloemerase activity of rat ovarian granulosa cells, and to explore its function in promoting follicular development. Methods Femal S. D. rats were used as a model and ovarian granulosa cells（GCs） were prepared. Two groups were set up： study group with rat serum contained Hes- houwu Yin decoction in different dosage and control group with normal serum. The telomerase activity of granulosa cells were measured by telomeric repeat amplification protocol-enzyme linked immunoadsordent assay （ TRAP - ELISA） ; the percentage of apoptosis and death of granulosa cells were quantitatively assessed by flow cytometry （FCM） analysis. Results The serum with medicine could increase the activity of telomerase of GCs ; compared with blank serum group, medium and high dosage se- rum groups could increase the activity of telomerase of GC obviously （ all P 〈 0.05 ）. The percentage of apoptosis of GCs in study group decreased with the dosage of medicine in serum increased, and the percentage of death of GCs in study group was same as that in blank serum group; compared with blank serum group, medium and high dosage serum groups could decrease the apoptosis of GCs significantly. Conclusion Heshouwu Yin decoction may increase the activity of telomerase of cultured rat ovarian granulosa cells, it has the effect on the function of ovarian follicle and could improved ovarian function.