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目的:观察不同浓度血管活性肠肽(VIP)对新生4 d大鼠卵巢体外培养过程中原始卵泡存活和生长发育的影响。方法:取新生4 d大鼠的卵巢72个,随机分为6组:新鲜组、基础培养组和不同浓度VIP组(10-9~10-6 mol/L)。新鲜组不经培养、其余各组体外培养14 d行组织形态学检查、增殖细胞核抗原(PCNA)免疫组织化学和TUNEL凋亡分析。结果:①各培养组的1级(早期初级)卵泡百分比明显高于新鲜组(P<0.05),且VIP 10-7 mol/L组最高。②各培养组PCNA阳性率与新鲜组比较均有统计学差异(P<0.001);不同浓度VIP组与基础培养组比较有统计学差异(P<0.05);VIP 10-7 mol/L组卵泡PCNA阳性率最高,与其他VIP浓度组比有显著差异(P<0.05),③各培养组卵泡凋亡率均显著高于新鲜组(P<0.001),培养组中以VIP 10-7 mol/L组较低,与VIP 10-8 mol/L组和VIP 10-9 mol/L组间有统计学差异(P<0.05)。结论:在卵巢体外培养过程中,VIP可以促进新生4 d大鼠的原始卵泡向初级卵泡转化;不同浓度的VIP均可降低卵巢组织卵泡细胞的凋亡,提高卵泡体外存活能力,VIP10-7 mol/L较其他浓度更明显地促进卵泡细胞的增殖,降低卵泡细胞的凋亡。
OBJECTIVE: To observe the effects of different concentrations of vasoactive intestinal peptide (VIP) on the survival and growth of primordial follicles during the in vitro culture of ovariectomized rats of 4 days old. Methods: Seventy-two ovaries of 4-day-old rats were randomly divided into 6 groups: fresh group, basal culture group and VIP group (10-9 ~ 10-6 mol / L). The other groups were cultured in vitro for 14 days for histomorphology, PCNA immunohistochemistry and apoptosis analysis of TUNEL. Results: ① The percentage of follicles of grade 1 (early primary) in each culture group was significantly higher than that in fresh group (P <0.05), and the highest in VIP 10-7 mol / L group. ② The positive rate of PCNA in each culture group was significantly higher than that in fresh group (P <0.001). There was significant difference between VIP group and basic culture group (P <0.05). VIP 10-7 mol / L follicle (P <0.05). ③The apoptosis rate of follicles in each culture group was significantly higher than that in fresh group (P <0.001). In the culture group, VIP 10-7 mol / L group was lower than that in VIP 10-8 mol / L group and VIP 10-9 mol / L group (P <0.05). CONCLUSION: VIP can promote the primary follicles to transform into primary follicles in ovariectomized vitro. VIP at different concentrations can reduce the apoptosis of ovarian follicles and enhance the in vitro survival of ovarian follicles. VIP10-7 mol / L than other concentrations more significantly promote follicular cell proliferation, reduce apoptosis of follicle cells.