摘要
为了探明促卵泡素(Follicle stimulating hormone,FSH)和促黄体素(Luteinizing hormone,LH)对体外培养的牦牛卵泡颗粒细胞凋亡及甾体激素分泌的影响,揭示促性腺激素调控卵泡发育的机理,采用细胞培养和显微观察技术,建立了牦牛卵泡颗粒细胞体外培养体系,观察分析牦牛卵泡颗粒细胞体外培养的生长特征和凋亡的形态特征;采用流式细胞技术(Flow cytometry,FCM)和放射免疫测定技术(Radioimmunoassay,RIA),分析了添加不同浓度的FSH和LH对牦牛卵泡颗粒细胞凋亡和雌二醇(Estradiol,E2)、孕酮(Progesterone,P)分泌的影响。结果显示,体外培养的牦牛卵泡颗粒细胞呈现出典型的上皮样细胞生长特性,具有典型的'S'形生长曲线,体外培养的牦牛颗粒细胞偶尔可见细胞核浓缩、边集化、染色质固缩、内质网疏松等凋亡的形态特征。在添加0.050~5.000IU·mL-1FSH,随着浓度的增加,凋亡细胞比例逐渐降低,E2和P水平呈上升趋势;在添加浓度达到5.000IU·mL-1时,凋亡细胞比例最低降到7.3%,与对照组差异极显著(P<0.01)。低浓度的LH(0.050~0.500IU·mL-1),可以抑制颗粒细胞凋亡,促使E2和P分泌增加;高浓度的LH(5.000IU·mL-1)则促使颗粒细胞凋亡,对颗粒细胞分泌E2和P的影响不明显。这一结果表明,FSH和LH可通过调节牦牛颗粒细胞的凋亡及分泌功能,在调控卵泡发育及排卵的过程中发挥重要作用。
This experiment was conducted to study the effect of follicle stimulating hormone(FSH),luteinizing hormone(LH)on apoptosis and steroids hormone secretion of yak's granulosa cells cultured in vitro,reveal the mechanism of gonadotropin regulating follicle development.In vitro culture system was established and the growth characteristics,apoptotic morphology were observed using cell culture and micro examination technology.The effect of FSH,LH on apoptosis and estradiol(E2),progesterone(P)secreting of yak's granulosa cells cultured in vitro was analyzed using flow cytometry(FCM)and radioimmunoassay(RIA).The results showed that:Cultured yak's granulosa cells demonstrated typical epithelioid growth characteristic and had'S'shape growth curve.Occasionally,the apoptotic morphological characteristics such as nuclear pyk-notic and margination,endoplasmic reticulum loosen can be found in the cell culture dishes.With the increase of FSH concentration from 0.050 to 5.000IU·mL-1,the apoptotic rate decreased and the E2,P level be on the rise.When FSH reached 5.000IU·mL-1,the apoptotic rate down to 7.3% which was very significantly from control group(P<0.01).The apoptosis of granulosa cells were inhibited and the E2,P secreting were prompted when LH was added in a small quantity(0.050-0.500IU·mL-1),on the other hand,high concentration of LH(5.000IU·mL-1)prompted the apoptosis of granulosa cells and inconspicuously effected the E2,P secreting.These results indicated that FSH and LH played an important role in the regulation of follicle development and ovulation through adjust the apoptosis and steroids hormone secretion of granulosa cells in yak.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2015年第6期932-939,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金面上项目(31272616)
甘肃省青年科技基金计划项目(1208RJYA081)
