经后增殖方含药血清对超排卵大鼠卵巢颗粒细胞GDF9表达及凋亡的调控机制

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中文摘要：[目的]观察益气血法经后增殖方含药血清对超排卵大鼠卵巢颗粒细胞生长分化因子9(GDF9)表达及凋亡的调控机制.[方法]制备超排卵大鼠血清(空白血清)和经后增殖方灌胃的超排卵大鼠血清(含药血清).建立控制性超排卵(COH)大鼠模型,收集卵巢颗粒细胞.实验分为5组:空白血清组,含药血清组,含药血清+ SB203580[p38丝裂原活化蛋白激酶(p38MAPK)抑制剂]组,含药血清+PDTC[核转录因子κB(NF-κB)抑制剂]组,含药血清+SB203580+PDTC组.采用实时定量聚合酶链反应(qRT-PCR)法检测p38MAPK、酪蛋白激酶2(CK2)、核转录因子κB抑制因子α(IκBα)、NF-κB、GDF9 mRNA表达水平,蛋白免疫印迹(Western Blot)法检测GDF9蛋白表达水平,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测卵巢颗粒细胞凋亡.[结果]经后增殖方含药血清降低COH大鼠卵巢颗粒细胞的p38MAPK和NF-κB mRNA表达,升高CK2和IκBα mRNA表达,提高GDF9 mRNA和蛋白的表达水平,降低卵巢颗粒细胞的凋亡率.单独添加p38MAPK抑制剂SB203580与单独添加NF-κB抑制剂PDTC均能促进GDF9 mRNA和蛋白表达、降低卵巢颗粒细胞凋亡率.[结论]益气血法经后增殖方含药血清可促进超排卵大鼠卵巢颗粒细胞GDF9表达,抑制卵巢颗粒细胞凋亡,其机制可能与调控p38MAPK和NF-κB双信号通路基因表达有关.

外文标题：Regulatory Mechanism of Drug-Containing Serum of Jinghou Zengzhi Prescription on GDF9 Expression and Apoptosis of Ovarian Granulosa Cells in Rats with Controlled Ovarian Hyperstimulation

外文摘要：Objective To observe the regulatory mechanism of drug-containing serum of Jinghou Zengzhi Prescription based on qi and blood replenishing method on the expression of growth and differentiation factor 9(GDF9)and apoptosis of ovarian granulosa cells in rats with controlled ovarian hyperstimulation(COH).Methods Serum of COH rats(blank serum)and serum of COH rats gavaged by the Jinghou Zengzhi Prescription were prepared.A COH rat model was established and ovarian granulosa cells were collected.The experiment was divided into 5 groups:blank serum group,drug-containing serum group,drug-containing serum+SB203580[p38 mitogen-activated protein kinase(p38MAPK)inhibitor]group,drug-containing serum + PDTC[nuclear transcription factor κB(NF-κB)inhibitor]group,drug-containing serum + SB203580 + PDTC group.The mRNA expression levels of p38MAPK,casein kinase 2(CK2),nuclear transcription factor κB inhibitor α(IκBα),NF-κB and GDF9 were detected by real-time quantitative polymerase chain reaction(qRT-PCR),and GDF9 protein expression level was detected by Western Blot,and ovarian granulosa cell apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL).Results The drug-containing serum of Jinghou Zengzhi Prescription decreased the mRNA expressions of p38MAPK and NF-κB,elevated the mRNA expressions of CK2 and IκBα,increased the mRNA and protein expression levels of GDF9,and decreased the apoptosis rate of ovarian granulosa cells in COH rats.The addition of p38MAPK inhibitor SB203580 alone and the addition of NF-κB inhibitor PDTC alone both promoted the mRNA and protein expressions of GDF9 and reduced the apoptosis rate of granulosa cells.Conclusion The drug-containing serum of Jinghou Zengzhi Prescription based on qi and blood replenishing method can promote the expression of GDF9 and inhibit the apoptosis of ovarian granulosa cells in rats with COH,and its mechanism may be related to the regulation of the expression of genes of the dual signaling pathways of p38MAPK and NF-κB.

外文关键词：

qi and blood replenishing methodJinghou Zengzhi Prescriptioncontrolled ovarian hyperstimulation(COH)apoptosisgrowth differentiation factor 9(GDF9)p38 mitogen-activated protein kinase(p38MAPK)nuclear transcription factor κB(NF-κB)granulosa cells

作者：

杨贞、陈小燕、江少如、叶淑珠、方晓宏、邓伟民、郭新宇

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作者单位：

揭阳市人民医院,广东揭阳 522000

南部战区总医院,广东广州 510010

前海人寿广州总医院,广东广州 511325

关键词：

益气血法经后增殖方控制性超排卵(COH) 凋亡生长分化因子9(GDF9) p38丝裂原活化蛋白激酶(p38MAPK) 核转录因子κB(NF-κB) 颗粒细胞

基金：

广东省自然科学基金资助项目广东省中医药局科研基金资助项目

项目编号：

2020A151501117520201412

出版年：

2024

DOI：

10.13359/j.cnki.gzxbtcm.2024.03.031

广州中医药大学学报

广州中医药大学

广州中医药大学学报

CSTPCD

影响因子：1.055

ISSN：1007-3213

年,卷(期)：2024.41(3)

参考文献量22