【摘要】 为探究添加梓醇（CAT）对体外培养猪卵巢中颗粒细胞（GCs）增殖及其相关基因表达的影响，本试验通过体外分离培养猪GCs，用免疫荧光鉴定促卵泡激素受体（FSHR）表达情况，并确认细胞纯度，采用不同浓度（0、5、10和20μmol/L）CAT处理猪GCs 24 h后，用CCK-8法检测细胞活力，进一步使用荧光定量PCR法检测猪GCs中细胞周期相关基因细胞周期蛋白B1(CCNB1)、细胞周期蛋白D1(CCND1)及增殖相关基因增殖细胞核抗原（PCNA）的mRNA表达量。结果表明：体外培养的猪GCs呈典型的梭形或不规则状生长，符合GCs的形态特征；分离出的细胞多数呈现绿色荧光表达FSHR，说明GCs分离纯度较高；10μmol/L CAT能够显著提高猪GCs的活力（P<0.05）并显著上调CCNB1、CCND1和PCNA的mRNA表达量（P<0.05）。综上所述，CAT可通过调控增殖相关基因表达促进猪卵巢GCs的体外增殖，为优化GCs培养体系及提高猪繁殖效率提供理论依据。更多还原

【Abstract】 The study aimed to investigate the effects of Catalpol(CAT) on the proliferation of porcine ovarian granulosa cells(GCs)and the expression of related genes in vitro. Porcine GCs were isolated and cultured, with cell purity confirmed by immunofluorescence detection of follicle-stimulating hormone receptor(FSHR) expression. The GCs were treated with various concentrations of CAT(0, 5,10, and 20 μmol/L) for 24 hours. Cell viability was assessed using the CCK-8 assay, and the mRNA expression levels of cell cycle-related genes(CCNB1 and CCND1) and a proliferation-related gene(PCNA) were measured using quantitative real-time PCR(q PCR). The results indicated that the porcine GCs cultured in vitro exhibited typical spindle-shaped or irregular morphology. The majority of the isolated cells tested positive for FSHR, demonstrating high cell purity. Treatment with 10 μmol/L CAT significantly enhanced GCS viability(P<0.05) and significantly upregulated the mRNA expression levels of CCNB1, CCND1, and PCNA(P<0.05).In conclusion, CAT promotes the in vitro proliferation of porcine ovarian GCs by modulating the expression of proliferation-related genes. The findings provide a theoretical basis for optimizing GCs culture systems and improving porcine reproductive efficiency.更多还原