【摘要】 滑液囊支原体(Mycoplasma synoviae, MS)是禽类的重要病原体，感染后常引起关节炎、滑膜炎以及蛋壳顶端异常等症状，蛋鸡(Gallus gallus domesticus)感染后产蛋率下降，严重影响养殖业的经济效益。卵泡颗粒细胞作为产蛋过程中的关键因素，通过分泌激素维持卵泡正常发育与排卵过程。本研究旨在构建MS感染鸡卵泡颗粒细胞的稳定体外模型，研究MS对鸡卵泡颗粒细胞增殖和凋亡的的调控作用。通过菌落计数法(colony-forming unit, CFU)、CCK-8和病变效应(cytopathic effect, CPE)确定MS的最佳感染复数(multiplicity of infection, MOI)和感染时间；通过免疫荧光技术观察MS在细胞中的定位情况；用ELISA法检测MS感染细胞后孕酮分泌量的变化；通过qPCR法检测细胞凋亡相关基因的表达。结果显示，当MS培养32 h后，以1 000 MOI的剂量感染鸡卵泡颗粒细胞12 h时，MS可成功黏附于细胞表面，且MS感染显著降低了鸡卵巢颗粒细胞孕酮的分泌量，凋亡相关标志基因半胱氨酸天冬氨酸蛋白酶3(cysteine-aspartic protease 3, Caspase-3)、肿瘤蛋白p53(tumor protein p53, p53)、Bcl-2相关X蛋白(Bcl-2-associated X protein, Bax)的表达上调。以上结果表明，MS能够感染鸡卵泡颗粒细胞，通过促进鸡卵泡颗粒细胞凋亡，抑制孕酮分泌，进而影响卵泡功能。本研究为进一步揭示MS的致病机制提供参考依据。更多还原

【Abstract】 Mycoplasma synoviae(MS) is a significant pathogen in poultry, its infection frequently causes symptoms such as arthritis, synovitis, and eggshell apex abnormalities. Infection in laying hens(Gallus gallus domesticus) leads to a decreased egg production rate, which seriously affects the economic benefits of the poultry industry. Follicular granulosa cells are crucial in the egg-laying process, primarily through their role in hormone secretion, which is essential for maintaining normal follicular development and ovulation. This study aimed to establish a stable in vitro model of MS-infected chicken follicular granulosa cells and to investigate the regulatory effects of MS on the proliferation and apoptosis of chicken follicular granulosa cells. The optimal multiplicity of infection(MOI) and infection duration for MS were determined through colonyforming unit(CFU) counting, CCK-8 assays, and observation of cytopathic effects(CPE) in cells.Immunofluorescence was employed to observe the localization of MS in infected cells, while changeed in progesterone secretion post-MS infection were assessed using ELISA. The results indicated that when MS was cultured for 32 h, infection at a multiplicity of infection(MOI) of 1 000 for 12 h facilitated the successful adhesion of MS to the cell surface, resulting in a significant reduction in progesterone secretion by chicken follicular granulosa cells. Furthermore, qPCR analysis showed an upregulation of apoptosis-related genes,including cysteine-aspartic protease 3(Caspase-3), tumor protein p53(p53) and Bcl-2-associated X protein(Bax). These results suggested that MS could infect chicken follicular granulosa cells and affect follicular function by promoting apoptosis and inhibiting progesterone secretion. This study provides a reference for further elucidating the pathogenic mechanisms associated with MS.更多还原