摘要
目的 研究PI3K/Akt信号通路调节蛋白表达与硫酸镍(NiSO_(4))诱导大鼠睾丸间质(Leydig)细胞睾酮合成障碍的联系。方法 使用清洁级8周龄雄性Wistar大鼠,采用胶原酶消化、Percoll密度梯度离心法,经体外差速贴壁培养获得纯化大鼠Leydig细胞。设立对照组[DMEM+1 IU/ml人绒毛膜促性腺激素(hCG)],低、中和高剂量NiSO_(4)组(250、500和1 000μmol/L NiSO_(4)+1 IU/ml hCG),处理Leydig细胞24 h,使用酶联免疫吸附试验(ELISA)检测睾酮含量,Western blot法检测睾酮相关合成酶和PI3K/Akt信号通路相关蛋白水平,实时荧光定量PCR测定PI3K/Akt信号通路Pik3r1、Akt1 mRNA表达。结果 与对照组相比,NiSO_(4)组睾酮分泌量(F=56.518,P<0.05)、StAR(F=11.121,P<0.05)和Cyp11a1(F=9.084,P<0.05)降低,Pik3r1 mRNA表达上升(F=61.992,P<0.05),Akt1 mRNA表达量明显降低(F=8.538,P<0.05),NiSO_(4)组p-PI3K p85/PI3K p85磷酸化比率降低(F=5.926,P<0.05)。结论 NiSO_(4)可能通过抑制PI3K/Akt信号通路,降低Leydig细胞睾酮合成相关酶StAR、Cyp11a1的表达进而抑制睾酮分泌。
Objective To investigate the role of PI3K/Akt signaling pathway in nickel sulfate(NiSO4)induced testosterone synthesis impairment in rat Leydig cells.Methods Testicular tissue was isolated from Wistar rats,gradient centrifugation and adherent culture were used to get purified Leydig cells.Leydig cells were randomly allocated into control group(1 IU/ml human chorionic gonadotropin,hCG)or experimental groups.In experimental groups,the low,medium,and high dose groups were treated with O,250,500 and 1000μmol/L NiSO4,respectively,with 1 IU/ml hCG.Testosterone secretion was detected using enzyme-linked immunosorbent assay.Western blot was used to detect testosterone-related synthase and PI3K/Akt signaling pathway-related protein levels,and real-time reverse transcription-polymerase chain reaction was used to determine the transcript level of PI3K/Akt signaling pathway.Results NiSO,decreased the testosterone level(F=56.518,P<0.05),the expression of StAR(F=11.121,P<0.05)and Cypllal(F=9.084,P<0.05).The expression of Pik3rl mRNA increased(F=61.992,P<0.05),Aktl mRNA significantly decreased in NiSO,treated cells(F=8.538,P<0.05),Nis04 p-PI3K p85/PI3K p85 phosphorylation ratio(F=5.926,P<0.05).Conclusion NisO,can inducedysfunction of testosterone secretion in Leydig cells by inhibiting PI3K/Akt signaling pathway.
作者
李瑞芬
俄倩男
王彩霞
韦琳誉
张莉
苏莉
LI Rui-fen;E Qian-nan;WANG Cai-xia;WEI Lin-yu;ZHANG Li;SU Li(Department of Maternal,Child and Adolescent Health,School of Public Health,Lanzhou University,Lanzhou Gansu 730000,China;Department of Infection Management,Gansu Provincial People's Hospital,Lanzhou Gansu 730000,China;Medical Experimental Center,School of Public Health,Lanzhou University,Lanzhou Gansu 730000,China;School of Public Health,Sun Yat-sen University(Shenzhen),Shenzhen Guangdong 515100,China)
出处
《毒理学杂志》
CAS
2023年第2期153-157,165,共6页
Journal of Toxicology
