磷酸酶和张力蛋白同源物在睾丸缺血再灌注损伤中对GC-1细胞焦亡的影响及作用机制  

Effect and mechanism of phosphatase and tensin homologs on pyroptosis of GC-1 cells in testicular ischemia-reperfusion injury

作　　者：宁金卓[1] 何凯翔 许立哲 蒋焜 Ning Jinzhuo;He Kaixiang;Xu Lizhe;Jiang Kun(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2022年第6期1079-1081,共3页Chinese Journal of Experimental Surgery

基　　金：中央高校基本科研业务专项资金课题(2042019kf0059)。

摘　　要：目的探讨磷酸酶和张力蛋白同源物(PTEN)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞焦亡的作用及其机制。方法建立睾丸GC-1细胞(美国ATCC细胞库)缺氧复氧模型,采用定量实时聚合酶链反应(qRT-PCR)检测不同复氧损伤时间点PTEN的表达变化;分别采用免疫荧光实验、蛋白质印迹法(Western blot)检测转染PTEN后对GC-1细胞焦亡相关蛋白NLRP3和Caspase-1表达水平的影响。组间比较采用t检验,多组间比较采用单因素方差分析。结果对照组及缺氧复氧组(0、6、12、24 h)PTEN的mRNA表达水平分别为(1.00±0.21、3.48±0.35、3.75±0.38、4.25±0.40、4.06±0.37)。与未缺氧GC-1细胞的对照组比较,随着复氧损伤时间的增加,PTEN表达明显增高,并在复氧12 h达到峰值(F=43.21,P<0.05)。Western blot结果显示,缺氧复氧组中NLRP3和Caspase-1表达水平(0.52±0.04、0.41±0.04)高于对照组(0.24±0.03、0.20±0.03,t=9.700,P<0.05;t=7.275,P<0.05)。过表达PTEN组中NLRP3和Caspase-1表达水平(0.75±0.06、0.62±0.05)高于其对照组(0.53±0.04、0.42±0.03)明显(t=5.284,P<0.05;t=5.941,P<0.05)。沉默PTEN组中NLRP3和Caspase-1表达水平(0.36±0.03、0.32±0.03)低于对照组(0.52±0.04、0.41±0.03)明显(t=-5.543,P<0.05;t=-3.674,P<0.05)。结论PTEN在GC-1细胞中呈高表达,其可能通过改变GC-1细胞焦亡水平,从而影响睾丸IRI的发生发展。Objective To investigate the effects of phosphatase and tensin homologs(PTEN)on pyroptosis of GC-1 cells in testicular ischemia reperfusion injury(IRI)and its mechanism.Methods The testicular GC-1 cell hypoxia-reperfusion model was established,and quantitative real-time PCR was used to detect the changes of PTEN expression at different time points of reperfusion injury.Immunofluorescence assay and Western blotting were used to detect the effects of PTEN transfection on pyroptosis of GC-1 cells.The expression levels of NLRP3 and Caspase-1 were examined by immunofluorescence assay and Western blotting respectively.T-test was used for inter group comparison,and one-way ANOVA was used for multi group comparison.Results qRT-PCR results showed that the mRNA expression levels of PTEN were(1.00±0.21,3.48±0.35,3.75±0.38,4.25±0.40,4.06±0.37)in control and hypoxic-reoxygenated groups at 0,6,12,24 h,respectively.The PTEN expression was significantly higher with increasing time of reoxygenation injury and peaked at 12 h of reoxygenation compared to the control group of non-hypoxic GC-1 cells(F=43.21,P<0.05).Immunofluorescence results showed that the expression levels of NLRP3 and Caspase-1 were significantly increased in the hypoxic reoxygenation group as compared with the control group.The expression levels of NLRP3 and Caspase-1 were significantly increased in the overexpression PTEN group as compared with its control group.The expression levels of NLRP3 and Caspase-1 were significantly decreased in the silenced PTEN group as compared with its control group.Western blotting results showed that the expression levels of NLRP3 and Caspase-1 were significantly increased in the hypoxic-reoxygenated group(0.52±0.04,0.41±0.04)as compared with the control group(0.24±0.03,0.20±0.03,t=9.700,P<0.05,t=7.275,P<0.05).The expression levels of NLRP3 and Caspase-1 were increased in the overexpression PTEN group(0.75±0.06,0.62±0.05)as compared with its control group(0.53±0.04,0.42±0.03,t=5.284,P<0.05;t=5.941,P<0.05).NLRP3 and Ca

关 键 词：睾丸缺血再灌注损伤 磷酸酶和张力蛋白同源物 焦亡 

分 类 号：R697.22[医药卫生—泌尿科学]