香烟烟雾通过激活NLRP3炎症小体导致睾丸细胞焦亡

Cigarette smoke causes testicular cell pyroptosis by activating the NLRP3 inflammasome

目的:探讨香烟烟雾通过激活NLRP3炎症小体诱导细胞焦亡导致大鼠睾丸损伤的作用及机制。方法:40只SPF级健康雄性SD大鼠随机分为对照组、低剂量(10支/d)吸烟组、中剂量(20支/d)吸烟组与高剂量(30支/d)吸烟组,吸烟组采用呼吸道静式染毒的方法吸入香烟烟雾8周后处死动物,检测睾丸组织病理学改变及细胞焦亡情况,同时检测大鼠睾丸组织中活性氧(ROS)水平及血浆中炎症相关因子IL-1β、IL-18、TGF-β1、MCP-1水平,RT-PCR法和Western blot法检测大鼠睾丸细胞焦亡相关因子NLRP3、ASC、Caspase-1、GSDMD、IL-1β与IL-18 mRNA和蛋白表达水平。结果:香烟烟雾暴露8周后,大鼠睾丸组织出现出明显的病理损伤,低、中、高剂量吸烟组大鼠睾丸组织ROS水平、细胞焦亡水平和血浆中炎症相关因子IL-1β、IL-18、TGF-β1、MCP-1水平均显著高于对照组(P<0.05)。随着香烟烟雾暴露剂量的增加,大鼠睾丸组织中NLRP3、ASC、Caspase-1、GSDMD、IL-1β与IL-18 mRNA与蛋白表达水平逐渐升高。结论:香烟烟雾暴露可导致大鼠睾丸炎性损伤,其机制可能与NLRP3炎症小体激活导致的睾丸细胞焦亡有关。

Objective:To explore the effect and mechanism of cigarette smoke-induced testicular injury in rats by activating NLRP3 inflammasome to induce pyroptosis.Methods:40 SPF healthy male Sprague-Dawley rats were randomly divided into a control group,a low-dose smoking group(10 cigarettes/day),a medium-dose smoking group(20 cigarettes/day),and a high-dose smoking group(30 cigarettes/day).The rats in the smoking group were sacrificed after 8 weeks of inhalation of cigarette smoke by respiratory static method.At the same time,the level of ROS in testicular tissue and the levels of inflammatory factors IL-1β,IL-18,TGF-β1,and MCP-1 in plasma were tested.The mRNA and protein expression levels of pyroptosis-related factors NLRP3,ASC,Caspase-1,GSDMD,IL-1β,and IL-18 were detected by RT-PCR and western blot.Results:After 8 weeks of cigarette smoke exposure,the testicular tissue of rats showed obvious pathological damage.The levels of ROS and pyroptosis in testicular tissue and the levels of IL-1β,IL-18,TGF-β1,and MCP-1 in plasma of rats in low-,medium-and high-dose smoking groups were significantly higher than those in the control group(P<0.05).With the increase of cigarette smoke exposure dose,the mRNA and protein expression levels of NLRP3,ASC,Caspase-1,GSDMD,IL-1β,and IL-18 in rat testis tissue increased gradually.Conclusion:Cigarette smoke exposure can cause testicular inflammatory injury in rats,and the mechanism may be relat-ed to pyroptosis caused by activation of NLRP3 inflammasome.