摘要
目的 探讨I型糖尿病(T1DM)睾丸组织发生泛凋亡(PANoptosis)的可能机制.方法 SPF级SD大鼠50只随机分为对照组(n=15)和I型糖尿病组(n=35).I型糖尿病大鼠造模成功 8 周后,取睾丸组织检测PANoptosis相关指标以及PI3K、AKT和NF-κB.GC-1 spg细胞分为Control组、高糖组(HG)和LY294002+高糖组(LY294002+HG);CCK-8 实验确定PI3K的抑制剂LY294002 最佳干预浓度和时间;检测细胞内PANoptosis相关指标以及PI3K、AKT、NF-κB等.结果 与Control相比,T1DM组大鼠睾丸组织PANoptosis相关的免疫组化指标、mRNA和蛋白表达均较正常组显著升高;T1DM组大鼠睾丸组织内PI3K/AKT/NF-κB信号通路相关指标表达升高.Western blotting实验证明HG组GC-1 spg细胞PANoptosis相关蛋白和PI3K、AKT、NF-κB蛋白表达升高;LY294002+HG组与HG组相比上述蛋白表达下降.结论 I型糖尿病大鼠睾丸组织中PANoptosis的发生可能是由PI3K/AKT/NF-κB信号通路激活介导的.
Abstract
Objective To explore the possible mechanism of PANoptosis ocurrence in testicular tissue of type Ⅰ diabetes mellitus(T1DM).Methods Fifty SPF-grade SD rats were randomly divided into the control group(n=15)and type 1 diabetic group(n=35).After 8 weeks of successful modeling,testicular tissues were collected for detecting PANoptosis-related indexes as well as PI3K,AKT and NF-κB expressions.GC-1 spg cells were divided into a control group,a hyperglycemic group(HG),and a LY294002 and hyperglycemic group(LY294002+HG);CCK-8 experiments were used to determine the optimal concentration and time of PI3K inhitbitor(LY294002);Intracellular PANoptosis-related indexes and PI3K,AKT,NF-κB expressions were examined,respectively.Results For in vivo animal experiments,PANoptosis-related immunohistochemical indexes,mRNA and protein expressions in testicular tissues in the T1DM group were all significantly higher than those in control group;Immunohistochemical indexes,mRNA and protein expressions related to PI3K/AKT/NF-κB signaling pathway were also elevated in testicular tissues in the T1DM group.For in vitro cell tests,caspase-1 expression within the GC-1 spg cells was upregulated in the HG group compared with the control group;The expressions of PANoptosis-related proteins and PI3K,AKT and NF-κB were increased in GC-1 spg cells of the HG group;But the expressions of the above proteins were all decreased in the LY294002+HG group compared with the HG group.Conclusion The occurrence of PANoptosis in testicular tissue of type I diabetic rats may be mediated by the activation of PI3K/AKT/NF-κB signaling pathway.
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