Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisp...Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.展开更多
基金supported by grants from the National Natural Science Foundation of China(8170060495 and 82170682)。
文摘Background:Cisplatin triggers Gasdermin E(GSDME)cleavage,causing membrane bubble formation,content release,and inflammation.Caspase-3 activation initiates GSDME cleavage,and thus inhibiting this pathway mitigates cisplatin-induced pyroptosis in hepatocytes.This study aimed to delve into how cisplatin induces liver injury via pyroptosis.Methods:For animal experiments,C57BL/6J mice were divided into three groups:control,liver injury model group,and Ac-DMLD-CMK(caspase-3 inhibitor)intervention group.The liver histology was evaluated by hematoxylin and eosin staining,immunohistochemistry,immunofluorescence and TUNEL staining.The mRNA and protein levels were detected by real-time polymerase chain reaction(PCR)and Western blot analysis.For in vitro experiments,HL-7702 cells were treated with cisplatin or GSDME siRNA.Cell pyroptosis was determined via cellular morphology,cytotoxicity and viability detection,flow cytometric assay,and Western blot detection for the expression of pyroptosis-related proteins.Results:Cisplatin-induced distinct liver morphological changes,hepatocellular injury,and inflammation in mice,along with elevated serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and increased pro-inflammatory cytokine expression.Heightened macrophage infiltration and hepatocellular death indicated cisplatin-induced hepatotoxicity.Cisplatin upregulated GSDME activation,along with Bax-mediated caspase-3 cleavage both in vivo and in vitro,implicating caspase-3/GSDME-dependent pyroptosis in liver injury.Treatment with Ac-DMLD-CMK ameliorated cisplatin-induced liver injury,reducing hepatocellular lesions,serum ALT and AST levels,cytokine expression,macrophage infiltration,and hepatocyte death.Ac-DMLD-CMK also attenuated GSDME-dependent pyroptosis post-cisplatin induction,as evidenced by decreased GSDME expression,Bax upregulation,and cleaved caspase-3 activation.For HL-7702 cells,GSDME siRNA transfection reduced GSDME expression,attenuated typical signs of cisplatin-induced pyroptosis,partially restored cell viability,and significantly inhibited cytotoxicity and a decrease in the proportion of propidium iodide-positive cells,indicating protection against cisplatininduced hepatocyte pyroptosis.Conclusions:Our study underscores the role of the caspase-3/GSDME signaling pathway in mediating cisplatin-induced hepatotoxicity,particularly in cases of excessive or cumulative cisplatin exposure.These findings suggest that targeting GSDME could represent a promising therapeutic approach to mitigate cisplatin-induced liver damage.
文摘目的检测PP1A与GSDME在结直肠癌组织中的表达与CD8^(+)T淋巴细胞丰度,探讨PP1A与GSDME介导焦亡的相关性和临床意义。方法应用GEPIA数据库分析PP1A与GSDME在结直肠癌组织与正常组织中mRNA的表达。采用Western blot法检测结直肠癌组织与对应癌旁正常黏膜中PP1A蛋白表达水平,运用免疫组化法检测107例结直肠癌与癌旁正常黏膜中PP1A、GSDME蛋白的表达和CD8^(+)T淋巴细胞丰度。利用Spearman等级相关性分析PP1A、GSDME和CD8^(+)T淋巴细胞丰度的相关性。结果GEPIA数据库检索显示,PP1A与GSDME的mRNA在结直肠癌组织和正常组织中的表达差异有统计学意义(P<0.05)。Western blot法检测结果显示,结直肠癌组织中PP1A相对表达量明显高于癌旁组织(0.937 vs 0.643,P<0.001)。免疫组化结果显示,结直肠癌组织中PP1A的表达明显高于正常黏膜,而GSDME的表达明显低于正常黏膜(P<0.05),GSDME表达与结直肠癌患者年龄、临床分期和错配修复蛋白密切相关(P<0.05);CD8^(+)T细胞在癌浸润前沿的分布明显高于癌旁正常黏膜,且CD8^(+)T细胞在癌组织中的分布与pT分期、临床分期及淋巴结转移相关。Spearman相关性分析显示,PP1A与GSDME表达呈负相关(r=-0.196,P<0.05)。PP1A阳性结直肠癌患者的总生存期低于PP1A阴性患者(P<0.05),患者预后与分化程度、淋巴结转移、pT分期和临床分期相关。PP1A表达、肿瘤分化程度、临床分期、pT分期和淋巴结转移均是影响结直肠癌患者预后的独立危险因素。结论PP1A在结直肠癌中高表达,与GSDME介导的细胞焦亡呈负相关,两者表达差异性与结直肠癌的发生、发展及预后均密切相关,可作为判断结直肠癌患者预后的潜在指标,CD8^(+)T细胞的差异性分布可能与GSDME介导的细胞焦亡及肿瘤的发展相关。
