双硫仑抑制细胞焦亡改善脓毒症大鼠神经细胞损伤的作用机制  

Disulfiram ameliorates neuronal injury through inhibiting cell pyroptosis in septic rats

作　　者：丁张纳 翁海旭 王晓娜 徐安聪 管磊 何国鑫 DING Zhangna;WENG Haixu;WANG Xiaona;XU Ancong;GUAN Lei;HE Guoxin(Department of Critical Care Medicine,the Third Affiliated Hospital of Wenzhou Medical University,Wenzhou 325200,China)

机构地区：[1]温州医科大学附属第三医院重症医学科,325200

出　　处：《浙江医学》2025年第7期678-681,687,I0003,I0004,共7页Zhejiang Medical Journal

基　　金：浙江省麻醉学重点实验室开放基金项目(KLAZJ2107);温州市科研项目(Y20210697)。

摘　　要：目的探讨双硫仑(DSF)抑制细胞焦亡改善脓毒症大鼠神经细胞损伤的作用机制。方法将58只无特定病原体的成年雄性SD大鼠按随机数字表法分为3组:假手术(Sham)组8只、盲肠结扎穿孔(CLP)组25只、DSF组25只,Sham组开腹后仅游离盲肠并直接回纳,其余两组均采用CLP法建立脓毒症模型。建模后DSF组腹腔注射DSF注射液(浓度为10 mg/mL)2 mL/次、2次/d,而Sham组、CLP组注射0.9%氯化钠注射液2 mL。建模7 d后采用断头法处死大鼠,取海马组织并观察病理学变化,采用蛋白质印迹法检测裂解的(cleaved)胱天蛋白酶(caspase)-1、消皮素D(GSDMD)、NOD样受体家族含pyrin结构域蛋白(NLRP)3、白细胞介素(IL)-1β蛋白表达水平,免疫荧光法检测caspase-1、GSDMD、NLRP3、IL-1β含量,对所测得结果进行组间比较。结果CLP组大鼠海马神经细胞核深染,核固缩,胞体空泡或皱缩,神经细胞明显损伤;Sham组大鼠神经细胞无明显损伤;DSF组大鼠部分海马神经细胞存在损伤,较CLP组神经细胞损伤相对较少,损伤程度较轻。CLP组大鼠海马组织cleaved caspase-1、GSDMD、NLRP3、IL-1β蛋白表达水平以及caspase-1、GSDMD、NLRP3、IL-1β含量均较Sham组明显升高(均P<0.05),而DSF组上述各项指标均较CLP组明显降低(均P<0.05)。结论脓毒症大鼠存在海马神经细胞损伤以及细胞焦亡激活,而DSF可以通过抑制细胞焦亡从而起到保护神经细胞的作用。Objective To investigate the effect of disulfiram(DSF)on neuronal injury in septic rats and its mechanism.Methods A total of 58 specific pathogen free adult male Sprague-Dawley rats were randomly divided into 3 groups:the sham group(n=8),the model group(n=25),and the DSF group(n=25).The sepsis model was induced by cecal ligation and puncture(CLP)in model and DSF groups,and sham operation was performed in Sham group.After modeling,the DSF group and was intraperitoneally injected with 2 mL DSF(10 mg/mL),2 times a day for 7 days,while the Sham group and the model group were injected with 2 mL of 0.9%sodium chloride injection.Seven days after modeling,the rats were sacrificed by decapitation.The hippocampal tissues were taken,and the pathological changes were observed.The protein expression of cleaved caspase-1,gasdermin D(GSDMD),NOD-like receptor family pyrin domain-containing(NLRP)3,and interleukin(IL)-1βin hippocampal tissues was detected by Western blotting and immunofluorescence,respectively,and the results were compared among groups.Results In the model group,the hippocampal neurons of the rats showed deep-stained nuclei,nuclear pyknosis,vacuoles or shrinkage of the cell bodies,and obvious nerve cell injury.There was no obvious nerve cell injury in the rats of the Sham group.In the DSF group,some hippocampal nerve cells were damaged,and the nerve cell injury was ameriolated and the degree of injury was milder compared with that in the model group.The protein expression levels and the contents of cleaved caspase-1,GSDMD,NLRP3,and IL-1βin the hippocampal tissues of the model group were significantly higher than those in the Sham group(all P<0.05).In the DSF group,the expression and contents of above proreins were significantly lower than those in the model group(all P<0.05).Conclusion In sptic rats,there is hippocampal nerve cell injury and activation of pyroptosis,and DSF can inhibit pyroptosis and ameriolate neoronal injury.

关 键 词：双硫仑 脓毒症 细胞焦亡 海马组织 神经细胞 

分 类 号：R459.7[医药卫生—急诊医学]