延胡索总生物碱对慢性癫痫大鼠皮层NLRP3/caspase-1/GSDMD介导的细胞焦亡的影响  

Effect of total alkaloids of Rhizoma Corydalis on NLRP3/caspase-1/GSDMD-mediated pyroptosis in epileptic rats

作　　者：齐雅芝 唐娅 李君[1] 曹睿 翟燕玲 韩玉生[1] 徐强[1] QI Yazhi;TANG Ya;LI Jun;CAO Rui;ZHAI Yanling;HAN Yusheng;XU Qiang(Heilongjiang University of Chinese Medicine,Harbin 150040,China;School of Jiamusi,Heilongjiang University of Chinese Medicine,Jiamusi 154007,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学佳木斯学院,黑龙江佳木斯154007

出　　处：《中国病理生理杂志》2023年第11期1938-1946,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金面上项目(No.82074530);黑龙江中医药大学科研基金项目(No.X200902)。

摘　　要：目的:研究延胡索总生物碱(TAC)对戊四氮(PTZ)所诱导的慢性癫痫大鼠的治疗作用,探讨其对大脑皮层核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)/caspase-1/消皮素D(GSDMD)介导的细胞焦亡的影响及可能机制。方法:清洁级雄性SD大鼠30只,随机抽取6只大鼠作为正常组,其余大鼠采用PTZ(35 mg/kg)腹腔连续注射28 d复制慢性癫痫模型。将造模成功的18只大鼠随机分为模型组、低剂量TAC组和高剂量TAC组,每组6只。低、高剂量TAC组大鼠分别灌胃给予7和14 mg/kg的TAC,连续给药14 d,正常组和模型组大鼠灌胃给予等体积生理盐水。观察记录大鼠全面强直阵挛发作(GTCS)和极小阵挛发作(MCS)的潜伏期;采用HE染色观察颞叶皮层的病理学变化;TUNEL法检测神经细胞凋亡;免疫组化和Western blot法检测皮层组织核因子κB(NF-κB)、NLRP3、caspase-1和GSDMD蛋白表达;免疫组化检测脑组织中肿瘤坏死因子α(TNF-α)、白细胞介素18(IL-18)和IL-1β蛋白表达;RT-qPCR检测各组NLRP3、caspase-1和GSDMD的mRNA表达。结果:与正常组相比较,模型组大鼠GTCS和MCS的潜伏期显著缩短(P<0.01);模型组大鼠皮层神经细胞凋亡数量显著增加,皮层组织中TNF-α、IL-18和IL-1β蛋白表达均显著增加,NF-κB、NLRP3、caspase-1 p20和GSDMD N端片段(GSDMD-N)的mRNA及蛋白表达也显著增加(P<0.05或P<0.01);与模型组相比,各剂量TAC组大鼠GTCS和MCS的潜伏期显著延长,皮层神经细胞凋亡数量显著减少,皮层组织中TNF-α、IL-18和IL-1β蛋白表达均显著减少,NF-κB、NLRP3、caspase-1 p20和GSDMD的mRNA及蛋白表达也均显著减少(P<0.01)。结论:TAC可能通过NLRP3/caspase-1/GSDMD信号通路,减少炎症因子分泌,抑制神经细胞焦亡,从而对PTZ诱导的慢性癫痫大鼠起到治疗作用。AIM:To study the therapeutic effect of total alkaloids of Rhizoma Corydalis(TAC)on pentetrazol(PTZ)-induced chronic epileptic rats,and to explore its effect on nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1/gasdermin D(GSDMD)-mediated pyroptosis in the cerebral cortex and its possible mechanism.METHODS:Six rats were randomly selected from 30 SPF male SD rats as the normal group,and the remaining rats were intraperitoneally injected with PTZ(35 mg/kg)for 28 d to replicate the chronic epilepsy model.The 18 successfully modeled rats were randomly divided into model,low-dose TAC(TAC-L)and high-dose TAC(TAC-H)groups,with 6 rats in each group.The rats in TAC-L and TAC-H groups received 7 and 14 mg/kg of TAC by continuous lavage for 14 d,while those in normal and model groups were given equal volume of saline.The latency of generalized tonic-clonic seizures(GTCS)and minimal clonic seizures(MCS)was recorded.The pathological changes of the temporal lobe cortex were observed by HE staining.Neuronal apoptosis was detected by TUNEL staining.The protein levels of NF-κB,NLRP3,caspase-1 and GSDMD in the cortex were detected by immunohistochemical staining and Western blot.The expression levels of TNF-α,interleukin-18(IL-18)and IL-1βin brain tissue were detected by immunohistochemical staining.The mRNA expression levels of NLRP3,caspase-1 and GSDMD in the cerebral cortex were detected by RT-qPCR.RESULTS:Com-pared with normal group,the latency of GTCS and MCS in the rats of model group was significantly shortened(P<0.01).Compared with normal group,the number of apoptotic cortical neurons in model group was significantly increased.The protein expression levels of TNF-α,IL-18 and IL-1βin the cortical tissues were significantly increased,and the mRNA and protein expression levels of NF-κB,NLRP3,caspase-1 and GSDMD were also significantly increased(P<0.05 or P<0.01).Compared with model group,the latency of GTCS and MCS in TAC treament groups was significantly prolonged,the number of apoptotic

关 键 词：癫痫 延胡索总生物碱 NLRP3/caspase-1 GSDMD信号通路 细胞焦亡 

分 类 号：R742.1[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]