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 cortical neurons in rats was significantly reduced,the protein expression levels of TNF-α,IL-18 and IL-1βin cortical tissue was reduced to varying degrees,and the mRNA and protein expression of NF-κB,NLRP3,caspase-1 and GSDMD also showed varying degrees of reduction(P<0.01).CONCLUSION:The TAC may reduce the secretion of inflammatory factors and inhibit the pyroptosis of nerve cells through the NLRP3/caspase-1/GSDMD signaling pathway,thus playing a therapeutic role in chronic epileptic rats induced by PTZ.
作者
齐雅芝
唐娅
李君
曹睿
翟燕玲
韩玉生
徐强
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)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
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)。
