摘要
目的探讨鞣花酸(EA)减少大鼠癫痫发作的作用机制。方法选取6周龄SPF级健康雄性Sprague-Dawley大鼠,将大鼠分为6组:对照组(n=12)、模型组(n=11)、EA低剂量组(n=11)、EA中剂量组(n=11)、EA高剂量组(n=11)、激动剂组(n=11)。对照组为正常SD大鼠,其他组均为戊四唑(PTZ)(35 mg/kg)诱导建模成功的癫痫模型大鼠。对照组和模型组大鼠腹腔注射生理盐水;EA低、中和高剂量组大鼠分别腹腔注射剂量为25、50和100 mg/kg/d的EA;激动剂组大鼠同时腹腔注射剂量为100 mg/kg/d的EA和20μg/kg/d的重组大鼠白介素-6(rrIL-6)。各组大鼠均给药或生理盐水4周。末次给药24 h后,再次腹腔注射PTZ(35 mg/kg),记录惊厥潜伏期及持续时间。通过HE和Nissl染色观察大鼠海马神经元损伤。采用ELISA法检测IL-6、IL-1β和肿瘤坏死因子-α(TNF-α)含量。采用免疫荧光染色检测大鼠海马胶质纤维酸性蛋白(GFAP)的表达。采用Western blot检测大鼠海马GFAP、磷酸化-Janus激酶2(p-JAK2)、JAK2、磷酸化-信号转导和转录激活因子3(p-STAT3)和STAT3的表达。结果与对照组比较,模型组大鼠出现明显惊厥,神经元出现明显损伤,Nissl+神经元计数减少,海马IL-6、IL-1β和TNF-α含量升高,海马CA1区GFAP相对荧光强度和蛋白相对水平升高,海马p-JAK2和p-STAT3蛋白相对水平升高(P<0.05)。与模型组比较,EA低、中和高剂量组大鼠的惊厥潜伏期延长,持续时间缩短,神经元损伤明显减轻,Nissl+神经元计数增加,海马IL-6、IL-1β和TNF-α含量降低,海马CA1区GFAP相对荧光强度和蛋白相对水平降低,海马p-JAK2和p-STAT3蛋白相对水平降低(P<0.05)。与EA高剂量组比较,激动剂组大鼠的惊厥潜伏期缩短,持续时间延长,神经元损伤加重,Nissl+神经元计数减少,海马IL-6、IL-1β和TNF-α含量升高,海马CA1区GFAP相对荧光强度和蛋白相对水平升高,海马p-JAK2和p-STAT3蛋白相对水平升高(P<0.05)。结论EA通过抑制JAK2/STAT3信号通路抑制神经炎症和星形胶质细胞的过度活化,从而减少大鼠癫痫发作。
Objective To explore the mechanism of ellagic acid(EA)in reducing epileptic seizures in rats.Methods Rats were divided into 6 groups:control group(n=12),model group(n=11),EA low dose group(n=11),EA medium dose group(n=11),EA high dose group(n=11)and agonist group(n=11).Control group was normal SD rats,and other groups were epileptic rats induced by pentylenetetrazol(PTZ)(35 mg/kg).Control group and model group were intraperitoneally injected with normal saline,while EA low dose group,EA medium dose group and EA high dose group were intraperitoneally injected with 25,50 and 100 mg/kg/d EA,respectively.The rats in agonist group were intraperitoneally injected with EA of 100 mg/kg/d and 20μg/kg/d recombinant rat interleukin-6(rrIL-6).The drug was given for 4 weeks.24 hours after the last administration,PTZ(35 mg/kg)was injected intraperitoneally again,and the latency and duration of convulsion were recorded.Hippocampal neuronal damage was observed by HE and Nissl staining.The contents of IL-6,IL-1βand tumor necrosis factor-α(TNF-α)were measured by ELISA method.The expression of glial fibrillary acidic protein(GFAP)in hippocampus was detected by immunofluorescence staining.The expressions of GFAP,phosphorylated-Janus kinase 2(p-JAK2),JAK2,phosphorylated signal transducer and activator of transcription 3(p-STAT3)and STAT3 in hippocampus were detected by Western blot.Results Compared with control group,model group showed obvious convulsion,obvious neuronal damage,Nissl+neuron count decreased,hippocampus IL-6,IL-1βand TNF-αcontent increased,hippocampus CA1 region GFAP relative fluorescence intensity and protein relative level increased,and the relative levels of p-JAK2 and p-STAT3 proteins in hippocampus increased(P<0.05).Compared with the model group,the convulsion latency of the EA low dose group,EA medium dose group and EA high dose group were prolonged,the duration was shortened,neuronal damage was significantly reduced,Nissl+neuron counts increased,hippocampal IL-6,IL-1βand TNF-αcontents decreased,hippocampus CA1 region GFAP relative fluorescence intensity and protein relative level decreased,and the relative protein levels of hippocampal p-JAK2 and p-STAT3 decreased(P<0.05).Compared with the EA high dose group,the convulsion latency of the agonist group was shortened,the duration was prolonged,neuronal damage was aggravated,Nissl+neuron count decreased,hippocampus IL-6,IL-1βand TNF-αcontent increased,hippocampus CA1 region GFAP relative fluorescence intensity and protein relative level increased,and the relative levels of p-JAK2 and p-STAT3 proteins in hippocampus increased(P<0.05).Conclusion EA inhibits neuroinflammation and excessive activation of astrocytes by inhibiting JAK2/STAT3 signal pathway,thus reducing epileptic seizures in rats.
作者
刘雨东
张垚
李凌
吴镇宇
杨欣伟
LIU Yudong;ZHANG Yao;LI Ling;WU Zhenyu;YANG Xinwei(Department of Pediatrics,The First Affiliated Hospital of Airforce Medical University,Xi'an 710032,China;Department of Pediatrics,Shenmu Second Hospital,Shenmu 719313,Shanxi,China)
出处
《西部医学》
2025年第7期981-987,共7页
Medical Journal of West China
基金
国家自然科学基金青年科学基金项目(81701487)。
