加味银翘散对反复热性惊厥幼...大鼠免疫功能及脑损伤的影响_曾洁.pdf

引用:曾洁，周姗，谢静，邓羿駃，帅云飞，李鑫.加味银翘散对反复热性惊厥幼龄大鼠免疫功能及脑损伤的影响J.湖南中医杂志，2023，39(4):173178.加味银翘散对反复热性惊厥幼龄大鼠免疫功能及脑损伤的影响曾洁1，2，周姗1，2，谢静1，2，邓羿駃1，2，帅云飞1，2，李鑫3(1.湖南中医药大学第一附属医院，湖南长沙，410007;2.湖南中医药大学中医学国内一流建设学科，湖南长沙，410208;3.湖南中医药大学中医诊断学湖南省重点实验室，湖南长沙，410208)摘要 目的:观察加味银翘散对反复热性惊厥(FS)幼龄大鼠免疫功能及脑损伤的影响。方法:对SPF 级幼龄 SD 大鼠采用热水浴方法建立 FS 模型。将热水浴中 5 min 内出现惊厥的大鼠随机分为模型组、丙戊酸钠组、加味银翘散大剂量组(JYPH 组)、加味银翘散小剂量组(JYPL 组)，每组各 10 只。另设空白组10 只。灌胃时，JYPL 组及 JYPH 组分别为 0.252、1.008 g/ml 的浓度给药，丙戊酸钠组为 9.45 g/ml 浓度给药，空白以及模型组给予等容积蒸馏水，连续干预 20 d。HE 染色及尼氏染色检测大鼠海马组织病理学变化;使用流式细胞术检测大鼠外周血 T 淋巴细胞亚群;采用透射比浊法分析免疫球蛋白 A(IgA)、免疫球蛋白 G(IgG)、免疫球蛋白 M(IgM)水平。结果:与空白组相比，模型组大鼠均有明显惊厥发作(P0.01);其CD4+水平、CD4+/CD8+比值降低(P001)，CD8+水平升高(P001)，IgA、IgG 水平降低(P001)。与模型组相比，丙戊酸钠组、JYPH 组及 JYPL 组肛温下降，惊厥潜伏时间延长，惊厥级别降低，IgA、IgG 水平升高，差异均有统计学意义(P001，P005);丙戊酸钠组及 JYPH 组 CD4+水平、CD4+/CD8+比值升高(P001)，CD8+水平降低(P001)。与模型组比较，各药物组大鼠海马 CA1 区、CA3 区的神经元数目增多;JYPH 组的CA1 区、CA3 区的神经元数目多于丙戊酸钠组、JYPL 组。结论:加味银翘散治疗幼龄大鼠热性惊厥反复发作的作用机制可能与调节免疫功能及保护脑损伤有关。关键词 反复热性惊厥;加味银翘散;免疫功能;脑损伤 中图分类号 285.5 文献标识码 ADOI:1016808/jcnkiissn10037705202304038基金项目:湖南省自然科学基金项目(2020JJ4478);湖南中医药大学中医学国内一流建设学科(49010200002002);湖南中医药大学校级科研基金项目(ZYYDX201750)第一作者:曾洁，女，医学硕士，医师，研究方向:中医儿科临床通信作者:谢静，女，医学博士，副教授，研究方向:中医儿科临床，Email:982100692 qqcomEffect of Jiawei Yinqiao powder on immune function and brain injury inyoung rats with recurrent febrile seizuresZENG Jie1，2，ZHOU Shan1，2，XIE Jing1，2，DENG Yijue1，2，SHUAI Yunfei1，2，LI Xin3(1.The First Affiliated Hospital of Hunan University of Chinese Medicine，Changsha 410007，Hunan，China;2.The Domestic Firstclass Construction Discipline of Chinese Medicine in Hunan University ofChinese Medicine，Changsha 410208，Hunan，China;3.Hunan Provincial Key Laboratory of Diagnostics of Traditional Chinese Medicine，Hunan University ofChinese Medicine，Changsha 410208，Hunan，China)Abstract Objective:To investigate the effect of Jiawei Yinqiao powder on immune function and brain injury inyoung rats with recurrent febrile seizures(FS).Methods:Specific pathogenfree young SpragueDawley rats were used toestablish a model of FS using the method of hot water bath.The rats with FS within 5 minutes of hot water bath were ran-371第 39 卷第 4 期2023 年 4 月HUNAN JOUNAL OF TADITIONAL CHINESE MEDICINEVol.39No.4Apr.2023domly divided into model group，valproic acid group，highdose Jiawei Yinqiao powder group(JYPH group)，and lowdose Jiawei Yinqiao powder group(JYPL group)，with 10 rats in each group，and a blank group with 10 rats was also es-tablished.The rats in the JYPL and JYPH groups were given Jiawei Yinqiao powder at a concentration of 0.252 and1.008 g/ml，respectively，by gavage，those in the valproic acid group were given the drug at a concentration of 9.45 g/ml，and those in the blank group and the model group were given an equal volume of distilled water，for 20 consecutivedays.HE staining and Nissl staining were used to observe histopathological changes of the hippocampus;flow cytometry wasused to measure peripheral blood T lymphocyte subsets;turbidimetric immunoassay was used to measure the levels of im-munoglobulin A(IgA)，immunoglobulin G(IgG)，and immunoglobulin M(IgM).esults:Compared with the blankgroup，the model group had obvious seizures(P0.01)，with asignificant reductions in the level of CD4+T lymphocytesand the ratio of CD4+/CD8+(P0.01)，a significant increase in the level of CD8+T lymphocytes(P0.01)，and signifi-cant reductions in the levels of IgA and IgG(P0.01)Compared with the model group，the groups of valproic acid，JYPHand JYPL had a significant reduction in rectal temperature，a significant increase in seizure latency，and a significant re-duction in the severity of seizures，the differences were statistically significant(P0.01，P0.05)，the groups of valproicacid and JYPH had a significant increases in the level of CD4+T lymphocytes and the ratio of CD4+/CD8+(P0.01)，asignificant reduction in the level of CD8+T lymphocytes(P0.01).Compared with the model group，the administrationgroups had an increased number of neurons in the hippocampal CA1 and CA3 regions，and compared with the valproic acidgroup and the JYPL group，the JYPH group had a higher number of neurons in the hippocampal CA1 and CA3 regionsCompared with the model group，the administration groups had an increased number of neurons in the hippocampal CA1and CA3 regions，and compared with the valproic acid group and the JYPL group，the JYPH group had a higher number ofneurons in the hippocampal CA1 and CA3 regions.Conclusion:Jiawei Yinqiao powder exerts a therapeutic effect on youngrats with recurrent FS possibly by regulating immune function and protecting against brain injury.Keywords recurrent febrile seizures;Jiawei Yinqiao powder;immune function;brain injury热性惊厥(febrile seizure，FS)是儿科常见的惊厥性危急重症，好发于 6 个月至 5 岁小儿。本病大多预后良好，但复发率较高1。临床研究发现惊厥反复发作容易损伤大脑的发育，部分患儿可继发癫痫，甚至导致智力低下，极大地影响了患儿的预后和生长发育2，对家长造成较大的心理压力和经济负担。本病病因复杂，发病机制尚不清楚，有研究表明其可能与高温、感染、海马神经元损伤、惊厥易感性、免疫功能等因素相关3。本病西医治疗可取得一定效果，但长期用药可能产生蓄积作用，不良反应较多4。中医学认为热性惊厥属于“急惊风”范畴，通常具备痰、热、惊、风四证，应以清热疏风、豁痰镇惊为基本治疗法则。通过前期临床观察发现，加味银翘散可在 FS 间歇期发挥其预防作用，从而减少患儿FS 的发作，但其作用机制尚不清楚。故本实验研究通过建立