痰热清注射液对实验性变态反应性脑脊髓炎小鼠炎性因子及炎性损伤影响的研究

Study on effects of TanReQing injection on inflammatory cytokines and injuries with EAE mice

目的观察痰热清注射液对实验性变态反应性脑脊髓炎(experimental allergic encephalomyelitis,EAE)小鼠炎性损伤的影响,应用HE染色、ELISA方法检测炎性浸润程度及炎性因子含量,从组织、分子层面揭示痰热清注射液治疗多发性硬化的科学内涵,为中医治疗多发性硬化提供科学依据。方法 SPF级C57BL/6雌性小鼠100只,将小鼠随机分为正常组,模型组,激素组,痰热清注射液大剂量组,痰热清注射液小剂量组。模型及治疗组注射MOG_(35-55)抗原乳剂及PTX诱导EAE。免疫当天痰热清注射液组进行腹腔注射治疗,激素组在发病后进行激素灌胃治疗。其余均以生理盐水代替。观察小鼠行为学的变化,在急性期及缓解期取小鼠大脑组织,HE观察病理组织学改变ELISA检测IL-17A等炎性因子的表达。结果(1)造模各组小鼠从免疫第8天开始发病,发病率为100%,痰热清注射液小剂量组发病潜伏期低于痰热清注射液大剂量组(P<0.05)。造模各组在第21天达到高峰,模型组最高,神经功能评分达到2.35。痰热清注射液大剂量组及激素组评分最低,与模型组相比有统计学意义(P<0.01)。(2)HE染色光镜下显示EAE模型各组在第21天可见大脑内小血管周围炎性细胞浸润和神经细胞核固缩,以第三脑室及侧脑室周围为主,大量的淋巴细胞聚集在血管周围,并形成典型的袖套状改变。第40天炎症浸润较第21天炎症浸润有所缓和。(3)痰热清注射液对EAE小鼠炎性因子的影响:ELISA统计结果发现:痰热清注射液小剂量组可以有效的抑制IL-6、IL-10、IL-17A、IL-23、TGF-β1多种细胞因子的表达(P<0.05)。结论痰热清可以有效的减少EAE的炎症反应及炎性因子的表达,从而抑制EAE的炎性进程。

Objective To study the effects of TanReQing injection on inflammatory injuries with ex-perimental autoimmune encephalomyelitis（EAE）mice,we used HE stain and ELISA to detect the inflammatory infiltration and cytokines. This study revealed the scientific connotation of TanReQing injection to treat multiple sclerosis （MS）disease. Methods One hundred SPF C57BL/6 female mice were randomly divided into normal group,model group,the PA group,TanReQing injection high-dose group and low-dose group. The EAE groups were induced by MOG35-55 and PTX. Intraperitoneal injection of TanReQing were carried out on the day of im-munization,PA group began to use gastric lavage when clinical symptoms appeared. The rest groups were injec-ted normal saline instead. The behavioral changes of mice were recorded and HE and ELISA were observed on day 21 and 40. Results （1）The EAE groups began to appear clinical symptoms on day 8,the incidence was 100%. The neurological scores of EAE groups peaked on day 21 and the model group had the highest point of 2.35. The scores of TanReQing injection high-dose group and PA group were lower than the model group （P〈0.01）. （2）HE stain showed EAE groups had inflammatory infiltration around the small blood vessels in the brain and nerve cells nucleus pycnosis,mainly around the third ventricle and the lateral ventricle. The inflammato-ry infiltration was remitted on day 40 compared with 21.3. TanReQing low-dose injection could inhibit the cytokines expression of IL-6,IL-10,IL-17a,IL-23 and TGF-β1 with EAE mice（P〈0.05）. Conclusion The TanReQing injection could effectively reduce EAE inflammatory reaction and the expression of inflammatory cytokines.