Mechanism of action of Zhuyu Annao pill in mice with cerebral intrahemorrhage based on TLR4

Objective:To explore the protective effect and possible mechanism of action of Zhuyu Annao pill in mice with intracerebral hemorrhage(ICH).Methods:Sixty mice were divided into the control group,hemorrhage group,drug-treated group(after hemorrhage),TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group(after hemorrhage) with 12 in each group.Model of autologous ICH was established in all groups.After drilling and 12 h of fasting,models in the control group hemorrhage group and TLR4-knockout hemorrhage group were all drenched with 10 mL/kg distilled water by intragastric administration.Models in the drug-treated group and TLR4-knockout hemorrhage + drugtreated group were drenched with 6.25 g/kg of Zhuyu Annao pill.All groups were treated for 7 d.Longa scoring method was used to measure the neurological defect scores and determine the brain water contents of all groups;ELISA was employed to detect the inflammatory factor interleukin(IL)-6,tumor necrosis factor- α(TNF- α) and IL-1β in brain tissues;and Western blot was applied to test the expression quantities of apoptotic protein Bax and anti-apoptotic protein Bcl-2 in brain tissues.Results:At day 3 and7,compared with the hemorrhage group,the neurological defect scores of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group decreased significantly(P<0.05) Compared with the hemorrhage group,the brain water contents of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group reduced significantly(P<0.05) Compared with the hemorrhage group,the inflammatory factor IL-6,TNF-α and IL-1β of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage + drug-treated group decreased significantly(P<0.05).Compared with the hemorrhage group,the expression of apoptotic protein Bax of the drug-treated group,TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+ drug-treated group decreased significantly and the expression of anti-apoptotic protein Bcl-2 increased significantly(P<0.05).Conclusions:Zhuyu Annao pill can alleviateencephaledema for mice with ICH and reduce inflammatory responsesandnerve cell apoptosis.TLR4 can mediate inflammatory injury induced by ICH.Thus,Zhuyu Annao pill can play a protective role for brains by decreasing the expression of TLR4.

逐瘀安脑丸对高血压性脑出血患者临床疗效及炎症因子影响

目的:观察逐瘀安脑丸对高血压性脑出血患者临床疗效及炎症因子的影响。方法:将2015年5月-2017年10月我院收治的84例出血量在15~40mL的高血压脑出血患者,随机分为治疗组和对照组,最后完成本研究的有治疗组39例,对照组41例。对照组给予常规西药治疗,治疗组加用逐瘀安脑丸,运用双抗体夹心ELISA法分别检测治疗前后两组患者空腹的血清IL-6、IL-1β、TNF-α水平,并记录两组治疗前、治疗后7d、治疗后28d的NIHSS评分。结果:治疗组总有效率达92.3%,优于对照组的68.3%(P<0.05);两组患者治疗后NIHSS评分明显降低,治疗组显著优于对照组,差异具有统计学意义(P<0.05);治疗7、14d后两组患者的血清IL-6、IL-1β、TNF-α浓度均较治疗前下降,治疗组显著低于对照组,差异具有统计学意义(P<0.05)。结论:逐瘀安脑丸结合常规西药与单纯西药治疗高血压性脑出血相比,其对改善患者的神经功能、降低炎症因子水平和提高临床疗效作用更显著。

逐瘀安脑丸对脑出血小鼠TLR4介导的神经炎症反应的影响

目的:探讨逐瘀安脑丸对脑出血小鼠的保护作用及可能的作用机制。方法:将正常WT小鼠分为对照组、出血组(ICH组)和出血后药物处理组(ICH+药物组),TLR4敲除小鼠分为TLR4敲除出血组(TLR4+ICH组)和TLR4敲除组出血+药物处理组(TLR4+ICH+药物组),每组各15例。自体脑出血造模,对照组钻孔后不注射股动脉血,禁食12小时后灌胃器灌服蒸馏水;ICH组和TLR4+ICH组造模成功后,禁食12小时后灌胃器灌服蒸馏水;ICH+药物组、TLR4+ICH+药物组造模成功后,禁食12小时后灌胃器灌服6.25g/kg逐瘀安脑丸,各组均治疗7天。Longa评分法测定各组神经功能缺损评分,测量脑含水量测定,酶联免疫吸附法(ELISA)测定脑组织中炎症因子IL-6、IL-1β、TNF-α。结果:第3天和第7天,与ICH组比较,ICH+药物组、TLR4+ICH组、TLR4+ICH+药物组神经功能缺损评分显著降低(P<0.05)。与ICH组比较,ICH+药物组、TLR4+ICH组、TLR4+ICH+药物组脑含水量显著降低(P<0.05)。与对照组比较,ICH组炎症因子IL-6、IL-1β和TNF-α显著增加(P<0.05);与ICH组比较,ICH+药物组、TLR4+ICH组、TLR4+ICH+药物组炎症因子IL-6、IL-1β和TNF-α和显著降低(P<0.05)。结论:逐瘀安脑丸能够减轻脑出血小鼠脑水肿,降低炎症反应;TLR4能够介导小鼠脑出血引起的炎症损伤,逐瘀安脑丸可能通过降低TLR4表达发挥脑保护作用。

逐瘀安脑丸对模型大鼠脑出血的保护作用研究

目的:研究逐瘀安脑丸对模型大鼠脑出血的保护作用。方法:实验分为6组,即空白、模型、安宫牛黄丸及逐瘀安脑丸高、中、低剂量组。以大鼠自体血凝块复制脑血肿模型,用积分法记录神经功能缺损的变化,用干湿比重法测定脑组织水分,同时观察血液流变学、凝血四项与脑组织形态学的变化。结果:与模型组比较,逐瘀安脑丸高、中、低剂量组脑含水量与神经功能缺损积分无显著变化;凝血四项中的凝血酶原时间(PT)显著延长(P<0.05),血液流变学中全血还原黏度显著降低(P<0.05)、红细胞压积显著降低(P<0.05);凝血时间无显著变化;脑组织形态学显著改善。结论:逐瘀安脑丸对急性脑出血有积极的保护作用。