逐瘀安脑丸对脑出血小鼠TLR4介导的神经炎症反应的影响
|
摘要
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。
Objective: To explore the protective effect and possible mechanism of action of Zhuyu Annao Pill in mice with intracerebral hemorrhage. Methods: The normal WT mice were divided into controlled group,ICH group and ICH+drug group,TLR4 knockout mice were divided into TLR4+ICH group and TLR4+ICH+drug group,with 15 cases in each group. Model of autologous ICH was established in all groups. After drilling and 12 h of fasting,models in the controlled group,the ICH group and the TLR4+ICH group were all drenched with 10 ml/kg distilled water by intragastric administration,the ICH+drug group and the TLR4+ICH+drug group were drenched with 6.25 g/kg of Zhuyu Annao Pill by intragastric administration. All groups were treated for 7 d. Longa scoring method was used to measure theneurological defect scores and determine the brain water contents of all groups. ELISA was employed to detect the inflammatory factor IL-6,IL-1βand TNF-a in brain tissues. Results: At day 3 and 7,compared with the ICH group,the neurological deficit score of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group decreased significantly(P<0.05). Compared with the ICH group,the brain water contents of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group reduced significantly(P<0.05). Compared with the controlled group,the inflammatory factor IL-6,IL-1β,TNF-a of the ICH group increased significantly(P<0.05). Compared with the ICH group,the inflammatory factor IL-6,IL-1β,TNF-a of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group decreased significantly(P<0.05). Conclusion: Zhuyu Annao Pill can alleviate encephaledema for mice with ICH and reduce inflammatory responses. TLR4 can mediate inflammatory injury induced by ICH. Thus,Zhuyu Annao Pill can play a protective role for brains bydecreasing the expression of TLR4.
Objective: To explore the protective effect and possible mechanism of action of Zhuyu Annao Pill in mice with intracerebral hemorrhage. Methods: The normal WT mice were divided into controlled group,ICH group and ICH+drug group,TLR4 knockout mice were divided into TLR4+ICH group and TLR4+ICH+drug group,with 15 cases in each group. Model of autologous ICH was established in all groups. After drilling and 12 h of fasting,models in the controlled group,the ICH group and the TLR4+ICH group were all drenched with 10 ml/kg distilled water by intragastric administration,the ICH+drug group and the TLR4+ICH+drug group were drenched with 6.25 g/kg of Zhuyu Annao Pill by intragastric administration. All groups were treated for 7 d. Longa scoring method was used to measure theneurological defect scores and determine the brain water contents of all groups. ELISA was employed to detect the inflammatory factor IL-6,IL-1βand TNF-a in brain tissues. Results: At day 3 and 7,compared with the ICH group,the neurological deficit score of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group decreased significantly(P<0.05). Compared with the ICH group,the brain water contents of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group reduced significantly(P<0.05). Compared with the controlled group,the inflammatory factor IL-6,IL-1β,TNF-a of the ICH group increased significantly(P<0.05). Compared with the ICH group,the inflammatory factor IL-6,IL-1β,TNF-a of the ICH+drug group,the TLR4 group and the TLR4+ICH+drug group decreased significantly(P<0.05). Conclusion: Zhuyu Annao Pill can alleviate encephaledema for mice with ICH and reduce inflammatory responses. TLR4 can mediate inflammatory injury induced by ICH. Thus,Zhuyu Annao Pill can play a protective role for brains bydecreasing the expression of TLR4.
引文
[1] Zhou L,Deng L,Chang N B,et al.Cell apoptosis and proliferation in rat brains after intracerebral hemorrhage:role of Wnt/β-catenin signaling pathway[J].Turkish Journal of Medical Sciences,2014,44(6):920
[2] 杨莎莎,田清友,周洪霞,等.TLR4和NF-κB在脑出血大鼠炎症损伤中的表达及意义[J].重庆医学,2014,43(05):584～586
[3] 王大永,徐翔,孙立倩,等.亚低温治疗对脑出血大鼠TLR4/NF-κB介导的神经炎症反应的影响[J].神经解剖学杂志,2015,31(03):309～314
[4] 陶蕾,王高翔.TLR4及NF-kB在脑出血大鼠血肿周围组织的表达[J].贵州医科大学学报,2017,42(07):812～816
[5] Wang J,Lin D,Peng H,et al.Cancer-derived immunoglobulin G promotes LPS-induced proinflammatory cytokine production via binding to TLR4in cervical cancer cells.[J].Oncotarget,2014,20(5):9727～9743
[6] 吴林,李文娅,张其瑞,等.逐瘀安脑丸对大鼠实验性脑出血后水通道蛋白4、铁离子表达及脑水肿的作用研究[J].时珍国医国药,2010,21(02):383～385
[7] 郑福奎,吴林,陈炜,等.逐瘀安脑方治疗急性期脑出血临床研究[J].中国中医急症,2011,7(10):1555～1556
[8] Wang J,Grishin AV,Ford HR.Experimental Anti-Inflammatory Drug Semapimod Inhibits TLR Signaling by Targeting the TLR Chaperonegp96[J].J Immunol,2016,196(12):5130～7
[9] Ciaramelli C,Calabrese V,Sestito SE,et al.Glycolipid-based TLR4Modulators and Fluorescent Probes:Rational Design,Synthesis,and Biological Properties[J].Chem Biol Drug Des,2016,88(2):217～29
[10] Hu QP,Mao DA.Histone deacetylase inhibitor SAHA attenuates post-seizure hippocampal microglia TLR4/MYD88signaling and inhibits TLR4gene expression via histone acetylation[J].BMC Neurosci,2016,17(1):22
[11] 秦龙,姜宁,张爱忠,等.Toll样受体4在机体免疫中的作用机制及一些营养因子对其的影响[J].动物营养学报,2017,29(09):3075～3082
[12] Church J S,Kigerl K A,Lerch J K,et al.TLR4Deficiency Impairs Oligodendrocyte Formation in the Injured Spinal Cord[J].Journal of Neuroscience the Official Journal of the Society for Neuroscience,2016,36(23):6352～6364
[2] 杨莎莎,田清友,周洪霞,等.TLR4和NF-κB在脑出血大鼠炎症损伤中的表达及意义[J].重庆医学,2014,43(05):584～586
[3] 王大永,徐翔,孙立倩,等.亚低温治疗对脑出血大鼠TLR4/NF-κB介导的神经炎症反应的影响[J].神经解剖学杂志,2015,31(03):309～314
[4] 陶蕾,王高翔.TLR4及NF-kB在脑出血大鼠血肿周围组织的表达[J].贵州医科大学学报,2017,42(07):812～816
[5] Wang J,Lin D,Peng H,et al.Cancer-derived immunoglobulin G promotes LPS-induced proinflammatory cytokine production via binding to TLR4in cervical cancer cells.[J].Oncotarget,2014,20(5):9727～9743
[6] 吴林,李文娅,张其瑞,等.逐瘀安脑丸对大鼠实验性脑出血后水通道蛋白4、铁离子表达及脑水肿的作用研究[J].时珍国医国药,2010,21(02):383～385
[7] 郑福奎,吴林,陈炜,等.逐瘀安脑方治疗急性期脑出血临床研究[J].中国中医急症,2011,7(10):1555～1556
[8] Wang J,Grishin AV,Ford HR.Experimental Anti-Inflammatory Drug Semapimod Inhibits TLR Signaling by Targeting the TLR Chaperonegp96[J].J Immunol,2016,196(12):5130～7
[9] Ciaramelli C,Calabrese V,Sestito SE,et al.Glycolipid-based TLR4Modulators and Fluorescent Probes:Rational Design,Synthesis,and Biological Properties[J].Chem Biol Drug Des,2016,88(2):217～29
[10] Hu QP,Mao DA.Histone deacetylase inhibitor SAHA attenuates post-seizure hippocampal microglia TLR4/MYD88signaling and inhibits TLR4gene expression via histone acetylation[J].BMC Neurosci,2016,17(1):22
[11] 秦龙,姜宁,张爱忠,等.Toll样受体4在机体免疫中的作用机制及一些营养因子对其的影响[J].动物营养学报,2017,29(09):3075～3082
[12] Church J S,Kigerl K A,Lerch J K,et al.TLR4Deficiency Impairs Oligodendrocyte Formation in the Injured Spinal Cord[J].Journal of Neuroscience the Official Journal of the Society for Neuroscience,2016,36(23):6352～6364
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |