清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤MyD88、TRAF6 mRNA表达的影响
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摘要
目的:观察清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤髓样分化因子88(MyD88)和肿瘤坏死因子受体相关因子6(TRAF6)mRNA表达的影响。方法:将大鼠随机分为两组,每组再分为空白(A)组、假手术(B)组、脑缺血再灌注(C)组、清热化瘀Ⅱ号方(D)组。B、C、D组按照缺血2h后,再灌注3、6、12、24、48h 5个时间点分为5个亚组,每组6只。测定各组大鼠神经功能缺损评分,各组实验大鼠取材后,一组行常规镜下观察实验大鼠脑组织的病理形态学变化;另一组取材后应用荧光定量PCR法检测实验大鼠的海马区MyD88和TRAF6mRNA表达变化。结果:与A组比较,C组神经评分显著升高(P<0.05),而D组较C组有明显改善(P<0.05)。A、B组大鼠脑组织病理形态学无明显改变;C、D组大鼠脑组织病理结构均明显受损,D组较C组在各时间点神经元受损显著减轻。A、B组My D88、TRAF6 m RNA均有少量表达,差异无统计学意义;C组My D88、TRAF6 m RNA表达含量较A组显著升高(P<0.05,P<0.01);与C组相比,D组MyD88、TRAF6 mRNA表达含量显著降低(P<0.05,P<0.01)。结论:清热化瘀Ⅱ号方可抑制大鼠脑缺血再灌注后MyD88、TRAF6的表达来减轻脑缺血再灌注损伤,而发挥脑保护作用。
Objective: To observe the effects of Qingre Huayu Decoction Ⅱ on mRNA expression levels of MyD88 and TRAF6 of rats with cerebral ischemia reperfusion. Methods: All SD rats were randomly divided into two groups, then each group was randomly divided into four groups: the normal control group(group A), sham operation group(group B), ischemiareperfusion group(group C) and Qingre Huayu Decoction Ⅱ group(group D). Except the normal control group, other groups were given 2 h ischemia, and they were further divided into 5 subgroups according to 3 h, 6 h, 12 h, 24 h and 48 h reperfusion, with six rats in each group. The rats in each group were sacrificed. The pathological morphological changes of brain tissues of rats in one group were observed under conventional microscope. Fluorescence quantitative PCR was used to detect the mRNA expression changes of MyD88 and TRAF6 in the hippocampal area of rats in the other group. Results: Compared with group A, the neurologic impairment score in group C significantly increased(P<0.05), and that in group D improved than in group C(P<0.05). There was no significant change in histopathological morphology of brain tissue in group A and group B. The pathological structure of brain tissue of rats in group C and group D was obviously damaged. The damage degree of neurons in group D was significantly lightened compared with group C at all same time points. Small amounts of MyD88 and TRAF6 mRNA in group A and group B were both expressed, and there was no statistical difference. The mRNA expression levels of MyD88 and TRAF6 in group C began to increase, and they were significantly higher in group C than those in group D(P<0.05, P<0.01). Compared with group C, the mRNA expression levels of MyD88 and TRAF6 in group D decreased significantly(P<0.05, P<0.01). Conclusion: Qingre Huayu Decoction Ⅱ can inhibit the expression levels of My D88 and TARF6 of rats with cerebral ischemia reperfusion injury to alleviate cerebral ischemia reperfusion injury, playing a role of brain protection.
Objective: To observe the effects of Qingre Huayu Decoction Ⅱ on mRNA expression levels of MyD88 and TRAF6 of rats with cerebral ischemia reperfusion. Methods: All SD rats were randomly divided into two groups, then each group was randomly divided into four groups: the normal control group(group A), sham operation group(group B), ischemiareperfusion group(group C) and Qingre Huayu Decoction Ⅱ group(group D). Except the normal control group, other groups were given 2 h ischemia, and they were further divided into 5 subgroups according to 3 h, 6 h, 12 h, 24 h and 48 h reperfusion, with six rats in each group. The rats in each group were sacrificed. The pathological morphological changes of brain tissues of rats in one group were observed under conventional microscope. Fluorescence quantitative PCR was used to detect the mRNA expression changes of MyD88 and TRAF6 in the hippocampal area of rats in the other group. Results: Compared with group A, the neurologic impairment score in group C significantly increased(P<0.05), and that in group D improved than in group C(P<0.05). There was no significant change in histopathological morphology of brain tissue in group A and group B. The pathological structure of brain tissue of rats in group C and group D was obviously damaged. The damage degree of neurons in group D was significantly lightened compared with group C at all same time points. Small amounts of MyD88 and TRAF6 mRNA in group A and group B were both expressed, and there was no statistical difference. The mRNA expression levels of MyD88 and TRAF6 in group C began to increase, and they were significantly higher in group C than those in group D(P<0.05, P<0.01). Compared with group C, the mRNA expression levels of MyD88 and TRAF6 in group D decreased significantly(P<0.05, P<0.01). Conclusion: Qingre Huayu Decoction Ⅱ can inhibit the expression levels of My D88 and TARF6 of rats with cerebral ischemia reperfusion injury to alleviate cerebral ischemia reperfusion injury, playing a role of brain protection.
引文
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[11]刘洋,黎瀚,关俊宏,等.TLR4及MyD88在大鼠局灶性脑缺血耐受过程中的作用.中华神经外科疾病研究杂志,2013,12(5):405-409
[12]Raffaella Gesuete,Steven G Kohama,Mary Stenzel-Poore.Toll-like receptors and ischemic brain injury.J Neuropathol Exp Neurol,2014,73(5):378-386
[13]祝美珍,齐玉洁,刘泰,等.清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤TLR4、TRIF mRNA表达的影响.中华中医药杂志,2017,32(2):529-532
[2]Longa E Z,Weinstein P R,Carlson S,et al.Reversible middle cerebral artery occulsio-n without craniectomy in rats.Stroke,1989,20(1):84-91
[3]贺石林,王键,王净净.中医科研设计与统计学.长沙:湖南科学技术出版社,2001:48
[4]尤晓欣,裴建.Toll样受体信号通路与缺血性中风炎症反应机制的研究进展.国际神经病学神经外科学杂志,2014,4(2):138-141
[5]陈莹莹,孙广玲,兰倩,等.Toll样受体与脑缺血.现代生物医学进展,2014,12(14):2369-2372
[6]Wang L,Liu X H,Chen H,et al.PicrosideⅡprotects rat kidney against ischemia/reperfusion-induced oxidative stress and inflammation by the TLR4/NF-κB path-way.Exp Ther Med,2015,9(4):1253-1258
[7]蔡炳冈,朱进,汪茂荣.Toll样受体4信号通路研究进展.医学研究生学报,2015,28(11):1228-1232
[8]Liang Y,Lei Z,Zhang H,et al.Toll-like receptor 4 isassociated with seizures following ischemia with hyperglycemia.Brain Res,2014,8993(14):1229-1233
[9]Moraga A,Pradillo J M,Cuartero M I,et al.Toll-like receptor 4modulates cell migration and cortical neurogenesis after focal cerebral ischemia.FASEB J,2014,28(11):4710-4718
[10]Song Z,Jin R,Yu S,et al.Crucial role of CD40 singnaling in vascular all cells in neointimal formation and vascular remodeling after vascular intervention.Arterioscler Thromb Vasc Biol,2012,32(1):50-64
[11]刘洋,黎瀚,关俊宏,等.TLR4及MyD88在大鼠局灶性脑缺血耐受过程中的作用.中华神经外科疾病研究杂志,2013,12(5):405-409
[12]Raffaella Gesuete,Steven G Kohama,Mary Stenzel-Poore.Toll-like receptors and ischemic brain injury.J Neuropathol Exp Neurol,2014,73(5):378-386
[13]祝美珍,齐玉洁,刘泰,等.清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤TLR4、TRIF mRNA表达的影响.中华中医药杂志,2017,32(2):529-532