僵蚕提取液促进小胶质细胞极化改善全脑缺血再灌注大鼠神经元损伤及行为学功能  

Bombyx Batryticatus extract promotes microglia polarization to improve neuron injury and behaviors of cerebral ischemia/reperfusion rats

作　　者：侯培媚 徐昊 李泽康[2,4] 周豪 王珊珊 葛金文[1,2] HOU Pei-mei;XU Hao;LI Ze-kang;ZHOU Hao;WANG Shan-shan;GE Jin-wen(College of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Key Laboratory of Hunan Province for Integrated Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases,Hunan University of Chinese Medicine,Changsha 410208,China;College of Acupuncture&Tuina and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China;School of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南中医药大学,中西医结合防治心脑血管疾病湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学针灸推拿与康复学院,湖南长沙410208 [4]湖南中医药大学药学院,湖南长沙410208

出　　处：《中国中药杂志》2023年第6期1589-1596,共8页China Journal of Chinese Materia Medica

基　　金：湖南省自然科学基金项目(2020JJ5424);湖南省教育厅青年基金项目(21B30386);湖南中医药大学中西医结合“双一流”学科开放基金项目(2020ZXYJH08);湖南省大学生创新创业训练项目(S202210541036)。

摘　　要：探讨僵蚕提取液对全脑缺血再灌注(I/R)大鼠行为学改善的作用及可能的机制。全自动血凝仪检测僵蚕提取液干预人血浆凝血四项对提取液进行质控。4周龄雄性SD大鼠60只,分为假手术组,模型组,阳性药物(肝素)组,僵蚕提取液低、中、高剂量组。除假手术组外,其余5组大鼠行双侧颈总动脉反复夹闭-再通手术,构建I/R大鼠模型。僵蚕提取液低、中、高剂量组予僵蚕提取液(0.45、0.9、1.8 mg·g^(-1)·d^(-1))腹腔注射,阳性药物(肝素)组予肝素注射剂(900 IU·kg^(-1))腹腔注射,假手术组、模型组予等体积生理盐水腹腔注射,共计7 d;平衡木(BBT)检测大鼠行为学;苏木素-伊红(HE)染色检测脑组织形态学变化,免疫荧光法检测脑皮质(CC)区白细胞共同抗原(CD45)、白细胞分化抗原(CD11b)、精氨酸酶1(Arg-1);酶联免疫吸附试验法(ELISA)检测白细胞介素-1β(IL-1β)、白细胞介素-4(IL-4)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)蛋白表达水平;非靶向代谢组学检测僵蚕提取液、干预后大鼠血浆、干预后大鼠脑皮质代谢物水平。质控结果显示僵蚕提取液有效延长了人血浆活化部分促凝血酶原激酶时间(activated partial thromboplastin time,APTT)、凝血酶原时间(prothrombin time,PT)、凝血酶时间(thrombin time,TT),与往期提取所得僵蚕提取液抗凝作用相似。行为学检测结果显示与假手术组比较,模型组大鼠BBT得分增加;与模型组相比,僵蚕提取液干预后小鼠BBT评分降低。组织形态学结果显示与假手术组比较,模型组大鼠CC脑区可见大量神经细胞形态改变;与模型组相比,僵蚕提取液干预后大鼠CC脑区异常形态神经细胞减少。免疫荧光结果显示与假手术组比较,模型组大鼠CC脑区CD45、CD11b平均荧光强度增加;与模型组相比,僵蚕提取液低剂量组大鼠CC脑区CD11b平均荧光强度减少,Arg-1平均荧光强度增多;僵蚕提取液中、高剂量组大鼠CThis study aims to investigate the effect of Bombyx Batryticatus extract(BBE)on behaviors of rats with global cerebral ischemia reperfusion(I/R)and the underlying mechanism.The automatic coagulometer was used to detect the four indices of human plasma coagulation after BBE intervention for quality control of the extract.Sixty 4-week-old male SD rats were randomized into sham operation group(equivalent volume of normal saline,ip),model group(equivalent volume of normal saline,ip),positive drug group(900 IU·kg^(-1)heparin,ip),and low-,medium-,and high-dose BBE groups(0.45,0.9,and 1.8 mg·g^(-1)·d^(-1)BBE,ip).Except the sham operation group,rats were subjected to bilateral common carotid artery occlusion followed by reperfusion(BCCAO/R)to induce I/R.The administration lasted 7 days for all the groups.The behaviors of rats were examined by beam balance test(BBT).Morphological changes of brain tissue were observed based on hematoxylin-eosin(HE)staining.Immunofluorescence method was used to detect common leukocyte antigen(CD45),leukocyte differentiation antigen(CD11b),and arginase-1(Arg-1)in cerebral cortex(CC).The protein expression of interleukin-1β(IL-1β),interleukin-4(IL-4),interleukin-6(IL-6),and interleukin-10(IL-10)was detected by enzyme-linked immunosorbent assay(ELISA).The non-targeted metabonomics was employed to detect the levels of metabolites in plasma and CC of rats after BBE intervention.The results of quality control showed that the BBE prolonged the activated partial thromboplastin time(APTT),prothrombin time(PT),and thrombin time(TT)of human plasma,which was similar to the anticoagulation effect of BBE obtained previously.The results of behavioral test showed that the BBT score of the model group increased compared with that of the sham operation group.Compared with the model group,BBE reduced the BBT score.As for the histomorphological examination,compared with the sham operation group,the model group showed morphological changes of a lot of nerve cells in CC.The nerve cells with abnormal morphol

关 键 词：缺血性脑卒中 缺血-再灌注损伤 僵蚕提取液 小胶质细胞 

分 类 号：R285.5[医药卫生—中药学]