叶下珠提取物对大鼠脑缺血损伤的神经保护作用  

Neuroprotective effect of Phyllanthus urinaria extract on ischemic brain injury in rats

作　　者：温小龙 李骏鹏 刘鑫鑫 李宗慧 熊芸雲 虞道锐 WEN Xiao-long;LI Jun-peng;LIU Xin-xin;LI Zong-hui;XIONG Yun-yun;YU Dao-rui(Department of Pharmacy,the Second Affiliated Hospital of Hainan Medical University,Haikou 570311,China;School of Clinical Medicine,Hainan Medical University,Haikou 571199,China;School of Basic and Life Science,Hainan Medical University,Haikou 571199,China)

机构地区：[1]海南医学院第二附属医院药学部,海南海口570311 [2]海南医学院临床学院,海南海口571199 [3]海南医学院基础医学与生命科学学院,海南海口571199

出　　处：《海南医学院学报》2022年第7期486-492,共7页Journal of Hainan Medical University

基　　金：海南省自然科学基金项目(817132)。

摘　　要：目的:考察叶下珠提取物对缺血性脑卒中的神经保护作用及其初级机制。方法:选用成年SD大鼠为研究对象,应用改良线栓法建立右侧大脑中动脉梗塞大鼠模型(MCAO)。模型制备成功后24 h、48 h和72 h分别观察神经功能缺损评分,之后用TTC染色法检测脑梗死面积,脑组织中超氧化物歧化酶(SOD)、一氧化氮(NO)和内皮细胞型NOS(eNOS)含量;免疫荧光法检测脑组织半胱氨酸天冬氨酸蛋白酶3(caspase-3)阳性细胞表达;Western blot法测定脑组织PI3K、AKT蛋白的表达。72只大鼠随机平均分为4组,假手术组,模型组(MCAO)、叶下珠提取物低剂量组(PuL 5 g/kg)、高剂量组(PuL 10 g/kg)。连续给药7d后制作MCAO模型,并继续灌注药液1次/d直至取材。结果:假手术组未见神经功能缺损,造模的24、48、72 h显示,叶下珠提取物两个剂量组和MCAO组大鼠的神经功能受损较假手术组严重(P<0.01),但随着造模时间的延长,叶下珠提取物两个剂量组的神经功能评分较MCAO组大鼠低,72 h最为显著(P<0.01);叶下珠提取物两个剂量组大鼠的梗死面积的大小比MCAO组的低(P<0.01),两组无剂量依赖性;与假手术组相比,MCAO组的SOD水平降低、NO和eNOS水平升高(P<0.05),与MCAO组相比,两给药组SOD水平升高、NO和eNOS水平降低,差异具有统计学意义(P<0.05);两给药组的caspase-3阳性细胞表达虽高于假手术组但明显低于MCAO组(P_(低)<0.05,P_(高)<0.01);MCAO组大鼠脑组织PI3K、AKT蛋白表达显著低于假手术组(P<0.05),叶下珠提取物高、低剂量组大鼠脑组织PI3K、AKT蛋白表达显著高于MCAO组(P<0.05)。结论:叶下珠提取物能够改善神经功能损伤及大鼠脑梗死面积,并降低MCAO大鼠caspase-3阳性细胞表达,继而提高PI3K、AKT蛋白表达水平来保护缺血性脑损伤。Objective:To investigate the neuroprotective effect and its primary mechanism of Phyllanthus(P.)urinaria extract on ischemic stroke.Methods:Adult SD rats were selected,and the rat model of right middle cerebral artery infarction was established by the modified suture method(MCAO).The neurological deficit score was observed at 24 h,48 h and 72 h after the model was successfully prepared,then TTC staining method was used to detect the area of cerebral infarction,the content of superoxide dismutase(SOD),nitric oxide(NO)and endothelial NOS(eNOS)in brain tissue.Immunofluorescence method was used to detect the expression of caspase-3 positive cells in brain tissue.Western blot method was used to detect the expression of PI3K and AKT protein in brain tissue.A total of 72 rats were randomly divided into 4 groups:the sham operation group,the model group(MCAO),the extract of P.urinaria low dose group(PuL 5 g/kg),and the extract of P.urinaria high dose group(PuL10 g/kg).The MCAO model was continuously administered for 7 days,and continued to infuse the medicine once a day until the material was obtained.Results:No neurological deficits were observed in the sham operation group.The modelling at 24 h,48 h and 72 h showed that the neurological impairment of the two doses of extract of P.urinaria and the MCAO group was severer than that of the sham operation group(P<0.01).However,with the extension of modeling time,the neurological function scores of the two doses of extract of P.urinaria were lower than those of the MCAO group,and it was the most significant at 72 h(P<0.01).The infarct size of the rats in the two dose groups of extract of P.urinaria was lower than that of the MCAO group(P<0.01),and the two groups were not dose-dependent.The content of SOD in the MCAO group was reduced,and the content of NO and eNOS was increased compared with the sham operation group(P<0.05).Compared with the MCAO group,the two administration groups significantly increased the content of SOD and decreased the content of NO and eNOS(P<0.05).Alt

关 键 词：叶下珠提取物 MCAO 大鼠 神经功能 

分 类 号：R285.5[医药卫生—中药学]