沙棘多糖对脑胶质瘤大鼠的抑瘤作用及对HMGB1、MGMT及TLR4表达的影响  

The Effect of Hippophae Rhamnoides Polysaccharide on Glioma and Expression of HMGB1,MGMT,and TLR4 in Rats

作　　者：徐大伟[1] 刁玉领[1] 惠磊[1] 周文科[1] 汲乾坤 XU Dawei;DIAO Yuling;HUI Lei;ZHOU Wenke;JI Qiankun(The First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,Henan,China)

机构地区：[1]新乡医学院第一附属医院,河南新乡453100

出　　处：《中西医结合心脑血管病杂志》2023年第1期51-56,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：2019年河南省医学科技攻关计划联合共建项目(No.LHGJ20190445)。

摘　　要：目的:探讨沙棘多糖(HRP)对脑胶质瘤大鼠的抑瘤作用及对高迁移率族蛋白1(HMGB1)、O6-甲基鸟嘌呤-DNA-甲基转移酶(MGMT)和Toll样受体4(TLR4)表达的影响。方法:选取60只SD大鼠,脑部接种C6脑胶质瘤细胞,建立脑胶质瘤模型。造模成功后随机分为模型组(15只)、HRP低剂量组(14只)、HRP高剂量组(14只)、替莫唑胺组(14只)。接种5 d后,HRP低剂量组、HRP高剂量组分别灌胃沙棘多糖100 mg/kg、200 mg/kg,替莫唑胺组灌胃替莫唑胺20 mg/kg,模型组灌胃等量生理盐水,每日1次,连续10 d。磁共振成像(MRI)检查并记录肿瘤大小,测量肿瘤体积,计算抑瘤率;苏木精-伊红(HE)染色观察脑胶质瘤形态学变化情况;流式细胞仪检测肿瘤组织细胞凋亡和细胞周期;实时定量聚合酶链式反应(PCR)检测肿瘤组织中HMGB1、MGMT和TLR4 mRNA水平;蛋白免疫印迹法(Western Blot)检测肿瘤组织HMGB1、MGMT和TLR4蛋白表达。结果:与模型组比较,HRP低剂量组、HRP高剂量组、替莫唑胺组肿瘤体积减小,Sub-G1期、G0/G1期细胞增加,S期、G2/M期细胞减少,凋亡率升高,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量降低,差异均有统计学意义(P<0.05);与HRP低剂量组比较,HRP高剂量组、替莫唑胺组肿瘤体积减小,抑瘤率升高,Sub-G1期、G0/G1期细胞增加,S期、G2/M期细胞减少,凋亡率升高,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量降低,差异均有统计学意义(P<0.05);与HRP高剂量组比较,替莫唑胺组抑瘤率降低,Sub-G1期、G0/G1期细胞减少,S期、G2/M期细胞增加,凋亡率降低,HMGB1、MGMT和TLR4 mRNA及蛋白相对表达量升高,差异均有统计学意义(P<0.05)。HE染色显示,模型组脑胶质瘤内肿瘤细胞生长活跃,数量丰富,排列密集,核大深染,肿瘤细胞异型性明显,核浆比例高,核分裂现象多见,呈浸润性生长,无明显细胞萎缩坏死现象;HRP低剂量组、HRP高剂量组和替莫唑胺组细胞数量较模型组减少,细胞生长密度�Objective:To investigate the anti-tumor effect of Hippophae rhamnoides polysaccharide on rats glioma and its effect on the expression of high mobility group box-1(HMGB1),O6-methylguanine-DNA-methyltransferase(MGMT),and Toll-like receptor 4(TLR4).Methods:Sixty SD rats were inoculated with C6 glioma cells to establish glioma model.After successfully modeling,the rats were randomly divided into model group(15 rats),Hippophae rhamnoides polysaccharide low-dose group(14 rats),Hippophae rhamnoides polysaccharide high-dose group(14 rats),and temozolomide(TMZ)group(14 rats).After 5 days of inoculation,the rats in Hippophae rhamnoides polysaccharide low-dose group,Hippophae rhamnoides polysaccharide high-dose group,and TMZ group were given Hippophae rhamnoides polysaccharide 100 mg/kg,200 mg/kg,and TMZ 20 mg/kg respectively,once daily,for 10 days.The rats in model group were intragastrically administered with the same amount of saline.Magnetic resonance imaging(MRI)was used to examinate,tumor size,and tumor volume,and finally calculate tumor inhibition rate.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of glioma.Apoptosis and cell cycle of tumor tissue were annalyzed by flow cytometry.The mRNA levels of HMGB1,MGMT,and TLR4 in tumor tissues were detected by real-time quantitative polymerase chain reaction(PCR).The expression levels of HMGB1,MGMT,and TLR4 protein were assayed by Western Blot.Results:Compared with model group,the tumor volume in Hippophae rhamnoides polysaccharide low-dose group,high-dose group,and TMZ group was decreased,the cells in Sub-G1 phase and G0/G1 phase were increased,the cells in S phase and G2/M phase were decreased,the apoptosis rate was increased,and the mRNA and protein expression levels of HMGB1,MGMT,and TLR4 were decreased(P<0.05).Compared with Hippophae rhamnoides polysaccharide low-dose group,the tumor volume of high-dose group and TMZ group was decreased,the tumor inhibition rate was increased,the cells in Sub-G1 and G0/G1 phases were increased,the cells in S

关 键 词：脑胶质瘤 沙棘多糖 抑瘤作用 高迁移率族蛋白1 O6-甲基鸟嘌呤-DNA-甲基转移酶 Toll样受体4 实验研究 

分 类 号：R285.5[医药卫生—中药学]