摘要
目的 探究桃红四物汤多糖的结构特征以及其对缺血性脑卒中大鼠的抗炎作用,为后续药效学研究提供基础。方法 采用水提醇沉法提取桃红四物汤多糖,苯酚—硫酸法测定多糖的总糖含量,凝胶渗透色谱测定桃红四物汤多糖的分子量,分析其单糖组成及红外吸收特征。采用线栓法建立大脑中动脉阻塞模型,24 h后进行神经功能评分,将大鼠随机分为7组:假手术组,模型组,桃红四物汤多糖低、中、高剂量组,桃红四物汤组和尼莫地平组。给药7 d后采用Berderson评分进行神经功能评分;TTC染色法检测脑梗死率,ELISA法检测血清白细胞介素(interleukin, IL)-1β、IL-6、肿瘤坏死因子-α (tumor necrosis factor-α,TNF-α)水平,苏木精—伊红染色观察脑组织病理损伤情况;免疫组织化学法和Western blot检测脑组织中炎症因子的表达水平。结果 桃红四物汤多糖中总糖的含量为75.25%,分子量为2.47×104Da,是由甘露糖、半乳糖醛酸、鼠李糖、葡萄糖、半乳糖和岩藻糖组成的杂多糖。红外光谱结果显示,桃红四物汤多糖的结构中有多糖的特征吸收。动物实验结果表明,桃红四物汤多糖干预后,大鼠脑组织损伤得到改善,神经功能评分和脑梗死率有不同程度的下降,血清和脑组织中IL-6、IL-1β和TNF-α的水平也明显降低。结论 桃红四物汤多糖通过抑制炎症反应发挥对缺血性脑卒中大鼠的脑保护作用。
Objective To investigate the structural features of Taohong Siwu Decoction polysaccharide(THSWDP)and its anti-inflammatory effect in rats with ischemic stroke,and to provide a basis for subsequent pharmacodynamic studies.Methods THSWDP was extracted by the water-extraction and alcohol-precipitation method,the phenol-sulfuric acid method was used to measure the content of total sugar in THSWDP,and gel permeation chromatography was used to measure the molecular weight of THSWDP.Monosaccharide composition and infrared absorption features were also analyzed.The suture method was used to establish a model of middle cerebral artery occlusion,and neurological score was determined after 24 hours.The rats were randomly divided into sham-operation group,model group,Taohong Siwu Decoction group,nimodipine group,and low-,middle-,and high-dose THSWDP groups.After 7 days of administration,Berderson score was used to evaluate neurological function;TTC staining was used to measure cerebral infarction rate;ELISA was used to measure the serum levels of interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factor-α(TNF-α);HE staining was used to observe the pathological injury of brain tissue;immunohistochemistry and Western blotting were used to measure the expression of inflammatory factors in brain tissue.Results The content of total sugar was 75.25%in THSWDP,and THSWDP had a molecular weight of 2.47×104 Da and was a heteropolysaccharide composed of mannose,galacturonic acid,rhamnose,glucose,galactose,and fucose.Infrared results showed the characteristic absorption of polysaccharides in the structure of THSWDP.The results of animal experiment showed that after THSWDP intervention,there was an improvement in brain tissue injury,varying degrees of reductions in neurological score and cerebral infarction rate,and significant reductions in the levels of IL-6,IL-1β,and TNF-αin serum and brain tissue.Conclusion THSWDP exerts a cerebral protection effect on a rat model of ischemic stroke by inhibiting inflammatory response.
作者
张亚南
赵梦蝶
唐林峰
侯欣宇
张亚中
韩岚
彭代银
ZHANG Yanan;ZHAO Mengdie;TANG Linfeng;HOU Xinyu;ZHANG Yazhong;HAN Lan;PENG Daiyin(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Anhui Institute for Food and Drug Control,Anhui Hefei 230051,China;Key Laboratory of Chinese Medicinal Formula of Anhui Province,Anhui Hefei 230012,China;MOE-Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials,Anhui Hefei 230012,China)
出处
《安徽中医药大学学报》
CAS
2023年第6期79-85,共7页
Journal of Anhui University of Chinese Medicine
基金
国家自然科学基金项目(8207142977,81903953)
安徽省高校自然科学研究项目(KJ2020A0420)。
关键词
桃红四物汤
多糖结构
单糖组成
缺血性脑卒中
Taohong Siwu Decoction
Polysaccharide structure
Monosaccharide composition
Ischemic stroke
作者简介
张亚南(1991-),女,硕士研究生;通信作者:彭代银(1963-),男,教授,博士研究生导师,pengdaiyin@163.com。
