基于NF-κB信号通路探讨当归多糖对LPS介导的SD大鼠骨性关节炎作用研究  

Effect of Angelica Sinensis Polysaccharide on LPS Mediated Osteoarthritis in SD Rats Based on NF-κB Signaling Pathway

作　　者：武艳芝[1] 姚远[1] 杨五小[1] 吴宁 周敏 黄振飞 WU Yanzhi;YAO Yuan;YANG Wuxiao;WU Ning;ZHOU Min;HUANG Zhenfei(Shanxi Provincial People's Hospital,Taiyuan 030000,Shanxi,China;The First Affiliated Hospital of Nanjing Medical University,Nanjing 210000,Jiangsu,China)

机构地区：[1]山西省人民医院,山西太原030000 [2]南京医科大学第一附属医院,江苏南京210000

出　　处：《中华中医药学刊》2024年第11期136-140,I0026-I0030,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82202697)。

摘　　要：目的探究当归多糖(Angelica sinensis polysaccharides,ASP)对骨关节炎软骨细胞的软骨保护和治疗作用。方法通过体外脂多糖(lipopolysaccharide,LPS)诱导来构建关节炎软骨细胞,使用CCK-8法(Cell Counting Kit-8 assay,CCK-8)筛选出实验组当归多糖的剂量,通过活死细胞染色、DNA和糖胺聚糖(glycosaminoglycan,GAG)含量检测、实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测、酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)、免疫荧光染色、番红O固绿及免疫组化染色等来探究当归多糖对关节炎性软骨细胞的保护和治疗效果。结果CCK-8成功筛选出25μg/mL的当归多糖作为实验组。活死细胞染色中实验组细胞活力较模型组好,且实验组DNA含量和GAG分泌量均显著高于模型组(P<0.05)。PCR中实验组的炎症相关基因相对表达量较模型组显著下调(P<0.05),而软骨相关基因相对表达量显著上调(P<0.05)。Elisa显示实验组肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)的分泌水平低于模型组(P<0.05),体内免疫组化染色进一步验证了当归多糖实验组的P65(NF-κB P65)、iNOS及MMP13免疫组化中实验组的阳性表达显著低于模型组。结论当归多糖通过抑制NF-κB信号通路的表达对LPS诱导的骨性关节炎具有一定的治疗作用。Objective To explore the protective and therapeutic effects of Angelica sinensis polysaccharide on osteoarthritis chondrocytes.Methods Arthritic chondrocytes were constructed by lipopolysaccharide(LPS)induction in vitro,and the dose of Angelica sinensis polysaccharide in the experimental group was screened by CCK-8 method.The protective and therapeutic effects of Angelica sinensis polysaccharide on articular inflammatory chondrocytes were investigated by live dead cell staining,DNA and glycosaminoglycan(GAG)content detection,quantitative real-time polymerase chain reaction(qRT-PCR),enzyme-linked immunosorbent assay(ELISA),immunofluorescence staining,saffron solid green staining and immunohistochemical staining.Results Angelica sinensis polysaccharide(25μg/mL)was successfully selected as the experimental group by CCK-8.The cell viability of the experimental group was better than that of the model group,and the DNA content and GAG secretion of the experimental group were significantly higher than those of the model group(P<0.05).The relative expressions of inflammation-related genes in the experimental group were significantly down-regulated compared with those in the model group(P<0.05),while the relative expression of chondrogene-related genes was significantly up-regulated(P<0.05).ELISA showed that the secretion levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the experimental group were lower than those in the model group(P<0.05).Immunohistochemical staining further confirmed that the positive expressions of P65,iNOS and MMP13 in the experimental group were significantly lower than those in the model group.Conclusion Angelica sinensis polysaccharide has a certain therapeutic effect on osteoarthritis by inhibiting the expression of NF-κB signaling pathway.

关 键 词：骨关节炎 脂多糖 当归多糖 软骨细胞 NF-ΚB 

分 类 号：R285.5[医药卫生—中药学]