浙江中医药大学学报2025，Vol.49Issue(9)：1094-1104,11.DOI:10.16466/j.issn1005-5509.2025.09.002
六味地黄汤通过TLR4/NF-κB途径治疗小鼠实验性自身免疫性脑脊髓炎
Treatment of Liuwei Dihuang Decoction in Experimental Autoimmune Encephalomyelitis in Mice Through TLR4/NF-κB Path-way
摘要
Abstract
[Objective]To elucidate the role and molecular mechanisms of Liuwei Dihuang Decoction(LWDHD)in the treatment of experimental autoimmune encephalomyelitis(EAE).[Methods]Sixty male C57BL/6 mice were randomly divided into normal group,model group,positive control group[prednisone acetate(PA)],LWDHD low-dose group(7.7 g·kg-1),medium-dose group(15.4 g·kg-1)and high-dose group(30.8 g·kg-1).Mice EAE model was established by using myelin oligodendrocyte glycoprotein 35-55(MOG 35-55)combined with pertussis toxin(PTX)and tuberculin.The mice in LWDHD groups were given LWDHD by gavage for 20 consecutive days,and the mice in positive control group were given PA of 6 mg·kg-1 by gavage.Neurological damage was evaluated by neurological functional impairment scores,hematoxylin-eosin(HE)staining was used to detect pathological changes in brain tissue,interleukin-1β(IL-1β)and interleukin-6(IL-6)levels were detected by enzyme-linked immunosorbent assay(ELISA),and Western blot was utilized to examine the expression of proteins in the Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway.T lymphocytes were extracted from the spleen of mice and treated with LWDHD containing serum.Flow cytometry was used to evaluate the proportion of CD4+and CD8+T lymphocytes and Western blot was used to detect the effect of LWDHD on the expression of proteins in the TLR4/NF-κB pathway.[Results]Different concentrations of LWDHD significantly improved the neurological function scores in EAE mice(P<0.01),reduced the abnormal proliferation of microglia in the cerebral cortex,and decreased neuronal cell death in the hippocampus.ELISA showed that LWDHD significantly inhibited the secretion of IL-1β and IL-6(P<0.01).Western blot analysis indicated that LWDHD markedly suppressed the expression and activation of proteins in the TLR4/NF-κB pathway(P<0.01).In cellular experiments,LWDHD containing serum significantly reduced the proportion of CD4+and CD8+T lymphocytes,and inhibited the expression and activation of proteins in the TLR4/NF-κB pathway(P<0.01).[Conclusion]LWDHD significantly inhibits brain inflammatory responses in EAE mice and reduces abnormal proliferation of microglia,which may be associated with the inhibition of protein activation in brain tissue and T lymphocytes via the TLR4/NF-κB pathway.关键词
实验性自身免疫性脑脊髓炎/多发性硬化/六味地黄汤/T淋巴细胞亚群/TLR4/NF-κB信号通路/神经功能损伤评分/IL-1β/IL-6Key words
experimental autoimmune encephalomyelitis/multiple sclerosis/Liuwei Dihuang Decoction/T lymphocyte subsets/TLR4/NF-κB signaling pathway/neurological functional impairment scores/IL-1β/IL-6分类
医药卫生引用本文复制引用
林翔,刘郑霞,屠友权,周云,黄明海,方春艳..六味地黄汤通过TLR4/NF-κB途径治疗小鼠实验性自身免疫性脑脊髓炎[J].浙江中医药大学学报,2025,49(9):1094-1104,11.基金项目
浙江省中医药科技计划项目(2022ZA163) Zhejiang Traditional Chinese Medicine Science and Technology Project(2022ZA163) （2022ZA163）
