| 高鹏,周凤蕊,刘俊华,等.刺五加多糖对自身免疫性肝炎小鼠肝损伤的改善作用及机制研究[J].浙江中医药大学学报,2021,45(7):705-712. |
| 刺五加多糖对自身免疫性肝炎小鼠肝损伤的改善作用及机制研究 |
| Effect of Acanthopanax Senticosus Polysaccharide on Liver Injury in Mice with Autoimmune Hepatitis and Its Mechanism |
| DOI：10.16466/j.issn1005-5509.2021.07.004 |
| 中文关键词: 刺五加多糖 自身免疫性肝炎 肝损伤 炎症因子 病理变化 Toll样受体4 髓样分化因子88 核因子-κB |
| 英文关键词: Acanthopanax senticosus polysaccharide autoimmune hepatitis liver injury inflammatory factors pathological changes Toll like receptor 4 myeloid differentiation factor 88 nuclear factor-κB |
| 基金项目:河南省医学科技攻关计划项目(2018020849) |
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| 中文摘要: |
| [目的]研究刺五加多糖(Acanthopanax senticosus polysaccharide，ASPS)对自身免疫性肝炎(autoimmune hepatitis，AIH)小鼠肝损伤的改善作用，并探讨相关分子机制。[方法]从120只C57BL/6小鼠中选取60只用于免疫剂的制备，其余50只腹腔注射免疫剂，建立AIH模型。将42只建模成功的小鼠随机分为AIH组(10只)、ASPS低剂量组(10只)、ASPS高剂量组(11只)和泼尼松龙组(11只)，其余10只小鼠不进行处理，设为健康组。ASPS低、高剂量组分别采用50、100mg·kg-1的ASPS灌胃，泼尼松龙组采用9mg·kg-1泼尼松龙灌胃，健康组与AIH组均以等量0.9%氯化钠溶液灌胃。检测各组小鼠血清天门冬氨酸氨基转移酶(aspartate aminotransferase，AST)、丙氨酸氨基转移酶(alanine aminotransferase，ALT)、白介素-22(interleukin-22，IL-22)、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白介素-6(interleukin-6，IL-6)水平；以多参数流式细胞术检测外周血辅助性T细胞22(T-helper cell 22，Th22)的比例；以苏木精-伊红(hematoxylin-eosin，HE)染色观察肝组织病理变化；Western blot检测肝组织Toll样受体4(Toll like receptor 4，TLR4)、髓样分化因子88(myeloid differentiation factor 88，MyD88)、核因子-κB(nuclear factor-κB，NF-κB)、磷酸化核因子-κB(phosphorylation-nuclear factor-κB，p-NF-κB)蛋白相对表达量。[结果]与健康组比较，AIH组血清AST、ALT、IL-22、TNF-α、IL-6水平，外周血CD4＋T淋巴细胞中Th22细胞比例均升高(P＜0.05)；与AIH组比较，ASPS低、高剂量组和泼尼松龙组血清AST、ALT、IL-22、TNF-α、IL-6水平，外周血CD4＋T淋巴细胞中Th22细胞比例均降低(P＜0.05)；与ASPS低剂量组比较，ASPS高剂量组和泼尼松龙组血清AST、ALT、IL-22、TNF-α、IL-6水平，外周血CD4＋T淋巴细胞中Th22细胞比例均降低(P＜0.05)。HE染色结果显示，AIH组肝细胞明显水肿，肝小叶结构被破坏，可观察到大面积小灶样坏死，且存在大量炎性细胞浸润；ASPS低、高剂量组和泼尼松龙组干预后，肝细胞小灶样坏死面积缩小，肝小叶结构较为完整，炎性细胞浸润减少。与健康组比较，AIH组肝组织TLR4、MyD88蛋白相对表达量，p-NF-κB p65/NF-κB p65均升高(P＜0.05)；与AIH组比较，ASPS低、高剂量组和泼尼松龙组肝组织TLR4、MyD88蛋白相对表达量，p-NF-κB p65/NF-κB p65均降低(P＜0.05)；与ASPS低剂量组比较，ASPS高剂量组、泼尼松龙组肝组织TLR4、MyD88蛋白相对表达量，p-NF-κB p65/NF-κB p65均降低(P＜0.05)。[结论]ASPS可保护AIH小鼠肝功能，调节Th22细胞比例，减轻炎症反应，缓解肝组织病理变化，且呈剂量依赖性，其分子机制可能与抑制TLR4/MyD88信号通路有关。 |
| 英文摘要: |
| [Objective] To study the effect of Acanthopanax senticosus polysaccharide(ASPS) on liver injury in autoimmune hepatitis(AIH) mice， and explore related molecular mechanisms. [Methods] A total of 120 C57BL/6 mice were taken， 60 of which were selected for preparation of immunization agent， and 50 were intraperitoneally injected with immunization agent to establish an AIH model. The 42 successfully modeled mice were randomly divided into AIH group(10 mice)， ASPS low dose group(10 mice)， ASPS high dose group(11 mice)， and prednisolone group(11 mice). The remaining 10 mice were left untreated and set as healthy group. ASPS low and high dose groups were given 50， 100mg·kg-1 ASPS by gavage， prednisolone group was given 9mg·kg-1 prednisolone， healthy group and AIH group were given the same amount of 0.9% sodium chloride solution. The levels of aspartate aminotransferase(AST)， alanine aminotransferase(ALT)， interleukin-22(IL-22)， tumor necrosis factor-α(TNF-α)， interleukin-6(IL-6) levels in serum were detected. The T-helper cell 22(Th22) proportion in peripheral blood was detected by multiparameter flow cytometry. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of liver tissue. The relative expression of Toll like receptor 4(TLR4)， myeloid differentiation factor 88(MyD88)， and nuclear factor-κB(NF-κB)， phosphorylation-nuclear factor-κB(p-NF-κB) protein in liver tissue were detected by Western blot. [Results] Compared with healthy group， the serum AST， ALT， IL-22， TNF-α， IL-6 levels， the proportion of Th22 cells in CD4+ T lymphocytes in peripheral blood in AIH group were all higher(P＜0.05).Compared with AIH group， the serum AST， ALT， IL-22， TNF-α， IL-6 levels， the proportion of Th22 cells in CD4+ T lymphocytes in peripheral blood in ASPS low， high dose group and prednisolone group were all lower(P＜0.05). Compared with ASPS low dose group， the serum AST， ALT， IL-22， TNF-α， IL-6 levels， the proportion of Th22 cells in CD4+ T lymphocytes in peripheral blood in ASPS high dose group and prednisolone group were all lower(P＜0.05). The results of HE staining showed that the liver cells in AIH group had obvious edema， the structure of liver lobules was destroyed， large areas of small focal necrosis could be observed， and there was a large amount of inflammatory cell infiltration. After intervention， the area of small focal necrosis in ASPS low， high dose group and prednisolone group were reduced， the structure of liver lobules was relatively complete， and the infiltration of inflammatory cells was reduced. Compared with healthy group， the relative expressions of TLR4， MyD88 protein， p-NF-κB p65/NF-κB p65 in liver tissue in AIH group were all higher(P＜0.05). Compared with AIH group， the relative expressions of TLR4， MyD88 protein， p-NF-κB p65/NF-κB p65 in liver tissue in ASPS low， high dose group and prednisolone group were all lower(P＜0.05). Compared with ASPS low dose group， the relative expressions of TLR4， MyD88 protein， p-NF-κB p65/NF-κB p65 in liver tissue in ASPS high dose group and prednisolone group were all lower(P＜0.05). [Conclusion] ASPS can protect the liver function of AIH mice， regulate the proportion of Th22 cells， reduce inflammation and pathological changes in liver tissue， and the effect is dose-dependent. Its molecular mechanism may be related to the inhibition of TLR4/MyD88 signaling pathway. |
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