首页|鸡血藤乙酸乙酯提取物对NK细胞免疫杀伤功能的影响

鸡血藤乙酸乙酯提取物对NK细胞免疫杀伤功能的影响

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探讨鸡血藤乙酸乙酯提取物(Spatholobi Caulis extract from ethyl acetate，SEA)在生理状态下对自然杀伤(natural kil-ler，NK)细胞的免疫调节作用及其潜在机制。利用C57BL/6J小鼠及NK-92细胞系为研究对象，设置空白组(NC)、给药组(SEA)的动物实验分组及0、25、50、100 μg·mL-1 SEA给药浓度的细胞实验分组。实验过程中及结束后获取小鼠体质量及免疫器官指数并进行差异分析;应用LDH法检测SEA对NK-92细胞的毒性及小鼠NK细胞对其靶细胞YAC-1的杀伤活性;应用CCK-8法检测SEA对NK-92细胞增殖的影响;应用流式细胞术检测小鼠外周血中NK细胞数目及NK细胞膜表面自然杀伤细胞2族成员A和D(natural killer group 2 member A、D，NKG2A、NKG2D)的表达;应用ELISA法检测小鼠血清中干扰素y(interferon gamma，IFN-γ)分泌的改变;应用半定量PCR法检测小鼠脾细胞中NKG2A、NKG2D、IFN-γ mRNA的表达;应用Western blot 法对磷脂酰肌醇 3-激酶(phosphoinositide 3-kinase，PI3K)/细胞外调节蛋白激酶 1(extracellular regulated protein ki-nases 1，ERK1)信号通路进行SEA药效机制的初步分析。实验结果证实，SEA对机体无明显毒副影响，可以显著增加小鼠体内NK细胞的数目及NK-92细胞对YAC-1的杀伤功能，可以抑制NKG2A受体表达并增强NKG2D受体表达，加强IFN-γ的分泌，加强PI3K/ERK信号通路蛋白表达。该研究结果提示，SEA具备增强NK细胞免疫识别杀伤功能，这一药效可能与增加NK细胞数目、重塑功能性受体表达稳态，调动PI3K/ERK信号通路并增强IFN-γ的分泌有关。
Effect of Spatholobi Caulis extract from ethyl acetate on immune killing function of NK cells
This study aims to investigate the regulatory effect of the Spatholobi Caulis extract from ethyl acetate(SEA)on natural killer(NK)cells under physiological conditions and elucidate the underlying mechanism.The C57BL/6 mice were randomized into NC and SEA groups,and NK-92 cells were respectively treated with 0,25,50,and 100 μg·mL-1 SEA.The body weight and immune or-gan index of the mice were compared between groups.The lactate dehydrogenase(LDH)assay was employed to examine the cytotoxici-ty of NK-92 cells treated with SEA and the killing activity of mouse NK cells against YAC-1 cells.The cell-counting kit-8(CCK-8)was used to examine the impact of SEA on the proliferation of NK-92 cells.Flow cytometry was employed to measure the number of NK cells in the peripheral blood as well as the expression levels of natural killer group 2 member A(NKG2A)and natural killer group 2 member D(NKG2D).The enzyme-linked immunosorbent assay(ELISA)was performed to determine the interferon(IFN)-γ secre-tion in the serum.Semi-quantitative PCR was conducted to determine the mRNA levels of NKG2A,NKG2D,and IFN-γ in spleen cells.Western blot was employed to investigate the involvement of phosphoinositide 3-kinase(PI3K)/extracellular regulated protein ki-nase 1(ERK1)signaling pathway.The results showed that SEA exhibited no adverse effects on the body,while significantly enhance the number of NK cells and augment the cytotoxicity of NK-92 cells against YAC-1 cells.Moreover,it suppressed the expression of NKG2A,enhanced the expression of NKG2D,promoted IFN-γ secretion,and upregulated the protein levels of PI3K and ERK.The findings suggest that SEA has the potential to enhance the immune recognition and effector function of NK cells by increasing the cell number,modulating the expression of functional receptors,and promoting IFN-γ secretion via the PI3K/ERK signaling pathway.

Spatholobi Caulisnatural killer(NK)cellsNKG2DNKG2AIFN-γ

杨丽娜、张阳、杜欣珂、刘丽、张径、杨庆、孙立东、朱晓新、李琦

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中国中医科学院中药研究所,北京 100700

鸡血藤 自然杀伤(NK)细胞 NKG2D NKG2A IFN-γ

国家自然科学基金北京市自然科学基金国家自然科学基金培育项目中国中医科学院科技创新工程项目中国中医科学院科技创新工程项目中国中医科学院优秀青年科技人才(创新类)培养专项

822741297232302ZXKT20016CI2021A05105CI2021B015ZZ13-YQ-044

2024

10.19540/j.cnki.cjcmm.20231123.402

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子：1.718
ISSN：1001-5302
年,卷(期)：2024.49(5)
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