人参果胶醇沉组分对小鼠骨髓源性树突状细胞表型及功能的影响

doi:10.13481/j.1671-587X.20260209

摘要/Abstract

摘要：

目的探讨人参果胶醇沉组分（WGPA）对小鼠骨髓源性树突状细胞（BMDCs）表型和功能的影响。方法采用水提、乙醇沉淀和离子交换层析法提取3种WGPA，分别为30%醇沉组分（WGPA₃₀）、50%醇沉组分（WGPA₅₀）和70%醇沉组分（WGPA₇₀）。原代培养小鼠BMDCs，并分为未成熟BMDCs和成熟BMDCs。采用细胞计数试剂盒8（CCK-8）法检测不同浓度WGPA作用下成熟BMDCs增殖活性，确定有效WGPA及其适宜浓度。对有效醇沉组分进行相对分子质量、红外光谱和单糖组成分析。将未成熟BMDCs和成熟BMDCs均随机分为对照组、脂多糖（LPS）组（加入LPS干预）和WGPA组（加入有效WGPA干预）。采用流式细胞术检测各组成熟BMDCs表面CD80、CD86和主要组织相容性抗原-Ⅱ（MHC-Ⅱ）表达水平，荧光微球吞噬实验检测各组未成熟BMDCs吞噬效率，酶联免疫吸附试验（ELISA）方法检测各组成熟BMDCs培养上清中IL-6、IL-12和TNF-α水平。结果 CCK-8法，与对照组比较，800和1 600 mg·L^-1 WGPA₃₀组细胞增殖活性均明显升高（P<0.01），WGPA₅₀和WGPA₇₀作用下各组细胞增殖活性无明显变化，差异无统计学意义（P>0.05）；选用800 mg·L^-1 WGPA₃₀进行后续实验。结构分析，WGPA₃₀相对分子质量约为24 230，是由α-构型糖苷键连接而成的吡喃糖，主要单糖组成包括葡萄糖醛酸、木糖、葡萄糖和半乳糖。流式细胞术，与对照组比较，LPS组和WGPA₃₀组成熟BMDCs表面MHC-Ⅱ、CD86及CD803表达水平均明显升高（P<0.01）；与LPS组比较，WGPA₃₀组成熟BMDCs表面MHC-Ⅱ、CD86和CD803表达水平均明显降低（P<0.05）。与对照组比较， LPS组和WGPA₃₀组未成熟BMDCs吞噬活性均明显降低（P<0.05）。ELISA法，与对照组比较，LPS组和WGPA₃₀组成熟BMDCs上清中IL-6、IL-12及TNF-α水平均明显升高（P<0.01）；与LPS组比较，WGPA₃₀组成熟BMDCs上清中IL-12和TNF-α水平明显降低（P<0.01），IL-6水平比较差异无统计学意义（P>0.05）。结论 WGPA₃₀是由α-构型糖苷键连接而成的吡喃糖，包含葡萄糖、半乳糖和葡萄糖醛酸等主要活性成分，相对分子质量约为24 230。WGPA₃₀可通过促进小鼠BMDCs增殖和细胞因子IL-6、 IL-12及TNF-α释放，提高BMDCs表面分子MHC-Ⅱ、CD86及CD80表达以及降低BMDCs吞噬效率等途径，促进BMDCs成熟，调节BMDCs活性。

关键词: 人参, 人参果胶, 醇沉组分, 骨髓源性树突状细胞, 脂多糖

Abstract:

Objective To investigate the effect of whole ginseng pectin alcohol-precipitated polysaccharide fraction （WGPA） on the phenotype and function of mouse bone marrow-derived dendritic cells （BMDCs）. Methods Three WGPA fractions were extracted using water extraction， ethanol precipitation， and ion-exchange chromatography： the 30% ethanol-precipitated fraction （WGPA₃₀）， the 50% ethanol-precipitated fraction （WGPA₅₀）， and the 70% ethanol-precipitated fraction （WGPA₇₀）. The primary mouse BMDCs were cultured and categorized into immature BMDCs and mature BMDCs. Cell counting kit-8 （CCK-8） method was used to detect the proliferation activities of mature BMDCs in various groups treated with different concentrations of WGPA to determine the effective WGPA fraction and its appropriate concentration. The effective alcohol-precipitated fraction was analyzed for relative molecular mass， infrared spectrum， and monosaccharide composition. Both immature and mature BMDCs were randomly divided into control group， lipopolysaccharide （LPS） group （treated with LPS）， and WGPA group （treated with effective WGPA）. Flow cytometry was used to detect the expression levels of CD80， CD86， and major histocompatibility complex class Ⅱ （MHC-Ⅱ） on the surface of mature BMDCs in various groups. Fluorescent microsphere phagocytosis assay was used to detect the phagocytic efficiency of immature BMDCs in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-6 （IL-6）， interleukin-12 （IL-12）， and tumor necrosis factor-α （TNF-α） in the culture supernatants of mature BMDCs in various groups. Results The results of CCK-8 method showed that compared with control group， the proliferation activities of mature BMDCs in 800 and 1 600 mg·L^-1 WGPA₃₀ groups were significantly increased （P<0.01），while the proliferation activities of mature BMDCs after treated with WGPA₅₀ or WGPA₇₀ showed no significant change （P>0.05）. Therefore， 800 mg·L^-1 WGPA₃₀ was selected for subsequent experiments.The structural analysis results revealed that WGPA₃₀ had a relative molecular mass of approximately 24 230， and it was a pyranose connected by α-configuration glycosidic bond whose main monosaccharide composition included glucuronic acid， xylose， glucose， and galactose. The flow cytometry results showed that compared with control group， the expression levels of MHC-Ⅱ， CD86， and CD80 on the surface of mature BMDCs in LPS group and WGPA₃₀ group were significantly increased （P<0.01）. Compared with LPS group， the expression levels of MHC-Ⅱ， CD86， and CD80 on the surface of mature BMDCs in WGPA₃₀ group were significantly decreased（P<0.05）. Compared with control group， the phagocytic activities of immature BMDCs in LPS group and WGPA₃₀ group were significantly decreased（P<0.05）.The ELISA results showed that compared with control group， the levels of IL-6， IL-12， and TNF-α in the supernatants of mature BMDCs in LPS group and WGPA₃₀ group were significantly increased （P<0.01）. Compared with LPS group， the levels of IL-12 and TNF-α in the supernatant of mature BMDCs in WGPA₃₀ group were significantly decreased （P<0.01）， while the level of IL-6 showed no statistically significant difference （P>0.05）. Conclusion WGPA₃₀ is a pyranose with α-configuration glycosidic bonds which contains primary active components such as glucose， galactose， and glucuronic acid， and has a relative molecular mass of approximately 24 230. WGPA₃₀ promotes the maturation and modulates the activity of BMDCs by enhancing their proliferation， stimulating the release of cytokines IL-6， IL-12， and TNF-α， upregulating the expression of surface molecules MHC-Ⅱ， CD86， and CD80， and decreasing their phagocytic efficiency.

Key words: Ginseng, Ginseng pectin, Alcohol-precipitated fraction, Bone marrow-derived dendritic cell, Lipopolysaccharide

中图分类号:

R392.12

杨如意,王煜灵,杨巍. 人参果胶醇沉组分对小鼠骨髓源性树突状细胞表型及功能的影响[J]. 吉林大学学报(医学版), 2026, 52(2): 375-383.

Ruyi YANG,Yuling WANG,Wei YANG. Effects of ginseng pectin alcohol-precipitated fraction on phenotype and function of mouse bone marrow-derived dendritic cells[J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 375-383.

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Group	Proliferation activity
Group	Concentration (mg·L^-1) 100	200	400	800	1 600
Control	0.45±0.02	0.44±0.01	0.44±0.01	0.45±0.01	0.45±0.01
WGPA₃₀	0.45±0.01	0.45±0.01	0.46±0.01	0.57±0.03^*	0.62±0.02^*
WGPA₅₀	0.44±0.02	0.45±0.02	0.45±0.02	0.43±0.01	0.43±0.02
WGPA₇₀	0.45±0.01	0.44±0.01	0.45±0.01	0.47±0.03	0.48±0.02
F	0.770	0.440	1.270	35.110	163.800
P	0.520	0.730	0.320	<0.001	<0.001

Group	MHC-Ⅱ	CD86	CD80
Control	17.67±0.87	11.73±0.60	2.63±0.06
LPS	28.53±0.55^*	20.77±1.30^*	5.33±0.15^*
WGPA₃₀	24.27±0.45^*△	17.90±0.87^*△	3.77±0.15^*△
F P	153.839 <0.001	61.285 <0.001	219.318 <0.001

Group	IL-12	IL-6	TNF-α
Control	18.3±3.1	12.3±3.1	123.3±5.8
LPS	594.3±20.0^*	2 861.9±301.1^*	1 940.0±20.0^*
WGPA₃₀	448.7±16.8^*△	2 811.9±86.1^*	1 670.0±26.5^*△
F P	3 767.333 <0.01	917.265 <0.01	5 635.235 <0.01