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基于改善肠道微生物群和激活RAW264.7细胞对玉竹活性成分的免疫调节作用

Immunological regulation of the active fraction from Polygonatum sibiricum F. Delaroche based on improvement of intestinal microflora and activation of RAW264.7 cells.

作者信息

He Yanfei, Huang Lin, Jiang Ping, Xu Guangpei, Sun Taotao

机构信息

College of Biotechnology and Pharmaceutical Engineering of West Anhui University, Lu'an, 237012, PR China.

College of Biotechnology and Pharmaceutical Engineering of West Anhui University, Lu'an, 237012, PR China.

出版信息

J Ethnopharmacol. 2022 Jul 15;293:115240. doi: 10.1016/j.jep.2022.115240. Epub 2022 Mar 31.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Polygonatum sibiricum, known as "Huangjing" in Chinese traditional medicine, of which functions include invigorating Qi and nourishing Yin, tonifying spleen and kidney, which are considered to replenish energy, and strengthen immunity. However, both the active components and mechanism of the immune-enhancing effect of Polygonatum sibiricum have not been clarified.

AIM OF THE STUDY

To evaluate the immunoregulation effects of PSE30 (Polygonatum sibiricum ethanol 30) and PSE75 (Polygonatum sibiricum ethanol 75). The gut microbial and activation of RAW264.7 cells were also evaluated for exploring the mechanism of PSE75.

MATERIALS AND METHODS

Female ICR mice were randomly divided into different groups, which were pretreated with 0.9% saline, Yupingfeng granules, different dosage of PSE30 or PSE75. And the immunosuppressed mice model was constructed using cyclophosphamide. And the total duration of the experiment was 15 d. After that, the serum Immunoglobulins G (IgG) and Immunoglobulins M (IgM) antibody, regular blood testing, assessment of natural killer cell activity, and histological observation of spleen in immunosuppressed mice were measured to evaluate the immunoregulation effects of PSE30 and PSE75. Besides, effects of PSE75 on gut microbial were evaluated using 16s rRNA sequence. And the mRNA expression and cytokine secretion of RAW264.7 cell were evaluated to analyze the immunoregulation mechanism of PSE75.

RESULTS

The content of serum IgG, IgM was significantly elevated by PSE75 (P<0.05, P<0.001). The NK cells killing activity in splenocytes against K562 cells induced by PSE30, and PSE75 was pronounced higher than that of the model group (P < 0.05). Both mRNA expression of Th1 molecular markers including interleukin (IL)-2, interferon (IFN)-γ, and signal transducers and activators of transcription (STAT) 4, and Th2 molecular markers including IL-4 in splenocytes were markedly enhanced by PSE30, and PSE75 (P < 0.05, P < 0.01, or P < 0.001). Besides, the result of 16s rRNA sequence indicated that PSE75 could recover the gut microbial community disturbed by cyclophosphamide. PSE75 could markedly promote the secretion of IL-6, IL-10, and IL-12 p40 from RAW264.7 cell (P<0.01, or P<0.001).

CONCLUSIONS

PSE75 was proved to be a more promising immunomodulation agent, of which may enhance the immunity of immunosuppressed mice by improving gut microbial and activating macrophages. And PSE75 could be developed as a good immune booster in the future.

摘要

民族药理学相关性

黄精在传统中药中被称为“黄精”,其功能包括补气养阴、健脾补肾,被认为能补充能量并增强免疫力。然而,黄精的活性成分及其免疫增强作用机制尚未阐明。

研究目的

评估30%黄精乙醇提取物(PSE30)和75%黄精乙醇提取物(PSE75)的免疫调节作用。还评估了肠道微生物群以及RAW264.7细胞的激活情况,以探索PSE75的作用机制。

材料与方法

将雌性ICR小鼠随机分为不同组,分别用0.9%生理盐水、玉屏风颗粒、不同剂量的PSE30或PSE75进行预处理。使用环磷酰胺构建免疫抑制小鼠模型。实验总时长为15天。之后,测量免疫抑制小鼠血清中的免疫球蛋白G(IgG)和免疫球蛋白M(IgM)抗体、血常规检测、自然杀伤细胞活性评估以及脾脏组织学观察,以评估PSE30和PSE75的免疫调节作用。此外,使用16s rRNA序列评估PSE75对肠道微生物群的影响。评估RAW264.7细胞的mRNA表达和细胞因子分泌,以分析PSE75的免疫调节机制。

结果

PSE75显著提高了血清IgG、IgM的含量(P<0.05,P<0.001)。PSE30和PSE75诱导的脾细胞中NK细胞对K562细胞的杀伤活性明显高于模型组(P < 0.05)。PSE30和PSE75均显著增强了脾细胞中Th1分子标志物包括白细胞介素(IL)-2、干扰素(IFN)-γ以及信号转导和转录激活因子(STAT)4,和Th2分子标志物包括IL-4的mRNA表达(P < 0.05,P < 0.01或P < 0.001)。此外,16s rRNA序列结果表明,PSE75可恢复被环磷酰胺扰乱的肠道微生物群落。PSE75可显著促进RAW264.7细胞分泌IL-6、IL-10和IL-12 p40(P<0.01或P<0.001)。

结论

PSE75被证明是一种更有前景的免疫调节剂,它可能通过改善肠道微生物群和激活巨噬细胞来增强免疫抑制小鼠的免疫力。未来PSE75有望被开发成为一种良好的免疫增强剂。

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