红景天苷对卡介苗感染的巨噬细胞免疫功能的影响

doi:10.3969/j.issn.1674-8115.2025.04.004

上海交通大学学报（医学版） ›› 2025, Vol. 45 ›› Issue (4): 426-433.doi: 10.3969/j.issn.1674-8115.2025.04.004

红景天苷对卡介苗感染的巨噬细胞免疫功能的影响

倪书奕¹^,², 姜钊¹^,², 汪中涛³, 何树梅¹^,²()

^1.西藏民族大学医学院高原生物医学大数据挖掘与生物信息学分析实验室，咸阳 712082
^2.西藏民族大学医学院西藏高原相关疾病分子遗传机制与干预研究省级重点实验室，咸阳 712082
^3.西藏军区总医院感染病科，拉萨 850007

收稿日期:2024-09-18 接受日期:2024-12-26 出版日期:2025-04-28 发布日期:2025-04-28
通讯作者: 何树梅 E-mail:smhe@xzmu.edu.cn
作者简介:倪书奕（1999—），女，硕士生；电子信箱：610598532@qq.com。
基金资助:
2023年西藏民族大学科研项目(23MDY01);西藏自治区自然科学基金[XZ2019ZR G-37(z)];2024年西藏民族大学研究生科研创新与实践项目(Y2024010)

Effect of salidroside on the immune function of BCG-infected macrophages

NI Shuyi¹^,², JIANG Zhao¹^,², WANG Zhongtao³, HE Shumei¹^,²()

^1.Plateau Biomedical Big Data Mining and Bioinformatics Analysis Laboratory, School of Medicine, Xizang Minzu University, Shaanxi Xianyang 712082, China
^2.Key Laboratory for Molecular Genetic Mechanisms and Intervention Research on High Altitude Disease of Xizang Autonomous Region, School of Medicine, Xizang Minzu University, Shaanxi Xianyang 712082, China
^3.Department of Infectious Diseases, General Hospital of Tibet Military Command, Xizang Lhasa 850007, China

Received:2024-09-18 Accepted:2024-12-26 Online:2025-04-28 Published:2025-04-28
Contact: HE Shumei E-mail:smhe@xzmu.edu.cn
Supported by:
Xizang Minzu University Research Project 2023(23MDY01);Natural Science Foundation of Xizang Autonomous Region [XZ2019ZR G-37(z)];Xizang Minzu University Postgraduate Research Innovation and Practice Programme 2024(Y2024010)

摘要/Abstract

摘要：

目的·研究红景天苷（salidroside，SAL）对卡介苗（Bacillus Calmette‑Guérin，BCG）感染巨噬细胞免疫功能的影响。方法·使用BCG感染RAW264.7小鼠巨噬细胞建立感染模型，实验分组为空白对照组（BCG组）、红景天苷组（SAL+BCG组）、异烟肼（isoniazid，INH）组（INH+BCG组）以及红景天苷+异烟肼组（SAL+INH+BCG组）。用MTT比色法检测SAL和INH对RAW264.7细胞增殖的影响，筛选出SAL和INH实验浓度。建立感染BCG的巨噬细胞模型后，用SAL进行预处理，通过细菌铺板计数及流式细胞术检测胞内细菌存活率，酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测各细胞培养组细胞上清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、干扰素-γ（interferon-γ，IFN-γ）、白介素-6（interleukin-6，IL-6）及IL-10细胞因子的水平；流式细胞术检测各组细胞的凋亡水平。结果·筛选出SAL用药浓度为800 μmol/L，INH用药浓度为10 μmol/L。与BCG组相比，SAL预处理36h作用效果最佳，给药组巨噬细胞内BCG生长均受抑制；SAL与INH联用对BCG胞内存活的抑制作用效果明显，差异具有统计学意义。与BCG组相比，SAL使BCG感染的巨噬细胞TNF-α、IFN-γ、IL-6和IL-10的分泌水平降低但无显著性差异，然而SAL与INH联合应用使细胞上清中TNF-α、IL-6和IL-10的含量显著降低（均P<0.05）。与BCG组比较，在感染早期SAL和INH能够显著降低BCG感染的RAW264.7细胞的凋亡水平（P=0.008，P=0.032），在感染后期SAL与INH联合应用能够显著增强BCG感染的RAW264.7细胞的凋亡水平（P=0.001）。结论·红景天苷在感染BCG的巨噬细胞中可以通过改善巨噬细胞免疫功能，抑制胞内细菌存活，减轻BCG感染导致的炎症反应，从而发挥其宿主抗分枝杆菌作用。

关键词: 红景天苷, 卡介苗（BCG）, 巨噬细胞, 免疫功能, 抗结核作用

Abstract:

Objective ·To Investigate the effects of salidroside (SAL) on the immune response of macrophages infected with BCG. Methods ·RAW264.7 mouse macrophages were infected with BCG, and the experiments were divided into blank control (BCG), salidroside (SAL+BCG), isoniazid (INH+BCG), and salidroside+isoniazid (SAL+INH+BCG) groups. The effects of SAL and INH on the proliferation of RAW264.7 cells were detected by MTT colorimetric assay, and the experimental concentrations of SAL and INH were screened. After the establishment of the BCG-infected macrophage model, intracellular bacterial survival was detected by bacterial plate count and flow cytometry following salidroside pretreatment. The levels of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-6 (IL-6) , and interleukin-10 (IL-10) in the culture supernatants were measured by enzyme-linked immunosorbent assay, and apoptosis levels were assessed by flow cytometry. Results ·SAL at a concentration of 800 μmol/L and INH at 10 μmol/L were selected for subsequent experiments. Compared with the BCG group, SAL pretreatment for 36 h had the best effect, and the growth of BCG in macrophages in the administered group was inhibited. The inhibitory effect of the combined application of SAL and INH was obvious and statistically significant. Compared with the BCG group, SAL alone led to decreased secretion of TNF-α, IFN-γ, IL-6, and IL-10, although the differences were not statistically significant. However, the combination of SAL and INH caused a significant decrease in the levels of TNF-α, IL-6 and IL-10 in the supernatants of the cells (all P<0.05). Compared with the BCG group, in the early stage of the infection, SAL and INH were able to significantly reduce the apoptotic levels of BCG-infected RAW264.7 cells (P=0.008, P=0.032), and the combined application of SAL and INH significantly enhanced the apoptosis levels of BCG-infected RAW264.7 cells in the late stage of infection (P=0.001). Conclusion ·In BCG-infected macrophages, SAL can exhibit antimycobacterial effects on the host by enhancing macrophage immune function, suppressing intracellular bacterial survival, and concurrently mitigating the inflammatory response elicited by BCG infection.

Key words: salidroside, Bacillus Calmette-Guérin(BCG), macrophage, immune function, anti-tuberculosis effect

中图分类号:

R392.1

倪书奕, 姜钊, 汪中涛, 何树梅. 红景天苷对卡介苗感染的巨噬细胞免疫功能的影响[J]. 上海交通大学学报（医学版）, 2025, 45(4): 426-433.

NI Shuyi, JIANG Zhao, WANG Zhongtao, HE Shumei. Effect of salidroside on the immune function of BCG-infected macrophages[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(4): 426-433.

图/表 5

图1 SAL和INH对RAW264.7细胞增殖的影响Note： A. MTT assays were used to detect the proliferation of RAW264.7 cells with SAL treatment. B. MTT assays were used to detect the proliferation of RAW264.7 cells with INH treatment. ①P=0.043,②P=0.024, compared with the control group.

Fig 1 Effect of SAL and INH on the proliferation of RAW264.7 cells

图 2 不同时间SAL预处理对BCG感染的巨噬细胞胞内细菌存活率的影响Note： A. Images show CFU of BCG survival in macrophages after 24 h and 36 h of SAL pretreatment. B. Statistical analysis of the relative amount of BCG survival in macrophages after 24 h and 36 h of SAL pretreatment. ①P<0.001, compared with the control group.

Fig 2 Effect of SAL pretreatment at different time points on intracellular bacterial survival in BCG-infected macrophages

图 3 SAL对BCG感染巨噬细胞胞内细菌存活率的影响Note： A. Representative results of flow cytometry detected intracellular bacterial survival in each group of BCG-infected macrophages. B. Statistical analysis of intracellular bacterial survival in A. ①P=0.042, ②P=0.041, ③P<0.001, compared with the rate of BCG-GFP of the BCG group.

Fig 3 Effect of SAL on intracellular bacterial survival in BCG-infected macrophages

图 4 SAL对BCG感染巨噬细胞细胞因子分泌的影响Note： A, B, C, and D respectively represent the levels of cytokines TNF-α、IL-6、IL-10 and IFN-γ in cell culture supernatants at 4 h and 24 h. ①P=0.003, ②P=0.016, ③P=0.029, ④P=0.036, ⑤P=0.047, compared with the levels of cytokines of the BCG group.

Fig 4 Effect of SAL on cytokine secretion in BCG-infected macrophages

图5 SAL对BCG感染巨噬细胞凋亡的影响Note： A. Representative results of flow cytometry detected the apoptosis rate of each group of BCG-infected macrophages at 4 h. B. Representative results of flow cytometry detected the apoptosis rate of each group of BCG-infected macrophages at 24 h. C. Statistical analysis of the total apoptosis rate of cells in A. D. Statistical analysis of the total apoptosis rate of cells in B. ①P=0.008, ②P=0.032,③P=0.003,④P=0.005,⑤P=0.001, compared with the total apoptosis rate of the BCG group.

Fig 5 Effect of SAL on apoptosis of BCG-infected macrophages

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红景天苷对卡介苗感染的巨噬细胞免疫功能的影响

Effect of salidroside on the immune function of BCG-infected macrophages

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