基于LPS炎症模型探究贝母素甲与贝母素乙对巨噬细胞Raw264.7的抗炎作用

doi:10.3969/j.issn.1674-7968.2024.10.014

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摘要贝母素甲与贝母素乙为百合科草本植物浙贝母(Fritillaria thunbergii)中的代表性生物碱类药用成分,对浙贝母抗炎功效发挥重要作用。为从细胞水平上进一步探究这两类生物碱的抗炎功效,本研究以脂多糖(lipopolysaccharide, LPS)诱导的小鼠(Mus musculus)单核巨噬细胞(Raw264.7细胞)为炎症模型,采用Griess法、ELISA、Western blot、qRT-PCR等方法分析贝母素甲和贝母素乙对LPS诱导的Raw264.7细胞分泌促炎因子一氧化氮(nitric oxide, NO)、肿瘤坏死因子-α (tumour necrosis factor-α,TNF-α)、白介素-6 (interleukin-6, IL-6)、IL-1β、一氧化氮合酶(induciblenitric oxide synthase,iNOS)、环氧合酶-2 (cyclooxygenase-2, COX-2)的影响。结果显示,贝母素甲与贝母素乙均能抑制Raw264.7细胞分化,减少LPS刺激产生的伪足,贝母素甲与贝母素乙中、高剂量组Raw264.7细胞上清液中NO、TNF-α、IL-6、IL-1β浓度显著降低(P<0.05, P<0.01)。贝母素甲与贝母素乙对Raw264.7细胞iNOS、COX-2蛋白表达均有抑制作用,且对于iNOS蛋白,贝母素乙的抑制作用高于贝母素甲,对于COX-2蛋白,贝母素甲的抑制作用高于贝母素乙。贝母素甲和贝母素乙均能显著降低Raw264.7细胞IL-6和IL-1β表达量。综上,贝母素甲和贝母素乙均有良好的抗炎效果。本研究为生物碱类物质对炎症的治疗提供一定的数据支持。

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	金泽兰
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	郭焕茹
	严灿龙
	王忠华

关键词 ：贝母素甲, 贝母素乙, Raw264.7, 抗炎活性

Abstract：Peimine and Peiminine are representative alkaloid medicinal component in the lily family herb Fritillaria thunbergii. They play an important role in the anti-inflammatory effect of F. thunbergii. In order to further investigate the anti-inflammatory effects of these 2 types of alkaloids at the cellular level, this study used lipopolysaccharide (LPS) induced Raw264.7 cells as an inflammatory model, Griess, ELISA, Western blot and qRT-PCR were used to analyze the effects of peimine and peiminine on LPS-induced secretion of pro-inflammatory factors nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in Raw264.7 cells, the results showed that both peimine and peiminine could inhibit the differentiation of Raw264.7 cells and reduce the pseudopodia produced by LPS stimulation, NO, TNF-α, IL-6 and IL-1β in Raw264.7 cell supernatant of medium and high dose groups of peimine and peiminine significantly decreased (P<0.05, P<0.01). Peimine and peiminine inhibited the expression of iNOS and COX-2 protein in Raw264.7 cells, and the inhibition of peiminine on iNOS protein was higher than that of peiminine on COX-2 protein, the inhibitory effect of peiminine a was higher than that of peiminine. The expression of IL-6 and IL-1β in Raw264.7 cells was significantly decreased by both peimine and peiminine. In conclusion, both peimine and peiminine have good anti-inflammatory effect, which provides some data support for the treatment of inflammation by alkaloids.

Key words： Peimine Peiminine Raw264 Anti-inflammatory activity

收稿日期: 2023-09-06

中图分类号： S326

基金资助:浙江省中药材新品种选育重大科技专项(2021C02074); 宁波市重点研发计划暨“揭榜挂帅”项目(2023Z120)

通讯作者: ** wang1972@zwu.edu.cn

作者简介: * 同等贡献作者

引用本文:

金泽兰, 董云哲, 李青, 董莉莉, 郭焕茹, 严灿龙, 王忠华. 基于LPS炎症模型探究贝母素甲与贝母素乙对巨噬细胞Raw264.7的抗炎作用[J]. 农业生物技术学报, 2024, 32(10): 2361-2370.
JIN Ze-Lan, DONG Yun-Zhe, LI Qing, DONG Li-Li, GUO Hua-Ru, YAN Can-Long, WANG Zhong-Hua. Exploring the Anti-inflammatory Effects of Peimine and Peiminine on Macrophage Raw264.7 Based on LPS Inflammation Model. 农业生物技术学报, 2024, 32(10): 2361-2370.

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https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2024.10.014 或 https://journal05.magtech.org.cn/Jwk_ny/CN/Y2024/V32/I10/2361

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