新冠肺炎寒湿疫小鼠模型的建立及其评价
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1. 广州中医药大学中药学院,广州 510006;2. 东莞市中医院,广东 东莞 523000; 3. 东莞广州中医药大学研究院, 广东 东莞 523808

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R-33

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Establishment and evaluation of a mouse model of COVID-19 as “cold-dampness epidemic”
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1. School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, China. 2. Dongguan Hospital of Traditional Chinese Medicine, Dongguan 523000. 3. Dongguan Institute of Guangzhou University of Chinese Medicine,Dongguan 523808

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    摘要:

    目的 为中医理论下新冠肺炎归属“寒湿疫”提供病理模型,本实验通过脂多糖诱导小鼠引发细胞因子风暴,结合寒湿刺激,旨在建立合理、实用的寒湿疫小鼠模型。 方法 24 只雄性 KM 小鼠随机分为两个造模组和正常组。造模组 1(脂多糖组)为腹腔注射 5 mg / kg 脂多糖生理盐水溶液,造模组 2(脂多糖+寒湿刺激组)为腹腔注射 5 mg / kg 脂多糖生理盐水溶液+寒湿刺激,正常组给予等量生理盐水。造模 8 d 后,评价模型小鼠在中医理论下的相关外观行为表征,以及肺指数、脾指数、胸腺指数,ELISA 检测肺组织细胞因子的水平,检测环氧化酶 2 (COX-2)、瞬时受体电位通道蛋白 V1 和 A1(TRPV1 和 TRPA1)的表达量。 结果 与正常组比较,脂多糖+寒湿刺激组小鼠的外在状态和行为表现与中医理论定义的寒湿证相一致,包括活跃程度下降、精神状态不佳。模型组的肺指数,胸腺指数,脾指数高于正常组;脂多糖+寒湿刺激组 TNF-α,IFN-γ,IL-6 和 PGE2 的水平明显高于正常组及脂多糖组;HE 染色显示,模型组肺组织损伤明显。 模型组中 COX-2 和 TRPA1 蛋白表达显著升高,TRPV1 蛋白表达显著降低。 结论 脂多糖+寒湿刺激小鼠模型可作为构建中医理论下新冠肺炎引起的“寒湿疫”病证较为理想的方法,模型组小鼠指标变化符合文献报导的新冠肺炎造成的机体损伤。

    Abstract:

    Objective To establish a reasonable and practical pathological mouse model for the attribution of coronavirus disease 2019 (COVID-19) as “ cold-dampness epidemic” under the theory of Traditional Chinese Medicine (TCM). Methods Twenty-four male KM mice were randomly divided into three groups: normal group and two model groups. The mice in model group 1 (lipopolysaccharide [LPS] group) were intraperitoneally injected with 5 mg / kg of LPS saline solution, those in model group 2 (“ LPS + cold-dampness stimulation” syndrome group) were intraperitoneally injected with 5 mg / kg of LPS saline solution and cold dampness stimulation, and those in the normal group were injected with the same amount of normal saline. After 8 days, the appearance and behavior of model mice were evaluated under the theory of TCM; additionally, the lung index, spleen index, and thymus index were evaluated, and the levels of inflammatory factors in the lung tissues were detected by ELISA, and the protein expression levels of COX-2, TRPV1 and TRPA1 were detected. Results Mice in the LPS + “cold-dampness stimulation” syndrome group exhibited an appearance and behavior similar to those of cold-dampness syndrome, e. g. , decreased activity and poor mental state. The lung index, thymus index, and spleen index of model groups were higher than those of the normal group. The levels of TNF-α, IFN-γ, IL-6 and PGE2 in the LPS + “ cold-dampness stimulation” syndrome group were significantly higher than those in the normal group or the LPS group. HE staining revealed that the extent of injury in the model groups were more severe than that in the normal group. The expression levels of COX-2 and TRPA1 protein were significantly higher in the model groups, whereas the expression level of TRPV1 protein was significantly lower. Conclusions The LPS + “ cold-dampness stimulation” syndrome mouse model can be used as an ideal method to construct the syndrome of “HanShiYi” caused by COVID-19 under the theory of TCM, and the changes of indexes of mice in the LPS + “ cold-dampness stimulation” syndrome model group are consistent with the body damage caused by COVID-19 reported in the literature.

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朱淑敏,宁为民,董明国,欧健钊,黄海阳,侯少贞,高 洁.新冠肺炎寒湿疫小鼠模型的建立及其评价[J].中国比较医学杂志,2022,32(6):62~68.

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  • 收稿日期:2022-03-03
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  • 在线发布日期: 2022-08-01
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