目的 筛选赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对炎症反应的抑制作用最强的流份。 方法 使用MTT法确定壮药赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对RAW264.7细胞安全给药浓度范围，通过ELISA法测定赪桐乙酸乙酯部位及不同洗脱梯度二氯甲烷-甲醇部位对LPS诱导RAW264.7细胞分泌NO、TNF-ａ、IL-12、IL-6、IL-1β含量，筛选对炎症反应的抑制作用最强的流份。 结果 赪桐乙酸乙酯部位及流份在0.06~2.0 mg/ml浓度范围内，赪桐乙酸乙酯部位及流份对细胞活力的抑制作用逐渐增强，浓度在0.5 mg/ml以上具有明显的细胞毒性，在0.5 mg/ml以下对细胞活力具有增强作用。二氯甲烷-甲醇洗脱部位高剂量能抑制炎症因子IL-12、IL-6、TNF-ａ、IL-1β释放，对NO的分泌没有抑制作用。 结论 赪桐乙酸乙酯部位及不同洗脱梯度的二氯甲烷-甲醇部位抗炎机制是通过抑制NO、TNF-ａ、IL-12、IL-6、IL-1β炎症因子的分泌，二氯甲烷-甲醇（50:1）洗脱部位和二氯甲烷-甲醇（30:1）洗脱部位的抗炎能力较强。
Abstract
Objective To screen the fractions with the strongest inhibitory effect on the inflammatory response from the ethyl acetate part of Clerodendrum japonicum (Thunb.) Sweet and the dichloromethane-methanol parts with different elution gradients. Methods The MTT method was used to determine the safe dose range of the Clerodendrum japonicum (Thunb.) Sweet ethyl acetate site and different elution gradients of dichloromethane-methanol to RAW264.7 cells, and the Clerodendrum japonicum (Thunb.) Sweet. ethyl acetate site and different elution gradient dichloride were determined by the ELISA method. The secretion of NO, TNF-a, IL-12, IL-6, and IL-1β were detected in RAW264.7 cells induced by LPS, and screened the fraction with the strongest inhibitory effect on the inflammatory response. Results The ethyl acetate parts and fractions of Clerodendrum japonicum (Thunb.) Sweet were within the concentration range of 0.06~2 mg/mL. The inhibitory effect of the ethyl acetate sections and fractions of Clerodendrum japonicum (Thunb.) Sweet on cell viability was gradually stronger, and the concentration above 0.5 mg/mL had obvious cytotoxicity. Below 0.5 mg/mL could enhance cell viability. The high dose of dichloromethane-methanol elution site could inhibit the release of inflammatory factors IL-12, IL-6, TNF-a, IL-1β, but had no inhibitory effect on the secretion of NO. Conclusions The anti-inflammation mechanism of the Clerodendrum japonicum (Thunb.) Sweet ethyl acetate site and the dichloromethane-methanol site with different elution gradients is through the inhibition of the secretion of NO, TNF-a, IL-12, IL-6, IL-1β inflammatory factors of cells, dichloride Methane-methanol (50:1) elution site and dichloromethane-methanol (30:1) elution site have strong anti-inflammatory ability.
关键词
赪桐； /
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乙酸乙酯部位； /
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流份部位； /
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RAW264.7； /
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抗炎
Key words
Clerodendrum japonicum (Thunb.) Sweet; /
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Ethyl acetate site; /
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Flow part; /
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RAW264.7; /
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Anti-inflammatory
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基金
国家自然科学基金（No.81660659）；广西自然科学基金项目（No.2019GXNSFAA245090、2018GXNSFAA050141）；2019年度广西高校中青年教师科研基础能力提升项目(No.2019KY0341)；广西壮瑶药重点实验室开放课题(No.GXZYKF2020A-08)；2020年广西国际壮医医院科研项目（No.GZ202001）；2019年广西中医药大学青年基金项目(No.2019QN036)。
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