云南白药在P.g-LPS诱导的炎症环境下对人牙髓细胞分泌TNF-α、IL-6、IL-1β的影响

黄松靖; 雷雅燕; 佘睿; 吴剑花; 刘暑梅; 张明珠

云南白药在P.g-LPS诱导的炎症环境下对人牙髓细胞分泌TNF-α、IL-6、IL-1β的影响

1. 昆明医科大学附属口腔医院牙体牙髓科
2. 昆明医科大学附属口腔医院牙周科

基金项目:

基金：云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目 (2013FB178, 2014FZ039);

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出版历程
- 收稿日期: 2017-04-10

Effect of Yunnan Baiyao on the Secretion of TNF-α, IL-6, IL-1β from Human Dental Pulp Cells under LPS-PG induced Inflammatory Environment

Funds:

基金：云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目 (2013FB178, 2014FZ039);

摘要

摘要: 目的研究云南白药在P.g-LPS诱导的炎症环境下对人牙髓细胞分泌IL-6、IL-lβ、TNF-α的影响, 探索云南白药对牙髓炎抑制的影响.方法采用改良组织块法体外培养人牙髓细胞, 免疫组化鉴定细胞来源, 10μg/m L的P.g-LPS刺激人牙髓细胞24 h后, 分3组, 空白对照组1%FBS培养液, 阳性对照组10μg/m L P.g-LPS, 白药组10μg/m L云南白药.于刺激结束, 换液后1 d、3 d时ELISA检测培养上清中IL-6、IL-lβ、TNF-α水平, 同时换液3 d时定量real-time PCR检测TNF-α、IL-6、IL-lβmRNA的表达.结果成功培养HDPCs, 波形丝蛋白表达阳性, 角蛋白为阴性.P.g-LPS刺激人牙髓细胞1 d时, 3组之间TNF-α、IL-6、IL-1β蛋白表达无差别, 换液后1 d和3 d, 空白对照组和白药组低于阳性对照组 (P<0.05) , 白药组与空白对照组差异无统计学意义 (P>0.05) .然而换液后3 d白药组TNF-α、IL-6、IL-1βmRNA的表达低于空白对照组 (P<0.05) .结论在P.g-LPS诱导的炎症环境下, 云南白药作用3 d时, 对HDPCs的TNF-α、IL-6、IL-1β蛋白分泌没有明显抑制作用, 但抑制TNF-α、IL-6、IL-1β mRNA的表达.
- 牙龈卟啉单胞菌脂多糖 /
- 人牙髓细胞 /
- TNF-α /
- IL-6 /
- IL-1β
Abstract: Objective To investigate the effect of Yunnan Baiyao on the secretion of TNF-α, IL-6, IL-1βfrom human dental pulp cells ( h DPCs) under lipopolysaccharide from Porphyromonas gingivalis (LPS-PG) induced inflammatory environment, and investigate the inhibition effect of Yunnan Baiyao on pulpitis. Methods HDPCs were cultured by the modified tissue explant culture technique. Cultured h DPCs were identified and verified by the immunocytochemical staining. HDPCs were exposed to 10 μg/m L LPS-PG solution for 1 day, then the LPS-PG containing culture media were removed and collected, the h DPCs were divided into3 groups cultured with different new culture media, the control group was cultured with 1% FBS MEM, the LPS-PG group was cultured with 10 μg/m L LPS-PG, the Baiyao group was cultured with 10μg/m L Yunnan Baiyao. Culturing media were collected on exposure finishing, 1st and 3rd day after the medium changing for detection of IL-6, IL-1β and TNF-αproteins by ELISA, and on 3rd day the quantitative Real-Time PCR (q RT-PCR) was performed for IL-6, IL-1β and TNF-α gene expression of the h DPCs.Results HDPCs culturing showed a positive vimentin expression and a negative keratin expression. 3 groups showed no difference in TNF-α, IL-6 and IL-1β secretions immediately after the LPS-PG exposure for 1 day by ELISA, and the LPS-PG group showed higher secretions of TNF-α, IL-6 and IL-1β than other groups on1 st and 3rd day (P<0.05) . Baiyao group and control group showed no differences (P>0.05) . However, q RT-PCR showed lower gene expressions of TNF-α, IL-6, IL-1β in the Baiyao group than other groups at 3rd day (P <0.05) .Conclusion Under inflammatory environment induced by LPS-PG, the Yunnan Baiyao shows no inhibitory effect on secretion of TNF-α, IL-6 and IL-1β on3 rd day after the medium changing, but can downregulate the m RNA expression.
- Porphyromonas gingivali Lipopolysaccharide /
- Human dental pulp cells /
- TNF-α /
- IL-6 /
- IL-1β

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