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);

计量
- 文章访问数: 2542
- HTML全文浏览量: 972
- PDF下载量: 136
- 被引次数: 0
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

HTML全文

参考文献(26)

[1]周丽珍, 郭玲.组织工程技术应用于牙髓-牙本质再生的研究进展[J].海南医学, 2007, 18 (6) :128-129.
[2]史俊南, 金岩.关于"牙本质-牙髓复合体"一词的译名[J].牙体牙髓牙周病学杂志, 2004, 14 (7) :410.
[3]梁静, 顾令婉, 王凯.转化生长因子和牙本质牙髓复合体[J].中国医药指南, 2015, 21 (4) :54-54, 55.
[4]	[4]LAI WY, KAO C T, HUNG C J, et al.An evaluation of the inflammatory response of lipopolysaccharide-treated primary dental pulp cells with regard to calcium silicate-based cements[J].International Journal of Oral Science, 2014, 6 (2) :94-98.
[5]	[5]EMIOLA A, ANDREWS S S, HELLER C, et al.Crosstalk between the lipopolysaccharide and phospholipid pathways during outer membrane biogenesis in Escherichia coli[J].Proceedings of the National Academy of Sciences of the United States of America, 2016, 113 (11) :3108-3113.
[6]肖水清, 汪竹平, 王铎.牙髓炎中的内毒素检测及其与临床表现的关系[J].牙体牙髓牙周病学杂志, 2002, 12 (4) :219.
[7]	[7]YAMADA H, NAKAJIMA T, DOMON H, et a1.Endoplasmic reticulumstress response and bone loss in experimental periodontitis inmice[J].Pericxlnntal Res, 2015, 50 (4) :500-508.
[8]武海春, 许尧生, 李彬, 等.LPS刺激对体外培养人牙周膜细胞产生IL-8和MCP-1的影响[J].牙体牙髓牙周病学杂志, 2013, 23 (8) :485-488.
[9]杨俊, 陈兴兴, 方煌, 等.LPS对人牙周膜细胞Toll样受体4及下游炎症因子表达的影响[J].临床口腔医学杂志, 2014, 14 (11) :689-691.
[10]	[10]NAKANE A, YOSHIDA T, NAKATA K, et al.Effects of lipopolysaccharides on human dental pulp cells[J].Journal of Endodontics, 1995, 21 (3) :128-130.
[11]	[11]HE W, QU T, YU Q, et al.LPS induces IL-8 expression through TLR4, My D88, NF-kappa B and MAPK pathways in human dental pulp stem cells[J].International Endodontic Journal, 2013, 46 (2) :128-136.
[12]	[12]PINERO G J, KIATPONGSAH S, HUTCHINS M O, et al.The effect of endotoxin on the synthesis of connective tissue matrix components by pulp fibroblasts in vitro[J].Journal of Endodontics, 1983, 9 (1) :2-7.
[13]王凤明, 胡涛, 周学东.脂多糖对人牙髓细胞增殖和碱性磷酸酶活性的影响[J].上海口腔医学, 2005, 14 (1) :77-80.
[14]	[14]KADONO H, KIDO J I, KATAOKA M, et al.Inhibition of osteoblastic cell differentiation by lipopolysaccharide extract from Porphyromonas gingivalis[J].Infection and Immunity, 1999, 67 (6) :2841-2846.
[15]杨柳, 杨安民, 杜俊蓉.云南白药含漱剂对口腔常见细菌抑制作用的体外研究[J].中成药, 2010, 32 (10) :1785-1788.
[16]胡幼芝, 杨如美.云南白药盖髓剂的动物实验及临床应用[J].江苏医药, 2000, 26 (9) :701-702.
[17]何飞, 谭颖徽, 张纲.人牙髓干细胞的体外培养和鉴定[J].华西口腔医学杂志, 2005, 23 (1) :75-78.
[18]蔡洪桢, 贺慧霞, 王飞翔, 等.人牙周膜干细胞与人牙髓干细胞的表型及生长特性比较[J].中华老年口腔医学杂志, 2017, 15 (2) :91-96.
[19]杨一新, 李桂源.LPS所介导的信号转导通路研究进展[J].中南大学学报 (医学版) , 2006, 31 (1) :141-145.
[20]罗琴, 方梅飞, 刘贞敏, 等.脂多糖对人牙龈上皮细胞β-防御素表达的影响及其机制研究[J].广西医科大学学报, 2017, 34 (2) :173-177.
[21]	[21]KUMAR A, BHATTACHARYYA S, RATTAN V.Effect of uncontrolled freezing on biological characteristics of human dental pulp stem cells.[J].Cell and Tissue Banking, 2015, 16 (4) :513-522.
[22]	[22]TRAN HLE B, DOAN V N.Human dental pulp stem cells cultured onto dentin derived scaffold can regenerate dentin-like tissue in vivo[J].Cell and Tissue Banking, 2015, 16 (4) :559-568.
[23]	[23]ASUTAY F, POLAT S, GL M, et al.The effects of dental pulp stem cells on bone regeneration in rat calvarial defect model:Micro-computed tomography and histomorphometric analysis[J].Archives of Oral Biology, 2015, 60 (12) :1729-1735.
[24]	[24]SABRAH A H, YASSEN G H, LIU W C, et al.The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm[J].Clinical Oral Investigations, 2015, 19 (8) :2059-2066.
[25]侯建霞.牙龈卟啉单胞菌超声提取物对人牙周膜干细胞骨向分化的影响[J].现代口腔医学杂志, 2007, 21 (6) :603-605.
[26]	[26]COIL J, TAM E, WATERFIELD J D.Proinflammatory cytokine profiles in pulp fibroblasts stimulated with lipopolysaccharide and methyl mercaptan[J].Journal of Endodontics, 2004, 30 (2) :88-91.

相关文章(20)

[1]	赵小青, 肖嘉欣, 林勇, 张剑锋, 王文辉, 张文杰. 不同毒力型的幽门螺旋杆菌对胃内微环境的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240221
[2]	石苒羲, 杨涛, 程庆, 赵梁辰, 郭嘉晖, 王利梅. 基于网络药理学与分子对接探讨铁皮石斛抗炎性衰老的作用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20231101
[3]	杨金润, 赵欢欢, 吴志聪, 余戈蕊, 马岚, 陈玉芬, 解谩伊, 董玉. 化湿润睛汤通过调控COX-2、TNF-α和 IL-6蛋白改善去势雄性干眼病大鼠的炎症反应, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20231106
[4]	世文彪, 马彩良, 赵亚林, 王峰, 汪菊林, 张米慧. 妊娠期高血压疾病患者外周血IL-1β、IL-2、IL-4、IL-5、IL-6的变化与意义, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230909
[5]	鲁智英, 王美芬, 彭俊超, 杜曾庆, 罗云娇, 马薇. 小儿轮状病毒和诺如病毒性肠炎血清PCT、IL-6、SAA水平测定及临床特征的差异, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220823
[6]	张莉, 余丹, 王雨婷, 苟志梅, 杨仁华, 何波, 沈志强, 柴文英, 陈鹏. 灯盏乙素对OX-LDL损伤的RAW264.7细胞中PKC和TNF-α表达的影响, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20211040
[7]	龙熙翠, 刘贝贝, 卢绍波, 李志红, 金文娇, 陆金芝, 韩雪松. 细胞焦亡因子Caspase-1、IL-1β与IL-18在子宫内膜息肉组织中的表达和意义, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210917
[8]	莽源祎, 李立, 冉江华, 升宁, 刘静, 刘剑, 高杨, 李来邦. 特利加压素联合FK-409通过减少IL-10和TNF-α释放保护移植肝的机制, 昆明医科大学学报.
[9]	杨金凤, 蒙国懿, 黄兆奎, 洪仕君, 李利华, 邢豫明, 赵永娜. 甲基苯丙胺对大鼠海马5-HT、IL-6及NF-κB表达的影响, 昆明医科大学学报.
[10]	汪君民, 吴晓虹, 王丽, 雷德鸿, 邱良武. 游泳运动对Fetuin-A诱导的胰岛素抵抗大鼠血清IL-2、IL-6的影响, 昆明医科大学学报.
[11]	聂祖庆, 张琼, 王鹏飞, 杨丽娟, 杨雯虹, 王若花, 马林昆, 杨明. 外伤性视神经损伤后大鼠视网膜中细胞因子的变化和来源, 昆明医科大学学报.
[12]	蒙国懿, 隋念含, 杨金凤, 边立功, 洪仕君, 李利华, 赵永娜. 甲基苯丙胺对大鼠海马TNF-α、IL-1β及CD22表达的影响, 昆明医科大学学报.
[13]	刘城秀. 云南汉族人群TNF-α基因和ALCAM基因多态性与HCV慢性感染的相关性, 昆明医科大学学报.
[14]	李俊杰. 脑缺血-再灌注损伤大鼠脑组织中TNF-α，IL-6和IL-1β的表达, 昆明医科大学学报.
[15]	艾宝. 多发性硬化患者血清褪黑激素和TNF-α水平与EDSS评分的相关性, 昆明医科大学学报.
[16]	李玉环. TNF-α在单纯性脑震荡大鼠皮质中的表达变化, 昆明医科大学学报.
[17]	孙鹥. Th17相关细胞因子在丙型肝炎患者血清中的变化及意义, 昆明医科大学学报.
[18]	谭小兵. 牙周基础治疗从不同途径对伴中重度牙周炎冠心病患者的影响, 昆明医科大学学报.
[19]	. 严重创伤大鼠TNF-α、p-EP及IL-1O的动态变化, 昆明医科大学学报.
[20]	. 严重创伤大鼠TNF-α、β-EP及IL-10的动态变化, 昆明医科大学学报.