黄芩苷对LPS诱导人牙髓细胞TLR4表达及TNF-α分泌的影响

摘要

目的:研究脂多糖(LPS)诱导人牙髓细胞(HDPCs)炎症信号受体--TOll样受体4(TLR4)表达、肿瘤坏死因子-α(TNF-α)分泌情况以及黄芩苷对其的影响,并探讨黄芩苷可能的细胞保护作用机理.方法:采用组织块法原代培养人牙髓细胞,免疫组化技术鉴定细胞来源,流式细胞术(Fc)与酶联免疫法(ELISA)分别检测TLR4与TNF-α的表达.结果:正常HDPCs微量表达TLR4与TNF-α;100μg/mL LPS诱导HDPCs表达TLR4与TNF-μ显著增加(P<0.01);与对照组比较,10μg/mL黄芩苷诱导HDPCs表达rrLR4与TNF-α,差异无统计学意义(P>0.05);0.001～20μg/mL黄芩苷均可抑制LPS诱导HDPCs表达TLR4与TNF-α,随着黄芩苷浓度升高,TLR4与TNF-α表达逐渐降低,且不同浓度组间比较,抑制作用有显著性差异(P<0.01).结论:LPS可能通过TLR4信号受体参与牙髓组织炎症过程.黄芩苷可抑制由LPS诱导的TLR4表达及TNF-α分泌,对LPS引起的牙髓炎可能具有潜在的治疗作用.%AIM:To study the effects of baicalin on the expression of Toll - like receptor 4(TLR4) and the secretion of tumor necrosis factor alpha (TNF -a) in human dental pulp cells(HDPCs) induced by lipopolysaccharide (IPS) , and the mechanism of the effects. METHODS: HDPCs were primarily cultured in vitro and their phenotype was identified by immunohistochemcial staining. Flow Cytometry(FC) was used to analyse the expression of TLR4, and en zyme - labeled immunosorbent assay (ELISA) was used to measure the secretion of TNF - a. RESULTS: TLR4 and TNF - a were detected weakly in normal HDPCs , but their expessions significantly increased after being stimulated with LPS at 100|μg/mL( P < 0.01) . The expressions of TLR4 and TNF - a had no statistical differences between the control group and the group of 10μg/mL baicalin ( P > 0.05 ). Baicalin at the concentrations from 0.001 μg/mL to 20μg/mL could significantly inhibit the activity of TLR4 and TNF - a induced by LPS in a dose - dependent manner. Among the different baicalin concentration groups ,the differences of expression were statistically significant (P<0.01). CONCLUSION; LPS may participate in the inflammation of pulp tissue by TLR4. Baicalin may play a role in the treatment of pulpitis induced by LPS with the help of inhibiting the expression of TLR4 and secretion of TNF - a.