高剂量脂多糖影响舌苔形成机制初探

Mechanism of high-dose lipopolysaccharide influencing formation of tongue fur

目的探讨脂多糖(lipopolysaccharide,LPS)影响舌苔形成的信号通路。方法将0.1、1、5、105、0、100 mg/L LPS作用于体外培养的舌鳞癌TCA-8113细胞163、2、48 h,MTT法检测细胞增殖活性。10、501、00 mg/L LPS作用TCA-8113细胞16、324、8 h,细胞流式细胞仪检测细胞周期分布。10、501、00 mg/L LPS作用TCA-8113细胞8 h,细胞免疫组化法研究p38-MAP kinasec、-myc、c-fos和NF-κB在TCA-8113细胞中的表达水平。结果与对照组相比,LPS作用TCA-8113细胞16 h,对细胞增殖活性影响不明显(P>0.05);作用32 h5,0、100 mg/L组明显促进细胞增殖活性(P<0.05),而0.1、15、、10 mg/L组效应不明显(P>0.05);作用48 h0,.11、、5、10 mg/L组抑制细胞增殖活性(P<0.05),而50、100 mg/L组效应不明显(P>0.05)。10、501、00 mg/L LPS对TCA-8113细胞周期分布和凋亡影响不明显1,6 h G1/G0期细胞比例下降,32 h促进S期细胞比例下降4,8 h细胞周期分布无明显变化(P>0.05)。10、50、100 mg/L LPS作用舌鳞癌细胞8 h,明显下调p38-MAPkinase、c-myc、c-fos和NF-κB p50表达水平,而上调NF-κB p65表达水平,统计学差异显著(P<0.05)。结论高剂量LPS可以影响TCA-8113的增殖活性,是参与舌苔形成的重要外源性因素,对NF-κB信号通路具有重要作用。

Objective To investigate the signal pathway of lipopolysaccharide （LPS） influencing the formation of tongue fur. Methods The squamous carcinoma cell line （TCA-8113 ） cultured in vitro were acted by LPS in the doses of 0. 1, 1,5, 10, 50 and 100 mg/L for 16, 32 and 48 hours respectively and the proliferative activity was detected by using MTT method. TCA-8113 ceils were acted by LPS in the doses of 10, 50 and 100 mg/L for 16, 32 and 48 hours respectively and the distribution of cell cycle was detected by using flow cytometer. TCA-8113 cells were acted by LPS in the doses of 10, 50 and 100 mg/ L for 8 hours and the expressions of p38-MAP kinase, c-myc, c-fos and natural factor-κB （NF-κB） were studied by applying immunohistochemistry. Results Compared with the control group, LPS had no significant influence on the proliferative activity of TCA-8113 cells after LPS acting for 16 hours （P 〉 0. 05）. In the 50 mg/L group and 100 mg/L group the proliferative activity was significantly improved （P〈0.05） after LPS acting for 32 hours, but the effectiveness was not significant in the 0. 1 mg/L group, 1 mg/L group, .5 mg/L group and 10 mg/L group （ P 〉 0.05 ）. The proliferative activity was inhibited in theO. 1 mg/L group, 1 mg/L group, 5 mg/L group and 10 mg/L group （P 〈0.05）, but the effectiveness was not obvious in the 50 mg/L group and 100 mg/L group （P 〉0. 05） after LPS acting for 48 hours. LPS in the doses of 10, 50 and 100 mg/L had no significant influences on the distribution of cell cycle and apoptosis of TCA-8113, and after acting for 16 hour the proportion of G1/GO-phase cells decreased, after acting for 32 hours the proportion of S-phase cells decreased, and after acting for 48 hours the distribution of cell cycle had no significant changes （P 〉 O. 05 ）. LPS in the doses of 10, 50 and 100 mg/L significantly regulated downward the expressions of p38-MAP kinase, c-myc, c-fos and NF-κB p50, but regulated upward the expression of NF-κB p65 after acting for 8 hours （ P 〈 O. 05 ）. Conclusion LPS in high dose can influence the proliferative activity of TCA-8113. It is an important exogenic factor in the formation of tongue fur and plays an key role in NF-κB signal pathway.