吸烟个体血清在具核梭杆菌侵入KB细胞及分泌MMP-1中的作用

The effects of smoking-individual-serum on the invasion of F. Nucleatum to KB cells and MMP-1 expression of KB cells

目的:探讨慢性牙周炎患者中吸烟个体血清在具核梭杆菌(Fn)侵入KB细胞及在侵入过程中对KB细胞分泌基质金属蛋白酶-1(MMP-1)的作用。方法:提取慢性牙周炎吸烟个体10例(Y组)、慢性牙周炎非吸烟个体10例(N组)、牙周健康非吸烟个体5例(H组)的血清。分别将各组血清200、400、800μl添加至Fn感染KB细胞模型,作用24 h后计数细菌菌落。用人酶联免疫吸附测定试剂盒检测各组上清液中MMP-1的浓度。用单因素方差分析进行数据分析。结果:3组800μl血清引起Fn侵入KB细胞的侵入率(%)分别为12.59±1.27、8.03±0.075和7.99±0.14(P<0.05),KB细胞分泌MMP-1的浓度(μg/L)分别为400.04±21.02、252.57±18.89、262.47±35.29(P<0.05)。200μl和400μl血清组与N组、H组比较,Fn侵入率及KB细胞MMP-1分泌量无统计学差异(P>0.05)。结论:较高剂量吸烟个体血清有利于Fn侵入KB细胞并促进KB细胞分泌MMP-1。

Objective： To investigate the effects of serum from smoking individuals with periodontitis in the process of Fusobocterium.Nucleatum（Fn） invading KB cells and the expression of matrixmetalloproteinase-1（MMP-1） of KB cells. Methods： Serum was prepared from 10 smokers with periodontitis（group Y）,10 non-smokers with periodontitis（group N） and 5 periodontal healthy non-smokers（group H）. Serum of 200,400 and 800 μl from the subjects was added to the model of Fn invading KB cells respectively and cultured for 24 hours,the cfu of cell-invaded bacteria was estimated by colony counting. MMP-1 protein level in culture supernatant was measured by ELISA. Results： In the 800 μl serum groups,the percentage of invaded cfu was 12. 59 ± 1. 27,8. 03 ± 0. 075 and 7. 99± 0. 14 in group Y,N and H（P〈0. 05） respectively,the concentration（μg / L） of MMP-1 in the cultrue supernatant of group Y,N and H was 400. 04 ± 21. 02,252. 57 ± 18. 89 and 262. 47 ± 35. 29（P〈0. 05） respectively. In the 200 μl and 400 μl serum groups,no significant difference was detected in invasion cfu of Fn in KB cells and MMP-1 secrition of KB cells among Y,N and H groups（P〉0. 05）. Conclusion： Higher doses of smoking-serum might play a role in the course of Fn invading KB cells and promote the expression of MMP-1 secretion from KB cells.