白细胞介素-1β对牙周膜细胞向破骨细胞分化的影响

Effect of interleukin-1βon differentiation of periodontal membrane cells into osteoclasts

[目的]探究白细胞介素-1β对牙周膜细胞向破骨细胞分化的影响。[方法]选取需要牙齿矫正患者的第一前磨牙和第二前磨牙,取牙周膜组织培养后按照1×10^(8)/mL的密度,对照组向培养基中加入0μg/L白细胞介素-1β或实验组向培养基中加入20μg/L白细胞介素-1β,对比分析两组牙周膜细胞体外增殖能力和TRAP阳性细胞数量;RT-PCR法分别测定牙周膜细胞OPG、RANKL、ODF、OCIF、CTSK、MMP-9和CAⅡmRNA表达水平,Western Blot法测定OPG和RANKL蛋白表达水平。[结果]与培养第1 d相比,培养第4、7、11、14和21 d的各组牙周膜细胞体外增殖能力增加(P<0.05),其中对照组细胞体外增殖能力低于实验组(21 d:0.45±0.03 vs 1.68±0.37)(P<0.05)。与培养第1 d相比,培养第4 d、7 d、11 d、14 d和21 d的各组牙周膜细胞TRAP阳性细胞数量增加(P<0.05),且在第11 d达到最大值(40.36±4.52%),随后出现下降趋势,其中对照组细胞TRAP阳性细胞数量低于实验组(P<0.05)。实验组牙周膜细胞OPG mRNA表达低于对照组(0.42±0.12 vs 1.14±0.19),而RANKL mRNA表达高于对照组(4.65±0.11 vs 2.31±0.27)(P<0.05)。实验组牙周膜细胞OPG蛋白表达低于对照组(0.78±0.17 vs 1.43±0.14),而RANKL蛋白表达高于对照组(2.92±0.17 vs 1.21±0.08)(P<0.05)。实验组牙周膜细胞ODF和OCIF mRNA表达高于对照组(ODF:2.56±0.85 vs 0.71±0.16;OCIF:4.04±0.77 vs 0.99±0.23)(P<0.05)(P<0.05)。实验组牙周膜细胞CTSK、MMP-9、CAⅡmRNA表达高于对照组(CTSK:2.57±0.23 vs 1.05±0.01;MMP-9:2.34±0.18 vs 1.04±0.03;CAII:3.79±0.12 vs 1.06±0.03)(P<0.05)。[结论]白细胞介素-1β可有效诱导牙周膜细胞分化为有骨吸收能力的破骨细胞,其机制是通过上调RANKL表达和下调骨保护素表达来介导。

[Objective]To investigate the effect of interleukin-1βon the differentiation of periodontal membrane cells to osteoclasts.[Method]The first premolar and the second premolar of patients needing orthodontic treatment were selected.Periodontal membrane tissues were cultured at a density of 1×108/mL,and then interleukin-1βwas added into the culture medium at 0(control group)or 20μg/L(experimental group).The proliferation ability of periodontal membrane cells in vitro and the number and positive rate of TRAP positive cells in the two groups were compared and analyzed.The mRNA expression levels of OPG and RANKL,ODF and OCIF,CTSK,MMP-9 and CAⅡin periodontal membrane cells were determined by RT-PCR,and the protein expression levels of OPG and RANKL were determined by Western Blot.[Result]Compared with 1st day,the in vitro proliferation of periodontal membrane cells was increased at 4th,7 th,11th,14th and 21th(P<0.05),and the proliferation of periodontal membrane cells in the control group was lower than that in the experimental group(21th day:0.45±0.03 vs 1.68±0.37)(P<0.05).Compared with the first day of culture,the number of TRAP positive cells in periodontal membrane cells in the 4,7,11,14 and 21 days increased(P<0.05),and reached the maximum value(40.36%±4.52%)on the 11th day,followed by a downward trend,and the number of TRAP positive cells in the control group was lower than that in the experimental group(P<0.05).The expression of OPG mRNA in periodontal membrane cells in experimental group was lower than that in control group(0.42±0.12 vs 1.14±0.19),while the expression of RANKL mRNA was higher than that in control group(4.65±0.11 vs 2.31±0.27)(P<0.05).The expression of OPG protein in the experimental group was lower than that in the control group(0.78±0.17 vs 1.43±0.14),while the expression of RANKL protein was higher than that in the control group(2.92±0.17 vs 1.21±0.08)(P<0.05).The expression of ODF and OCIF mRNA in periodontal membrane cells in experimental group was higher than that in control group(ODF:2.56±0.85 vs 0.71±0.16;OCIF:4.04±0.77 vs 0.99±0.23)(P<0.05).The mRNA expressions of CTSK,MMP-9 and CAⅡin periodontal membrane cells in the experimental group were higher than those in the control group(CTSK:2.57±0.23 vs 1.05±0.01;Mmp-9:2.34±0.18 vs 1.04±0.03;CAⅡ:3.79±0.12 vs 1.06±0.03)(P<0.05).[Conclusion]Interleukin-1βcan effectively induce periodontal membrane cells to differentiate into osteoclasts with bone resorption ability.The mechanism is mediated by up-regulation of RANKL expression and down-regulation of osteoprotectin expression.