白介素-35促进牙髓干细胞骨向分化的体外实验研究

In vitrostudy on interleukin-35 in the promotion of DPSCs osteogenic differentiation

目的探究白介素-35(interleukin-35, IL-35)对人牙髓干细胞(dental pulp stem cells, DPSCs)骨向分化的影响。方法从正畸患者的牙髓组织中分离培养DPSCs在成骨诱导液中培养21 d,RT-qPCR检测成骨相关基因(RUNX2,COL-1)以及IL-35受体(gp130)表达情况,茜素红染色鉴定矿化结节,用氯化十六烷基吡啶溶液定量分析染色情况;用浓度为100 ng/ml的IL-35刺激DPSCs的骨向分化,分别在有/无IL-35的情况下,比较成骨相关基因表达及茜素红染色定量分析结果。结果茜素红染色及成骨相关基因表达证明了DPSCs的骨向分化(P<0.05),分化过程伴有gp130(IL-35受体之一)的表达升高(P<0.05),在人重组IL-35作用下DPSCs显示出更强的茜素红染色(P<0.001),并伴有成骨相关基因表达升高(P<0.05)。结论 IL-35不仅具有抗炎作用,也可以促进DPSCs的骨向分化。

Objective Toexploretheeffectofinterleukin-35(IL-35)on osteogenicdifferentiation ofdentalpulp stemcells(DPSCs).Methods The DPSCs were isolated from pulp tissue of orthodontic patients and cultured in osteogenic induction medium for 21 days. The osteogenic related genes such as runt-related transcription factors(RUNX2) and collagen type 1(COL-1) and the expression of receptors of IL-35(glycoprotein 130, gp130) were detected by RT-qPCR. The calcium mineralized nodules were stained by alizarin red and the staining was quantitative analyzed by cetylpyridinium chloride(CPC). The osteogenic differentiation of DPSCs was stimulated by IL-35 with the concentration of 100 ng/ml. Under the condition with or without IL-35, the expression of osteogenic related gene and the quantitative analysis results of alizarin red staining were compared. Results The osteogenic differentiation of DPSCs was demonstrated by alizarinred staining and expression of osteogenic related genes(P<0.05). The differentiation process accompanied with increased expression of gp130(P<0.05). Under the action of human recombinant IL-35, the DPSCs showed strong alizarin red staining(P<0.001) and accompanied with increased expression of RUNX2 and COL-1(P<0.05). Conclusion The IL-35 can suppress inflammation and promote the osteogenic differentiation of DPSCs.