模拟微重力环境对人牙周膜成纤维细胞MMP8表达的影响

发布时间：2018-01-02 22:38

本文关键词：模拟微重力环境对人牙周膜成纤维细胞MMP8表达的影响　出处：《空军医学杂志》2016年06期 　论文类型：期刊论文

【摘要】：目的观察模拟微重力环境下人牙周膜成纤维细胞(human periodontal ligament fibroblasts,h PDLFs)中MMP8表达的变化。方法 (1)采用改良酶组织块法培养h PDLFs并对其来源进行免疫组织化学鉴定;(2)建立模拟微重力下h PDLFs培养模型;(3)建立药物诱导h PDLFs成骨向分化的模型,将实验分为骨向诱导组以及非骨向诱导组,并分别观察骨向诱导组以及非骨向诱导组中培养第1、2、3天,h PDLFs中ALP和OC表达的变化;(4)应用RT-PCR和Western-blot检测h PDLFs中MMP8在模拟微重力下表达量的变化。结果 (1)成功分离获得了h PDLFs,免疫组织化学染色结果为Keratin染色阴性,Vimentin染色阳性,细胞胞浆棕黄色着色,着色分布较为均匀;(2)成功地建立了模拟微重力下h PDLFs培养模型,通过生长曲线的绘制,选择3 d为实验观察最佳时间,h PDLFs在回转器中生长状态良好;(3)非诱导组第3天,ALP的分泌较正常重力对照组下降约20%,差异具有统计学意义(P0.05)。在骨向诱导组第3天,模拟微重力组ALP的分泌较正常重力对照组下降约16%,骨向诱导组中,正常重力对照组和模拟微重力组均可观察到ALP的分泌增加,具有时间依赖性,有统计学差异(P0.05),说明骨模拟微重力效应可降低h PDLFs中ALP和OC的分泌,但骨诱导剂可以减弱其降低的趋势;(4)在模拟微重力组中,h PDLFs中MMP8的m RNA和蛋白的表达均较正常重力组增加,并且MMP8的表达在骨向诱导组中高于非诱导组,第3天差异具有统计学意义(P0.05)。结论模拟微重力环境中降低h PDLFs骨向分化的能力,并且MMP8表达较正常重力组增加,表明MMP8参与调节模拟微重力环境对h PDLFs的成骨向分化。
[Abstract]:Objective to observe the human periodontal ligament fibroblasts of human periodontal ligament fibroblasts in simulated microgravity environment. Methods: h PDLFs was cultured by modified enzyme tissue block method and its origin was identified by immunohistochemistry. (2) establish h PDLFs culture model under simulated microgravity; The model of PDLFs osteogenic differentiation induced by drugs was established. The experiment was divided into osteoinduction group and non-osteotropic induction group, and the 1st stage of osteoinduction group and non-osteotropic induction group were observed respectively. The changes of ALP and OC expression in PDLFs at day 3; (4) RT-PCR and Western-blot were used to detect the expression of MMP8 in h PDLFs under simulated microgravity. H PDLFs was obtained successfully. The results of immunohistochemical staining showed that Keratin staining was negative and vimentin staining was positive, and the cytoplasm of the cells was brownish stained, and the staining distribution was more uniform. (2) the model of h PDLFs culture under simulated microgravity was successfully established. Through drawing the growth curve, 3 days was selected as the best time to observe the experiment. H PDLFs grew well in the gyroscope. The secretion of ALP in the non-induced group was about 20% lower than that in the normal gravity control group on the 3rd day, and the difference was statistically significant (P 0.05). The secretion of ALP in simulated microgravity group was about 16% lower than that in normal gravity control group. In the bone orientation induction group, normal gravity control group and simulated microgravity group could observe the increase of ALP secretion in a time-dependent manner. There was significant difference (P 0.05), which indicated that bone mimic microgravity could decrease the secretion of ALP and OC in h PDLFs, but bone inducer could weaken the decreasing trend. (4) in the simulated microgravity group, the expression of m RNA and protein of MMP8 in PDLFs was higher than that in the normal gravity group, and the expression of MMP8 in the group of bone orientation induction was higher than that in the non-induced group. On the 3rd day, the difference was statistically significant (P 0.05). Conclusion in the simulated microgravity environment, the ability of reducing the bone differentiation of h PDLFs is reduced, and the expression of MMP8 is higher than that of the normal gravity group. The results indicated that MMP8 was involved in regulating the osteogenic differentiation of h PDLFs in simulated microgravity environment.
【作者单位】：解放军第306医院全军口腔疾病诊治中心;
【基金】：航天医学基础与应用国家重点实验室开放课题(SMFA14K03) 国家自然科学基金课题(81341106)
【分类号】：R852.22
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