HTRA1在中老年人牙周膜组织中的表达及对细胞增殖的作用

发布时间：2018-02-13 00:12

本文关键词： 丝氨酸蛋白酶牙周膜细胞增殖　出处：《中国老年学杂志》2017年24期 　论文类型：期刊论文

【摘要】：目的探讨丝氨酸蛋白酶(HTRA)1在中老年人牙周膜组织中的表达及对细胞增殖的作用。方法收集该院口腔颌面外科50岁以上患者的患者前磨牙8颗和第三磨牙4颗,要求牙根结构完整,无明显牙周组织炎症。RT-PCR和免疫组化法分别检测人牙周膜组织中HTRA1 mRNA和蛋白表达情况;组织块法培养原代人牙周膜细胞,3-(4,5-二甲基噻唑-2)-2'5-二苯基四氮唑溴蓝(MTT)法检测增殖情况;慢病毒转染使细胞中HTRA1过表达和基因沉默,以空载体为对照,使用细胞增殖与活性检测试剂盒(CCK8)检测HTRA1对人牙周膜细胞增殖的影响。结果 RT-PCR显示HTRA1 mRNA在牙周膜组织中呈阳性表达;免疫组化染色显示,HTRA1在牙周组织中呈阳性表达,主要表达于牙周膜细胞和细胞外基质中。人牙周膜细胞经体外培养后生长曲线符合该细胞的生长特征。慢病毒转染成功干预HTRA1基因表达后,在培养1、3、5、7、9 d时,过表达组牙周膜细胞增殖速度明显低于过表达空载体对照组,基因沉默组牙周膜细胞增殖速度明显高于基因沉默空载体对照组(P0.05)。结论 HTRA1 mRNA及蛋白均表达于中老年人牙周膜组织中,并对牙周膜细胞增殖起到明显调节作用。
[Abstract]:Objective to investigate the expression of serine protease (HTRA1) in periodontal ligament and its effect on cell proliferation. Methods 8 premolars and 4 third molars were collected from patients over 50 years of age in oral and maxillofacial surgery. HTRA1 mRNA and protein expression in human periodontal ligament tissues were detected by RT-PCR and immunohistochemistry. The proliferation of primary human periodontal ligament cells was detected by tissue block method, and the HTRA1 overexpression and gene silencing were induced by lentivirus transfection, and the empty vector was used as control. The effect of HTRA1 on the proliferation of human periodontal ligament cells was detected by using the cell proliferation and activity test kit CCK8. Results RT-PCR showed that HTRA1 mRNA was positive in periodontal ligament tissue, and the expression of HTRA1 mRNA was positive in periodontal tissue by immunohistochemical staining. It was mainly expressed in periodontal ligament cells and extracellular matrix. The growth curve of human periodontal ligament cells in vitro was in line with the growth characteristics of human periodontal ligament cells. Lentivirus transfection successfully interfered with the expression of HTRA1 gene. The proliferation rate of periodontal ligament cells in overexpression group was significantly lower than that in empty expression vector control group. The proliferation rate of periodontal ligament cells in gene silencing group was significantly higher than that in blank gene silencing vector control group P0.05.Conclusion HTRA1 mRNA and protein are expressed in periodontal ligament tissue of middle-aged and elderly people and play a significant role in regulating the proliferation of periodontal ligament cells.
【作者单位】：锦州医科大学中国人民解放军第306医院研究生培养基地;
【基金】：北京市科委首都市民健康项目(No.Z151100003915091)
【分类号】：R781.4

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