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  本文關鍵詞： 絲氨酸蛋白酶 牙周膜 細胞增殖　出處：《中國老年學雜志》2017年24期 　論文類型：期刊論文

【摘要】：目的探討絲氨酸蛋白酶(HTRA)1在中老年人牙周膜組織中的表達及對細胞增殖的作用。方法收集該院口腔頜面外科50歲以上患者的患者前磨牙8顆和第三磨牙4顆,要求牙根結構完整,無明顯牙周組織炎癥。RT-PCR和免疫組化法分別檢測人牙周膜組織中HTRA1 mRNA和蛋白表達情況;組織塊法培養(yǎng)原代人牙周膜細胞,3-(4,5-二甲基噻唑-2)-2'5-二苯基四氮唑溴藍(MTT)法檢測增殖情況;慢病毒轉染使細胞中HTRA1過表達和基因沉默,以空載體為對照,使用細胞增殖與活性檢測試劑盒(CCK8)檢測HTRA1對人牙周膜細胞增殖的影響。結果 RT-PCR顯示HTRA1 mRNA在牙周膜組織中呈陽性表達;免疫組化染色顯示,HTRA1在牙周組織中呈陽性表達,主要表達于牙周膜細胞和細胞外基質中。人牙周膜細胞經體外培養(yǎng)后生長曲線符合該細胞的生長特征。慢病毒轉染成功干預HTRA1基因表達后,在培養(yǎng)1、3、5、7、9 d時,過表達組牙周膜細胞增殖速度明顯低于過表達空載體對照組,基因沉默組牙周膜細胞增殖速度明顯高于基因沉默空載體對照組(P0.05)。結論 HTRA1 mRNA及蛋白均表達于中老年人牙周膜組織中,并對牙周膜細胞增殖起到明顯調節(jié)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.
【作者單位】： 錦州醫(yī)科大學中國人民解放軍第306醫(yī)院研究生培養(yǎng)基地;
【基金】：北京市科委首都市民健康項目(No.Z151100003915091)
【分類號】：R781.4

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本文編號：1506882