【摘要】：[目的]檢測(cè)低濃度硝苯地平(nifedipine, NIF)作用下人牙齦上皮細(xì)胞(human Gin-gival epithelial cells, HGECs)凋亡相關(guān)基因B細(xì)胞淋巴瘤/白血病(B cell lym-Phoma/leukmia, Bcl-2)基因,Bcl-2同源促凋亡基因Bax (Bcl-2associated proteinX,13ax)及p53的表達(dá)變化,探尋低濃度NIF干預(yù)牙齦增生的可能作用。 [方法]牙周手術(shù)切除的健康牙齦組織,酶消化法分離培養(yǎng)HGECs;免疫組織化學(xué)方法對(duì)培養(yǎng)細(xì)胞進(jìn)行細(xì)胞鑒定；實(shí)時(shí)定量PCR法檢測(cè)不同濃度NIF(0.25μg/ml、0.5μg/ml、1.0μg/ml、1.5μg/ml、3.0μg/ml)處理下HGECs中Bcl-2、Bax及P53的mRNA表達(dá)水平。 [結(jié)果]酶消化法獲得的HGECs在體外培養(yǎng)中生長(zhǎng)良好；免疫組織化學(xué)顯示,HGECs抗角蛋白染色陽(yáng)性,抗波形蛋白染色陰性,證實(shí)原代培養(yǎng)細(xì)胞為外胚層來(lái)源的牙齦上皮細(xì)胞。0.25、0.5、1.0、1.5和3.Oμg/mlNIF分別作用HGECs24h后,HGECs Bcl-2mRNA表達(dá)水平與對(duì)照組相比均顯著降低(P0.05)。且Bcl-2表達(dá)呈先低后高變化,即在0.25-1gg/ml范圍內(nèi),伴隨NIF濃度升高,Bcl-2表達(dá)持續(xù)下降,而隨后續(xù)NIF濃度的升高,Bcl-2表達(dá)逐漸上升。在0.25NIF作用下,Bax及p53mRNA表達(dá)與對(duì)照組相比明顯升高(P0.05),而隨著NIF濃度升高,Bax及p53mRNA表達(dá)下調(diào)(P0.05)。 [結(jié)論]低濃度NIF顯著干預(yù)體外培養(yǎng)的HGECs凋亡相關(guān)基因中Bcl-2、Bax及P53的轉(zhuǎn)錄表達(dá)。NIF通過(guò)對(duì)P53的表達(dá)調(diào)控間接實(shí)現(xiàn)對(duì)Bax的表達(dá)調(diào)控。
[Abstract]:[objective] to detect the human gingival epithelial cells (human Gin-gival epithelial cells, HGECs) apoptosis-related gene B cell lymphoma / leukemia (B cell lym-Phoma/leukmia, Bcl-2) gene induced by low-concentration nifedipine (nifedipine, NIF). To explore the possible role of low concentration NIF in gingival hyperplasia, the expression changes of Bax (Bcl-2associated proteinX,13ax) and p53 in Bcl-2 homologous apoptotic gene. [methods] healthy gingival tissues were removed by periodontal surgery. HGECs; was isolated and cultured by enzyme digestion. The cultured cells were identified by immunohistochemical method. The mRNA expression of Bcl-2,Bax and p53 in HGECs treated with different concentrations of NIF (0.25 渭 g / ml, 0.5 渭 g / ml, 1.0 渭 g / ml, 1.5 渭 g / ml, 3.0 渭 g/ml) was detected by real-time quantitative PCR. [results] HGECs obtained by enzyme digestion grew well in vitro. Immunohistochemical staining showed that HGECs anti-keratin staining was positive, and anti-vimentin staining was negative, which confirmed that primary cultured gingival epithelial cells were derived from ectoderm. After HGECs24h was treated with 0.25 渭 g/mlNIF, 0.5 渭 g/mlNIF and 3.0 渭 g/mlNIF, respectively. The expression of HGECs Bcl-2mRNA was significantly lower than that of control group (P0.05). The expression of Bcl-2 decreased at first and then increased at the same time. In the range of 0.25-1gg/ml, the expression of Bcl-2 decreased with the increase of NIF concentration, and then the expression of Bcl-2 increased gradually with the increase of NIF concentration. The expression of Bax and p53mRNA in 0.25NIF group was significantly higher than that in control group (P0.05), but the expression of Bax and p53mRNA was down-regulated with the increase of NIF concentration (P0.05). [conclusion] low concentration NIF significantly interferes with the transcription and expression of Bcl-2,Bax and p53 in HGECs apoptosis-related genes cultured in vitro. NIF indirectly regulates the expression of Bax through the regulation of p53 expression.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 文海燕;束蓉;蔣少云;姜云濤;;硝苯地平對(duì)人牙齦上皮細(xì)胞bcl-2基因表達(dá)的影響[J];上�？谇会t(yī)學(xué);2010年01期

2 王文 ,劉力生 ,龔蘭生 ,張廷杰 ,吳寧;我國(guó)大樣本隨機(jī)臨床試驗(yàn)治療心腦血管病的效價(jià)評(píng)估[J];中華心血管病雜志;2003年01期

3 張廷杰;高血壓干預(yù)試驗(yàn)──硝苯地平與安慰劑隨機(jī)對(duì)照研究[J];中華心血管病雜志;1994年03期

，

本文編號(hào)：2336740