低浓度硝苯地平诱导人牙龈上皮细胞bcl-2,bax及P53表达的实验研究
发布时间：2018-11-17 06:26
【摘要】：[目的]检测低浓度硝苯地平(nifedipine, NIF)作用下人牙龈上皮细胞(human Gin-gival epithelial cells, HGECs)凋亡相关基因B细胞淋巴瘤/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的表达变化,探寻低浓度NIF干预牙龈增生的可能作用。 [方法]牙周手术切除的健康牙龈组织,酶消化法分离培养HGECs;免疫组织化学方法对培养细胞进行细胞鉴定；实时定量PCR法检测不同浓度NIF(0.25μg/ml、0.5μg/ml、1.0μg/ml、1.5μg/ml、3.0μg/ml)处理下HGECs中Bcl-2、Bax及P53的mRNA表达水平。 [结果]酶消化法获得的HGECs在体外培养中生长良好；免疫组织化学显示,HGECs抗角蛋白染色阳性,抗波形蛋白染色阴性,证实原代培养细胞为外胚层来源的牙龈上皮细胞。0.25、0.5、1.0、1.5和3.Oμg/mlNIF分别作用HGECs24h后,HGECs Bcl-2mRNA表达水平与对照组相比均显著降低(P0.05)。且Bcl-2表达呈先低后高变化,即在0.25-1gg/ml范围内,伴随NIF浓度升高,Bcl-2表达持续下降,而随后续NIF浓度的升高,Bcl-2表达逐渐上升。在0.25NIF作用下,Bax及p53mRNA表达与对照组相比明显升高(P0.05),而随着NIF浓度升高,Bax及p53mRNA表达下调(P0.05)。 [结论]低浓度NIF显著干预体外培养的HGECs凋亡相关基因中Bcl-2、Bax及P53的转录表达。NIF通过对P53的表达调控间接实现对Bax的表达调控。
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
本文编号：2336740
[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.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
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