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目的探讨黄连提取物对HaCat细胞凋亡及核因子κB(NF-κB)基因转录水平的影响。方法以41.6,20.8,10.4μg.mL-1黄连提取物作用于HaCat细胞,采用噻唑蓝(MTT)法测定HaCat细胞增殖情况,DNA Ladder法检测黄连提取物对HaCat细胞凋亡的影响,半定量逆转录聚合酶链反应(RT-PCR)测定给药后吸光度值,计算各组NF-κB/磷酸甘油醛脱氢酶(NF-κB/GAPDH)值,分析NF-κB基因转录水平变化。结果黄连提取物对HaCat细胞增殖具有抑制作用,抑制强度呈浓度依赖性,41.6μg.mL-1黄连提取物抑制作用最强(抑制率66.17%)。不同剂量黄连提取物导致HaCat细胞呈阶梯状DNA,呈明显凋亡特征。41.6和20.8μg.mL-1黄连提取物对NF-κB基因转录具有显著下调作用。结论黄连提取物可诱导HaCat细胞凋亡,并下调NF-κB基因转录水平。
Objective To investigate the effects of Coptidis Rhizoma extract on the apoptosis of HaCat cells and the transcriptional level of nuclear factor-κB (NF-κB). Methods HaCat cells were treated with extracts of Coptis chinensis at 41.6, 20.8 and 10.4μg.mL-1 respectively. The proliferation of HaCat cells was determined by MTT assay. The effect of Coptidis Rhizoma extract on the apoptosis of HaCat cells was detected by DNA Ladder method. The absorbance values after administration were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The NF-κB / GAPDH values were calculated and the changes of NF-κB gene transcription were analyzed. Results Coptis extract inhibited the proliferation of HaCat cells in a concentration-dependent manner. The extract of Coptidis Rhizoma with 41.6μg.mL-1 had the strongest inhibitory effect (inhibition rate of 66.17%). Different doses of Coptis extract led to ladder-like DNA HaCat cells showed obvious apoptotic characteristics. 41.6 and 20.8μg.mL-1 Coptis extract on NF-κB gene transcription was significantly down-regulated. Conclusions Coptis extract can induce apoptosis of HaCat cells and down-regulate NF-κB gene transcription.