目的通过研究天麻(普通粉、破壁粉、冻干粉)对戊四唑诱导癫痫大鼠海马组织B细胞白血病-淋巴瘤-2基因(B-cell leukemia-iymphoma-2,Bcl-2),B细胞白血病淋巴瘤相关蛋白基因(Bcl-2-associated X protein gene,Bax)、半胱氨酸天冬氨酸蛋白酶...目的通过研究天麻(普通粉、破壁粉、冻干粉)对戊四唑诱导癫痫大鼠海马组织B细胞白血病-淋巴瘤-2基因(B-cell leukemia-iymphoma-2,Bcl-2),B细胞白血病淋巴瘤相关蛋白基因(Bcl-2-associated X protein gene,Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和γ-氨基丁酸转运体1(GAT-1)基因和蛋白的表达影响对天麻神经保护作用机制进行初步探讨,并对三种加工方式的天麻效应差异性进行对比研究。方法以SD大鼠为研究对象,建立戊四唑癫痫模型。随机分为12组,正常组,戊四唑组,丙戊酸钠组,天麻(普通粉、破壁粉、冻干粉)分别对应的低、中、高剂量组(0.5、1.0、2.0 g·kg-1)连续灌胃10 d。用荧光定量聚合酶链式反应法(Real-time PCR)检测三种加工方式的天麻对戊四唑诱导癫痫海马组织Bcl-2、Bax、Caspase-3和GAT-1基因表达的情况;用蛋白免疫印迹法(Western Blot)检测天麻对戊四唑诱导癫痫海马组织Bax、Bcl-2、Caspase-3和GAT-1蛋白表达情况。结果模型组癫痫发作强度均为Ⅵ级,表明造模成功。与模型组比较,丙戊酸钠组及天麻普通粉、破壁粉、冻干粉各剂量组癫痫潜伏时间显著延长(P<0.05),癫痫持续时间显著缩短(P<0.05)。与丙戊酸钠组比较,天麻普通粉高剂量组、天麻破壁粉高剂量组癫痫潜伏时间无显著差异(P<0.05),而天麻冻干粉高剂量组癫痫潜伏时间显著延长(P<0.05),且持续时间与之无显著性差异(P<0.05)。与正常组比较,模型组Bax、Caspase3和GAT-1mRNA及蛋白相对表达量显著升高(P<0.05),Bcl-2 mRNA及蛋白相对表达量显著下降(P<0.05);与模型组比较,丙戊酸钠组和天麻各组可显著降低Bax、Caspase3和GAT-1 mRNA及蛋白相对表达量(P<0.05),显著升高Bcl-2 mRNA及蛋白相对表达量(P<0.05)。结论天麻冻干粉高剂量组效果最佳,天麻破壁粉高剂量组次之,最后天麻普通粉高剂量组。天麻对神经的保护具有剂量依赖性,并与天麻不同加工方式相关,其作用机制与调节Bcl-2、Bax、Caspase-3和GAT-1mRNA及蛋白表达水平有关。展开更多
The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein a...The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein and designated as MGABAT-G, was fished out from the library, the 5’ proximal region and nitron 1 were sequenced and analysed, and low homology was found in the above region between GAT-1 genes from mouse and human except some short conserved sequences. The DNA-protein interactions between DNA fragments containing the conserved sequences in the 5’ proximal region and nuclear proteins from different tissues of mouse were studied by means of gel-shift assay, and Southern-Western blot. The results indicate a possible positive-negative regulation mode controlling the expression of the mouse GAT-1 gene.展开更多
文摘The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein and designated as MGABAT-G, was fished out from the library, the 5’ proximal region and nitron 1 were sequenced and analysed, and low homology was found in the above region between GAT-1 genes from mouse and human except some short conserved sequences. The DNA-protein interactions between DNA fragments containing the conserved sequences in the 5’ proximal region and nuclear proteins from different tissues of mouse were studied by means of gel-shift assay, and Southern-Western blot. The results indicate a possible positive-negative regulation mode controlling the expression of the mouse GAT-1 gene.
