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目的从能量代谢方面探讨胃康宁治疗功能性消化不良的可能作用机制。方法人胃黏膜上皮细胞GES-1采用3条靶向电压依赖性阴离子通道蛋白1(VDAC1)的siRNA转染细胞后检测其沉默效率;转染DDK-Myc质粒、VDAC1-DDK-Myc质粒检测VDAC1的过表达效率。GES-1细胞以1×10~4个细胞铺96板,分为空白对照组、阳性对照组和胃康宁低、中、高剂量组,每组5个平行孔。空白对照组不加任何药物,阳性对照组细胞沉默或过表达后加多潘立酮片2.5μmol,胃康宁低、中、高剂量组细胞沉默或过表达后分别加0.025、0.100、0.300μg/μl。24 h后利用生物发光法检测各组沉默或过表达前后细胞ATP的含量。结果siRNA1转染能降低VDAC1基因和蛋白水平,DDK-Myc质粒、VDAC1-DDK-Myc质粒转染能均能升高VDAC1蛋白水平,与对照组比较差异有统计学意义(P<0.01)。各组细胞沉默VDAC1后ATP含量均较本组沉默前降低(P<0.05或P<0.01)。各组细胞过表达VDAC1后ATP含量较本组转染前明显升高(P<0.05);与空白对照组VDAC1-DDK-Myc转染后比较,胃康宁中剂量VDAC1-DDK-Myc转染后细胞中ATP含量明显升高(P<0.05)。结论胃康宁可能通过促进胃上皮细胞能量代谢治疗功能性消化不良。
Objective To investigate the possible mechanism of Weikangning’s treatment of functional dyspepsia from energy metabolism. Methods Human gastric mucosal epithelial cells GES-1 were transfected with three siRNAs targeting voltage-dependent anion channel protein 1 (VDAC1) and their silencing efficiencies were determined. After transfected with DDK-Myc plasmid and VDAC1-DDK-Myc plasmid, VDAC1 The over-expression efficiency. GES-1 cells were plated on 96 plates at 1 × 10 4 cells and divided into blank control group, positive control group and Wepening low, medium and high dose groups with 5 parallel wells in each group. The blank control group without any drugs, the positive control group cells were silenced or overexpressed more domperidone tablets 2.5μmol, Weike Corning low, medium and high doses of cells were silenced or overexpression were added 0.025,0.100,0.300μg / μl. After 24 h, the bioluminescence method was used to detect the content of ATP in the cells before and after silencing or overexpression. Results siRNA1 transfection reduced VDAC1 gene and protein levels. The transfection of DDK-Myc and VDAC1-DDK-Myc increased the level of VDAC1 protein, which was significantly different from the control (P <0.01). The content of ATP in each group of cells after silencing VDAC1 was significantly lower than that before the silence (P <0.05 or P <0.01). Compared with the control group VDAC1-DDK-Myc after transfection, the content of ATP in VDAC1-overexpression group was significantly higher than that in the control group (P <0.05) ATP content in cells was significantly increased (P <0.05). Conclusion Weikangning may treat functional dyspepsia by promoting the energy metabolism of gastric epithelial cells.