目的:考察厚朴汁炙远志炮制品含药血清对胃cajal间质细胞(interstitial cells of cajal,icc)活力及胞内nNOS表达的影响,从细胞水平阐释其降低胃动力障碍的生物学机制。方法:采用体外培养胃cajal闻质细胞,再加入含药血清与之共培养的方法开展实验。实验分为空白组、厚朴10g/kg组、远志20g/kg组、厚朴汁炙远志炮制品高剂量30∥kg组4个不同组,采用MTT比色法检测细胞活性、Western-Blot法分析各药对细胞内nNOS表达的影响。结果:远志20g/kg组ICC细胞的OD值明显低于空白组,且细胞突起变短,网状脱落;而厚朴10g/kg组、厚朴汁炙远志30g/kg组细胞OD值明显高于空白组和单味远志组。免疫印迹法检测结果显示,远志20g/,kg组的nNOS的表达与空白组比较明显增强,厚朴10g/kg组的表达明显下降,远志20g/kg组呈下降趋势。结论:远志能抑制胃动力起搏细胞活性,抑制胃间质细胞nNOS表达增强;而厚朴汁炙远志能明显促进细胞活性、下调nNOS表达从而缓解胃动力障碍。
Objective: To study the Polygalae decoction processed by Magnolia officinalis medicated production on cultured gastric interstitial cells of Cajal ( interstitial ceils of Cajal, ICC) of the expression of nNOS to explain the cellular mechanisms of NO gastrointestinal dysfunction i. Methods: ICCs were cultured in vitro, then adding method containing serum and co culture experiments. The experiment was divided into blank group, Magnolia 10 g/kg group, Polygala 20 g/kg group, Polygalae decoction processed by Magnolia 30g/kg group such four different groups, using the MTT colorimetric detection cell activity, Western-Blot method of intracellular the expression of nNOS analysis. Results: Compared with blank group, Polygala 20 g/kg group of cells'OD values was significantly lower (P 〈 0.05, P 〈 0.01 ) , and shorter cell processes, reticular off; while Magnolia 10 g/kg group, Polygala decoction processed by Magnolia products high-dose 30 g/kg group cells OD values were significantly higher( P 〈 0.05, P 〈 0.01 ). Western blot showed that compared with the blank group, the expression of nNOS of Polygala group was significantly enhanced, Magnolia 10 g/kg group was significantly decreased, Polygala decoction processed by Magnolia 30 g/kg group declined. Conclusion: Polygalae can inhibit tile activity of ICC motility, and the expression of nNOS; and Polygalae decoction processed by Magnolia can obviously promote the down-regulation of nNOS expression in order to ease the cell activity, gastric motility disorders.