养阴润肠方对小鼠结肠ICC凋亡、周期调控作用机制研究

发布时间：2019-05-24 13:49

【摘要】：目的：观察不同剂量养阴润肠方含药血清对离体培养的小鼠结肠Cajal间质细胞(Interstitial Cells of Cajal ICC)凋亡和周期调控的影响,为养阴润肠方治疗慢传输型便秘(slow transit constipation STC)提供理论依据。方法：将20只BALB/c小鼠随机分为四组,原代分离和培养每组小鼠结肠组织,通过细胞流式分选技术分选出成熟的ICC,用提前制备好的正常大鼠血清及高、中、低剂量养阴润肠方含药血清对ICC进行分组干预,干预2d后观察ICC数量及其形态学改变,并运用流式检测技术评定ICC凋亡机制和细胞周期调控作用。结果：各组培养所得细胞经相应的干预后,镜下观察后统计高、中、低及空白剂量组细胞平均计数分别为(13.94±2.64；10.17±1.33；8.17±1.47；6.33±1.63),高剂量和中剂量组细胞数量较空白组表达升高(P0.01),低剂量组与—表达无明显差异(P0.05),且低剂量组与高剂量组存在统计学差异(P0.01),中剂量组与高剂量组和低剂量组之间无统计学差异(P0.05)。在细胞凋亡作用上,早凋亡和晚凋亡之总和为总凋亡率,空白组、养阴润肠方含药血清高、中、低剂量组早期和晚期凋亡和为16.9%、9.5%、12.4%、12.6%,总凋亡率：空白组低剂量组中剂量组高剂量组。养阴润肠方血清能有效减少cajal细胞凋亡。细胞周期调控作用中,空白组、养阴润肠方含药血清高、中、低剂量组S期和G2期细胞总和所占比分别为36.52%、41.02%、43.86%、82.40%,含药血清组内正处于复制活动期的ICC数量所占比例高剂量组中剂量组低剂量组空白剂量组。结论：养阴润肠方能促进ICC数量的增长,并能有效地减少ICC凋亡数量和提高细胞分裂能力及活性,其通便机制可能为通过促进ICC的数量生长及减少其凋亡而达到促进肠动力,为治疗STC寻求一条新的途径。
[Abstract]:Objective: to observe the effect of different doses of Yangyin Runchang recipe containing serum on (Interstitial Cells of Cajal ICC) apoptosis and cycle regulation of mouse colon Cajal Leydig cells in vitro. It provides a theoretical basis for Yangyin Runchang recipe in the treatment of slow transit constipation (slow transit constipation STC). Methods: 20 BALB/c mice were randomly divided into four groups. The colon tissue of each group was isolated and cultured. Mature ICC, was separated by flow cytometry and normal rat serum was prepared in advance. The serum containing low dose Yangyin Runchang recipe was divided into groups to intervene ICC. After 2 days of intervention, the number and morphological changes of ICC were observed, and the apoptosis mechanism and cell cycle regulation of ICC were evaluated by flow cytometry. Results: after the corresponding intervention, the average cell count of the cultured cells in the middle, low and blank dose groups was (13.94 卤2.64) 鈮，

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