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目的:探讨常通口服液(CTOL)对体外培养的正常腹膜成纤维细胞(NF)及粘连腹膜成纤维细胞(AF)周期的影响。方法:雄性SD大鼠15只,随机分为正常血清组、CTOL中、高剂量组。大鼠灌服相应剂量的药物7d后采血,分离药物血清;采用组织块贴壁培养法进行成纤维细胞培养,本实验所用细胞为第3-8代;取对数生长期的NF及AF,分别加入正常对照血清,CTOL中、高剂量含药血清,培养12、24、48h后,流式细胞仪检测分析成纤维细胞周期。结果:与正常血清组比较,CTOL中剂量作用12、24、48h对NF的DNA合成前期(G1期)细胞比例无显著影响,而高剂量含药血清组的NF的G1期细胞比例显著增加(P<0.001);CTOL中、高剂量含药血清组的AF在12、24、48h的G1期细胞比例均显著升高(P<0.01)。而S期及G2期细胞比例呈下降趋势,但各组间无显著性差异。结论:CTOL可以抑制体外培养的NF和AF增殖,调整细胞周期。CTOL对成纤维细胞增殖的抑制率之间存在剂量依赖关系,而且,药物血清对AF作用更敏感。
Objective: To investigate the effects of Changtong oral liquid (CTOL) on the culture period of normal peritoneal fibroblasts (NF) and adherent peritoneal fibroblasts (AF). Methods: Fifteen male Sprague-Dawley rats were randomly divided into normal sera, CTOL medium and high dose groups. The rats were fed with the corresponding dose of the drug for 7 days and then blood was collected to separate the drug serum. The fibroblasts were cultured using the tissue block adherent culture method. The cells used in this experiment were from the 3rd to the 8th generation; NF and AF in the logarithmic growth phase were taken. The normal control serum, CTOL medium and high-dosage drug-containing serum were added to cultured cells. After 12, 24, and 48 hours of culture, the fibroblast cell cycle was analyzed by flow cytometry. RESULTS: Compared with the normal sera group, the CTOL medium dose effect at 12, 24, and 48h had no significant effect on the proportion of NF DNA in the early stage of DNA synthesis (G1 phase), while the high-dose drug serum group showed a significant increase in the proportion of NF G1 phase cells ( P<0.001); CTOL medium- and high-dose sera contained AF at the 12th, 24th, and 48th hour in the G1 phase (all P<0.01). The proportion of cells in S phase and G2 phase showed a downward trend, but there was no significant difference among the groups. Conclusion: CTOL can inhibit the proliferation of NF and AF cultured in vitro and regulate the cell cycle. There was a dose-dependent relationship between CTOL inhibition of fibroblast proliferation and drug serum was more sensitive to AF.