MTT法测定常通口服液对正常及粘连腹膜成纤维细胞增殖的影响

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)增殖的影响.方法 (1)药物血清的制备:雄性SD大鼠20只,随机分为4组:正常血清组,CTOL低、中、高剂量组.大鼠灌服相应剂量的药物7d后,腹主动脉采血,分离药物血清.(2)成纤维细胞的体外培养: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.(3)成纤维细胞的增殖:细胞以5×10~3/孔接种到96孔培养板,分别加入CTOL低、中、高剂量的含药血清和空白对照血清,培养24、48、72、96h,MTT法观察细胞增殖情况.结果 药物剂量与吸光度值之间存在负相关性,同时药物剂量与抑制率之间存在正相关性.与NF正常血清对照组比较,CTOL各剂量组在作用24、48h时,其OD值均无显著性变化(P>0.05),但当作用72、96 h时,其OD值均呈显著性降低(P<0.01);与AF正常血清对照组比较,CTOL低、中、高剂量组药物血清作用24、48、72、96h,其0D值均呈显著性降低(P<0.05),同一时间点比较,CTOL低剂量对AF的抑制率比对NF的抑制率高,但无统计学差异.CTOL中剂量在作用96h,对AF的抑制率与NF的抑制率比较,有统计学差异(P<0.05).CTOL高剂量在作用72、96 h,对AF的抑制率与NF的抑制率比较,均有统计学差异(P<0.05).结论 CTOL能够抑制体外培养的NF和AF增殖.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.

Abstract：

Objective To investigate the effect of Changtong oral liquid (CTOL) on the proliferation of cultured fibroblasts derived from normal peritoneum (NFs) and adhesive peritoneum (AFs) of rats. Methods Twenty male SD rats were randomized into 4 groups, including a normal serum group and 3 CTOL groups with CTOL treatment at low, medium or high doses. Serum samples were obtained from the abdominal arteries of the rats after oral treatment with CTOL for 7 days. The fibroblasts were isolated from the peritoneum by means of tissue culture, and the passage 3-8 cells were cultured with the sera of the normal control and CTOL groups for 24, 48, 72 and 96 h. MTT assay was used to observe the proliferation of the fibroblasts. Results The dose of CTOL was inversely correlated to the absorbance but positively to the growth inhibition rates.Compared with the NFs cultured in normal control rat serum, the NFs in serum from CTOL groups showed no obviously changes in the absorbance at 24 and 48 h, but displayed significant reduction at 72 and 96 h (P<0.01). Compared with the AFs in normal rat serum, the AFs in the 3 CTOL groups all showed significantly decreased absorbance at 24, 48, 72 and 96 h (P<0.05). At the same time point, the inhibition rate of AFs in low-dose CTOL group showed no significant difference from that in the normal control group, but CTOL at a medium dose resulted in a significantly higher inhibition rate of AFs at 72 h (P<0.05). High-dose CTOL produced significant differences in the inhibition rates of AFs and NFs (P<0.05). Conclusion CTOL can inhibit the proliferation of AFs and NFs in vitro. AFs appear to be more sensitive to CTOL, which has a dose-dependent inhibitory effect of AF proliferation.

Effect of Changtong oral liquid on fibroblast proliferation in normal and adhesive rat peritoneal tissues

Objective To investigate the effect of Changtong oral liquid (CTOL) on the proliferation of cultured fibroblasts derived from normal peritoneum (NFs) and adhesive peritoneum (AFs) of rats. Methods Twenty male SD rats were randomized into 4 groups, including a normal serum group and 3 CTOL groups with CTOL treatment at low, medium or high doses. Serum samples were obtained from the abdominal arteries of the rats after oral treatment with CTOL for 7 days. The fibroblasts were isolated from the peritoneum by m...