台盼蓝活细胞计数法检测中药复方对血管平滑肌抑制作用

Detection of Inhibiting Effect of Chinese Herbal Compound on Vascular Smooth Muscle by Trypan Blue Cell Counting Assay

目的 观察辛通畅络中药复方对慢性肾脏病心血管危险因素的改善作用。方法 大鼠按每天15 g/kg剂量,灌胃给药,连续灌服10 d,于最后一次给药后2~3h,股动脉放血,分离血清,即为中药血清,经56℃,30 min灭活后,分别配成含10、20、30%中药血清的培养液备用,以正常大鼠血清为对照。用含辛通畅络中药处方不同浓度血清培养血管平滑肌细胞,用台盼蓝活细胞计数法及四甲基偶氮唑蓝（MTT）法分别检测活细胞数目和细胞生长抑制率。结果 实验结果显示：24 h、72 h经不同药物浓度血清培养后,与对照组相比,培养24 h后,20%药物组、30%药物组均较其在统计学上有显著差异（P〈0.01）;培养72 h后培养24 h后,10%药物组较对照组在统计学上有显著差异（P〈0.05）,20%药物组、30%药物组均较其在统计学上有显著差异（P〈0.01）。结论 揭示辛通畅络中药复方抑制血管平滑肌细胞增殖,从而防治慢性肾脏病早期动脉粥样硬化所致的血管重构。

Objective： To observe the improving effect of Xintongchangluo TCM Compound on cardiovascular risk factors of chronic kidney disease. Methods： Rats were orally administered by 15g/kg dose daily for successive 10 days and at 2 to 3 hours after the last ad- ministration, their femoral arteries were fleamed to get their serum, i.e. TCM serum. After the inactivation at the 56℃ and in 30 min, the serum was respectively matched into nutrient solutions, containing 10% , 20% and 30% of TCM serum while the serum of normal rats were taken as a control. The different concentration serums were used to culture vascular smooth muscle cells and the number of viable cells and the cell growth inhibition rate were detected by Trypan blue cell counting assay and methyl thiazoly tetrazolium（MTF）. Results： The test results showed that when the different drug concentration serums cultured in 24 and 72 hours were compared with the control group, 24 - hour cultured serums, containing 20% and 30% of the drug had a statistically significant differences（P 〈 0. 01 ）, and 72 - hour and 24 - hour cultured serums containing 10% of the drug had a statistically significant difference （ P 〈 0. 05 ） when compared with the control group. Both 20% and 30% of the drug groups had a statistically significant difference （P 〈 0. 01 ）. Conclusion ： Xintongchangluo TCM Compound inhibits the proliferation of vascular smooth muscle cells, thus preventing the vascular remodeling caused by early atherosclerosis of chronic kidney disease.