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目的研究雷公藤内酯醇(Triptolide TP)和雷公藤内酯醇聚合物胶束(Triptolide loaded poly-meric micelles,TP-PM)对人结肠癌HT29细胞增殖及凋亡的作用及机制,评价聚合物胶束作为雷公藤内酯醇纳米载体的可行性。方法用含不同浓度的TP和TP-PM(1-100ng/mL)培养液孵育HT29细胞24h、48h和72h后,利用WST-1、BrdU试剂盒测定细胞的存活率、增殖率;通过测定LDH活性判断细胞膜完整性;利用流式细胞仪测定细胞死亡、存活和凋亡率;并通过Caspase-Glo3/7定量试剂盒测定细胞caspase3/7的表达情况。结果 TP和TP-PM对HT29细胞活性和细胞增殖的抑制作用显著,呈明显的剂量、时间依赖性,TP-PM对HT29的抑制作用优于TP(P<0.05)。TP和TP-PM对HT29细胞膜的破坏轻微。FDA-PI双染结果显示,两组中FDA阳性细胞均呈现时间、剂量依赖性的下降(P<0.01);PI阳性细胞率呈现时间、剂量依赖性的上升(P<0.01);FDA、PI双阴性细胞也明显上升,但不呈时间、剂量依赖关系。Caspase测定结果显示TP和TP-PM均可引起Caspase3/7活性增高,TP-PM组引起的凋亡作用强于TP组(P<0.05)。结论 TP和TP-PM可以通过诱导凋亡的方式抑制肿瘤细胞生长和增殖,且TP-PM的作用强于TP,聚合物胶束是一种极具前景的载药体系。
Objective To investigate the effects and mechanisms of triptolide TP and triptolide loaded poly-meric micelles (TP-PM) on proliferation and apoptosis of human colon carcinoma HT29 cells, Beam as a triptolide nanocarrier feasibility. Methods HT29 cells were incubated with different concentrations of TP and TP-PM (1-100ng / mL) for 24h, 48h and 72h. The survival rate and proliferation rate of HT29 cells were measured by WST-1 and BrdU kit. The activity of cell membrane was determined. The cell death, survival and apoptosis rate were determined by flow cytometry. Caspase-Glo3 / 7 kit was used to determine the expression of caspase 3/7. Results TP and TP-PM significantly inhibited HT29 cell proliferation and cell proliferation in a dose-and time-dependent manner. The inhibitory effect of TP-PM on HT29 was better than that of TP (P <0.05). TP and TP-PM destruction of HT29 cell membrane minor. The results of FDA-PI double staining showed that both the time and dose-dependent decrease of FDA-positive cells were observed in both groups (P <0.01); the rate of PI-positive cells increased in a dose- and time-dependent manner (P <0.01) Double-negative cells also significantly increased, but not in a time-and dose-dependent manner. The results of Caspase assay showed that both TP and TP-PM increased the activity of Caspase3 / 7, and the apoptosis induced by TP-PM was stronger than that of TP (P <0.05). Conclusion TP and TP-PM can inhibit the growth and proliferation of tumor cells by inducing apoptosis, and the effect of TP-PM is stronger than that of TP. Polymer micelles is a promising drug-carrying system.