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目的研究鬼臼毒素纳米脂质载体(POD-NLC)对永生化人宫颈上皮细胞(H8细胞)的凋亡诱导作用及机制。方法采用超声乳化法制备POD-NLC,分别以不同质量浓度(0.000 1、0.001、0.01、0.1、1μg/ml)的POD-NLC和鬼臼毒素(POD)处理H8细胞,采用MTT法检测细胞增殖抑制率,流式细胞术(FCM)检测细胞凋亡率,荧光显微镜和透射电镜观察细胞形态变化。结果 POD-NLC和POD干预后均能抑制H8细胞增殖,且呈时间和剂量依赖性,在同一时间相同浓度条件下,POD-NLC干预后细胞增殖抑制率明显高于POD干预后;0.01μg/ml的POD-NLC干预H8细胞24、48 h后细胞凋亡率均明显高于POD。POD-NLC和POD干预H8细胞48 h后,荧光显微镜和透射电镜下观察均出现核固缩、染色质高度凝聚、凋亡小体等典型的凋亡形态改变。结论与POD相比,POD-NLC对H8细胞具有更强的增殖抑制和凋亡诱导效果,其机制可能为POD经NLC包裹后对组织的黏附性增大,更易黏附于细胞表面,细胞内的药物浓度提高。
Objective To study the apoptosis-inducing effect of podophyllotoxin nanoliposome carrier (POD-NLC) on immortalized human cervical epithelial cells (H8) cells and its mechanism. Methods POD-NLC was prepared by phacoemulsification. H8 cells were treated with POD-NLC and podophyllotoxin (POD) with different concentrations (0.000 1, 0.001, 0.01, 0.1 and 1 μg / ml), respectively. Inhibition rate, flow cytometry (FCM) detection of apoptosis rate, fluorescence microscopy and transmission electron microscopy morphological changes. Results POD-NLC and POD could inhibit the proliferation of H8 cells in a time and dose-dependent manner. Under the same concentration of POD-NLC and POD-NLC, the inhibition rate of POD-NLC and POD was significantly higher than that of POD-NLC and 0.01μg / ml POD-NLC intervention H8 cells 24,48 h after apoptosis were significantly higher than the POD. After POD-NLC and POD interfered with H8 cells for 48 h, typical morphological changes of apoptosis such as nuclear pyknosis, highly condensed chromatin and apoptotic bodies were observed under fluorescence microscope and transmission electron microscope. Conclusion Compared with POD, POD-NLC has stronger inhibition on proliferation and induction of apoptosis in H8 cells. The possible mechanism is that the adhesion of POD to tissues increases after POD is encapsulated by NLC, and it is more likely to adhere to the cell surface and intracellular Drug concentration increased.