姜黄素纳米粒制剂对髓母细胞瘤Daoy细胞的影响

发布时间：2018-04-19 08:20

本文选题：姜黄素纳米粒制剂 + 髓母细胞瘤　；参考：《重庆医科大学》2014年硕士论文

【摘要】：目的：髓母细胞瘤（Medulloblastoma，，MB）是儿童常见恶性程度极高的中枢神经系统肿瘤之一，临床上以手术治疗为主，但疗效欠佳，病死率高，预后差。已有报道指出姜黄素(Curcumin, Cur)对髓母细胞瘤细胞有较强的抑制效果且基本无毒副作用。但由于姜黄素水溶性低，稳定性差，发挥药效所需用量大的缺点阻碍了其在临床的应用和推广。药学研究证实姜黄素纳米粒制剂（Curcumin nanoparticles，Cur-N）较单纯姜黄素具有更高生物利用度，更好稳定性而用药量更低，但其对髓母细胞瘤的生物学活性是否有所增强尚不明确。本文主要研究姜黄素固体脂质纳米粒制剂对髓母细胞瘤Daoy细胞株的抑制效应，并将其与单纯姜黄素进行对比，以明确其是否具有更高的药效和安全性。方法：体外培养髓母细胞瘤Daoy细胞株并分为4组给予不同的干预措施，分别为空白对照组(不加任何处理的Daoy细胞株)，原药组（加入单纯姜黄素处理），制剂对照组（加入空白纳米微粒载体溶剂处理），制剂组（加入姜黄素纳米粒制剂处理），药物浓度梯度分别为2.5、5.0、10.0、20.0、40.0μM/L，处理时相点为24h、48h、72h,处理后用MTT检测髓母瘤细胞增殖情况的变化；划痕实验和Transwell小室模型检测细胞迁移，流式细胞术检测细胞凋亡率、细胞周期和细胞内钙离子浓度变化。结果：MTT、划痕实验和Transwell小室模型显示：原药组（加入单纯姜黄素处理）及制剂组（加入姜黄素纳米粒复合制剂处理）在不同浓度下作用不同时间对细胞的增殖、迁移有不同程度的抑制效应，在作用48h时对细胞增殖的抑制效应最明显，IC50分别为37.3μM/L、5.2μM/L，与对照组相比差异存在统计学意义（P0.05）；流式细胞术显示：原药组及制剂组对Daoy细胞有明显的抑制效应，可不同程度促进Daoy细胞的凋亡（P0.05），使细胞内钙离子浓度明显升高（P0.05）；抑制效应表现在细胞周期上可见到更多细胞停留在S/G2期（P0.05），甚至出现G2/M期阻滞。其中全部实验均显示制剂组较原药组的抑制效应明显增强，差异具有统计学意义（P0.05）；制剂对照组（加入空白纳米微粒载体溶剂处理）和空白对照组(不加任何处理的Daoy细胞株)相比对细胞的影响无明显变化，差异不具有统计学意义（P0.05）。结论：姜黄素纳米粒制剂明显提高了姜黄素对髓母细胞瘤Daoy细胞的抑制效应，较姜黄素能更大程度的促进细胞凋亡，抑制其增殖和迁移，具有更好的抗肿瘤生物学活性和安全性，也许今后能成为姜黄素的替代品更广泛的应用在髓母细胞瘤的临床治疗上。
[Abstract]:Objective: Medulloblastomaomb (Medulloblastomaomb) is one of the most common malignant central nervous system tumors in children. The main clinical treatment is surgical treatment, but the curative effect is poor, the fatality rate is high, and the prognosis is poor.It has been reported that curcumin (Curcumin, Curcumin) has a strong inhibitory effect on medulloblastoma cells and has no toxic side effects.However, because curcumin has low water solubility and poor stability, the large dosage of curcumin hinders its clinical application and popularization.Pharmacological studies confirmed that curcumin nanoparticles-N- (Curcumin nanoparticles-N) had higher bioavailability, better stability and lower dosage than curcumin. However, it was not clear whether curcumin nanoparticles could enhance the biological activity of curcumin nanoparticles-N in medulloblastoma.The inhibitory effect of curcumin solid lipid nanoparticles on myeloblastoma Daoy cell line was studied and compared with curcumin alone to determine whether curcumin has higher efficacy and safety.Methods: Daoy cell lines of medulloblastoma were cultured in vitro and divided into 4 groups.Blank control group (Daoy cell line without any treatment), raw drug group (adding curcumin treatment alone), preparation control group (adding solvent treatment of blank nanoparticles carrier, preparation group (adding curcumin nanoparticles preparation)After treatment, the gradient of drug concentration was 2.5 ~ 5.0 ~ 10.0 ~ 20.0 渭 m / L, and the phase point was 24 h ~ 48 h ~ 72 h. After treatment, the proliferation of medulloblastoma cells was detected by MTT.Cell migration was detected by scratch assay and Transwell chamber model. Apoptosis rate, cell cycle and intracellular calcium concentration were detected by flow cytometry.Results the cell proliferation of the original drug group (treated with curcumin alone) and the preparation group (treated with curcumin nanoparticles compound) at different concentrations and at different time were found by the scratch test and Transwell chamber model.Migration has different inhibitory effects,The inhibitory effects of IC50 on cell proliferation were 37.3 渭 M / L ~ 5.2 渭 M / L, respectively, which were significantly different from those of the control group (P 0.05). Flow cytometry showed that the inhibitory effect on Daoy cells was obvious in the raw drug group and the preparation group.It could promote the apoptosis of Daoy cells to some extent and increase the intracellular calcium concentration of P0.05G, and the inhibitory effect was that more cells could stay in S/G2 phase P0.05N, and even G2 / M phase arrest could be seen in the cell cycle.All the experiments showed that the inhibitory effect of the preparation group was significantly stronger than that of the raw drug group.The difference was statistically significant (P 0.05), but there was no significant difference between the preparation control group (adding solvent treatment with blank nanoparticles carrier) and the blank control group (Daoy cell line without any treatment), and the difference was not statistically significant (P 0.05).Conclusion: curcumin nanoparticles can significantly increase the inhibitory effect of curcumin on Daoy cells of medulloblastoma. Curcumin can promote apoptosis, inhibit the proliferation and migration of myeloblastoma cells to a greater extent than curcumin.It may be used as a substitute for curcumin in clinical treatment of medulloblastoma.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.4

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