新型姜黄素纳米粒影响Lewis肺癌增殖的体内外研究

发布时间：2018-05-19 02:39

  本文选题：姜黄素 + 纳米粒　； 参考：《第三军医大学》2015年硕士论文

【摘要】：背景肺癌是发病率及死亡率最高的恶性肿瘤,且其发病率有增加趋势。化疗药物的耐药性及副作用造成目前肺癌化疗的效果不理想。近年来从传统中药姜黄中提取的姜黄素(curcumin,Cur)成为研究热点,初步研究证实Cur对肺癌、乳腺癌、肝癌等癌症有抑制作用,而且具有副作用小、价格低廉的优点。然而Cur因不稳定和其水溶性差的特点,限制了其临床应用。采用药物制剂技术,制成纳米粒,可以提高Cur的水溶性及稳定性。本研究采用新型姜黄素纳米粒(curcumin nanoparticles,Cur-NPs)作为研究对象,在体内外考察其对Lewis肺癌增殖的影响。目的通过体内外实验考察Cur-NPs对Lewis肺癌增殖的影响,并初步探讨Cur诱导Lewis肺癌凋亡的可能机制。研究方法1.MTT法测定不同浓度的Cur-NPs及Cur对Lewis细胞的增殖抑制作用。2.流式细胞术测定Cur-NPs及Cur对Lewis细胞周期、凋亡、细胞内钙离子浓度、细胞内活性氧及线粒体膜电位的影响。3.建立C57BL/6J小鼠Lewis肺癌模型,考察Cur-NPs及Cur对Lewis肺癌的抑瘤作用。4.Western blot测定Cur在体内外对Lewis肺癌中Akt、Fox O1及Bim蛋白的影响。结果1.Cur-NPs及Cur均可抑制Lewis肺癌的增殖,24h的IC50分别为:10.65μmol/L及34.91μmol/L。2.Cur组和Cur-NPs组抑制Lewis细胞周期于S期比例分别为(81.57±9.15)%、(92.20±7.65)%。Cur组和Cur-NPs诱导Lewis肺癌细胞凋亡比例分别为(7.43±1.13)%、(67.69±9.86)%。Cur组和Cur-NPs组作用Lewis肺癌细胞后细胞内活性氧浓度分别为(2088.01±238.19)、(2568.67±361.85)。Cur组和Cur-NPs组作用Lewis肺癌细胞后细胞内钙离子浓度分别为(52.11±11.28)、(97.85±15.68)。Cur组和Cur-NPs组作用Lewis肺癌细胞后线粒体膜电位分别为(128.97±18.56)、(94.00±12.11)。因此Cur-NPs及Cur均可阻滞Lewis肺癌细胞周期于S期,诱导Lewis肺癌细胞凋亡,升高细胞内钙离子浓度,升高细胞内活性氧浓度,降低细胞线粒体膜电位,且Cur-NPs作用更强(P0.05)。3.给药14天后,处死小鼠,解剖肿瘤称重后,Ct组、Cur组、Cur-NPs和NPs组的瘤重分别为(4.10±0.31)g、(2.50±0.27)g、(1.40±0.19)g和(4.04±0.32)g。Cur组、Cur-NPs组的抑瘤率分别为(39.02±7.63)%和(65.85±10.81)%。因此Cur-NPs及Cur均可抑制小鼠Lewis肺癌的生长,且Cur-NPs抑制生长作用更强(P0.05)。4.Cur在体内外抑制Akt的表达,促进Fox O1及Bim的表达(P0.05)。结论一、在体外Cur-NPs可以明显地增强Cur抑制Lewis肺癌细胞增殖、阻滞细胞周期于S期、诱导凋亡、增高细胞内钙离子浓度、增高细胞内活性氧及降低细胞线粒体膜电位的能力,在体内Cur-NPs可以明显地增强Cur抑制Lewis肺癌生长的能力。Cur-NPs可增强Cur对Lewis肺癌细胞增殖和肿瘤生长的抑制作用。二、Cur可能通过PI3K/Akt-FoxO1-Bim信号通路诱导Lewis肺癌凋亡。
[Abstract]:Background Lung cancer is a malignant tumor with the highest morbidity and mortality. The drug resistance and side effects of chemotherapeutic drugs cause the effect of chemotherapy in lung cancer is not ideal. Curcumin (Curcumin Curcumin) extracted from traditional Chinese medicine turmeric has become a research hotspot in recent years. Preliminary studies have proved that Cur has inhibitory effect on lung cancer, breast cancer and liver cancer, and has the advantages of low side effect and low price. However, the clinical application of Cur is limited by its instability and poor water solubility. The water solubility and stability of Cur can be improved by using pharmaceutical preparation technology to prepare nanoparticles. The effect of curcumin nanoparticles-Cur-NPs on the proliferation of Lewis lung cancer was investigated in vitro and in vivo. Objective to investigate the effect of Cur-NPs on the proliferation of Lewis lung cancer in vitro and in vivo, and to explore the possible mechanism of Lewis lung cancer apoptosis induced by Cur. Methods 1.MTT assay was used to determine the inhibitory effects of Cur-NPs and Cur on the proliferation of Lewis cells. The effects of Cur-NPs and Cur on the cell cycle, apoptosis, intracellular calcium concentration, intracellular reactive oxygen species and mitochondrial membrane potential were determined by flow cytometry. C57BL/6J mouse model of Lewis lung cancer was established to investigate the inhibitory effect of Cur-NPs and Cur on Lewis lung cancer. 4. Western blot was used to determine the effect of Cur on the expression of AkttFox O1 and Bim protein in Lewis lung cancer in vitro and in vivo. Results both 1.Cur-NPs and Cur could inhibit the proliferation of Lewis lung cancer for 24 hours, and the ratio of inhibition of Lewis cell cycle in S phase was 81.57 卤9.15 卤7.65)%.Cur and Cur-NPs induced Lewis lung cancer cell apoptosis was 7.43 卤1.1367.69 卤9.86)%.Cur and Cur-NPs respectively in the group of 10. 65 渭 mol/L and 34.91 渭 mol/L.2.Cur and Cur-NPs. The intracellular concentration of reactive oxygen species (Ros) was 2088.01 卤238.19 (2568.67 卤361.85).Cur) and (52.11 卤11.28) (97.85 卤15.68).Cur) in Lewis lung cancer cells and 128.97 卤18.56 (12.11 卤12.11) in Cur-NPs group respectively. Therefore, both Cur-NPs and Cur can block the cell cycle of Lewis lung cancer cells in S phase, induce apoptosis of Lewis lung cancer cells, increase intracellular calcium concentration, increase intracellular reactive oxygen species concentration, and decrease mitochondrial membrane potential of Lewis lung cancer cells, and Cur-NPs has a stronger effect on Lewis lung cancer cells. After 14 days of administration, the mice were killed. The tumor inhibition rates of Cur-NPs and NPs in curr group and NPs group were 4.10 卤0.31, 2.50 卤0.27, 1.40 卤0.19g and 4.04 卤4.04 卤0.32)g.Cur, respectively. The tumor inhibition rates of Cur-NPs group were 39.02 卤7.63% and 65.85 卤10.81%, respectively. Therefore, both Cur-NPs and Cur could inhibit the growth of mouse Lewis lung cancer, and Cur-NPs inhibited the expression of Akt more strongly in vitro and in vivo, and promoted the expression of Fox O1 and Bim. Conclusion: in vitro, Cur-NPs can significantly enhance the ability of Cur to inhibit the proliferation of Lewis lung cancer cells, block cell cycle in S phase, induce apoptosis, increase intracellular calcium concentration, increase intracellular reactive oxygen species and decrease mitochondrial membrane potential. In vivo, Cur-NPs significantly enhanced the ability of Cur to inhibit the growth of Lewis lung cancer. Cur-NPs enhanced the inhibitory effect of Cur on the proliferation and tumor growth of Lewis lung cancer cells. Cur may induce apoptosis of Lewis lung cancer through PI3K/Akt-FoxO1-Bim signaling pathway.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

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