胡椒碱脂质体的制备及对于胃癌细胞体外抗肿瘤活性的作用研究

doi:10.12092/j.issn.1009-2501.2018.11.007

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (11): 1240-1245.doi: 10.12092/j.issn.1009-2501.2018.11.007

胡椒碱脂质体的制备及对于胃癌细胞体外抗肿瘤活性的作用研究

郭丽¹，韩晨阳²

¹嘉兴市第二医院中心实验室，²药学部，嘉兴 314001，浙江

收稿日期:2018-07-09 修回日期:2018-09-13 出版日期:2018-11-26 发布日期:2018-11-22
通讯作者: 韩晨阳，男，硕士，药师，研究方向：药理学。 E-mail：691513770@qq.com
作者简介:郭丽，女，硕士，主管技师，研究方向：药理学，分子生物学。 E-mail：taishanglg@126.com
基金资助:
浙江省科技厅公益类实验动物项目(2017C37174)

Preparation of piperine liposomes and its antitumor activity of gastric cancer cells in vitro

GUO Li¹, HAN Chenyang ²

¹ Central Laboratory, ²Department of Pharmacy, the Second Hospital of Jiaxing，Jiaxing 314001，Zhejiang, China

Received:2018-07-09 Revised:2018-09-13 Online:2018-11-26 Published:2018-11-22

摘要/Abstract

摘要：

目的: 研究胡椒碱脂质体的制备方法及胡椒碱脂质体体外抗胃癌细胞活性及作用机制。方法: 利用旋转薄膜冻融法制备胡椒碱脂质体，采用透射电镜（TEM）观察脂质体的形态，粒径分析仪和Zeta电位测定仪分析胡椒碱脂质体的粒径和Zeta电位，进行脂质体的稳定性实验、体外释药特性考察。体外培养人胃肿瘤MKN45细胞，分为正常组、对照组、实验组，对照组使用50 μmol/L的胡椒碱干预，实验组使用等剂量的胡椒碱脂质体干预。采用CCK-8法检测细胞活力，流式细胞术检测细胞凋亡水平，蛋白免疫印迹法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3、Caspase-9的表达水平，Hoechst 33342染色观察细胞染色。结果: 胡椒碱脂质体形态为类球状，平均粒径为(92.32±2.10) nm，Zeta电位为(-49.8±3.5) mV，体外释药实验显示胡椒碱脂质体释药速率显著高于胡椒碱原料药；细胞实验结果显示，对照组和实验组细胞活力相比正常组显著降低，具有时间依赖性（P<0.05），而实验组相比对照组具有统计学意义（P<0.05）；流式细胞术检测，对照组和实验组细胞凋亡率高于正常组，而实验组显著高于对照组（P<0.05）；对照组和实验组中Bcl-2水平显著下调，而Bax、Caspase-3、Caspase-9的表达显著上调，与正常组比较，差异具有统计学意义（P<0.05）。结论: 胡椒碱脂质体释药速率高于原料药，说明脂质体对于胡椒碱起到了增溶作用，而胡椒碱脂质体体外诱导MKN45凋亡能力高于原料药。

关键词: 脂质体, 胡椒碱, 胃癌, 抗肿瘤活性

Abstract:

AIM: To study the preparation method of piperine liposomes and the mechanism on anti-tumor activity of gastric cancer cells in vitro. METHODS: Piperine liposomes were prepared by freezing and thawing method. The morphology of liposomes was observed by transmission electron microscopy (TEM). Particle size analyzer and Zeta potential analyzer were used to analyze the particle size and Zeta potential of piperine liposomes, and to investigate the stability of liposomes and the special release of drug in vitro. Human gastric cancer cell MKN45 cells were cultured in vitro and divided into normal group, control group and experimental group. The control group was treated with 50 μmol/L piperine, the experimental group was treated with the liposomes of piperine, the cell viability was detected by CCK-8, the level of apoptosis was detected by flow cytometry, the expression level of apoptosis related protein Bcl-2, Bax, Caspase-3, Caspase-9 was detected by protein immunoblotting, and Hoechst 33342 staining was used to observe the details. RESULTS: The morphology of piperine liposomes was spheroid, the average particle size was （92.32+2.10）nm, and the Zeta potential was（-49.8+3.5）mV. The release rate of piperine liposomes was significantly higher than that of piperine raw material in vitro. The results of cell test showed that the cell viability of the control group and the experimental group was significantly lower than that in the normal group in time dependent manner (P<0.05), the experimental group had statistical significance compared with the control group (P<0.05). The apoptosis rate of the control group and the experimental group was higher than that of the normal group, but the experimental group was significantly higher than that of the control group (P<0.05). The levels of Bcl-2 in the control group and the experimental group were significantly decreased, while the expressions of Bax, Caspase-3 and Caspase-9 were significantly increased. Compared with the normal group, it was statistically significant (P<0.05). CONCLUSION: The release rate of piperine liposomes is higher than that of the drug, which shows that liposomes increase the solubility of piperine, and the ability of piperine liposome to induce MKN45 apoptosis in vitro is higher than that of the drug.

Key words: liposomes, piperine, gastric cancer, antitumor activity

中图分类号:

R965.2

郭丽，韩晨阳. 胡椒碱脂质体的制备及对于胃癌细胞体外抗肿瘤活性的作用研究[J]. 中国临床药理学与治疗学, 2018, 23(11): 1240-1245.

GUO Li, HAN Chenyang. Preparation of piperine liposomes and its antitumor activity of gastric cancer cells in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(11): 1240-1245.

参考文献

［1］Atal S, Atal S, Vyas S, et al.Bio-enhancing effect of piperine with metformin on lowering blood glucose level in alloxan induced diabetic mice［J］. Pharmacognosy Res，2016, 8(1):56-60.
［2］Toyoda T, Shi L, Takasu S, et al.Anti-inflammatory effects of capsaicin and piperine on helicobacter pylori-induced chronic gastritis in mongolian gerbils［J］.Helicobacter,2016, 21(2):131-142.
［3］Liu J, Chen M, Wang X, et al. Piperine induces autophagy by enhancing protein phosphotase 2A activity in a rotenone-induced Parkinson's disease model［J］.Oncotarget,2016, 7(38):60823-60843.
［4］Dhivya V, Priya LB, Chirayil HT, et al. Piperine modulates isoproterenol induced myocardial ischemia through antioxidant and anti-dyslipidemic effect in male Wistar rats［J］.Biomed Pharmacother，2017, 87:705-713.
［5］Johnson JJ, Nihal M, Siddiqui IA, et al.Enhancing the bioavailability of resveratrol by combining it with piperine［J］. Mol Nutr Food Res, 2011, 55(8):1169-1176.
［6］Lambert JD, Hong J, Kim DH, et al.Piperine enhances the bioavailability of the tea polyphenol (-)-epigallocatechin-3-gallate in mice［J］. J J Nutr, 2004, 134(8):1948-1952.
［7］Zununi VS, Salehi R, Davaran S, et al. Liposome-based drug co-delivery systems in cancer cells［J］. Mater Sci Eng C Mater Biol Appl, 2016, 71:1327-1341.
［8］Xing Y, Wen C, Li S, et al. Non-viral liposome-mediated transfer of brain-derived neurotrophic factor across the blood-brain barrier［J］. Neural Regen Res，2016, 11(4):617-622.
［9］Tak JK, Lee JH, Park JW. Resveratrol and piperine enhance radiosensitivity of tumor cells［J］. Bmb Reports, 2012, 45(4):242-246.
［10］Li C, Wang Z, Wang Q, et al. Enhanced anti-tumor efficacy and mechanisms associated with docetaxel-piperine combination-in vitro and in vivo investigation using a taxane-resistant prostate cancer model［J］. Oncotarget, 2018, 9(3):3338-3352.
［11］Selvendiran K, Banu SM, Sakthisekaran D. Protective effect of piperine on benzo( a )pyrene-induced lung carcinogenesis in Swiss albino mice［J］. Clin Chim Acta，2004, 350(1/2):73-78.
［12］Sehgal A, Kumar M, Jain M, et al.Modulatory effects of curcumin in conjunction with piperine on benzo(a)pyrene-mediated dna adducts and biotransformation enzymes［J］.Nutr Cancer，2013, 65(6):885-890.
［13］Khatri S, Awasthi R.Piperine containing floating microspheres: an approach for drug targeting to the upper gastrointestinal tract.［J］. Drug Deliv Transl Res, 2016, 6(3):1-9.
［14］刘庆生，俞建顺，蔡丹莉，等.大鼠胃癌前病变形成过程中胃黏膜Sonic Hedgehog信号通路的变化［J］. 中国临床药理学与治疗学, 2018, 23(7): 761-769.
［15］陈淑慧, 阳帆帆, 何君, 等.胃癌靶向治疗研究最新进展［J］.中国临床药理学与治疗学, 2015, 20(8): 950-955.

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胡椒碱脂质体的制备及对于胃癌细胞体外抗肿瘤活性的作用研究

Preparation of piperine liposomes and its antitumor activity of gastric cancer cells in vitro

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