Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells

ZHANG Liming; MA Yage; NIU Ruonan; ZHANG Xi; ZHAO Shenglan; CHEN Chaoyin

doi:10.13386/j.issn1002-0306.2021070177

Volume 43 Issue 5

Feb. 2022

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Science and Technology of Food Industry > 2022 > 43(5): 230-236. > DOI: 10.13386/j.issn1002-0306.2021070177

ZHANG Liming, MA Yage, NIU Ruonan, et al. Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells[J]. Science and Technology of Food Industry, 2022, 43(5): 230−236. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021070177.

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Purification of Proanthocyanidins from Binchuan Grape Seed and Its Effects on Proliferation of HepG2 Cells

1.
School of Chinese Materia Medica, Yunnan University of Traditional Chinese Medicine, Kunming 650500, China
2.
School of Medicine, Yunnan School of Economics and Management, Kunming 650106, China

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Received Date: July 18, 2021
Available Online: December 24, 2021

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Abstract

Abstract

Objective: To study the purification of grape seed proanthocyanidins from Binchuan, Dali, Yunnan and their effects on the proliferation of HepG2 cells. Method: The effects of sample concentration, pH, flow rate, and ethanol volume fraction on the adsorption and resolution of grape seed proanthocyanidins were investigated, and the conditions for the AB-8 type macroporous adsorption resin to purify grape seed proanthocyanidins from Binchuan, Dali, Yunnan were studied. Grape seed proanthocyanidins of different polarities obtained by the ethanol gradient elution during the purification process were applied to HepG2 cells, and the effects of different concentrations of grape seed proanthocyanidins of different polarities on the proliferation of liver cancer HepG2 cells were detected by the CCK8 method. Results: The best process for AB-8 resin to purify grape seed proanthocyanidins was the pH of the sample solution 4.0, the mass concentration was 6.0 mg/mL, the flow rate was 1.0 BV/h, the volume fraction of the eluent was 30%, and the elution volume was 4 BV. This purification increased the content of proanthocyanidins in the grape seed extract from 22.77%±0.32% to 94.73%±0.6429%, with a recovery rate of 80.55%±1.6499%. The polar parts of grape seed proanthocyanidins eluted with 10%, 20%, 30% and 40% ethanol had inhibitory effects on the proliferation of liver cancer HepG2 cells, and the unpurified part (that is, the extract had not been purified by macroporous resin) that inhibitory effect was most significant when the administration concentration was 200 μg/mL (P<0.001). The IC₅₀ of the 10% ethanol group was 25.09 μg/mL, which showed a better inhibitory effect. Conclusion: The grape seed proanthocyanidins in Binchuan, Dali, Yunnan are purified with AB-8 type macroporous resin, and the method is feasible. The grape seed proanthocyanidins have a good inhibitory effect on the proliferation of liver cancer HepG2 cells.
- grape seed,
- proanthocyanidin,
- purification,
- hepatocarcinoma HepG2 cell,
- proliferation

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References

[1]	赵坤生, 时倩, 李秀昌. 葡萄籽化学成分与药理作用研究进展[J]. 泰山医学院学报,2019,40(9):718−720. [ZHAO K S, SHI Q, LI X C. Research progress on the chemical constituents and pharmacological effects of grape seeds[J]. Journal of Taishan Medical College,2019,40(9):718−720. doi: 10.3969/j.issn.1004-7115.2019.09.022
[2]	BAGCHI D, BAGCHI M, STOHS S J, et al. Free radicals and grape seed proanthocyanidin extract: Importance in human health and disease prevention[J]. Toxicology,2000,148(2):187−197.
[3]	QI Y, ZHANG H, WU G, et al. Mitigation effects of proanthocyanidins with different structures on acrylamide formation in chemical and fried potato crisp models[J]. Food Chemistry,2018,250:98−104. doi: 10.1016/j.foodchem.2018.01.012
[4]	GJ A, MM A, PMB C, et al. Mechanisms of endothelium-dependent vasorelaxation induced by procyanidin B₂ in venous bypass graft[J]. Journal of Pharmacological Sciences,2020,142(3):101−108. doi: 10.1016/j.jphs.2019.11.006
[5]	刘丹, 邓建军, 张超, 等. 原花青素防治Ⅱ型糖尿病的研究进展[J]. 食品科学,2019,40(9):302−308. [LIU D, DENG J J, ZHANG C, et al. Research progress of proanthocyanidins in the prevention and treatment of type Ⅱ diabetes[J]. Food Science,2019,40(9):302−308. doi: 10.7506/spkx1002-6630-20180131-441
[6]	WANG E H, YU Z L, PING G F, et al. Grape seed procyanidin extract attenuate sodium fluoride-induced oxidative damage and apoptosis in rat kidneys[J]. Biomedical and Environmental Sciences,2020,33(6):454−457.
[7]	GONZÁLEZ-QUILEN C, RODRÍGUEZ-GALLEGO E, BELTRÁN-DEBÓN R, et al. Health-promoting properties of proanthocyanidins for intestinal dysfunction[J]. Nutrients,2020,12(1):130. doi: 10.3390/nu12010130
[8]	GONG X, XU L, FANG X, et al. Protective effects of grape seed procyanidin on isoflurane-induced cognitive impairment in mice[J]. Pharmaceutical Biology,2020,58(1):200−207. doi: 10.1080/13880209.2020.1730913
[9]	WEI S, ZHENG Y, ZHANG M, et al. Grape seed procyanidin extract inhibits adipogenesis and stimulates lipolysis of porcine adipocytes in vitro[J]. J Anim Sci,2018,96(7):2753−2762. doi: 10.1093/jas/sky158
[10]	CAO J, YU X, DENG Z, et al. Chemical compositions, antiobesity, and antioxidant effects of proanthocyanidins from lotus seed epicarp and lotus seed pot[J]. J Agric Food Chem,2018,66(51):13492−13502. doi: 10.1021/acs.jafc.8b05137
[11]	VAZQUEZ-FLORES A A, MARTINEZ-GONZALEZ A I, ALVAREZ-PARRILLA E, et al. Proanthocyanidins with a low degree of polymerization are good inhibitors of digestive enzymes because of their ability to form specific interactions: A hypothesis[J]. Journal of Food Science,2018,83(10-12):2895−2902.
[12]	刘洁. 葡萄籽提取物原花青素对人膀胱癌细胞生长的抑制作用及机制研究[D]. 武汉: 华中科技大学, 2009. LIU J. Study on the inhibitory effect and mechanism of grape seed extract proanthocyanidins on the growth of human bladder cancer cells[D]. Wuhan: Huazhong University of Science and Technology, 2009.
[13]	徐春明, 庞高阳, 李婷. 花青素的生理活性研究进展[J]. 中国食品添加剂,2013(3):205−210. [XU C M, PANG G Y, LI T. Research progress on the physiological activities of anthocyanins[J]. China Food Additives,2013(3):205−210. doi: 10.3969/j.issn.1006-2513.2013.03.028
[14]	TANG L, JIN X, HU X, et al. Glaucocalyxin a inhibits the growth of liver cancer focus and SMMC-7721 cells[J]. Oncol Lett,2016,11(2):1173−1178. doi: 10.3892/ol.2015.4002
[15]	ZHONG J H, LI H, XIAO N, et al. Hepatic resection is safe and effective for patients with hepatocellular carcinoma and portal hypertension[J]. Plos One,2014,9(9):e108755. doi: 10.1371/journal.pone.0108755
[16]	INGLE P V, SAMSUDIN S Z, CHAN P Q, et al. Development and novel therapeutics in hepatocellular carcinoma: A review[J]. Ther Clin Risk Manag,2016,12:445−455.
[17]	闫文杰, 李凯, 吕倩倩, 等. 海娇颜片中原花青素含量快速测定方法优化[J]. 食品安全质量检测学报,2020,11(3):841−846. [YAN W J, LI K, LU Q Q, et al. Optimization of rapid determination method for proanthocyanidin content in Haijiaoyan tablets[J]. Journal of Food Safety and Quality Testing,2020,11(3):841−846.
[18]	宋倩, 赵声兰, 刘芳, 等. 大孔吸附树脂分离纯化核桃壳总黄酮[J]. 食品与发酵工业,2012,38(12):180−184. [SONG Q, ZHAO S L, LIU F, et al. Separation and purification of total flavonoids from walnut shell by macroporous adsorption resin[J]. Food and Fermentation Industries,2012,38(12):180−184.
[19]	马小琴, 王晓敏, 周凡. 大孔吸附树脂法分离纯化葡萄籽中原花青素[J]. 食品研究与开发,2017,38(16):54−57. [MA X Q, WANG X M, ZHOU F. Separation and purification of proanthocyanidins from grape seeds by macroporous adsorption resin method[J]. Food Research and Development,2017,38(16):54−57. doi: 10.3969/j.issn.1005-6521.2017.16.012
[20]	郭方明, 李述刚, 宋关玲, 等. 葡萄籽提取物原花青素诱导人食管癌细胞ECA109凋亡[J]. 食品科学,2019,40(9):115−121. [GUO F M, LI S G, SONG G L, et al. Grape seed extract proanthocyanidins induce apoptosis in human esophageal cancer cells ECA109[J]. Food Science,2019,40(9):115−121. doi: 10.7506/spkx1002-6630-20180202-024
[21]	胡俊丽, 张月飞, 宋瑾, 等. 葡萄籽提取物对人鼻咽癌细胞HIF-1α表达的影响[J]. 广东医科大学学报,2017,35(4):356−359. [HU J L, ZHANG Y F, SONG J, et al. Effects of grape seed extract on the expression of HIF-1α in human nasopharyngeal carcinoma cells[J]. Journal of Guangdong Medical University,2017,35(4):356−359. doi: 10.3969/j.issn.1005-4057.2017.04.008
[22]	谢佳卿, 赵佳丽, 李爱芳, 等. 葡萄籽原花青素对人骨肉瘤143B细胞的作用及机制研究[J]. 中国细胞生物学学报,2017,39(12):1543−1549. [XIE J Q, ZHAO J L, LI A F, et al. Effects and mechanism of grape seed proanthocyanidins on human osteosarcoma 143B cells[J]. Chinese Journal of Cell Biology,2017,39(12):1543−1549.
[23]	于长水, 徐佳元, 王春雷, 等. 葡萄籽原花青素抑制人骨肉瘤MG63细胞增殖的机制研究[J]. 中华骨与关节外科杂志,2019,12(2):141−144. [YU C S, XU J Y, WANG C L, et al. Study on the mechanism of grape seed proanthocyanidins inhibiting the proliferation of human osteosarcoma MG63 cells[J]. Chinese Journal of Bone and Joint Surgery,2019,12(2):141−144. doi: 10.3969/j.issn.2095-9958.2019.02.13
[24]	张慧瑛, 邢芙玲, 罗光宏, 等. 葡萄籽原花青素抑制小鼠B16-F0黑色素瘤细胞黑色素生成的影响[J]. 中药药理与临床,2017,33(1):63−66. [ZHANG H Y, XING F L, LUO G H, et al. Effects of grape seed proanthocyanidins on melanogenesis in B16-F0 melanoma cells of mice[J]. Pharmacology and Clinics of Chinese Materia Medica,2017,33(1):63−66.
[25]	石焕焕, 吕程程, 朱运峰. 葡萄籽原花青素对结肠癌细胞抑制作用的初步研究[J]. 中国细胞生物学学报,2019,41(8):1523−1533. [SHI H H, LU C C, ZHU Y F. Preliminary study on the inhibitory effect of grape seed proanthocyanidins on colon cancer cells[J]. Chinese Journal of Cell Biology,2019,41(8):1523−1533.
[26]	王海, 连燕娜, 周游, 等. 低聚葡萄籽原花青素联合顺铂对人肺腺癌细胞A549凋亡的影响[J]. 食品科学,2017,38(19):177−181. [WANG H, LIAN Y N, ZHOU Y, et al. Effects of oligomeric grape seed proanthocyanidins combined with cisplatin on apoptosis of human lung adenocarcinoma cell A549[J]. Food Science,2017,38(19):177−181. doi: 10.7506/spkx1002-6630-201719028
[27]	孙英殊, 王雪峰, 何丽霞, 等. 葡萄籽原花青素联合顺铂对喉癌细胞Hep-2生物学作用及机制研究[J]. 临床军医杂志,2017,45(2):176−179. [SUN Y S, WANG X F, HE L X, et al. Study on the biological effects and mechanism of grape seed proanthocyanidins combined with cisplatin on Hep-2 laryngeal carcinoma cells[J]. Clinical Journal of Medical Officer,2017,45(2):176−179.
[28]	周世繁, 张娟. 葡萄籽原花青素联合吉西他滨对胰腺癌细胞增殖与凋亡的研究[J]. 中国临床药理学杂志,2018,34(15):1862−1864. [ZHOU S F, ZHANG J. Study of grape seed proanthocyanidin combined with gemcitabine on the proliferation and apoptosis of pancreatic cancer cells[J]. Chinese Journal of Clinical Pharmacology,2018,34(15):1862−1864.
[29]	张琳, 马龙, 刘涛, 等. 琐琐葡萄提取物多糖、黄酮对肝癌细胞HepG2的增殖抑制作用[J]. 新疆医科大学学报,2009,32(5):529−532. [ZHANG L, MA L, LIU T, et al. Inhibitory effects of Suosuo grape extract polysaccharides and flavonoids on the proliferation of hepatocellular carcinoma cell HepG2[J]. Journal of Xinjiang Medical University,2009,32(5):529−532. doi: 10.3969/j.issn.1009-5551.2009.05.005
[30]	朱芳成, 毕华强, 赵毅, 等. 乙型肝炎病毒核心蛋白触发人肝癌细胞异常卟啉代谢的机制[J]. 病毒学报,2019,35(4):592−598. [ZHU F C, BI H Q, ZHAO Y, et al. The mechanism of hepatitis B virus core protein triggering abnormal porphyrin metabolism in human liver cancer cells[J]. Acta Virology,2019,35(4):592−598.