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Purification of Polysaccharides from Orostachys cartilaginous Cell Cultures by Macroporous Resin Absorption and Bioactivities of the Purified Polysaccharides
Chemistry & Biodiversity ( IF 2.3 ) Pub Date : 2022-11-03 , DOI: 10.1002/cbdv.202200452 Ya-Nan Xu 1 , Xiao-Li An 1 , Lu Han 1 , Xuan-Chun Piao 1 , Yue-Jun Hao 1 , Wan Meng 2 , Mei-Lan Lian 1
Chemistry & Biodiversity ( IF 2.3 ) Pub Date : 2022-11-03 , DOI: 10.1002/cbdv.202200452 Ya-Nan Xu 1 , Xiao-Li An 1 , Lu Han 1 , Xuan-Chun Piao 1 , Yue-Jun Hao 1 , Wan Meng 2 , Mei-Lan Lian 1
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Orostachys cartilaginous is a traditional herbal medicine and its cell cultures contain large amounts of polysaccharides. To utilize the cultured O. cartilaginous cells, this study purified the crude polysaccharides of O. cartilaginous cells by macroporous resin absorption and optimized the purification process in the experiment of orthogonal design with four factors (sample concentration and volume, and eluent concentration and volume) and three levels; the antibacterial and anti-cancer effects of the purified polysaccharides (OTP) were further examined. The results showed that polysaccharide purity reached 95 % in the optimized group, i. e., 1.6 mg/mL of sample (crude polysaccharides) concentration, 3.0 bed volume (BV) of sample volume, 65 % eluent (ethanol) concentration, and 3.0 BV of eluent volume. In the antibacterial experiment, the growth of three bacterial species, i. e., Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis was inhibited by OTP, whereas that of Escherichia coli was not affected; the largest diameter of the inhibitory zone was found on B. subtilis; the extracellular alkaline phosphatase activity and the electrical conductivity, nucleic acid, and protein levels of B. subtilis increased after OTP treatment, indicating that the inhibition of B. subtilis growth was caused by the leakage of cell contents. In the anti-cancer experiment, OTP decreased the cell viabilities of the tested human cancer cells, i. e., AGS (gastric cancer), HCT116 (colon cancer), HepG2 (liver cancer), and HeLa (cervical cancer), and the highest inhibitory effect was on HCT116. OTP promoted HCT116 apoptosis and affected the expression of apoptosis-related proteins, i. e., the expression of B-cell lymphoma-2 decreased and that of bcl-2 associated X protein, cytochrome c, caspase 9 and caspase 3 increased. The findings of the present study suggest that O. cartilaginous cell cultures have a potential application in food or drug production.
中文翻译：
通过大孔树脂吸附和纯化多糖的生物活性从 Orostachys 软骨细胞培养物中纯化多糖
Orostachys cortilaginous是一种传统草药，其细胞培养物含有大量多糖。为了利用培养的O. 软骨细胞，本研究纯化了O. 软骨的粗多糖大孔树脂吸附细胞并在四因素（样品浓度和体积、洗脱液浓度和体积）和三个水平的正交设计实验中优化纯化工艺；进一步检验了纯化多糖 (OTP) 的抗菌和抗癌作用。结果表明，优化组多糖纯度达到95%，即。即，样品（粗多糖）浓度为 1.6 mg/mL，样品体积为 3.0 床体积 (BV)，洗脱液（乙醇）浓度为 65%，洗脱液体积为 3.0 BV。在抗菌实验中，三种细菌的生长，i。例如，铜绿假单胞菌、金黄色葡萄球菌和枯草芽孢杆菌被 OTP 抑制，而大肠杆菌不受影响；抑菌圈直径最大的是枯草芽孢杆菌；OTP 处理后B. subtilis的胞外碱性磷酸酶活性和电导率、核酸和蛋白质水平升高，表明B. subtilis的抑制生长是由细胞内容物的泄漏引起的。在抗癌实验中，OTP 降低了被测人类癌细胞的细胞活力，即。即AGS（胃癌）、HCT116（结肠癌）、HepG2（肝癌）、HeLa（宫颈癌），其中对HCT116的抑制作用最强。OTP促进HCT116细胞凋亡并影响细胞凋亡相关蛋白的表达，即。即，B细胞淋巴瘤-2的表达降低，bcl-2相关X蛋白、细胞色素c、半胱天冬酶9和半胱天冬酶3的表达增加。本研究的结果表明O. 软骨细胞培养物在食品或药物生产中具有潜在的应用。
更新日期：2022-11-03
中文翻译：
通过大孔树脂吸附和纯化多糖的生物活性从 Orostachys 软骨细胞培养物中纯化多糖
Orostachys cortilaginous是一种传统草药，其细胞培养物含有大量多糖。为了利用培养的O. 软骨细胞，本研究纯化了O. 软骨的粗多糖大孔树脂吸附细胞并在四因素（样品浓度和体积、洗脱液浓度和体积）和三个水平的正交设计实验中优化纯化工艺；进一步检验了纯化多糖 (OTP) 的抗菌和抗癌作用。结果表明，优化组多糖纯度达到95%，即。即，样品（粗多糖）浓度为 1.6 mg/mL，样品体积为 3.0 床体积 (BV)，洗脱液（乙醇）浓度为 65%，洗脱液体积为 3.0 BV。在抗菌实验中，三种细菌的生长，i。例如，铜绿假单胞菌、金黄色葡萄球菌和枯草芽孢杆菌被 OTP 抑制，而大肠杆菌不受影响；抑菌圈直径最大的是枯草芽孢杆菌；OTP 处理后B. subtilis的胞外碱性磷酸酶活性和电导率、核酸和蛋白质水平升高，表明B. subtilis的抑制生长是由细胞内容物的泄漏引起的。在抗癌实验中，OTP 降低了被测人类癌细胞的细胞活力，即。即AGS（胃癌）、HCT116（结肠癌）、HepG2（肝癌）、HeLa（宫颈癌），其中对HCT116的抑制作用最强。OTP促进HCT116细胞凋亡并影响细胞凋亡相关蛋白的表达，即。即，B细胞淋巴瘤-2的表达降低，bcl-2相关X蛋白、细胞色素c、半胱天冬酶9和半胱天冬酶3的表达增加。本研究的结果表明O. 软骨细胞培养物在食品或药物生产中具有潜在的应用。
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