黑果枸杞果实甲醇提取物对2,2'-偶氮二(2-脒基丙烷)二盐酸盐诱导的LLC-PK1细胞氧化应激的保护作用。
Protective effects of methanolic extract form fruits of Lycium ruthenicum Murr on 2,2'-azobis (2-amidinopropane) dihydrochloride-induced oxidative stress in LLC-PK1 cells.
作者信息
Song Jia-Le, Gao Yang, Xu Jianguo
机构信息
Department of Nutrition and Food Hygiene, School of Public Health, Guilin Medical University, Guangxi 541004, People's Republic of China ; Department of Food Science and Nutrition, Pusan National University, Busan 609-735, South Korea.
Department of Pharmacy, Northern Jiangsu People's Hospital Affiliated to Yangzhou University (Clinical Medical College of Yangzhou University), Yangzhou, Jiangsu 225001, People's Republic of China.
出版信息
Pharmacogn Mag. 2014 Oct;10(40):522-8. doi: 10.4103/0973-1296.141790.
BACKGROUND
Fruits of Lycium ruthenicum Murr is a health food and also used as a folk to treat heart disease, abnormal menstruation and menopause in Tibetan, China. However; whether L. ruthenicum Murr fruits methanolic extracts (LFME) protect LLC-PK1 porcine renal tubules cells from AAPH-induced oxidative damage has not been investigated.
OBJECTIVE
To investigate the protective effects of L. ruthenicum Murr fruits methanolic extracts (LFME) against 2, 2'- azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative damage in renal proximal tubule LLC-PK1 cells.
MATERIALS AND METHODS
LLC-PK1 cells were co-incubated with AAPH (1mM) and different concentrations of LFMW together for 24 h. Cell viability was determined by MTT assay. Total intercellular reactive oxygen species (ROS) levels and lipid peroxidation were measured using a fluorescent probe 2', 7'-dichlorfluorescein-diacetate (DCFH-DA) and the TBA reactive substance (TBARS) assay, respectively. The endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and intercellular glutathione (GSH) levels were determined using commercial assay kits according to the manufacturer's instructions.
RESULTS
LFME did not show a significant cytotoxic effect and increased the viability of LLC-PK1 cells in a concentration-dependent manner. LFME also decreased the total intercellular levels of ROS, reduced lipid peroxidation and increased the GSH levels as well as the activities of endogenous antioxidant enzymes to protect LLC-PK1 cells against AAPH-induced oxidative damage.
CONCLUSION
The results from the present study indicated that LFME is an effective ROS scavenger to protect LLC-PK1 cells against AAPH-induced oxidative damage through decreasing ROS generation, reducing lipid peroxidation and up-regulation of endogenous GSH levels and antioxidant enzymes.
背景
黑果枸杞果实是一种保健食品，在中国藏族地区也被用作民间治疗心脏病、月经不调和更年期的药物。然而，黑果枸杞果实甲醇提取物（LFME）是否能保护LLC-PK1猪肾小管细胞免受AAPH诱导的氧化损伤尚未见研究报道。
目的
研究黑果枸杞果实甲醇提取物（LFME）对2,2'-偶氮二异丁脒二盐酸盐（AAPH）诱导的肾近端小管LLC-PK1细胞氧化损伤的保护作用。
材料与方法
将LLC-PK1细胞与AAPH（1mM）和不同浓度的LFMW共同孵育24小时。采用MTT法测定细胞活力。分别使用荧光探针2',7'-二氯荧光素二乙酸酯（DCFH-DA）和TBA反应物质（TBARS）法测定细胞内总活性氧（ROS）水平和脂质过氧化。根据制造商的说明，使用商业检测试剂盒测定内源性抗氧化酶，包括过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-px）和细胞内谷胱甘肽（GSH）水平。
结果
LFME未显示出明显的细胞毒性作用，且以浓度依赖的方式增加了LLC-PK1细胞的活力。LFME还降低了细胞内ROS的总水平，减少了脂质过氧化，增加了GSH水平以及内源性抗氧化酶的活性，从而保护LLC-PK1细胞免受AAPH诱导的氧化损伤。
结论
本研究结果表明，LFME是一种有效的ROS清除剂，通过减少ROS生成、降低脂质过氧化以及上调内源性GSH水平和抗氧化酶活性，保护LLC-PK1细胞免受AAPH诱导的氧化损伤。
Zotero 插件