鹿茸水溶性蛋白对庆大霉素诱导HEK293细胞毒性的影响
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摘要
目的研究梅花鹿茸水溶性蛋白(Sika deer velvet anlter protein,SVPr)的体外抗氧化能力及其对庆大霉素(Gentamicin,GM)诱导HEK293细胞增殖、氧化应激水平和细胞形态的影响。方法通过SVPr对1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟基(OH)自由基清除力测定及Fe~(3+)还原力测定研究其体外抗氧化活性。以MTT法分别检测GM、SVPr及SVPr与GM联合使用对HEK293细胞活力影响,并测定了SVPr对HEK293细胞内乳酸脱氢酶(LDH)水平、谷胱甘肽(GSH)含量、丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性的影响。在光学显微镜下观察SVPr对GM致HEK293细胞形态改变的影响。结果 SVPr对DPPH·和·OH自由基的最大清除率分别达62.64%和66.35%,且对Fe~(3+)有较强的还原力。SVPr具有显著的促细胞增殖能力,当剂量达4.00 mg/ml水平时,HEK293细胞增殖率达164.71%(P<0.01),且其能明显改善GM诱导的HEK293细胞损伤现象,提升HEK293细胞存活率,调节细胞内源性抗氧化剂平衡,恢复受损细胞形态。结论研究结果表明,SVPr可有效预防GM诱导HEK293细胞的氧化应激损伤,促进细胞增殖,恢复细胞正常形态及功能,从而改善GM诱导HEK293细胞损伤,具有肾保护的潜力。
引文
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[19] 杨玉娈,袁美兰,陈丽丽,等.河蚬中水溶性蛋白的提取及其抗氧化性质[J].食品科学,2015,36(6):96-102.
[20] 张敏,沈建明.依达拉奉对庆大霉素诱导肾小管上皮细胞损伤的保护作用[J].武汉大学学报(医学版),2014,35(6):852-856.
[21] 刘佳,赵海平,张伟,等.鹿茸水溶性与脂溶性成分提取方法的优化及含量分析[J].中国实验方剂学杂志,2017,23(15):65-69.
[22] Yang H,Li W,Wang L,et al.The protective effects of Sika deer antler protein on cisplatin-induced nephrotoxicity[J].Cell Physiol Biochem,2017,43(1):395-404.
[23] Chashmi NA,Emadi S,Khastar H.Protective effects of hydroxytyrosol on gentamicin induced nephrotoxicity in mice[J].Biochem Biophys Res Commun,2017,482(4):1427-1429.
[2] Sui Z,Zhang L,Huo Y,et al.Bioactive components of velvet antlers and their pharmacological properties[J].J Pharm Biomed Anal,2014,87(1):229-240.
[3] 杨慧海,王露露,何晓凤,等.梅花鹿鹿茸总蛋白对顺铂诱导肾损伤小鼠的保护作用及其机制[J].中国药理学与毒理学杂志,2017,31(6):561-567.
[4] 李超华.鹿茸蛋白提取方法的建立和神经保护作用研究[D].吉林:吉林农业大学,2016.
[5] Yang H,Wang L,Sun H,et al.Anticancer activity in vitro and biological safety evaluation in vivo of Sika deer antler protein[J].J Food Biochem,2017,54(24):1-8.
[6] 简心韵,邓子新,孙宇辉.氨基糖苷抗生素庆大霉素:基础研究的新进展及应用研究的新潜力[J].生物工程学报,2015,31(6):829-844.
[7] Galdino PM,Alexandre LN,Pacheco LF,et al.Nephroprotective effect of Rudgea viburnoides (Cham.) Benth leaves on Gentamicin-Induced Nephrotoxicity in Rats[J].J Ethnopharmacol,2017,201:100-107.
[8] Liu Y,Lv H,Qin Y,et al.Gentamicin modified chitosan film with improved antibacterial property and cell biocompatibility[J].Int J Biol Macromol,2017,98:550-557.
[9] Ansari MA,Raish M,Ahmad A,et al.Sinapic acid mitigates gentamicin-induced nephrotoxicity and associated oxidative/nitrosative stress,apoptosis,and inflammation in rats[J].Life Sci,2016,165:1-8.
[10] Mahmoud YI.Kiwi fruit (Actinidia deliciosa) ameliorates gentamicin-induced nephrotoxicity in albino mice via the activation of Nrf2 and the inhibition of NF-κB (Kiwi & gentamicin-induced nephrotoxicity)[J].Biomed Pharmacother,2017,94:206-218.
[11] EI-Kashef DH,EI-Kenawi AE,Suddek GM,et al.Protective effect of allicin against gentamicin-induced nephrotoxicity in rats[J].Int Immunopharmacol,2015,29(2):679-686.
[12] Aldahmash BA,EI-Nagar DM,Ibrahim KE.Reno-protective effects of propolis on gentamicin-induced acute renal toxicity in swiss albino mice[J].Nefrologia,2016,36(6):643-652.
[13] Kandemir FM,Ozkaraca M,Yildirim BA,et al.Rutin attenuates gentamicin-induced renal damage by reducing oxidative stress,inflammation,apoptosis,and autophagy in rats[J].Ren Fail,2015,37(3):518-525.
[14] Fouad AA,Albuali WH,Zahran A,et al.Protective effect of naringenin against gentamicin-induced nephrotoxicity in rats[J].Environ Toxicol Pharmacol,2014,38(2):420-429.
[15] Subramanian P,Anandan R,Jayapalan JJ,et al.Hesperidin protects gentamicin-induced nephrotoxicity via Nrf2/HO-1 signaling and inhibits inflammation mediated by NF-κB in rats[J].J Funct Foods,2015,13(5):89-99.
[16] Jose SP,S A,Im K,et al.Nephro-protective effect of a novel formulation of unopened coconut inflorescence sap powder on gentamicin induced renal damage by modulating oxidative stress and inflammatory markers[J].Biomed Pharmacother,2017,85:128-135.
[17] Katary M,Salahuddin A.Ameliorative effect of gossypin against gentamicin-induced nephrotoxicity in rats[J].Life Sci,2017,176:75-81.
[18] 张静,王喜丰,李刚,等.过表达TRPM7蛋白对HEK293细胞氧糖剥夺再氧合损伤后NF-κB活化的影响[J].华中科技大学学报(医学版),2017,46(2):117-121.
[19] 杨玉娈,袁美兰,陈丽丽,等.河蚬中水溶性蛋白的提取及其抗氧化性质[J].食品科学,2015,36(6):96-102.
[20] 张敏,沈建明.依达拉奉对庆大霉素诱导肾小管上皮细胞损伤的保护作用[J].武汉大学学报(医学版),2014,35(6):852-856.
[21] 刘佳,赵海平,张伟,等.鹿茸水溶性与脂溶性成分提取方法的优化及含量分析[J].中国实验方剂学杂志,2017,23(15):65-69.
[22] Yang H,Li W,Wang L,et al.The protective effects of Sika deer antler protein on cisplatin-induced nephrotoxicity[J].Cell Physiol Biochem,2017,43(1):395-404.
[23] Chashmi NA,Emadi S,Khastar H.Protective effects of hydroxytyrosol on gentamicin induced nephrotoxicity in mice[J].Biochem Biophys Res Commun,2017,482(4):1427-1429.