采用脯氨酸-组氨酸-脯氨酸(Pro-His-Pro,PHP)分别处理人源肝细胞LO2和酒精性肝损伤小鼠,研究PHP是否能够缓解由肝细胞氧化损伤和食用酒精对小鼠造成的肝损伤。构建肝细胞氧化应激模型,通过不同浓度PHP处理后,检测抗氧化酶相关含量,评价氧化相关蛋白情况。构建小鼠酒精性肝损伤模型,观察肝组织病理形态学改变及生理生化和氧化相关指标。与偶氮二异丁脒盐酸盐(2,2′-azobis-2-methyl-propanimidamide,AAPH)处理组相比较,不同浓度的PHP处理组会导致抗氧化酶升高;在氧化通路方面,细胞核内核转录因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)蛋白增加,胞质Kelch ECH 相关蛋白1(Kelch like ECH associated protein 1,Keap1)蛋白增多。与食用酒精组相比较,PHP处理组小鼠肝组织细胞变性程度较轻,相关指标均恢复正常,抗氧化酶氧化升高。短肽PHP可以通过调控氧化相关指标和氧化通路关键蛋白Nrf2来达到预防酒精性肝损伤的目的。
Tripeptide Pro-His-Pro (PHP) was used to treat LO2 cells and alcoholic liver injury mice respectively. It was verified in vivo and in vitro to study whether PHP can alleviate the liver injury caused by 2,2′-azobis-2-methyl-propanimidamide (AAPH) to hepatocytes and edible alcohol to mice, and to provide relevant ideas for alleviating liver injury. Oxidative stress model was constructed with LO2 cells. The cells were divided into normal group and PHP treatment group. The contents of superoxide dismutase (SOD), catalase (CAT), glutathione reduction (GSH) and malondialdehyde (MDA) in cell lysate were detected. The oxidation related marker proteins were evaluated by Western Blot. Alcoholic liver model was established with C57BL/6J mice. The pathomorphological changes of liver were observed, the physiological and biochemical indexes and oxidation related indexes in liver were detected. The results showed that compared with the alcohol group, the liver tissue of PHP treatment group showed less mild degeneration of hepatocytes, the levels of ALT and AST in serum decreased, the levels of TC, TG and LDL-C decreased, and the levels of HDL-C increased. The increase of CAT, GSH and SOD levels, the decrease of MDA (P<0.05). Peptide PHP can alleviate liver injury by regulating oxidation related indexes and key protein Nrf2 of oxidation pathway.
[1] REHM J, BALIUNAS D, BORGES G L G, et al.The relation between different dimensions of alcohol consumption and burden of disease:An overview[J].Addiction, 2010, 105(5):817-843.
[2] CEDERBAUM A I, LU Y K, WU D F.Role of oxidative stress in alcohol-induced liver injury[J].Archives of Toxicology, 2009, 83(6):519-548.
[3] ROMAN S, ZEPEDA-CARRILLO E A, MORENO-LUNA L E, et al.Alcoholism and liver disease in Mexico:Genetic and environmental factors[J].World Journal of Gastroenterology, 2013, 19(44):7 972-7 982.
[4] CAI Z, SONG L H, QIAN B J, et al.Understanding the effect of anthocyanins extracted from purple sweet potatoes on alcohol-induced liver injury in mice[J].Food Chemistry, 2018, 245:463-470.
[5] POPOVA T A, PROKOFIEV I I, KHUSAINOVA G K, et al.Age-related changes in the functional indices of cardiac mitochondria in chronic alcohol intoxication in rats[J].Advances in Gerontology, 2019, 9(3):274-282.
[6] BOUTROS N, DER-AVAKIAN A, KESBY J P, et al.Effects of adolescent alcohol exposure on stress-induced reward deficits, brain CRF, monoamines and glutamate in adult rats[J].Psychopharmacology, 2018, 235(3):737-747.
[7] LU D, WANG W, LIU J F, et al.Peroxiredoxins in inflammatory liver diseases and ischemic/reperfusion injury in liver transplantation[J].Food and Chemical Toxicology, 2018, 113:83-89.
[8] REDDYVARI H, GOVATATI S, MATHA S K, et al.Therapeutic effect of green tea extract on alcohol induced hepatic mitochondrial DNA damage in albino wistar rats[J].Journal of Advanced Research, 2017, 8(3):289-295.
[9] IMLAY J A, LINN S.DNA damage and oxygen radical toxicity[J].Science, 1988, 240(4 857):1 302-1 309.
[10] LYU C, HE D, CHANG Y M, et al.Cobalt oxyhydroxide as an efficient heterogeneous catalyst of peroxymonosulfate activation for oil-contaminated soil remediation[J].Science of the Total Environment, 2019, 680:61-69.
[11] NIE W, ZHOU K, WANG Y, et al.Isolation and identification of bioactive peptides from Xuanwei ham that rescue oxidative stress damage induced by alcohol in HHL-5 hepatocytes[J].Food & Function, 2020, 11(11):9 710-9 720.
[12] 邹仙果. 亚麻籽环肽组成、氧化结构变化及其体外抗肿瘤抗炎活性研究[D].南昌:南昌大学, 2019.
ZOU X G.The profiles, structural changes during oxidation and in vitro antitumor and anti-inflammatory activities of linusorbs in flaxseed(Linum usitatissimum L.)[D].Nanchang:Nanchang University, 2019.
[13] 施用晖, 乐国伟.生物活性短肽在动物生产中的应用前景[J].饲料工业, 2001, 22(7):34-36.
SHI Y H, LE G W.Application prospect of bioactive short peptides in animal production[J].Feed Industry, 2001, 22(7):34-36.
[14] 孙金沅, 姜云松, 刘国英, 等.白酒酒醅中两种多肽的鉴定及其抗氧化和降血压功能评价[J].食品与发酵工业, 2019, 45(18):1-8.
SUN J Y, JIANG Y S, LIU G Y, et al.In vitro antioxidant and angiotensinz-converting enzyme inhibitory activities of two oligopeptides in Jiupei of Baijiu[J].Food and Fermentation Industries, 2019, 45(18):1-8.
[15] 霍嘉颖, 孙宝国, 郑福平, 等.白酒中一种三肽Arg-Asn-His的鉴定及其细胞内抗氧化活性[J].食品科学, 2018, 39(23):126-133.
HUO J Y, SUN B G, ZHENG F P, et al.Identification of a tripeptide Arg-Asn-His from Chinese Baijiu and its antioxidant activity[J].Food Science, 2018, 39(23):126-133.
[16] RANGASAMY T, CHO C Y, THIMMULAPPA R K, et al.Genetic ablation of Nrf2 enhances susceptibility to cigarette smoke-induced emphysema in mice[J].The Journal of Clinical Investigation, 2004, 114(9):1 248-1 259.
[17] KAYNAR M Y, HANCI M, KUDAY C, et al.Changes in the activity of antioxidant enzymes (SOD, GPX, CAT) after experimental spinal cord injury[J].The Tokushima Journal of Experimental Medicine, 1994, 41(3-4):133-136.
[18] 刘臻华, 康学东, 赵鲲鹏, 等.少阴固本调枢法对STZ诱导糖尿病大鼠血清SOD、MDA、GSH-Px的影响[J].中国中医药科技, 2018, 25(6):794-796;811.
LIU Z H, KANG X D, ZHAO K P, et al.Effect of Shaoyin kidney supporting and pivot adjusting method on serum SOD, MDA, GSH-Px in STZ-induced diabetic rats[J].Chinese Journal of Traditional Medical Science and Technology, 2018, 25(6):794-796;811.
[19] KOBAYASHI M, YAMAMOTO M.Molecular mechanisms activating the Nrf2-Keap1 pathway of antioxidant gene regulation[J].Antioxidants & Redox Signaling, 2005, 7(3-4):385-394.
[20] SUN J, HONG Z X, SHAO S, et al.Liver-specific Nrf2 deficiency accelerates ethanol-induced lethality and hepatic injury in vivo[J].Toxicology and Applied Pharmacology, 2021, 426:115617.
[21] XIONG Z E, DONG W G, WANG B Y, et al.Curcumin attenuates chronic ethanol-induced liver injury by inhibition of oxidative stress via mitogen-activated protein kinase/nuclear factor E2-related factor 2 pathway in mice[J].Pharmacognosy Magazine, 2015, 11(44):707-715.
[22] RODRIGUES N R, BATISTA J E D S, DE SOUZA L R, et al.Activation of p38MAPK and NRF2 signaling pathways in the toxicity induced by chlorpyrifos in Drosophila melanogaster:Protective effects of Psidium guajava pomífera L.(Myrtaceae) hydroalcoholic extract[J].Arabian Journal of Chemistry, 2019, 12(8):3 490-3 502.
[23] LISI V, MOULTON C, ANTONIONI A, et al.The role of extracellular vesicles in the regulation of redox homeostasis during exercise:A focus on Nrf2 and antioxidant enzymes[J].Free Radical Biology and Medicine, 2021, 165:21-22.
[24] YI Z W, LIU X F, LIANG L H, et al.Antrodin A from Antrodia camphorata modulates the gut microbiome and liver metabolome in mice exposed to acute alcohol intake[J].Food & Function, 2021, 12(7):2 925-2 937.
