石榴皮多酚有效部位单次给药毒性及对无水乙醇致大鼠胃溃疡的保护作用
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摘要
目的研究石榴皮多酚有效部位对小鼠单次给药毒性,评价其安全性,为新药研发和临床用药提供理论依据。观察石榴皮多酚有效部位对无水乙醇致大鼠胃溃疡的保护作用。方法选择健康昆明种小鼠50只,随机分为5组,每组10只,分别灌胃给予不同剂量的石榴皮多酚有效部位,连续观察14 d,每日记录各组小鼠一般情况、毒性反应及死亡情况,采用Bliss法计算半数致死量。选择健康大鼠70只,随机分为:正常组、模型组(等体积生理盐水)、三九胃泰颗粒组(1 850 mg·kg-1)、枸橼酸铋钾组(33 mg·kg-1)和石榴皮多酚有效部位低、中、高剂量组(430、852、1 704 mg·kg-1),连续灌胃10 d;d 10,除正常组外,其余各组采用无水乙醇灌胃(每只1.5 m L)致大鼠胃溃疡;计算各组胃溃疡指数、溃疡抑制率、观察胃黏膜组织形态学改变、测定胃黏膜组织中PGE2、NO、SOD、MDA含量。结果石榴皮多酚有效部位的半数致死量(LD50)为8 520.9 mg·kg-1,其95%的可信限范围为(7 291.2~9 914.4)mg·kg-1;病理学显示,给药剂量为16 000 mg·kg-1的小鼠脏器有不同程度的损伤。实验表明,与模型组比较,石榴皮多酚有效部位对胃黏膜损伤有明显修复作用,且明显升高胃溃疡大鼠胃黏膜NO含量、提高胃溃疡大鼠胃黏膜SOD活性及降低MDA含量、增加PGE2含量。结论石榴皮多酚有效部位给药剂量为5 063mg·kg-1未出现死亡,随着石榴皮多酚有效部位给药剂量的增加,给药剂量为16 000 mg·kg-1可造成小鼠的心、肝、肺、肾脏有不同程度的损伤,甚至导致死亡。石榴皮多酚有效部位的LD50为8 520.9 mg·kg-1,其95%的可信限范围为(7291.2~9 914.4)mg·kg-1。石榴皮多酚有效部位对无水乙醇致大鼠胃溃疡具有良好的保护作用,这种作用可能与促进胃溃疡上皮细胞合成、提高再生黏膜功能、增强抗氧化能力和诱导、促进NO合成有关。
Aims To study single dose toxicity of polyphenols effective parts from Punica granatum,to evaluate their safety,and thus to provide a theoretical basis for drug development and clinical use. To observe their protective effect on ethanol-induced gastric ulcer in rats. Methods 50 healthy Kunming mice were randomly divided into five groups and given different doses of polyphenols' effective parts from Punica granatum via intragastric administration. Toxicity and death in each group of mice were observed and recorded after administration for 14 d. The median lethal dose was calculated by Bliss method. 70 rats were randomly divided into normal group,model group( constant volume of normal saline),sanjiuweitai particles( 1 850 mg ·kg- 1) group,colloidal bismuth subcitrate( 33 mg·kg- 1) group and polyphenols effective parts from Punica granatum low-dose,medium-dose,high-dose( 430,852,1 704 mg · kg- 1) groups. On the 9th day of 10days' gavage,all except the normal group were fed ethanol( 1. 5 m L / only) to induce gastric mucosal injury in rats with acute gastric ulcer. Gastric ulcer index,the rate of ulcer inhibition were calculated for each group. The morphological changes of gastric mucosa were observed. The gastric mucosa levels of PGE2,NO,SOD and MDA were determined. Results The LD50 and 95% confidence limit of the polyphenols' effective parts from Punica granatum were 8 520. 9 mg·kg- 1and 7 291. 2 ~ 9 914. 4 mg ·kg- 1,respectively.Pathology showed that the organs receiving dose of16 000 mg · kg- 1had different degrees of damage.Compared with the model group,the extract from Punica granatum significantly repaired the gastric mucosa,and significantly increased the gastric mucosa levels of NO and reduced MDA content,and improved SOD content and the levels of PGE2. Conclousion The dose of 5 063 mg · kg- 1of polyphenols effective part from Punica granatum showed no death. The dose of16 000 mg · kg- 1of polyphenols effective parts from Punica granatum could cause varying degrees of damage in heart,liver,lung,kidney or the death of mice.The LD50 and 95% confidence limit of the polyphenols effective parts from Punica granatum were 8 520. 9 mg·kg- 1and 7 291. 2 ~ 9 914. 4 mg·kg- 1,respectively. The extract from Punica granatum plays a protective role against gastric mucosa damage induced by absolute ethanol,and the mechanism may be related to promoting ulcer epithelial cells synthesis,enhancing mucosal regeneration function,regulating NO content and enhancing antioxidant capacity.
Aims To study single dose toxicity of polyphenols effective parts from Punica granatum,to evaluate their safety,and thus to provide a theoretical basis for drug development and clinical use. To observe their protective effect on ethanol-induced gastric ulcer in rats. Methods 50 healthy Kunming mice were randomly divided into five groups and given different doses of polyphenols' effective parts from Punica granatum via intragastric administration. Toxicity and death in each group of mice were observed and recorded after administration for 14 d. The median lethal dose was calculated by Bliss method. 70 rats were randomly divided into normal group,model group( constant volume of normal saline),sanjiuweitai particles( 1 850 mg ·kg- 1) group,colloidal bismuth subcitrate( 33 mg·kg- 1) group and polyphenols effective parts from Punica granatum low-dose,medium-dose,high-dose( 430,852,1 704 mg · kg- 1) groups. On the 9th day of 10days' gavage,all except the normal group were fed ethanol( 1. 5 m L / only) to induce gastric mucosal injury in rats with acute gastric ulcer. Gastric ulcer index,the rate of ulcer inhibition were calculated for each group. The morphological changes of gastric mucosa were observed. The gastric mucosa levels of PGE2,NO,SOD and MDA were determined. Results The LD50 and 95% confidence limit of the polyphenols' effective parts from Punica granatum were 8 520. 9 mg·kg- 1and 7 291. 2 ~ 9 914. 4 mg ·kg- 1,respectively.Pathology showed that the organs receiving dose of16 000 mg · kg- 1had different degrees of damage.Compared with the model group,the extract from Punica granatum significantly repaired the gastric mucosa,and significantly increased the gastric mucosa levels of NO and reduced MDA content,and improved SOD content and the levels of PGE2. Conclousion The dose of 5 063 mg · kg- 1of polyphenols effective part from Punica granatum showed no death. The dose of16 000 mg · kg- 1of polyphenols effective parts from Punica granatum could cause varying degrees of damage in heart,liver,lung,kidney or the death of mice.The LD50 and 95% confidence limit of the polyphenols effective parts from Punica granatum were 8 520. 9 mg·kg- 1and 7 291. 2 ~ 9 914. 4 mg·kg- 1,respectively. The extract from Punica granatum plays a protective role against gastric mucosa damage induced by absolute ethanol,and the mechanism may be related to promoting ulcer epithelial cells synthesis,enhancing mucosal regeneration function,regulating NO content and enhancing antioxidant capacity.
引文
[1]周龙平,韦玉琼.消化性溃疡的药物治疗及进展[J].中外健康文摘,2009,6(24):42-4.[1]Zhou L P,Wei Y Q.Progress in pharmacotherapy of peptic ulcer[J].World Health Digest Med Period,2009,6(24):42-4.
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[4]张双霞,李光迪,于晓辉,张方信.根除幽门螺杆菌对慢性胃炎和胃溃疡组织中MIF蛋白表达的影响[J].西安交通大学学报:医学版,2009,30(6):724-8.[4]Zhang S X,Li G D,Yu X H,Zhang F X.Effect of Helicobacter pylori infection on macrophage migration inhibitory factor protein expression in patients with chronic gastritis and gastric ulcer precancerous lesions[J].J Xi’an Jiaotong Univ(Med Sci),2009,30(6):724-8.
[5]梅武轩,曾常春,余娜.健脾活血中药对胃溃疡大鼠H,KATP酶及壁细胞胃泌素受体作用的影响[J].时珍国医国药,2009,20(11):2778-80.[5]Mei W X,Ceng C C,Yu N.Effects of jianpi huoxue recipe on the activities of H,K-ATPase and gastrin receptor in gastric parietal cells of gastric ulcer in rats[J].Lishizhen Med Mat Med Res,2009,20(11):2778-80.
[6]Kim J J,Kim N,Lee B H,et al.Risk factors for development and recurrence of peptic ulcer disease[J].Korean J Gastroenterol,2010,56(4):220-8.
[7]涂朝勇,田徽,王建,等.路边菊水煎液抗实验性胃溃疡的研究[J].时珍国医国药,2009,20(10):2595-6.[7]Tu C Y,Tian H,Wang J,et al.Effect of water-decocted solution from Boltonia indica(L.)on experimental gastric ulcer[J].Lishizhen Med Mat Med Res,2009,20(10):2595-6.
[8]Okabe S,Pfeiffer C J.Chronicity of acetic acid ulcer in the rat stomach[J].Am J Dig Dis,1972,17(7):619-29.
[9]纪白慧,倪鑫炯,曹玉华.石榴皮抗氧化活性成分的提取及其组分的研究[J].天然产物研究与开发,2012,24(B12):17-22.[9]Ji B H,Ni X J,Cao Y H.Research antioxidant pomegranate peel extract active ingredients and components[J].J Nat Prod Res Development,2012,24(B12):17-22.
[10]Boussetta T,Raad H,Letteron P,et al.Punicic acid a conjugated linolenic acid inhibits TNFα-induced neutrophil hyperactivation and protects from experimental colon inflammation in rats[J].PLo S One,2009,4(7):e6458.
[11]Abdollahzadeh S H,Mashouf R,Mortazavi H,et al.Antibacterial and antifungal activities of punica granatum peel extracts against oral pathogens[J].Dentist Tehran Nivers Med Sci,2011,8(1):1-6.
[12]Jang A,Srinivasan P,Lee N Y,et al.Comparison of hypolipidemic activity of synthetic gallic acid-linoleic acid ester with mixture of gallic acid and linoleic acid,gallic acid,and linoleic acid on highfat diet induced obesity in C57BL/6 Cr Slc mice[J].Chem Biol Interact,2008,174(2):109-17.
[13]Vroegrijk I O,van Diepen J A,van den Berg S,et al.Pomegranate seed oil,a rich source of punicic acid,prevents diet-induced obesity and insulin resistance in mice[J].Food Chem Toxicol,2011,49(6):1426-30.
[14]Fuhrman B,Volkova N,Aviram M.Pomegranate juice inhibits oxidized LDL uptake and cholesterol biosynthesis in macrophages[J].Nutr Biochem,2005,16(9):570-6.
[15]Lewis S N,Brannan L,Guri A J,et al.Dietaryα-eleostearic acid ameliorates experimental inflammatory bowel disease in mice by activating peroxisome proliferator-activated receptor-γ[J].PLo S One,2011,6(8):e24031.
[16]邱红梅,赖舒,尚京川,周岐新.石榴皮提取物对大鼠幽门结扎所致胃损伤的保护作用研究[J].中国药房,2011,22(7):594-6.[16]Qiu H M,Lai S,Shang J S,Zhou Q X.Protective effect of the extracts of punica granatum on gastric damage induced by pylorus ligation in rats[J].China Pharm,2011,22(7):594-6.
[17]Lansky E P,Newman R A.Punica granatum(pomegranate)and its potential for prevention and treatment of inflammation cancer[J].Ethnopharmacol,2007,109(2):177-206.
[18]Ajaikumar K B,Asheef M,Babu B H,Padikkala J.The inhibition of gastric mucosal injury by Punica granatum L.(pomegranate)methanolic extract[J].Ethnopharmacol,2005,96(1-2):171-6.
[19]韦奇志,周智,陈邦树,刘廷快.胃乐胶囊的抗溃疡作用实验研究[J].广东药学院学报,2000,16(4):300-2.[19]Wei Q Z,Zhou Z,Chen B S,Liu T K.Experimental study of antiulcer effect Weile capsule[J].Academ J Guangdong Coll Pharm,2000,16(4):300-2.
[20]赖舒,周岐新,张颖,等.石榴皮对实验性胃损伤及相关机制的研究[J].中药药理与临床,2009,25(3):49-51.[20]Lai S,Zhou Q X,Zhang Y,et al.Effects and mechanisms of the acetone extract obtained from pomegranate bark on the experimental gastric damages[J].Pharmacol Clin Chin Mat Med,2009,25(3):49-51.
[21]Guth P H,Aures D,Paulsen G.Topical aspirin plus HCL gastric lesions in the rat[J].Gastroenterol,1979,76(1):88-93.
[22]符健,韩丽,邢桂兰,等.秋茄提取物抗大鼠实验性胃溃疡作用研究.[J].中国药理学通报,2007,23(10):1379-83.[22]Fu J,Han L,Xiang G L,et al.Study of gastric antiulcer effects on experimental rats of extracts of Kandelia candel[J].Chin Pharmacol Bull,2007,23(10):1379-83.
[2]古学文,廖永州.疏肝健脾活血法对胃溃疡患者胃黏膜表皮生长因子及受体表达的影响[J].广州中医药大学学报,2010,27(2):113-5.[2]Gu X W,Liao Y Z.Effect of therapy of soothing liver,strengthening spleen and activating blood on expression of epidermal growth factor and its receptor in patients with gastric ulcer[J].J Guangzhou Univ Tradit Chin Med,2010,27(2):113-5.
[3]Shay H,Komarov S A,Fels S S,et al.A simple method for the uniform production of gastric ulceration in the rat[J].Gastroenterol,1945,5(1):43-61.
[4]张双霞,李光迪,于晓辉,张方信.根除幽门螺杆菌对慢性胃炎和胃溃疡组织中MIF蛋白表达的影响[J].西安交通大学学报:医学版,2009,30(6):724-8.[4]Zhang S X,Li G D,Yu X H,Zhang F X.Effect of Helicobacter pylori infection on macrophage migration inhibitory factor protein expression in patients with chronic gastritis and gastric ulcer precancerous lesions[J].J Xi’an Jiaotong Univ(Med Sci),2009,30(6):724-8.
[5]梅武轩,曾常春,余娜.健脾活血中药对胃溃疡大鼠H,KATP酶及壁细胞胃泌素受体作用的影响[J].时珍国医国药,2009,20(11):2778-80.[5]Mei W X,Ceng C C,Yu N.Effects of jianpi huoxue recipe on the activities of H,K-ATPase and gastrin receptor in gastric parietal cells of gastric ulcer in rats[J].Lishizhen Med Mat Med Res,2009,20(11):2778-80.
[6]Kim J J,Kim N,Lee B H,et al.Risk factors for development and recurrence of peptic ulcer disease[J].Korean J Gastroenterol,2010,56(4):220-8.
[7]涂朝勇,田徽,王建,等.路边菊水煎液抗实验性胃溃疡的研究[J].时珍国医国药,2009,20(10):2595-6.[7]Tu C Y,Tian H,Wang J,et al.Effect of water-decocted solution from Boltonia indica(L.)on experimental gastric ulcer[J].Lishizhen Med Mat Med Res,2009,20(10):2595-6.
[8]Okabe S,Pfeiffer C J.Chronicity of acetic acid ulcer in the rat stomach[J].Am J Dig Dis,1972,17(7):619-29.
[9]纪白慧,倪鑫炯,曹玉华.石榴皮抗氧化活性成分的提取及其组分的研究[J].天然产物研究与开发,2012,24(B12):17-22.[9]Ji B H,Ni X J,Cao Y H.Research antioxidant pomegranate peel extract active ingredients and components[J].J Nat Prod Res Development,2012,24(B12):17-22.
[10]Boussetta T,Raad H,Letteron P,et al.Punicic acid a conjugated linolenic acid inhibits TNFα-induced neutrophil hyperactivation and protects from experimental colon inflammation in rats[J].PLo S One,2009,4(7):e6458.
[11]Abdollahzadeh S H,Mashouf R,Mortazavi H,et al.Antibacterial and antifungal activities of punica granatum peel extracts against oral pathogens[J].Dentist Tehran Nivers Med Sci,2011,8(1):1-6.
[12]Jang A,Srinivasan P,Lee N Y,et al.Comparison of hypolipidemic activity of synthetic gallic acid-linoleic acid ester with mixture of gallic acid and linoleic acid,gallic acid,and linoleic acid on highfat diet induced obesity in C57BL/6 Cr Slc mice[J].Chem Biol Interact,2008,174(2):109-17.
[13]Vroegrijk I O,van Diepen J A,van den Berg S,et al.Pomegranate seed oil,a rich source of punicic acid,prevents diet-induced obesity and insulin resistance in mice[J].Food Chem Toxicol,2011,49(6):1426-30.
[14]Fuhrman B,Volkova N,Aviram M.Pomegranate juice inhibits oxidized LDL uptake and cholesterol biosynthesis in macrophages[J].Nutr Biochem,2005,16(9):570-6.
[15]Lewis S N,Brannan L,Guri A J,et al.Dietaryα-eleostearic acid ameliorates experimental inflammatory bowel disease in mice by activating peroxisome proliferator-activated receptor-γ[J].PLo S One,2011,6(8):e24031.
[16]邱红梅,赖舒,尚京川,周岐新.石榴皮提取物对大鼠幽门结扎所致胃损伤的保护作用研究[J].中国药房,2011,22(7):594-6.[16]Qiu H M,Lai S,Shang J S,Zhou Q X.Protective effect of the extracts of punica granatum on gastric damage induced by pylorus ligation in rats[J].China Pharm,2011,22(7):594-6.
[17]Lansky E P,Newman R A.Punica granatum(pomegranate)and its potential for prevention and treatment of inflammation cancer[J].Ethnopharmacol,2007,109(2):177-206.
[18]Ajaikumar K B,Asheef M,Babu B H,Padikkala J.The inhibition of gastric mucosal injury by Punica granatum L.(pomegranate)methanolic extract[J].Ethnopharmacol,2005,96(1-2):171-6.
[19]韦奇志,周智,陈邦树,刘廷快.胃乐胶囊的抗溃疡作用实验研究[J].广东药学院学报,2000,16(4):300-2.[19]Wei Q Z,Zhou Z,Chen B S,Liu T K.Experimental study of antiulcer effect Weile capsule[J].Academ J Guangdong Coll Pharm,2000,16(4):300-2.
[20]赖舒,周岐新,张颖,等.石榴皮对实验性胃损伤及相关机制的研究[J].中药药理与临床,2009,25(3):49-51.[20]Lai S,Zhou Q X,Zhang Y,et al.Effects and mechanisms of the acetone extract obtained from pomegranate bark on the experimental gastric damages[J].Pharmacol Clin Chin Mat Med,2009,25(3):49-51.
[21]Guth P H,Aures D,Paulsen G.Topical aspirin plus HCL gastric lesions in the rat[J].Gastroenterol,1979,76(1):88-93.
[22]符健,韩丽,邢桂兰,等.秋茄提取物抗大鼠实验性胃溃疡作用研究.[J].中国药理学通报,2007,23(10):1379-83.[22]Fu J,Han L,Xiang G L,et al.Study of gastric antiulcer effects on experimental rats of extracts of Kandelia candel[J].Chin Pharmacol Bull,2007,23(10):1379-83.