【作者】 张丽；

【导师】 田颖刚；

【作者基本信息】 南昌大学 ， 营养与食品卫生学， 2013， 硕士

【摘要】 BP256是以一种特定食物来源的蛋白质为原料,经酶解、过滤、干燥等工艺制得的食源性肽。顺铂被广泛用于肿瘤化疗,属细胞非特异性药物,除了对肿瘤细胞有杀伤作用,对正常组织细胞也产生毒副作用。肾毒性是顺铂主要毒副作用,大大限制了其临床应用。寻找能够减轻顺铂肾毒性,同时不减弱其抗肿瘤效果的食品新资源,具有重要的意义。1.研究食源性肽BP256对顺铂所致小鼠肾毒性的防护作用。一次性腹腔注射20 mg/kg b.w.顺铂建立顺铂损伤动物模型,顺铂注射前3天及后3天连续给予不同剂量BP256灌胃7天。病理切片显示,顺铂模型组肾组织有明显炎性细胞浸润,肾小管上皮细胞脱落、坏死严重,BP256各防护组未见明显炎性细胞浸润,且肾小管上皮细胞脱落、坏死减少。顺铂模型组血清BUN、Scr含量升高,肾组织MDA含量升高、SOD、GSH-PX活力降低,BP256各防护组血清BUN、Scr含量降低,中剂量组降低效果好于低、高剂量组。BP256各组MDA含量均有不同程度的下降。SOD活力较顺铂模型组上升,中、高剂量组与CDDP模型组比较,有显著性差异(P<0.01)。BP256低剂量组肾组织GSH-PX活力有较大提高,与顺铂模型组比有显著性差异(P<0.01)。食源性肽BP256对顺铂所致小鼠肾毒性有防护作用。2.研究食源性肽BP256对顺铂损伤HUVEC-12细胞的保护作用。BP256与HUVEC-12细胞共同培养,观察其对HUVEC-12细胞自身增殖活力的影响。通过考察0.125、0.5、2.5、5、10、20μg/mL顺铂对HUVEC-12细胞的抑制率,筛选顺铂最适作用浓度、时间。MTT法、单细胞凝胶电泳法,研究BP256对顺铂损伤HUVEC-12细胞的保护作用。并测定各组细胞培养上清SOD活力变化。BP256对HUVEC-12细胞自身增殖活力无明显影响,选择顺铂2 μg/mL,作用48小时,建立顺铂损伤HUVEC-12细胞模型。MTT法结果显示BP256保护组细胞存活率较顺铂损伤组提高,单细胞凝胶电泳结果表明,顺铂损伤组细胞尾长、尾部DNA含量百分比、尾矩、Olive尾矩与正常对照组比,有显著性差异。BP256保护组HUVEC-12细胞DNA损伤减轻。3.探讨食源性肽BP256对顺铂抑制A549细胞的影响。考察不同细胞初始接种个数、不同顺铂浓度及作用时间对A549细胞抑制率的影响。BP256与A549细胞共同孵育,观察其对A549细胞生长的影响。BP256与不同浓度顺铂联用,对顺铂抑制A549细胞的影响。BP256对A549细胞生长有微弱的抑制作用,以A549细胞初始接种5×103个,0.125、0.25、0.5、1、2μg/mL顺铂作用48小时建立顺铂抑制A549细胞模型。BP256与各浓度顺铂联用,抑制率较相同浓度顺铂单用组高。BP256对顺铂抑制A549细胞有增敏作用。更多还原

【Abstract】 BP256 is a kind ofpeptide derived from food protein,with the process of enzymolysis.filtration and drying.Cis-platinum is a kind of drug that is nonspecific to cells,which is widely used cancer chemotherapy.And cis-platinum could cause injury to normal tissue cells while it makes effects on antitumor cells.Renal toxicity is a major side effects of cis-platinum,which greatly limits its clinical application.It is significant to find a kind of substance which could reduce the renal toxicity of cis-platinum and not abate its antitumor effect at the same time.1.The research of peptide derived from food protein BP256 was studied on the protective effecton renal toxicity caused by cis-platinum in mice.Cis-platinum injury animal model was set up by the injection of cis-platinum 20mg/kg b.w.one time.The experiment groups were fed with BP256 in different concentrations for 7 consecutive days after 3 days and 3 days before the cis-platinum injection in mice.Kidney tissue pathological slice showed that cis-platinum model group had obvious inflammatory cell infiltration.malpighian tube skin cell fall off,and severely necrotized.The group fed with BP256 did not see obvious inflammatory cell infiltration or cells necrosis.In the cis-platinum model group,it has been found that the content of the serum urea nitrogen(BUN)and the creatinine(Scr)rise.the MDA content increased in the kidney tissues and the activity of SOD and GSH-PX decreased.The result showed that the content of the BUN and the Scr decreased in the group fed with BP256.What’s more,there is a better effect in the middle dose group than the low-dose group and high-dose group.And the content of MDA in BP256 group dropped to a varying degree.The activity of the SOD increased compared with the cis-platinum model group.There was a significant difference between the CDDP model group and the medium and high dose group.And it has been found that the activity of the GSH-PX improved greatly in kidney tissue with the BP256 low dose group,and there was significant difference contrast with the CDDP model group.It has been demonstrated that food-borne peptide BP256 has protective effect on renal toxicity caused by cis-platinum in mice.2.The research of peptide derived from food protein BP256 was studied on the protective effect on cell injury caused by cis-platinum in HUVEC-12cell.The effect on the activity of the HUVEC-12 cell proliferation was observed when the HUVEC-12 cell was cultured with BP256.The optimal concentration and the function time have been screened of the inhibition ratio on HUVEC-12 with the cis-platinum content of 0.125、0.5、2.5、5、10、20 μg/mL.And the contents of the SOD activity were been determinate in each cell culture supernatant.There was no obvious effect on HUVEC-12 cell proliferation of the BP256.The HUVEC-12 cell model injury has been established caused by cis-platinum.The concentrate was 2 μg/mL and the function time was 48 hours.It has been found that the cell survival rate increased in cell group protected with BP256 compared with the cell model injury group caused by cis-platinum with the MTT method.Single cell gel electrophoresis results showed that there was a significant difference in cell percentage of tail length,tail moment,tail DNA content,olive tail moment when the cell model injury group compared with the normal group.It has been found BP256 could decrease the HUVEC-12 cell DNA damage.3.The influence of the BP256 on A549 cells has been investigated that was inhibited by cis-platinum.To study the influence of BP256 on A549,A549 cell has been cultured with BP256.The effect on A549 cell inhibition rate has been studied by observation of different initial inoculation cells,cis-platinum concentration and the action time.The A549 cells inhibited by cis-platinum have been studied by BP256 combined with different concentration cis-platinum.The A549 cells model inhibited by cis-platinum have been established with the initial inoculation cells 5×103and the cis-platinum for 48 hours that the concentrations were 0.125、0.25、0.5、1、2 pg/mL.The inhibition rate was higher than cis-platinum alone group in the same concentration,which was acted by BP256 combined with various concentrations cis-platinum.The BP256 could sensibilize A549 cells on the inhibition of the cis-platinum.更多还原