番茄红素对大鼠肺缺血再灌注损伤的保护作用研究

目的：研究番茄红素对大鼠肺缺血再灌注损伤的保护作用,并考察其作用机制。方法取120只SD大鼠随机分为假手术组、模型组、舒血宁注射液（4 mg/kg）组以及番茄红素5、10、20 mg/kg组,每组20只。除假手术组外采用夹闭左肺门45 min后松夹的方法制备大鼠肺缺血再灌注损伤模型。再灌注2 h后,分别测定各组大鼠肺组织湿质量/干质量比值;通过苏木精–尹红（HE）染色观察肺组织病理学改变;原位末端标记（TUNEL）法观察肺组织细胞凋亡并计算凋亡指数（AI）;检测血清中丙二醛（MDA）含量和髓过氧化物酶（MPO）活性;测定肺组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、过氧化氢酶（CAT）活性。结果与模型组比较,番茄红素10、20 mg/kg组肺组织湿质量/干质量比值显著降低（P＜0.05、0.01）,肺组织病变显著改善,肺组织细胞凋亡状况明显改善;血清中 MDA含量和 MPO活性显著降低（P＜0.05、0.01）,肺组织中SOD、CAT活性显著升高（P＜0.05、0.01）。其中番茄红素20 mg/kg组对肺缺血再灌注损伤大鼠肺组织病变改善最为显著、细胞凋亡状况改善以及凋亡指数降低效果最为显著,对血清中MDA含量和MPO活性显著降低效果更加显著、肺组织中GSH-Px活性显著升高（P＜0.01）。结论番茄红素能够有效降低肺组织湿质量/干质量比值,改善肺组织病变,抑制肺组织细胞凋亡、降低凋亡指数,提示番茄红素对大鼠肺缺血再灌注损伤具有剂量相关性的保护作用,其作用机制可能与番茄红素能够有效改善机体抗氧化酶活性、抑制氧化应激损伤有关。     

番茄中番茄红素提取工艺条件的优化研究

目的：优化筛选番茄红素的最佳提取工艺。方法：利用番茄酱为原料,通过方法学验证、单因素试验、正交试验等对番茄红素的提取工艺进行研究。结果：用乙酸乙酯提取番茄红素的最佳优化工艺为：提取温度50℃,提取时间40 min,乙酸乙酯浓度80％,固液比1?2（g·mL^-1）。在此工艺下,番茄红素的提取率达15.564 mg·100g^-1。结论：优化番茄红素的提取工艺,为番茄红素的进一步开发和利用提供试验基础。     

番茄红素对心肌细胞氧化应激损伤的保护作用

目的：探讨番茄红素（Lycopene）对心肌细胞氧化应激损伤的保护作用及机制。方法体外培养 H9c2心肌细胞,分为正常对照组（对照组）、番茄红素预处理组（Lycopene组）、模型组（H2 O2组）和番茄红素预处理后建立模型组（Lycopene加H2 O2组）。除对照组及Lycopene组外,其余各组用过氧化氢（H2 O2）200μmol／L处理6 h ,建立心肌细胞氧化应激损伤模型。Lycopene组建模前30 min加入5μmol／L的番茄红素。6 h后M T T观察H9c2心肌细胞存活率;分光光度计检测细胞培养液上清中乳酸脱氢酶（LDH）、肌酸激酶同工酶（CK-MB）活性和细胞中丙二醛（MDA）、超氧化物歧化酶（SOD）活性;激光共聚焦联合流式细胞仪观察细胞内活性氧（ROS）的产生和线粒体膜电位;流式细胞仪检测细胞凋亡;酶标仪检测细胞内ATP水平。结果与对照组比较,H2O2组LDH、CK-BM、ROS、MDA及细胞凋亡率显著增加,差异有统计学意义（P＜0．05）;H9c2心肌细胞存活率、SOD、ATP及线粒体膜电位显著降低,差异有统计学意义（P＜0．05）。与 H2O2组比较,Lycopene加 H2O2组LDH、CK-BM、ROS、MDA及细胞凋亡率显著降低,差异有统计学意义（P＜0．05）;H9c2心肌细胞存活率、SOD、ATP及线粒体膜电位显著增加,差异有统计学意义（P＜0．05）。结论番茄红素可以减轻氧化应激条件下H9c2心肌细胞的损伤,其机制可能与清除细胞内氧化应激产物,增强细胞内抗氧化酶活性及改善线粒体功能有关。     

解脂耶氏酵母异源合成番茄红素的初步研究

为了探究番茄红素在解脂耶氏酵母中的生物合成,利用两个解脂耶氏酵母整合表达质粒（单拷贝质粒pINA1269和多拷贝质粒pINA1312）,将成团泛菌来源的番茄红素生物合成基因（crtE、crtB、crtI）克隆到解脂耶氏酵母营养缺陷型宿主菌Po1f中实现多基因共表达,并成功构建了异源合成番茄红素的解脂耶氏酵母工程菌株。在hp4d启动子和XPR2终止子的控制下,本实验共构建了8个能产番茄红素的解脂耶氏酵母工程菌株;同时筛选了其中5个代表性菌株（Po1f 1269IBE 1312、Po1f 1312E 1269IB、Po1f 1312B 1269IE、Po1f 1312I 1269EB、Po1f 1312IBE 1269）基于摇瓶培养进一步研究其生长特性和产物合成情况。结果表明,工程菌Po1f 1312E 1269IB的培养可以获得最高番茄红素产量,相对于细菌干重其产量约为0.9 mg/g。     

微胶囊化番茄红素片对小鼠免疫功能的影响

目的研究微胶囊化的番茄红素片剂对小鼠免疫功能的影响。方法按照卫生部《保健食品检验与评价技术规范》中增强免疫力功能的检验方法,以某品牌微胶囊化番茄红素片为样品,将实验小鼠按体重随机分为5批,每批4组,每组10只,分别进行小鼠脾淋巴细胞转化试验和自然杀伤(NK)细胞活性测定、二硝基氟苯诱导小鼠迟发性变态反应(DTH)试验、抗体生成细胞检测和血清溶血素测定、小鼠碳廓清试验、小鼠腹腔巨噬细胞吞噬鸡红细胞试验。每批小鼠均分为溶剂(纯净水)对照组、低剂量组(83mg/kg b.w)、中剂量组(167mg/kg b.w)、高剂量组(500mg/kg b.w),各剂量组小鼠连续灌胃30d后测定各项指标,实验前后称体重,研究微胶囊化番茄红素片对小鼠免疫功能的影响。结果低、中、高剂量组小鼠的NK细胞活性均高于对照组(P0.05)。低、中剂量能促进ConA诱导的小鼠淋巴细胞转化能力(P0.01,P0.05),提高抗体生成细胞数(P0.05)。在受试剂量下番茄红素片对小鼠迟发型变态反应、半数溶血值、小鼠碳廓清能力、巨噬细胞吞噬鸡红细胞的能力无影响(P0.05)。结论微胶囊化的番茄红素片在适宜剂量下具有增强小鼠免疫力的功能。     

Effect of Lycopene and Rosmarinic Acid on Gentamicin Induced Renal Cortical Oxidative Stress,Apoptosis, and Autophagy in Adult Male Albino Rat

Gentamicin nephrotoxicity accounts for 10%–15% of all cases of acute renal failure. Several natural antioxidants were found to be effective against drug‐induced toxicity. The possible protective effects of lycopene (Lyc) and rosmarinic acid (RA) alone or combined on gentamicin (Gen) induced renal cortical oxidative stress, apoptosis, and autophagy were evaluated. Sixty‐three rats were randomly divided into seven groups named: control, group II received RA 50 mg/kg/day, group III received Lyc 4 mg/kg/day, group IV received Gen 100 mg/kg/day, group V (RA + Gen), group VI (Lyc + Gen), and group VII (RA + Lyc + Gen). At the end of the experiment, kidney functions were estimated then the kidneys were sampled for histopathological, immunohistochemistry, and biochemical studies. Administration of rosmarinic acid and lycopene decreased elevated serum creatinine, blood urea nitrogen, renal malondialdehyde and immunoexpression of the proapoptotic protein (Bax), autophagic marker protein (LC3/B), and inducible nitric oxide synthase (iNOS) induced by gentamicin. They increased reduced glutathione, glutathione peroxidase, superoxide dismutase, and immunoexpression of the antiapoptotic protein (Bcl2). They also improved the histopathological changes induced by gentamicin. The combination therapy of rosmarinic acid and lycopene shows better protective effects than the corresponding monotherapy. Anat Rec, 300:1137–1149, 2017. © 2016 Wiley Periodicals, Inc.     

5-ASA and lycopene decrease the oxidative stress and inflammation induced by iron in rats with colitis

Background Supplementation of 5-aminosalicylic acid (5-ASA) and of iron are among the principal therapies in patients with inflammatory bowel disease. Therapeutic iron, as well as heme iron from chronic mucosal bleeding, can increase iron-mediated oxidative stress in colitis. This study was designed to examine the influence of iron supplementation on histological expression and oxidative status relative to 5-ASA treatment and antioxidant treatment.Methods Colitis was induced using the iodoacetamide rat model, and rats were divided into different dietary groups of 6 rats each: 1, normal chow diet (control); 2, diet supplemented with iron; 3, iron supplementation and lycopene; 4, iron and -carotene; 5, 5-ASA; 6, 5-ASA and lycopene; 7, 5-ASA and iron; 8, 5-ASA, iron, and lycopene. The animals were killed after 3 days and the weight of the ulcerated area recorded. Mucosal specimens were histologically evaluated. Myeloperoxidase (MPO) was measured to evaluate inflammatory status (U/g). Malondialdehyde (MDA) was measured in colonic tissue (µmol/g) and superoxide dismutase (SOD) in erythrocytes to assess the degree of tissue oxidative stress.Results Significantly more severe colitis, including necrosis, ulceration, and hemorrhage, was seen in colonic biopsies of rats with colitis when iron was supplemented. This pathology was attenuated when iron was given in combination with 5-ASA and/or lycopene. There was no significant benefit from adding -carotene.Conclusions Iron supplementation can amplify the inflammatory response and subsequent mucosal damage in a rat model of colitis. We suggest that the resultant oxidative stress generated by iron supplementation leads to the extension and propagation of crypt abscesses, either through direct membrane disruption by lipid peroxidation or through the generation of secondary toxic oxidants. Simultaneous treatment with 5-ASA and/or lycopene minimizes the potential hazard of iron. Therefore, we suggest giving iron supplementation with 5-ASA or lycopene or both.     

Lycopene Treatment of Prostate Cancer Cell Lines Inhibits Adhesion and Migration Properties of the Cells

Background: Consumption of lycopene through tomato products has been suggested to reduce the risk of prostate cancer. Cellular adhesion and migration are important features of cancer progression and therefore a potential target for cancer interception. In the present study we have examined the in vitro effect of lycopene on these processes.Methods: Prostate cancer cell lines PC3, DU145 and immortalised normal prostate cell line PNT-2 were used. The adhesion assay consisted of seeding pre-treated cells onto Matrigel™, gently removing non-adherent cells and quantitating the adherent fraction using WST-1. Migratory potential was assessed using ibidi™ migration chamber inserts, in which a cell-free zone between two confluent areas was allowed to populate over time and the migration measured.Results: 24 hour incubation of prostate cell lines with 1.15µmol/l lycopene showed a 40% reduction of cellular motility in case of PC3 cells, 58% in DU145 cells and no effect was observed for PNT2 cells. A dose related inhibition of cell adhesion to a basement membrane in the form of Matrigel™ was observed in all three cell lines and it reached statistical significance for PC3 and PNT2 cells at lycopene concentrations ≥1.15µmol/l. However, in case of DU145, only a concentration of 2.3µmol/l showed a significant reduction.Conclusion: This in vitro investigation indicates that lycopene can influence the cell adhesion and migration properties of cancer cells at a dose which is arguably achievable in patients. The results of our study expand our understanding of a chemo preventive role of lycopene in prostate cancer.     

番茄红素对四氯化碳肝肾损伤的保护作用

番茄红素(Lycopene)属于烃类类胡萝卜素,存在于红色水果和蔬菜中,研究发现番茄红素具有淬灭单线态氧和清除自由基、阻断亚硝胺形成、抑制细胞增殖、诱导细胞分化、减少DNA损伤及诱导细胞间隙连接通讯等作用。番茄红素能够减缓动脉粥样硬化,防止冠心病的发生[1]。番茄红素的生物