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蓝莓花色苷对CP致脏器损伤的保护及抗衰老作用研究

Effects of Anthocyanins from Blueberry on the Protection of Cyclophosphamide-induced Viscera Damage and Anti-aging Function.

【作者】 宋德群

【导师】 孟宪军;

【作者基本信息】 沈阳农业大学 , 食品科学, 2013, 博士

【摘要】 蓝莓是杜鹃花科(Ericaceae)越橘属植物,在我国有丰富的资源。研究表明,蓝莓果实的主要活性成分为花色苷类物质,长期食用可以消除用眼疲劳,保护视力,并且具有抗炎、抗氧化、抗溃疡、抗衰老和抗癌等生理活性。本文对蓝莓花色苷进行了较系统的研究,在课题组研究的基础上,研究了蓝莓花色苷的提取工艺,通过溶剂法和酶法研究蓝莓花色苷的提取工艺;并研究了蓝莓花色苷在环磷酰胺所致大鼠心脏、肺与肝脏损伤模型的保护作用及抗衰老模型中的保健功效,为更好地开发和利用蓝莓资源提供理论支持。主要研究结论如下:1.采用酸性乙醇对蓝莓中花色苷进行提取,在单因素的基础上通过响应面试验方法确定蓝莓花色苷提取所需最佳提取条件为:提取液乙醇体积分数60.65%,料液比1g:20.65ml,提取时间122.53mm, pH3.0,提取温度50℃,提取两次。在此条件下进行提取,确定蓝莓冻果花色苷含量约为327.35mg/100g。2.通过环磷酰胺(CP)对大鼠进行造模,研究了蓝莓花色苷对CP所致心脏损伤的保护作用。本试验从血清心肌酶谱、组织病理学二个方面研究了蓝莓花色苷对CP所致心脏损伤作用的保护作用,通过对抗氧化酶SOD活性、MDA水平及凋亡蛋白的测定,探讨了保护机制。3.研究了蓝莓花色苷对环磷酰胺所致肺损伤的保护作用。探讨了蓝莓花色苷对CP所致的肺损伤的拮抗作用,实验结果显示:蓝莓花色苷改善了CP所致的肺损伤的病理变化,减轻了炎性渗出,减少了促炎因子的表达,增加了抑炎因子的活性,同时减轻了过氧化损伤,表明蓝莓花色苷对CP所致的肺损伤有一定的保护作用。4.研究了蓝莓花色苷对环磷酰胺所致肝损伤的保护作用。本试验从血清指标、组织病理学两个方面研究了蓝莓花色苷对CP所致肝脏损伤作用的保护作用。病理结果表明,CP导致大鼠肝脏中央静脉周围可见肝细胞出现弥漫性空泡变性,血清测试结果发现,CP导致大鼠ALT、AST与ALP值均显著升高,这与其它人的报导一致。与模型组相比较,蓝莓花色苷组病理变化、血清酶学指标都有所改善,证明了蓝莓花色苷对CP导致的肝脏损伤具有明显的保护作用。5.研究了低、高剂量组的蓝莓花色苷对小鼠学习能力的影响,发现两个剂量组均能够显著的改善实验中衰老小鼠的学习记忆障碍,提高小鼠的空间分辨能力,并能够延长衰老小鼠的游泳时间;延长小鼠跳台实验中小鼠的潜伏期,并减少错误次数;提高Y迷宫实验中大鼠的学习能力;并可缩短明暗箱实验中大鼠的潜伏期。这些结果表明低、高剂量的蓝莓花色苷能够有效的改善衰老小鼠和大鼠的学习记忆障碍,提高学习记忆能力。

【Abstract】 Blueberry belongs to Ericaceae Vaccinium.spp and is abundant over our country. It has been studied that there are abundant anthocyanins in blueberries and many physiological and officinal functions in blueberries such as eliminating eye fatigue, protecting the eyesight, antioxidant, anti-aging, anti-ulcer, anti-inflammatory and anti-cancer et al. In this paper, blueberry anthocyanins have been analyzed by pH differential method; the extraction technology of blueberry anthocyanins is optimized by response surface method with acidic alcohol as extractant.The main conclusions are as follows:1. The blueberry pigments is extracted by acidic alcohol. On the basis of single factor test, the optimization of various factors and their interaction effect is analyzed by response surface method. The best extraction parameters are:ethanol content is60.65%, material-solvent ratio is1g:20.65ml, extraction time is122.53min, the value of PH is3.0extraction temperature is50℃, extraction twice. Under this extraction condition, the determination of the blueberry anthocyanins is327.35mg/100g.2. The heart protective effects of the blueberry anthocyanins (20mg/kg and80mg/kg, ig) are investigated in CP-induced heart injury model (100mg/kg, ip). The protective role of the blueberry anthocyanins is demonstrated by the tests from two aspects—serum myocardial enzyme spectrum and histopathologic study. The blueberry anthocyanins show protective effect on heart damage. The mechanism is investigated by the change of SOD activity of antioxidant enzymes and MDA level, and the apoptosis protein of Bax and Bcl-2with westernblot, the results showed blueberry anthocyanins increased the SOD activities and lower MDA level, and decreased the proapoptosis protein Bax expression and increased the antiapotosis protein Bcl-2expression, the results indicate that antioxidative stress and antiapoptosis are involved in the protective function of blueberry anthocyanins.3. Blueberry anthocyanins have been studied on the lung injury caused by cyclophosphamide. The rats are the same with described in heart protective test. The experimental results show that:blueberry anthocyanins improve the pathological changes of lung injury caused by CP, reduce inflammatory exudation and the expression of proinflammatory factors, increase the activity of the suppresve inflammatory factors, and reduce the peroxidation damage at the same time, indicating that anthocyanins of blueberry possess certain protective effect in lung injury caused by CP.4. Blueberry anthocyanins have been studied in liver injury caused by cyclophosphamide. The protective effects of the test are from two aspects called serum index and histopathological study. The results show that the CP treated rat liver with visible diffuse vacuoles degeneration of hepatic cells around the central vein, serum test results show that ATL AST and ALP values are significantly increased in CP treated rats, which are consistent with other researchers’ results. Pathological changes of serum enzymology indexes are improved in blueberry anthocyanins groups, which demonstrat the blueberry anthocyanins have the outstanding clear protective role in liver injury.5. The influence of high and low dose group of blueberry anthocyanins on learning ability has been investigated in mice, the results show that two groups can significantly improve the performances of aging mice in Morris water maze experiment, and extends the diving platform incubation period, besides, it can decrease the data error, and improve the learning ability of rat in Y maze experiment and also; can shorten latent period of rats in the light-dark transition test. Blueberry anthocyanins can increase the aging mice swimming time. The results indicate that the blueberry anthocyanins can effectively improve the aging mice and rats learning and memory impairment, and increase the muscle function.

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