多代连续饮用四种饮水大鼠肝肾功能和骨骼生长发育比较研究

  本文关键词：多代连续饮用四种饮水大鼠肝肾功能和骨骼生长发育比较研究，由笔耕文化传播整理发布。

        水是人体最重要和最基本的营养素之一。随着水污染的日益严重和经济的快速发展以及大众消费观念的改变，饮水质量受到人们的高度重视，对饮用水的要求也越来越高。目前以纯净水、天然矿泉水和矿物质水为主的各类瓶（桶）装水在日常饮用水中的比例逐年增高。经过反渗透、纳滤、吸附等工艺深度加工处理的瓶（桶）装水在有害污染物的含量上的确得到了控制，但对人体健康有重要影响的水中天然矿物质含量也出现不同程度的下降。受研究过程中性别、水质、饮食、实验方法等因素的影响，长期饮用较低矿物质含量的水对健康有何利弊至今仍没有统一定论。本团队研究饮用软水对健康的影响已有10余年，前期研究显示，饮用纯净水的雌雄大鼠平均体重均持续显著性高于饮用凉开水。长期饮用纯净水后，雌鼠血清中镁含量显著降低，尿中镁含量雌雄均有下降趋势，钾、钠浓度均有升高趋势，铁、锌、铜元素含量显著降低，铬含量显著升高。大鼠骨骼的生长发育指标中股骨长度、骨生物力学性能及骨密度纯净水和自来水无显著差异。目前国内外在饮用水的生物学效应方面主要集中在上世纪五六十年代的人群流行病学调查和基于单代动物的实验研究，尚未见多代连续饮用不同饮水的相关生物学效应研究和报道。因此，我们选择自来水、天然水、矿物质水、纯净水进行比较研究，建立多代连续饮用四种饮水的大鼠模型，从一般生长发育、肝肾功能、机体酸碱状态、骨骼生长发育几方面研究和探讨多代连续饮用四种饮水与健康效应的关系，为进一步深入研究奠定基础。实验方法：1．多代连续饮用四种饮水的动物模型F0代：断乳SD雌鼠120只/雄鼠60只，均随机分为自来水组（Tap water，T组）、天然水组（Natural water，N组）、矿物质水组（Mineralized water，M组）、纯净水组（Purified water，P组）四组，每组30（雌性）/15（雄性）只。饮水至4月龄开始繁殖实验，雌雄2：1比例交配，产生约20只受孕F0代雌鼠。雄鼠处死，孕鼠保留至生产F1代仔鼠。F1代：各组F1代雌仔鼠30只，雄仔鼠15只。饮用与F0代相同饮水至4月龄，雌雄2：1比例交配，产生约20只受孕F1代雌鼠。雄鼠处死，孕鼠保留至生产F2代仔鼠。F2代：各组F2代雌仔鼠30只，雄仔鼠30只，与F1代相同饮水，采用饮水瓶给水，除饮水外其他条件完全相同，每日早上8点定时更换饮水和添加饲料。自来水源于重庆城市供水系统，天然水、矿物质水、纯净水均采用市售相同批次的瓶装饮用水。四种饮水经检测均符合相应水质标准。2．取材和指标饲喂期间，①每日观察各组动物的外观和行为，包括动物的饮食情况、精神和活动状况、毛发润泽度及排泄物情况等。②每月称量体重1次。③每月测定1次大鼠的24h饮水、摄食量。饲喂F2代大鼠至10月龄处死。采集腹主动脉血，一部分用血气分析仪检测机体酸碱状态指标，另一部分分离血清，用自动生化分析仪检测肝肾功能指标和用酶联免疫试剂盒测定骨代谢指标。采血后立即剖取雄鼠心、肝、主动脉弓，将器官周围的脂肪结缔组织去除，并将脏器表面血液吸干净，用电子天平称重，计算脏器系数，脏器系数以脏器重量（g）与体重（g）之比表示。同时雌鼠取右侧股骨、胫骨和门齿，剔除肌肉和软组织，用生理盐水浸湿的纱布包裹后，再用锡箔纸包裹，，置于-20℃低温冰箱保存，实验前24h取出标本，放置室温下解冻。股骨用于骨密度和生物力学检测，胫骨和门齿检测其中钙、镁、磷含量。3．统计学分析数据经SPSS13.0统计软件处理，实验数据均以均数±标准差(（?0±s）的形式表示，组间均数差异的显著性比较采用单因素方差分析。结果与讨论1．生长发育1.1一般生长发育情况F2代大鼠观察期内一般生长发育情况均未见显著性差异。提示大鼠在一般生长发育指标上无明显差别。1.2雄鼠重要脏器系数雄鼠心、肝、主动脉弓脏器系数的组间差异均无统计学意义。提示长期饮用四种饮水对雄鼠心、肝、主动脉弓无明显改变。2．肝肾功能2.1肝脏蛋白合成能力以及营养状况雌雄大鼠血清总蛋白（total protein，TP）、血清白蛋白（albumin，ALB）、血清球蛋白（globulin，GLB）、白球比（A/G）的组间差异均无统计学意义。与自来水组比较，雄鼠天然水组（P=0.006）和矿物质水组（P=0.037）前白蛋白（Prealbumin，PA）含量明显降低；雌鼠纯净水组PA含量明显升高（P=0.030）。提示雄鼠自来水组的营养状况较天然水组和矿物质水组好，雌鼠纯净水组营养状况较其余三组好。2.2肾脏功能饮用四种水的雄鼠血清尿酸（Serum uric acid，UA），雌鼠血清尿素（Urea）和尿酸组间差异均无统计学意义。雄鼠血清中尿素含量自来水组高于天然水组（P=0.015）和矿物质水组（P=0.012）；血清中肌酐（Serum creatinine，SCR）含量自来水组低于天然水组（P=0.050），纯净水组低于天然水组（P=0.000）和矿物质水组（P=0.004）；尿素肌酐比（U/C）自来水组高于天然水组（P=0.001）和矿物质水组（P=0.002），纯净水组高于天然水组（P=0.000）和矿物质水组（P=0.000）。雌鼠SCR含量自来水组低于天然水组（P=0.033）和矿物质水组（P=0.000），纯净水组低于矿物质水组（P=0.015）；U/C自来水组高于天然水组（P=0.002）、矿物质水组（P=0.003）和纯净水组（P=0.044）。提示天然水组雌雄大鼠和矿物质水组雌鼠较自来水组的肾小球滤过功能低；矿物质水组雌雄大鼠和天然水组雄鼠较纯净水组的肾小球滤过功能低。3．机体酸碱状态饮用四种水的雌雄大鼠机体酸碱状态指标[pH、PO2、PCO2、HCO3-act、HCO3-st、SO2%、TCO2、BE（B）、BE（ecf）]组间差异均无统计学意义。提示在观察期内饮用四种水均能维持机体酸碱平衡。4．骨矿物质含量天然水组胫骨中钙（calcium，Ca）含量较自来水组（P=0.001）和矿物质水组（P=0.005）降低，牙齿中Ca含量较自来水组（P=0.044）和矿物质水组（P=0.032）降低。纯净水组胫骨中Ca含量较自来水组（P=0.000）、天然水组（P=0.000）、矿物质水组（P=0.000）均降低。天然水组胫骨（P=0.048）和牙齿（P=0.001）中镁（magnesium，Mg）含量、纯净水牙齿中Mg含量（P=0.003）较自来水组均降低。胫骨和牙齿中磷含量各组间差异无统计学意义。提示水中钙、镁元素的含量对骨骼和牙齿的矿化有很大的影响，长期饮用低矿物质含量的纯净水和天然水可使骨骼和牙齿中钙镁的沉积减少，含量下降。5．骨密度饮用四种水的雌鼠股骨骨密度（bone mineral density，BMD）、骨矿物盐含量（bonemineral content，BMC）和面积组间差异均无统计学意义。6．骨生物力学性能与自来水组比较，大鼠股骨最大桡度天然水组（P=0.000）、矿物质水组（P=0.000）、纯净水组（P=0.000）均有显著降低；弹性桡度天然水组（P=0.001）、矿物质水组（P=0.022）均有显著降低；断裂应变天然水组（P=0.001）、矿物质水组（P=0.002）均有显著降低；杨氏模量天然水组明显升高（P=0.010）。提示自来水组骨骼整体生物力学性能优于其他三组，天然水组在维持骨骼材料的弹性上有一定优势。7．骨代谢水平血清中骨钙素（bone glutarny I protein，BGP）含量天然水组高于自来水组（P=0.005）、矿物质水组（P=0.002）和纯净水组（P=0.025）。I型前胶原羧基端前肽（Procollagen typeI caboxyterminal propeptide，PICP）含量自来水组高于天然水组（P=0.000）和纯净水组（P=0.000），矿物质水组高于天然水组（P=0.000）和纯净水组（P=0.000）。I型胶原交联羧基末端肽（Cross-linked-telopeptide of type I collagen，ICTP）含量天然水组低于自来水组（P=0.023）和纯净水组（P=0.000）。骨碱性磷酸酶（bone alkaline phosphatase，BALP）含量各组间差异无统计学意义。提示饮用自来水后大鼠的成骨细胞活性、合成胶原速度以及骨正常矿化速度比天然水和纯净水快，天然水组对破骨细胞活性的抑制作用强于自来水组和纯净水组。纯净水组成骨细胞活性低于天然水组，破骨细胞活性高于天然水组和自来水组，骨代谢动态平衡被打破的几率较自来水组和天然水组大。8．四种饮水的水质特征以及各项指标变化与水质关系的分析自来水、天然水、矿物质水、纯净水的总硬度（以CaCO3计）分别为200.3、69.6、2.3、0.8mg/L；TDS分别为229、87.2、10.9、1.2mg/L。四种水的化学耗氧量（Chemicaloxygen demand，COD）≤1.0mg/L，表明四种水在有机物水平方面没有本质区别，本研究中观察到的生物学效应改变主要归结于水质矿物元素水平上的差异。实验观察到四种水的钙镁元素含量以及钙镁浓度比值也存在差异，自来水钙镁比值为4.18:1，天然水钙镁比值为1.13:1，矿物质水钙镁比值为0.05:1，纯净水钙镁比值为2:1。自来水中锌含量高于其他三组。推测水中钙、镁、锌等元素的含量和钙镁比例可能是引起几种指标出现差异的主要原因。结论1．四种水质的优缺点自来水在维持雌鼠肾小球滤过功能、提升成骨细胞活性、增加骨骼系统钙镁沉积、提高骨骼整体的生物力学性能和维持雄鼠肝脏蛋白合成能力继而提升机体营养水平方面优于其他三种饮水。但饮用自来水在抑制破骨细胞活性和维持骨骼材料弹性方面较饮用天然水略显不足。天然水在维持骨骼材料弹性方面优于自来水；在抑制破骨细胞活性方面优于自来水和纯净水。但在肝脏蛋白合成能力、营养水平、肾小球滤过功能上雌雄大鼠较自来水均显著降低；雄鼠肾小球滤过功能较纯净水显著降低；雌鼠的骨矿物质含量、骨骼整体生物力学性能、成骨细胞活性较自来水均显著降低；骨钙含量较矿物质水也有降低。大鼠在饮水适应期出现饮用矿物质水的雄鼠饮食量较纯净水组明显减少，雌鼠摄食量较其余三组明显增加。矿物质水仅在较天然水的骨钙含量和成骨细胞活性，较纯净水的成骨细胞活性方面略显优势，其余各项指标均未发现有明显优劣势。纯净水在维持大鼠肾小球滤过功能优于矿物质水和天然水，在维持雌鼠肝脏蛋白合成能力继而提升机体营养水平优于其他三种饮水。但在骨骼生长发育的指标上均未发现有明显优势。2．生物效应肝脏蛋白合成能力和营养水平：雄鼠的自来水组优于天然水组和矿物质水组，雌鼠的纯净水组优于其余三组。维持肾小球的滤过功能：自来水组和纯净水组优于天然水组和矿物质水组。骨矿物质含量：自来水组优于天然水组和纯净水组，矿物质水组优于天然水组。骨代谢水平：自来水组和矿物质水组的成骨细胞活性优于天然水组和纯净水组，天然水组抑制破骨细胞活性的能力优于其余三组。骨生物力学性能：自来水组整体性能优于其余三组，但天然水组在维持骨骼自身的弹性上显示出明显优势。3．性别在肝脏蛋白合成能力和营养水平方面存在性别差异：雄鼠自来水组和雌鼠纯净水组营养状况处于较好水平。创新性发现1．本实验首次在雌性大鼠中发现，多代连续饮用矿物质含量丰富的自来水与以天然水、矿物质水、纯净水为代表的软水相比，对骨骼生长发育、骨生物力学性能和骨代谢更为有益。提示水中矿物元素含量较多的自来水使骨骼代谢维持较高水平，从而增加骨骼系统钙镁沉积，继而提升骨骼整体的生物力学性能。天然水组在维持骨骼材料弹性和抑制破骨细胞活性方面有明显优势，天然水中的钾含量较自来水组高且含有锶元素和偏硅酸，两者是否存在关联及其机制还有待进一步验证。2．大鼠在肝脏合成蛋白能力和机体营养水平方面存在性别差异。

    Water is one of the most important and most essential nutrients for human body. With therapidly economic development and the change of public consumption concept as well asincreasingly severe water pollution, People pay high attention to drinking water quality andrequirement is been increasing. At present, new drinking water, like natural water, mineralwater and pure water, are taking the place of tap water gradually, and being the main source ofdrinking water. The noxious pollutant is under control after deep processing by reverseosmosis, nanofiltration, adsorption, exchange, oxidation etc. However, the natural mineral inwater, what is very important for human body healthy, are on decline in different degree.There is no conclusion whether drinking different water is good at health or not for thedifference of gender, water quality, diet, experimental methods in the research of the relationbetween water quality and health.Our team had been researching the influence of drinking soft water to health for morethan ten years. Prophase research shows the average weight of SD rats drinking pure water islast significant higher than rats drinking cold tap water.After the rats drinking pure water for along times, we test the sixteen macroelements and microelements in serum. The Mg in femalerats serum is decline obviously, and the other elements are stable; In urine, the Mg of bothmale and female rats is on the decline, the K、Na density are increase, Fe、Zn、Cu decreasedmarkedly, Cr rose dramatically. The growth and development index of SD rats bones: femurlength、bone biomechanical standard and bone density, pure water is not different from tapwater obviously. At present, the biological effects of drinking water mainly focus on researchabout people and single generation animal. There is no research and report about biologicaleffects of multigeneration drinking different water continuously yet. Due to this, we chosethree soft water in different degree （nature water、 pure water、mineralized water） to docomparison research with tap water, build multigeneration SD rats model with continuousdrinking the four different water, study and discuss the influence of to health from growth and development、liver and kidney function、Body soda acid state、bone density、bonebiomechanical performance、bone mineral content and bone metabolism, to establishfoundation for further research.Methods:1．The model of multigeneration continuous drinking the four water in animal feedsF0:Weaning SD rat （120female,60male） were assigned into4group by therandomization method, group T fed with tap water、group N fed with natural water、group Mfed with mineralized water、group P fed with purified water.（30female,15male per group）At4month of age, two females were paired with one male. The male rat were deadimmediately. Keep the pregnant rat （≥20） until F1pup to be born.F1: Weaning F1rat （30female,15male per group） were allowed to pass on twoconsecutive generations. At4month of age, two females were paired with one male. The malerat were dead immediately. Keep the pregnant rat （≥20） until F2pup to be born.F2: Weaning F1rat （30female,15male per group） were allowed to pass on twoconsecutive generations.The other conditions are indentical besides drinking water, changedrinking water and add feed at eight every morning.The tap water is provided by ChongQing city water supply system, water quality fitnational sanitary standard for drinking water （GB5749-2006）. Disembogue left water in waterpipe about two minutes every morning, then change fresh water. Purified water、 mineralizedwater and nature water are from bottled water in market.2．Materials and indexIn feeding period,①Observe the appearance and behavior of each set SD rats everyday, include the drinking and eating situation、mental and action status、hair moist degree andexcreta etc.②Weighing monthly.③Check24h water intake and24h food intake monthly.Feed the rats and kill them ten months later. Gather abdominal aorta blood, some ofblood used for test Body soda acid state index. Isolate serum of left blood to test liver andkidney function indicatorand bone metabolism index.Heart、liver、aortic arch of male rat werepromptly removed and weighed. Meanwhile, collect right femur、tibia、incisor of female SDrat. Cut off muscles, to avoid bone surface damage, section attached soft tissue carefully.Wrap with carbasus soaked physiological saline first, then pack by tin foil paper. Keep in coolfreezer-20℃. Take the sample24hours before experiment, unfreeze under room temperature. Test bone density、bone biomechanical of femur, test K、Mg、P content of tibia and incisor.3．Statistical anlysisThe data is handled by SPSS13.0statistics software, experience data is expressed byform of average standard deviation (（?0±s）, adopt one-way ANOVA to anlysis the significancecomparison of mean difference.Results and Discussion1．Growth and development1.1common growth and developmentIn observation period,there is no remarked difference on the common growth anddevelopment situation、 water intake、food intake、weight increase of four sets rats.1.2male rats’ organ ratioDrink the four water make no statistical meaning to organ ratio of male SD rats’ heart、liver、aortic arch. Prompt: Drink the four water for long term make no change to male SDrats’ heart、liver、aortic arch.2．Liver and kidney function2.1liver protein synthesis capacity and nutrition statusDrink the four water make no statistical meaning to male and female SD rats’ TP、ALB、GLB、A/G. Compare to tap water set, the male SD rats’ PA of nature water set andmineral water set is decreased. The female SD rats’ PA of pure water set is increased. Prompt:The male SD rats’ nutrition of tap water set is better than nature water set and mineral waterset. The female SD rats’ nutrition of tap water is lower than pure water set.2.2kidney functionDrink the four water make no statistical meaning to male SD rats’UA、female SD rats’UREA.The UREA of male SD rats’, T set is, the difference has statistical meaning（P<0.05）; theCREA of T set is lower than N set（P=0.050）, P set is lower than N set（P=0.000）and Mset（P=0.004）; the U/C of T set is higher than N set （P=0.001）and M set（P=0.002）,and P set ishigher than N set （P=0.000）and M set（P=0.000）.The SCR of female SD rats’, T set is lowerthan N set （P=0.033）and M set（P=0.000）,P set is lower than M set（P=0.015）; the U/C of T setis higher than N set（P=0.002）、M set（P=0.003） and P set（P=0.044）.3．Body soda acid state Drink the four water mutilgenerationly and continuously make no statistical meaning tobody soda acid state index of rats. Prompt: In present observation period, the influence ofdrinking the four water made to body soda acid does not reach abnormal clinical state.4．Mineral content in boneThe Ca content in tibia of nature water set is lower than tap water set（P=0.001）、mineralwater set（P=0.005）,the Ca content in teeth of nature water set is lower than tap waterset（P=0.044）、mineral water set（P=0.032）. The Ca content in tibia of pure water set is lowerthan tap water set、nature water set、mineral water set（P=0.000）. Both the Mg content intibia（P=0.048） and teeth （P=0.001）of nature water set and pure water set（P=0.003）is lowerthan tap water set. The P content in tibia and teeth of each set’ difference has no statisticalmeaning. Prompt: The Ca and Mg content in water makes obvious influence to themineralization of boon and teeth, drinking pure water with little mineral can make the Ca andMg sedimentation in boon and teeth decrease. The ratio of Ca and Mg will affect thesedimentation of Ca、Mg in boon. The specific reason and pathogenesis is needed furtherresearch.5．Bone mineral densityThe influence of drinking the four water mutilgeneration and continuously made tofemale SD rats’BMD、BMC、area has no statistical meaning.6．Bone biomechanicsCompare to tap water set, the femur max radial degrees of nature water set（P=0.000）、mineral water（P=0.000）、pure water set（P=0.000） decreased dramatically; the elastic radialdegrees of nature water set（P=0.001）、mineral water set（P=0.022）decreased; the femurmaximum strain of tap water of both nature water set（P=0.001）and mineral waterset（P=0.002）is decreased.The young modulus of nature water set increasedobviously（P=0.010）. Prompt: The bone biomechanical standard of tap water set is better thanthe other three sets; the bone material elasticity of nature water.7．Bone metabolismThe BGP content in serum of T set is lower than N set（P=0.005）、M set（P=0.002）、Pset（P=0.025）. The PICP content of T set is higher than P set（P=0.000）and N set（P=0.000）、Mset is higher than N set （P=0.000）and P set（P=0.000）. The ICTP content of N set is lower thanT set（P=0.023）and P se（P=0.000）. The BALP content difference of each set has no statistical meaning. Prompt: After drunk tap water, the SD rats’ osteogenesis cell activity、composecollagen speed、normal boon mineralization speed is faster than nature water set and purewater set. The inhibition function to bone cell activity, the nature water set is stronger than tapwater set and pure water set. The osteoblast activity of pure water set is lower than naturewater set, osteoclast activity is higher than nature water set and tap water set. The probabilityof the bone metabolism dynamic balance be broken is higher than tap water set and naturewater set.8．Water quality characteristics and the indicators changeThe total hardness（CaCO3）of tap water、nature water、pure water、mineral water are200.3、69.6、0.8、2.3mg/L respectively; TDS are229、87.2、1.2、10.9mg/L respectively. CODof four water is less than or equal to1.0mg/L, it shows four water do not have materialdifference in organic matter level （COD more or less the same）. Therefore the change ofbio-effect observed in this essay is owe to the difference of water mineral element.The experiment shows that the Ca、Mg content and the concentration ratio is different inthose four set of water. The Ca、Mg ratio of tap water is4.18:1; of nature water is1.13:1; ofmineral water is0.05:1; of pure water is2:1. The Zn content of tap water is higher than theother three sets water. Thus it can infer that the content of Ca、Mg、Zn elements in water andthe Ca、Mg ratio are the main reason which lead to the differences of above index.Conclusion1．Advantages and disadvantages of four water qualityTap water to maintain the female glomerular filtration function, enhance osteoblastactivity and increased deposits of calcium and magnesium of skeletal system, improving theoverall biomechanical properties of bone and the maintenance of male rat liver proteinsynthesis activity and then upgrade nutritional levels than the other three kinds of drinkingwater. But drinking tap water in inhibiting osteoclast activity and maintain elasticity ofskeletal material, slightly less than natural water for drinking.Natural water better than tap water in maintaining elasticity of skeletal materials; ininhibiting osteoclast activity is better than tap water and bottled water. Liver protein synthesis,nutrient levels and Glomerular filtration function in male and female rats are significantlylower than tap water; male rat glomerular filtration function is significantly lower thanpurified water; female bone mineral content and bone biomechanics, Osteoblast activity are significantly lower than tap water; calcium content lower than the mineral water.Water adaptation period, rats drinking mineral water, male rats diet compared withdrinking pure water Group significantly reduced, female rats significantly increased foodconsumption than the other three groups. The mineral water only in the natural water bonecalcium content and osteoblast activeness, the pure water osteoblast active aspect reveals thesuperiority slightly, other each target discovery has not had the obvious superior inferiority.Pure water in the maintenance of rat glomerular filtration function better than mineralwater and natural water, to maintain the female liver protein synthesis and thus improve thenutritional level of the body is better than the other three drinking water. However, in bonegrowth and development indicators were not found to have obvious advantages.2．Biological effectLiver protein synthesis and nutrient levels: tap water group of male rats than naturalwater group and mineral water group, female group of purified water better than the otherthree groups. The glomerular filtration function: tap water group and the purified water groupwas superior to the natural water group and the mineral water group. Bone mineral content:tap water group was superior to the natural water group and the purified water group, themineral water group was superior to the natural water group. The level of bone metabolism:the tap water group and the mineral water group, osteoblast activity is better than the naturalwater group and the purified water group, the natural water group the ability to inhibit theactivity of osteoclasts better than the other three groups. Bone biomechanical properties: thetap water group overall performance is better than the other three groups, but the natural watergroup showed significant advantages in maintaining the elasticity of the bone itself.3. GenderThere are gender differences in liver protein synthesis and nutrient levels: male tap groupand group nutritional status of female pure water at a good level.Innovative discovery1．It was first found in female rats, multiple generations of continuous drinking tap waterin bone biomechanics performance and bone metabolism are more useful than natural water,mineral water, pure water. Prompt more mineral elements in tap water to maintain a high levelof bone metabolism, thereby increasing the deposition of calcium and magnesium in theskeletal system, and thus improve the biomechanical properties. Natural water group there are obvious advantages in maintaining bone material elasticity and inhibit osteoclast activity, thepotassium content of natural water than the tap water group and containing strontium andsilicic acid, the association and its mechanism remains to be further verified.2．There are gender differences in the protein synthesis function of the liver and thebody’s nutritional level in rats.

        多代连续饮用四种饮水大鼠肝肾功能和骨骼生长发育比较研究

英文缩写一览表4-6Abstract6-12摘要13-18前言18-21    技术路线20-21第一部分 多代连续饮用四种饮水大鼠生长发育、肝肾功能及机体酸碱状态比较研究21-37    材料与方法21-23    结果23-34    讨论34-36    小结36-37第二部分 多代连续饮用四种饮水雌性大鼠骨矿物质含量比较研究37-44    材料与方法37-40    结果40-42    讨论42-43    小结43-44第三部分 多代连续饮用四种饮水雌性大鼠骨生物力学性能和骨代谢水平比较研究44-56    材料与方法44-47    结果47-52    讨论52-55    小结55-56全文总结56-58致谢58-59参考文献59-64文献综述 饮用水中矿物元素对骨与骨代谢的影响64-72    参考文献69-72硕士在读期间发表的论文情况72-73附件一：四种饮水的水质检测报告73-77附件二：四种饮水矿物质含量检测报告77-78

  本文关键词：多代连续饮用四种饮水大鼠肝肾功能和骨骼生长发育比较研究，由笔耕文化传播整理发布。