CN114569607B - 5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用 - Google Patents
5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用 Download PDFInfo
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Abstract
本发明公开了5‑羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用，实验证明5‑羟色氨酸可以显著提高雌性机体密闭缺氧存活时间，并且可以显著改善低压低氧暴露所致的雌性生理周期紊乱。
Description
技术领域
本发明属于生物技术领域，具体涉及5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用。
背景技术
目前，全世界居住在高原高山地区人口约5亿多，而我国高原面积辽阔，海拔3000m以上的地区约占全国总面积的1/6，而居住在青藏高原的人口约1000万，是世界上居住高原高山地区人口最多的国家。高原是一个特殊环境，有很多对人体不利的因素，如低气压、低氧分压、寒冷、干燥和阳光辐射强等，其中低氧是最大的威胁，海拔4600米高原吸入气氧分压几乎只到海平面地区的一半，高原特有的自然环境因素对机体健康的影响凸显，相关医学保障的问题亟待解决。高原环境可对全身各个系统(神经内分泌免疫、循环、呼吸、消化、泌尿生殖、能量代谢)的生理功能造成明显影响。研究发现高原低氧可导致女性激素分泌功能减弱，女性第二性征推迟，女性激素分泌紊乱，雌激素与促性腺释放激素水平下降，卵泡发育迟缓，卵子成熟率降低，月经初潮推迟，月经周期紊乱(痛经、闭经和月经周期提前)，难以怀孕和孕期胎儿发育不良等，有的可能造成长期不孕不育。前期调研数据也显示，在高原低氧环境下，紧急入藏的女性出现了显著的月经失调。而且高原现场问卷调查也显示平原移居高原(海拔4000米)初期女性月经紊乱率高达70.40％。因此，寻求干预防治高原低氧雌性生理周期紊乱的干预措施势在必行。
5-羟色胺最早是从血清中发现的，又名血清素，而5-羟基色氨酸是血清素的前驱物质，在动物和人体实验中，5-羟基色氨酸可以促进儿茶酚胺的新陈代谢，特别是对多巴胺和去甲肾上腺素有促进作用，已有研究表明5-羟基色氨酸具有调节情绪及睡眠的作用。我们前期研究发现高原低氧暴露导致雌性大鼠生理周期紊乱，伴随黄体生成素的分泌异常，而5-羟色氨酸是否可以提高雌性机体抗缺氧能力，改善高原低氧雌性生理周期紊乱仍未见报道。
发明内容
本发明的目的是克服现有技术的不足，提供5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用。
本发明的技术方案概述如下：
5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱保健品或药物中的应用。
实验证明5-羟色氨酸可以显著提高雌性机体密闭缺氧存活时间，并且可以显著改善低压低氧暴露所致的雌性生理周期紊乱。
附图说明
图1为5-羟色氨酸对雌性小鼠摄食量的影响。
图2为5-羟色氨酸对雌性小鼠体重的影响。
图3为5-羟色氨酸对雌性小鼠常压密闭缺氧存活时间的影响(*p<0.05与空白对照组相比较)
图4为5-羟色氨酸对低氧暴露雌性大鼠摄食量的影响(*p<0.05与空白对照组相比较)。
图5为5-羟色氨酸对低氧暴露雌性大鼠摄食量的影响(*p<0.05与空白对照组相比较)。
图6为5-羟色氨酸对低氧暴露雌性大鼠生理周期的影响(*p<0.05与空白对照组相比较，#p<0.05与低氧对照组相比较)。
具体实施方式
以下通过实施例对本发明进一步说明。
以下实施例中的方法中如无特别说明，所有的方法均为常规方法，所用的生化试剂均为市售试剂。
实施例1 5-羟色氨酸对雌性小鼠常压密闭缺氧存活时间的影响
1.实验动物
SPF级昆明雌性小鼠20只，体重18～20g，购于北京维通利华实验动物技术有限公司提供。
2.实验药物
(1)0.5％羧甲基纤维素钠溶液：取羧甲基纤维素钠粉末5g，加超纯水至1000ml，用磁力搅拌器搅拌，加速溶解。
(2)5-羟色氨酸溶液：根据小鼠的体重称取5-羟色氨酸(Swanson)，溶解在0.5％羧甲基纤维素钠溶液中，制成5-羟色氨酸溶液。
3.实验动物分组及给药
20只18-20g的雌性小鼠适应性饲养7天。7天后根据小鼠的体重进行随机分组，每组10只，分别为空白对照组(0.5％羧甲基纤维素钠溶液)、5-羟色氨酸组(15mg/kg)。标准条件下饲养，实验过程中动物自由摄食和饮水。每天早上8：30给小鼠进行灌胃给药，每只给药0.5ml，连续给药15天，同时每天称量小鼠食物的重量，计算摄食量的变化，每隔3天称量一次小鼠的重量。
4.常压密闭缺氧实验
在250ml磨砂广口瓶的瓶盖上抹上凡士林，防止空气透过瓶口进入瓶中，制造严格的缺氧环境，同时在每个实验用的广口瓶底铺上5g碱石灰，其作用是吸收实验过程中小鼠在瓶中呼吸产生的水和二氧化碳，碱石灰上盖上一张圆形的滤纸，防止碱石灰遇水放热烫伤小鼠。最后一次灌胃给药30分钟后，将小鼠放入广口瓶中，迅速扭转瓶盖到不漏气，同时立刻用计数器开始计时，以小鼠最后一次呼吸(小鼠强烈挣扎后忽然瘫痪，胸部也不在发生起伏)为死亡特征，记录每只小鼠的存活时间。
5.结果
实验期间，各组动物均无咬伤现象，毛色正常。统计学分析结果表明，与空白对照组相比，5-羟色氨酸组雌性小鼠的摄食量无显著差别(P>0.05)，表明5-羟色氨酸对雌性小鼠的摄食量没有不良影响(图1)，同时研究发现5-羟色氨酸干预后的雌性小鼠的体重与空白对照组小鼠的体重相比无显著的统计学差异(P>0.05)，说明5-羟色氨酸对雌性小鼠的体重没有不良影响(图2)。灌胃给药两周后，检测雌性小鼠常压密闭缺氧条件下的存活时间，统计学分析结果显示，与空白对照组相比，5-羟色氨酸灌胃给药的雌性小鼠在常压密闭缺氧条件下的存活时间显著延长(P<0.05)(图3)。
实施例2 5-羟色氨酸对低氧暴露雌性大鼠生理周期的调控作用
1.实验动物
SPF级雌性SD大鼠36只，体重180～200g，由北京维通利华实验动物技术有限公司提供。
2.实验药物
(1)0.5％羧甲基纤维素钠溶液：取羧甲基纤维素钠粉末5g，加超纯水至1000ml，用磁力搅拌器搅拌，加速溶解。
(2)5-羟色氨酸溶液：根据小鼠的体重称取5-羟色氨酸(Swanson)，溶解在0.5％羧甲基纤维素钠溶液中，制成5-羟色氨酸溶液。
3.实验动物分组及给药
雌性SD大鼠适应性饲养7天，随后根据大鼠的体重随机分组，分为空白对照组(Control， 0.5％羧甲基纤维素钠溶液)、低氧对照组(Control，0.5％羧甲基纤维素钠溶液)，低氧+5-羟色氨酸组(10mg/kg)，每组12只，实验过程中动物自由摄食和饮水，低氧+5-羟色氨酸组灌胃给予10mg/kg/天的5-羟色氨酸1周，空白对照组及低氧对照组给予等量的溶剂1周，低氧对照组与低氧+5-羟色氨酸组随后置于低压舱内(模拟海拔6000m)暴露2周，期间继续灌胃给药，空白对照组整个实验过程置于正常标准条件下饲养。
4.动情周期观察
每天上午8：00－9：00、下午19∶00－20：00进行阴道脱落细胞涂片检查，根据阴道脱落细胞的形态学变化确定大鼠的动情周期。固定大鼠，暴露阴道后，用移液枪吸取100μL无菌生理盐水，打入雌性大鼠阴道内部并来回抽吸两次，均匀平铺到载玻片中央，自然风干，95％乙醇固定15min，室温下自然晾干，进行HE染色15min，蒸馏水冲洗干净后镜检。按照大鼠性周期阴道涂片的细胞变化特点来划分大鼠性周期的各个阶段：①动情前期:可见大量有核上皮细胞、少量角化上皮细胞；②动情期：可见满视野角化上皮细胞、少量有核上皮细胞；③动情后期：可见有核上皮细胞、角化上皮细胞及白细胞；④动情间期：可见大量白细胞和少量黏液。
5.实验结果
实验期间，各组动物均无咬伤现象，毛色正常。统计学分析结果表明，低氧+5-羟色氨酸组大鼠的摄食量与低氧对照组雌性大鼠的摄食量无明显差别(P>0.05)，说明5-羟色氨酸对雌性大鼠摄食量没有不良影响(图4)，同时研究发现在实验的中期和实验的末期，低氧+5-羟色氨酸组雌性大鼠的体重与低氧对照组雌性大鼠的体重相比无显著的统计学差异(P>0.05)，说明5-羟色氨酸对雌性大鼠的体重没有不良影响(图5)
雌性哺乳动物的动情周期一般分为4个阶段：动情前期、动情期、动情后期和动情间期。在雌性的非妊娠期间，动情状态呈周期性变化，卵子的形成和性激素的分泌也呈周期性波动，雌性大鼠的动情周期常被作为体内激素水平变化以及生殖器官如卵巢和子宫病理生理状态的重要标志物。雌性大鼠正常的生理周期约为4-5天，动情前期持续时间约17-21小时。动情期持续时间9-15小时；动情后期，以白细胞和角质化细胞为特征，持续时间10-14小时；动情间期持续时间60-70小时；阴道脱落细胞涂片检查结果发现：与空白对照组相比，低氧对照组雌性大鼠生理周期紊乱率显著升高(P<0.05％)，而5-羟色氨酸干预可以显著改善高原低氧暴露导致的雌性大鼠生理周期紊乱，差异有统计学差异(见表1和图6)。
表1.5-羟色氨酸对低氧暴露雌性大鼠生理周期的影响
*p<0.05vs对照组，#p<0.05vs低氧组。
Claims (1)
1.5-羟色氨酸在制备改善高原低氧所致雌性生理周期紊乱药物中的应用。
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