CN104027386A - 叶下珠在失眠症的新用途 - Google Patents
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Abstract
本发明涉及叶下珠的新用途，本发明采取的技术方案是：叶下珠在制备预防、治疗失眠症疾病药物中的应用。本发明优点在于：1、本发明对叶下珠发掘了新的医疗用途,开拓了一个新的应用领域。2、本发明的叶下珠安全无毒,药理作用强,预示着很好的药用前景。3、经大量动物实验研究证明，药物叶下珠减少小鼠自主活动的作用较明显，可用于预防及治疗失眠症疾病。
Description
技术领域
本发明涉及叶下珠的新用途,尤其涉及在制药领域中的应用。
背景技术
失眠已成为当今社会的一个发病率较高的疾病，严重影响人们的工作、生活质量，甚至导致失去生活信心而自杀。因此，失眠已成为许多国家广泛关注的社会公共卫生问题。调查研究显示，有33％的美国人、4％～22％的欧洲人患有失眠，而我国目前失眠的发病率高达10％～20％。
叶下珠为大戟科Euphorbiaceae叶下珠属Phyllanthus植物叶下珠的全草，最早记载于《植物名实图考》。《生草药性备要》称其为“日开夜闭草”，《临证指南》称为“阴阳草”，《江西民间草药》称为“夜合草”，《纲目拾遗》云“此草背有小珠，昼开夜闭，高三四寸，生人家墙脚下，处处有之”。叶下珠性味微苦、甘，凉，归肝、脾、肺经。功效有平肝明目、清热解毒、利尿通淋、除疳止痢。中国民间传统用于水肿、尿淋、泄泻、痢疾、黄疸、疳积、目赤肿痛、夜盲、毒蛇咬伤等。王祖堂认为其有清肝明目而柔滋足厥阴之虚热，泻火透气而外发手少阳之经气，临床配伍应用于肝郁化火、肝阳上亢、肝阳化风所致诸症。在古代美洲还用于癌症治疗。
现代研究发现叶下珠中已分离并鉴定的化合物共有二十多个，主要化学成分有：没食子酸、甲氧基糅花酸、卵谷莆醇、丁二酸、胡萝卜苷、山茶索、阿魏酸、术脂素、槲皮素、短叶苏术酸、柯里拉京、黄酮、糅质、生物碱、芸香营、糅料云实索、短叶苏木酸乙脂、老鹤草素、短叶苏木酚酸和去氢诃子次酸三甲脂等。其中叶下珠的化学成分中没食子酸的含量最高，具有抗病毒的作用；柯里拉京的含量很少并且无保肝作用，但是具有抗肿瘤的作用。
对叶下珠的临床应用及药理作用的现代研究显示其有：
1、抗HBV（乙肝病毒）作用：Thyagarajan等用氯仿提取叶下珠，将所得提取物经层析分离的部分分别配制成2％和1％的溶液，再与等量的HbsAg阳性人血清在37℃下混合，48h后粗提取物使血清中HbsAg全部灭活。刘庄等研究证实不同品种、剂型和添加成分的叶下珠提取物的抗HBV作用还提示其抑制病毒不仅在转录水平，可能对mRNA的翻译水平也有影响。中日友好医院以鸭体内实验证实叶下珠可降低鸭血清中DHBVDNA和DNA聚合酶。邓学龙等用先天感染DHBV的广州麻鸭作为鸭乙肝模型，发现台湾叶下珠有降低DHBV--DNA的作用。
2、对动物肝细胞损伤的防治作用：任丽娟等试验显示叶下珠能抑制CCl4和D一半乳糖所致的血清谷丙转氨酶、丙氨酸转氨酶升高和肝细胞坏死。周世文、陈晓红等动物实验证实叶下珠能明显降低CCl4引起的小鼠血清ALT的升高，抑制肝脏MDA(丙二醛)的生成，能阻止CCl4体内引起的大鼠肝细胞膜流动性的降低，抑制肝细胞ca2+的升高，提示叶下珠的抗CCl4肝损伤作用机制与其抑制脂质过氧化反应和阻止Ca2+升高有关，从而保护了肝细胞膜正常流动性和完整性。冯天保等的实验结果提示福建叶下珠能显著降低血清转氨酶活力和肝脾脏器指数，对肝细胞坏死有一定的改善作用，具有较好的抗小鼠免疫性肝损伤作用，这为临床应用福建叶下珠治疗病毒肝炎提供了一定的药理学依据。
3、治疗丙型病毒性肝炎：含叶下珠的健脾补肾方（由太子参、茯苓、三七、赤芍、丹参、杜仲、珍珠草、田基黄、菟丝子、五爪龙、郁金、何首乌等组方）和健脾活血方（由太子参、茯苓、五爪龙、白术、丹参、赤芍、三七、红花、珍珠草、楮实子组方）治疗慢性丙型病毒性肝炎，在恢复肝功能和抗纤维化方面有一定疗效。
4、对人肝癌细胞的影响：王昌俊等人用叶下珠提取物处理人肝癌细胞SMMC7221，观察到SMMC7221活力明显减弱，氚标胸腺嘧啶核甘渗入率明显降低，DNA合成抑制率与药物剂量成线性关系。说明叶下珠具有杀伤人肝癌细胞SMMC7221和抑制其增值作用。国外学者研究发现叶下珠对土拨鼠肝癌的发生有抑制作用，经叶下珠治疗后肝癌发生率由85％下降到20％，两组间有显著性差异。
5、抗单纯疱疹病毒Ⅱ型的作用：郭卫真等人采用地鼠肾细胞(BHK)和原代兔肾细胞对8种叶下珠属植物水提物进行了体外抗单纯疱疹病毒II型作用的研究。结果显示：叶下珠水提物在该两种细胞培养上均有不同程度抑制单纯疱疹病毒II型的作用。提示叶下珠属植物对单纯疱疹病毒II型感染有一定的治疗作用。
6、抗爱滋病毒：对爱滋病毒逆转录酶（HIV-IRT）的抑制作用，目前认为是预防爱滋病的新进展，天然产物已应用于HIV-IRT的抑制剂。
7、抑菌、抗菌作用：苦味叶下珠醇提取物具有抗菌作用，对肺炎克雷伯菌、金黄色葡萄球菌、大肠杆菌具有较强的抑菌作用，其最低抑菌浓度（MIC）分别为4.68mg/L，2.34mg/L，2.34mg/L；高浓度时甚至可杀死肺炎克雷伯菌，最低杀菌浓度（MBC）为9.28mg/L。
8、其它作用：叶下珠水、醇提取液经有机溶剂萃取的成分具有抗内毒素作用，质量浓度为0.12、0.24、0.48g/L时对内毒素的清除率分别为22.9%，45.6%，81.3%。水提取物还具有明显的抗血栓形成作用，其机制与阻抑血小板-中性粒细胞间相互作用密切相关，与阿司匹林或尿激酶比较，其出血的危险性明显减小。
9、叶下珠毒副作用：仲英等用叶下珠提取物(29．ml-1)，每鼠灌胃3ml(360／kg)，经7d未见动物有不良反应。李彩东用叶下珠复方“复方苦味叶下珠胶囊”的急性毒性试验结果显示，给小鼠灌服药物最大耐受量可达300g/kg，相当于成人临床用量的800倍，未见小鼠死亡；长期毒性试验中3个剂量组大鼠的一般情况、体重、血液学指标、血清生化指标及病理检查结果与对照组相比无显著性差异（ P＞0.05）。说明复方苦味叶下珠胶囊无明显毒性，临床应用安全、可靠。该药尚未发现因长期服用出现明显毒副作用。
迄今为止，对叶下珠的临床应用研究多从其抗肝炎病毒、防治肝损害出发，多家医院及专家报道了叶下珠及制剂在治疗乙肝、丙肝取得显著疗效。除上述药理及临床研究外，从叶下珠传统功效及应用进行现代临床研究的较少，也未发现其新的治疗领域。
发明内容
本发明的目的是针对现有技术中的不足，提供一种药物的新用途。
为实现上述目的，本发明采取的技术方案是：
叶下珠在制备预防、治疗失眠症疾病药物中的应用。
本发明优点在于：
1、本发明对叶下珠发掘了新的医疗用途,开拓了一个新的应用领域。
2、本发明的叶下珠安全无毒,药理作用强,预示着很好的药用前景。
3、经大量动物实验研究证明，药物叶下珠减少小鼠自主活动的作用较明显，可用于预防及治疗失眠症疾病。
具体实施方式
下面对本发明提供的具体实施方式作详细说明。
一、材料：
1. 叶下珠：由福建省三明市中西医结合医院药剂科提供。取100克叶下珠生药，加水1600ml，武火煎煮，煮沸后文火15min，倒出药液过滤，再加水1000ml进行第二次煎煮，倒出药液，二次合并过滤后浓缩至40ml（2.5g生药/ml）
2. 酢酱草：由上海雷允上药店提供。取100克酢酱草生药，加水1200ml，武火煎煮，煮沸后文火15min，倒出药液过滤，再加水800ml进行第二次煎煮，倒出药液，二次合并过滤后浓缩至66.7ml（1.5g生药/ml）
3. 落花生枝叶：由上海宝龙药业有限公司提供。取100克落花生枝叶生药，加水1200ml，武火煎煮，煮沸后文火15min，倒出药液过滤，再加水800ml进行第二次煎煮，倒出药液，二次合并过滤后浓缩至66.7ml（1.5g生药/ml）
4. 实验动物：SPF级，雄性，昆明种小鼠16—18g，由上海杰思捷实验动物有限公司提供。许可证号：SCXK（沪）2012—0006，合格证号：0078697。
5. 实验仪器：电脑一台；小鼠自发活动光电测试仪一套，由上海凯成电子技术研究所研制，型号：2001—HDX。
二、实验方法：
1. 分组：水对照组；叶下珠组；酢酱草组；落花生枝叶组。
2. 方法：采用光电管法电脑自动记数，记录小鼠的平行移动距离。将小鼠放入活动箱内先适应5min，再记录10min活动。以平移距离为筛选指标进行筛选，用药前10分钟活动次数高于1850或低于1000予以剔除。合格动物进行灌胃给药，放入活动箱内5min 后开始记录小鼠活动次数作为药理实验数据。并储存数据、分析数据。
3. 给药途径和方法：
1) 水对照组：随机分组，每组10只，将小鼠放入自发活动仪中观察15分钟后，按0.4ml/20g体重，灌胃给蒸馏水，灌胃后放入自发活动仪中观察40分钟。
2) 叶下珠组：随机分组，每组10只，灌胃给叶下珠提取液50g /kg。方法同上。
3) 酢酱草组：随机分组，每组10只，灌胃给酢酱草提取液30g /kg。方法同上。
4) 落花生枝叶组：随机分组，每组10只，灌胃给落花生枝叶提取液30g /kg。方法同上。
三、实验数据：
水对照组用药后小鼠活动次数数据见表1
叶下珠提取物组用药后小鼠活动次数数据见表2
酢酱草提取物组用药后小鼠活动次数数据见表3
落花生枝叶提取物组用药后小鼠活动次数数据见表4
四、统计方法和实验结果：
采用SPSS18.0统计软件LSD法做方差分析统计比较试验药物各时间段的活动差异。实验结果见下表5：
叶下珠。酢酱草提取物对小鼠自主活动的影响（光电管法）
注：各时段水对照组与用药组比较：* P<0.05；
结果：各给药组与水对照组比较：用药后各时间段叶下珠组、酢酱草组均有显著性差异（P<0.05）；酢酱草组仅灌胃后第一个时间段有差异，其余时间段无差异。
五、结论：
根据以上实验结果可见叶下珠（50g生药 /kg）、落花生枝叶（30g生药 /kg）减少小鼠自主活动的作用较明显，而酢酱草组无明显镇静催眠作用。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员，在不脱离本发明方法的前提下，还可以做出若干改进和补充，这些改进和补充也应视为本发明的保护范围。
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1.叶下珠在制备预防、治疗失眠症疾病药物中的应用。
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