CN111297885A - 老鹳草多糖在制备保肝药物中的应用 - Google Patents
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Abstract
本发明属于生物技术领域,具体涉及老鹳草(Geranium wilfordii Maxim.)多糖的制备以及其在开发保肝药物中的应用。本发明采用了刀豆球蛋白A诱导的免疫性肝损伤模型,考察其保肝活性。实验结果表明,老鹳草多糖能够显著降低肝损伤小鼠肝功能指标,可用来研发保肝药物,尤其是治疗免疫性肝损伤的药物。本次发明对老鹳草进一步开发利用具有重要意义。
Description
技术领域
本发明属于生物技术领域,具体涉及老鹳草多糖的制备方法及其在开发保肝药物中的应用。
背景技术
自身免疫性肝炎是由自身免疫反应介导的慢性肝炎,其临床特征为不同程度的血清转氨酶AST、ALT升高、高丙球蛋白血症、自身抗体阳性、组织学特征为以淋巴细胞、浆细胞浸润为主的界面性肝炎,严重者可发展成为肝硬化。目前治疗自身免疫性肝炎的药物为糖皮质激素等一些免疫抑制剂,其副作用明显一直是临床治疗的困扰之一。结合临床实际问题,寻找低毒且作用显著的保肝药物迫在眉睫。
多糖在植物、藻类、菌类及动物体内广泛存在,是自然界中含量最丰富的生物大分子之一。研究表明多糖类成分具有多种药理活性,如调节免疫、抗炎、抗氧化、抗肿瘤等,在药品及保健食品领域有很多的研究与应用。
老鹳草(Geranium wilfordii Maxim.)为牻牛儿苗科老鹳草属植物,是我国常用中药之一,具有祛风湿、通经络等功效。为了更好地开发利用其有效成分、我们将目标锁定于于其暂未报道过的多糖成分,发现其多糖成分确实具有显著的保肝活性,对老鹳草进一步开发利用具有重要意义。
发明内容
本发明的目的之一在于提供老鹳草多糖的制备方法;目的之二在于提供老鹳草多糖在制备低毒高效保肝药物中的应用;目的之三在于对老鹳草进一步开发利用。
为实现上述发明目的,本发明提供如下技术方案:
老鹳草多糖在制备保肝药物中的应用
老鹳草多糖的制备方法为:将老鹳草药材粉碎,经甲醇回流脱脂后以80℃水提、80%乙醇醇沉,醇沉后得到的粗多糖经sevege法除蛋白后,离心、弃去沉淀得到精制的老鹳草多糖。
老鹳草多糖通过降低血清的中丙氨酸转氨酶(ALT)值与天冬氨酸转氨酶(AST)值来达到保肝的作用
老鹳草多糖的剂量为每日2-3次,每次0.4-1.8g。
老鹳草多糖的剂量为每日3次,每次1g。
老鹳草多糖加入药学上可接受的辅料制备成片剂、硬胶囊、软胶囊、散剂、丸剂、颗粒剂。
老鹳草多糖作为活性成分或者是药用载体。
为证实上述老鹳草多糖的保肝活性,本发明采用了ConA诱导小鼠急性免疫性肝损伤模型,评价其对肝损伤的预防和治疗作用,实验结果表明:老鹳草多糖能够显著降低免疫性肝损伤小鼠的肝功能指标。
本发明的优点:
经本发明的制备方法,可以有效快捷地得到多糖成分。
发现了老鹳草多糖成分的保肝活性,可应用于低毒高效的保肝药物的研发,对老鹳草进一步开发利用。
附图说明
图1为老鹳草对ConA诱导的小鼠急性免疫性肝损伤模型肝功能指标(a:AST、b:ALT)的干预效果;
图中NC为正常对照组、MC为模型对照组、PC为联苯双酯阳性对照组、LL为老鹳草多糖低剂量组200mg/kg、LH为老鹳草多糖高剂量组600mg/kg。与NC组相比,“#”表示P<0.05,“##”表示P<0.01,“###”表示 P<0.001;与MC组相比,“*”表示 P<0.05,“**”表示P<0.01,“***”表示 P<0.001。
具体实施方式
下面结合附图及实施例对本发明作进一步说明,但不以任何方式对本发明加以限制,基于本发明教导所作的任何变换或改进,均落入本发明的保护范围。
老鹳草多糖在制备保肝药物中的应用
老鹳草多糖的制备方法为:将老鹳草药材粉碎,经甲醇回流脱脂后以80℃水提、80%乙醇醇沉,醇沉后得到的粗多糖经sevege法除蛋白后,离心、弃去沉淀得到精制的老鹳草多糖。
老鹳草多糖通过降低血清的中丙氨酸转氨酶(ALT)值与天冬氨酸转氨酶(AST)值来达到保肝的作用
老鹳草多糖的剂量为每日2-3次,每次0.4-1.8g。
老鹳草多糖的剂量为每日3次,每次1g。
老鹳草多糖加入药学上可接受的辅料制备成片剂、硬胶囊、软胶囊、散剂、丸剂、颗粒剂。
老鹳草多糖作为活性成分或者是药用载体。
实施例1
老鹳草粗多糖的制备:将老鹳草药材粉碎后,60℃下用甲醇回流脱脂3次。将脱脂后的老鹳草药材以80℃进行热水煮提,每次提取3小时,共提取2次,合并提取液,减压浓缩。将浓缩后的浸膏以80%乙醇醇沉,离心,弃去上清液,得到老鹳草粗多糖。
实施例2
老鹳草多糖的精制:将得到的老鹳草多糖用蒸馏水重溶,以sevege法除蛋白,离心,弃去沉淀,共5次,直至离心再无沉淀。最后将除蛋白后的多糖减压浓缩,冷冻干燥,以硫酸-苯酚法检测多糖的含量,最终得到精制的老鹳草多糖。
实施例3
老鹳草多糖的保肝药效研究
1.动物分组、造模及给药方法
老鹳草多糖由实施例2制备。
将ICR小鼠随机分为正常对照组、模型组、阳性组(联苯双酯150mg/kg)、老鹳草多糖低剂量组(200mg/kg)、老鹳草多糖高剂量组(600mg/kg),每组10只。灌胃给药,正常组及模型组给予生理盐水,其它组给予相应剂量药物,1天1次,连续7天。末次给药30min后,模型组及各给药组均静脉注射12mg/kgConA制备急性免疫性肝损伤模型,正常组静脉注射相等体积的生理盐水。禁食不禁水7h后,眼眶采血处死动物,收集肝脏。
2.血清中肝功能指标检测
眼球取血于1.5mL离心管中,常温离心4500r/min离心10min,留取上清液。按丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)试剂盒操作步骤处理,使用酶标仪测定血清中ALT、AST吸光度值(OD值)。通过查找标准曲线,根据OD值计算相应酶活力值。
实验结果见图1,与正常组相比,模型组AST、ALT值均显著升高,与模型组相比,阳性药及不同剂量的给药组AST、ALT均显著降低,且呈现剂量依赖性。结果表明老鹳草多糖对免疫性肝损伤小鼠具有保护作用。
在本发明所述的小鼠急性免疫性肝损伤试验中,老鹳草多糖的剂量为200-600mg/kg/d。换算为人(以成人体重60kg计算)剂量为1.2-3.6g/d。考虑到人的体重有不同,及各种其他病理情况,推荐剂量为每日2-3次,每次0.4-1.8g。最常用的剂量为每日3次,每次1g。
综上可以说明,老鹳草多糖具有显著的保肝活性,且具有毒副作用小的特点,在制备保肝类药品的开发上具有广泛的应用,尤其对于免疫性肝损伤疾病的治疗。对老鹳草进一步开发利用具重要意义。
Claims (7)
1.老鹳草多糖在制备保肝药物中的应用。
2.如权利要求1所述的应用,其特征在于,老鹳草多糖的制备方法为:将老鹳草药材粉碎,经甲醇回流脱脂后以80℃水提、80%乙醇醇沉,醇沉后得到的粗多糖经sevege法除蛋白后,离心、弃去沉淀得到精制的老鹳草多糖。
3.如权利要求1所述的应用,其特征在于,老鹳草多糖通过降低血清的中丙氨酸转氨酶(ALT)值与天冬氨酸转氨酶(AST)值来达到保肝的作用。
4.如权利要求1所述的应用,其特征在于,老鹳草多糖的剂量为每日2-3次,每次0.4-1.8g。
5.根据权利要求1所述的应用,其特征在于,老鹳草多糖的剂量为每日3次,每次1g。
6.一种权利要求1-5任一所述的老鹳草多糖的制剂,其特征在于,所述的老鹳草多糖加入药学上可接受的辅料制备成片剂、硬胶囊、软胶囊、散剂、丸剂、颗粒剂。
7.一种药物组合物,其特征是包含权利要求1-5任一所述的老鹳草多糖作为活性成分或者是药用载体。
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CN114605567A (zh) * | 2022-04-13 | 2022-06-10 | 云南中医药大学 | 一种老鹳草酸性多糖及其制备方法与应用 |
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CN114591449A (zh) * | 2022-04-13 | 2022-06-07 | 云南中医药大学 | 一种老鹳草中性多糖及其制备方法与应用 |
CN114605567A (zh) * | 2022-04-13 | 2022-06-10 | 云南中医药大学 | 一种老鹳草酸性多糖及其制备方法与应用 |
CN114605567B (zh) * | 2022-04-13 | 2023-01-10 | 云南中医药大学 | 一种老鹳草酸性多糖及其制备方法与应用 |
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