CN108096320A - 一种具有降血糖作用的藏荆芥提取物及其制备方法 - Google Patents
一种具有降血糖作用的藏荆芥提取物及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种具有降血糖作用的藏荆芥提取物及其制备方法,以藏荆芥为原料,经粉碎后,加乙醇回流提取,所得滤液合并,经回收乙醇,浸膏热水分散,用乙酸乙酯或正丁醇进行萃取,回收溶剂,即得到藏荆芥提取物。本发明制备的藏荆芥提取物可用于糖尿病的预防与治疗,可起到降低血糖水平,改善糖尿病相应症状的作用。
Description
技术领域
本发明属于医药技术领域,具体涉及的是一种具有降血糖作用的藏荆芥提取物及其制备方法。
背景技术
糖尿病是一种胰岛素缺乏或胰岛素生物功能缺失而引起的糖、脂肪和蛋白质代谢紊乱的代谢性疾病,可引起多种严重的并发症。近年来,随着人们生活水平的逐步提高以及人口老龄化的到来,糖尿病的发病率在全球范围内呈快速增长的发展趋势,我国已经成为全球糖尿病的第一大国。长期慢性的血糖升高可使一些器官或组织发生结构及功能的改变,导致各种并发症的发生,严重影响人们的生活质量。
目前,对糖尿病的治疗,主要通过注射胰岛素或口服降糖药(如二甲双胍、磺脲类)等途径,虽能改善糖尿病相关症状,但存在病人顺应性差、疗效低、不良反应较多等问题,无法从根本上治疗糖尿病。随着医学的发展,人们也逐渐认识到糖尿病的发病机制并非由单一生理机制所致,而是由多种因素联合作用的结果。中药、藏药等天然药物的多成分和作用的多靶点,使得其能够通过多通路、多效应器官发挥协同治疗作用,因此,中药、藏药等天然药物已成为降血糖、治疗糖尿病作用的研究热点。
藏荆芥(藏药名:萨都那保)为唇形科藏荆芥Nepeta angustifolia C.Y.Wu的全草,性凉,味辛、苦,具有息风止痉,消肿止痛的作用,用于癫痫抽搐、创伤肿痛、卒中、出血性脑梗死、创伤及疼痛等症。目前,藏荆芥已有抗炎、镇痛方面的研究报道,但其用于降血糖治疗未见有报道。
发明内容
针对上述情况,为克服现有技术之缺陷,本发明的目的是提供一种具有降血糖作用的藏荆芥提取物及其制备方法。
为了实现上述发明目的,本发明的技术方案如下:一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入5-10倍量(V/m)50%-90%乙醇,回流提取1-3次,每次提取1-3小时,滤过,滤液合并,在55-75℃条件下减压回收乙醇,得浸膏;将浸膏中加入3-5倍量(m/m)热水,搅拌分散均匀,用乙酸乙酯或正丁醇进行萃取,回收溶剂,得到乙酸乙酯萃取物或正丁醇萃取物,即得到藏荆芥提取物。
本发明有益效果:本发明采用西藏特有药材藏荆芥为原料,制备的提取物具有降血糖作用,可用于制备降血糖药物,开拓了藏荆芥的新用途,是藏药应用上的创新,也是降血糖药物的创新。试验证明:藏荆芥提取物可降低四氧嘧啶诱导糖尿病模型小鼠血糖水平,提高糖耐受量,改善“三多一少”症状,恢复体重正常增长;藏荆芥提取物能降低糖尿病模型小鼠血清NO及肝脏MDA含量,提高血清SOD及肝脏GSH-Px活性,说明藏荆芥提取物具有增强机体内源抗氧化系统活性,减轻体内氧化应激反应,缓解糖尿病发展的作用;此外,藏荆芥提取物还可降低糖尿病模型小鼠肝脏中TC、TG含量,说明藏荆芥提取物可改善四氧嘧啶性糖尿病模型小鼠脂质代谢紊乱状况。上述结果说明,本发明制备的藏荆芥提取物可用于糖尿病的预防与治疗,可起到降低血糖水平,改善糖尿病相应症状的作用。基于以上研究结果,本发明的另一个目的在于提供一种制药用途,即:上述的藏荆芥提取物在制备治疗糖尿病的药物中的应用。
具体实施方式
下面结合具体实施例进一步说明本发明的技术解决方案,这些实施例不能理解为是对技术方案的限制。
实施例1
一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入5倍量(V/m)90%乙醇,回流提取3次,每次提取2小时,滤过,滤液合并,在55℃条件下减压回收乙醇,得浸膏。将浸膏中加入3倍量(m/m)热水,搅拌分散均匀,用乙酸乙酯进行萃取,回收溶剂,得到乙酸乙酯萃取物,即得到藏荆芥提取物。
实施例2
一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入10倍量(V/m)70%乙醇,回流提取3小时,滤过,滤液在75℃条件下减压回收乙醇,得浸膏。将浸膏中加入5倍量(m/m)热水,搅拌分散均匀,用正丁醇进行萃取,回收溶剂,得到正丁醇萃取物,即得到藏荆芥提取物。
实施例3
一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入8倍量(V/m)50%乙醇,回流提取2次,每次提取1小时,滤过,滤液在65℃条件下减压回收乙醇,得浸膏。将浸膏中加入4倍量(m/m)热水,搅拌分散均匀,用乙酸乙酯进行萃取,回收溶剂,得到乙酸乙酯萃取物,即得到藏荆芥提取物。
实施例4
一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入9倍量(V/m)60%乙醇,回流提取3次,每次提取1.5小时,滤过,滤液在70℃条件下减压回收乙醇,得浸膏。将浸膏中加入5倍量(m/m)热水,搅拌分散均匀,用正丁醇行萃取,回收溶剂,得到正丁醇萃取物,即得到藏荆芥提取物。
实施例5
一种具有降血糖作用的藏荆芥提取物,以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入7倍量(V/m)60%乙醇,回流提取2次,每次提取3小时,滤过,滤液在60℃条件下减压回收乙醇,得浸膏。将浸膏中加入3倍量(m/m)热水,搅拌分散均匀,用乙酸乙酯行萃取,回收溶剂,得到乙酸乙酯萃取物,即得到藏荆芥提取物。
实施例6
藏荆芥提取物对四氧嘧啶诱导糖尿病模型小鼠的影响
1实验材料
四氧嘧啶(Sigma公司),血清超氧化物歧化酶(SOD)试剂盒、丙二醛(MDA)试剂盒、一氧化氮(NO)试剂盒、谷胱甘肽过氧化物酶(GSH-Px)试剂盒、总胆固醇(TC)试剂盒、甘油三酯(TG)试剂盒(南京建成生物工程研究所),盐酸二甲双胍片(北京中惠药业有限公司,批号:00120160307)
2实验方法
2.1动物模型建立及分组给药
清洁级雄性昆明种小鼠90只,体重20±2g,由青岛市实验动物和动物实验中心提供,适应性饲养一周后,随机分出10只为空白组,其余小鼠禁食不禁水12h后,腹腔注射四氧嘧啶200mg/kg(四氧嘧啶采用pH4.5柠檬酸缓冲液溶液配制),连续注射2d,空白组腹腔注射等容积生理盐水,60h后再禁食12h,剪尾尖取血,血糖仪测定空腹血糖值,选择血糖≥11.1mmol/L为成功模型纳入实验。将筛选合格的60只小鼠随机分为5组:模型组、阳性对照组(盐酸二甲双胍片,0.26g/kg/d)、实施例1制备得到的藏荆芥提取物组低剂量组(按照提取物换算的生药量给药,0.5g生药/kg/d)、中剂量组(1g生药/kg/d)、高剂量组(2g生药/kg/d),空白组及模型组给予等体积的生理盐水,连续给药21天,测定相关指标。
2.2小鼠体重测定
分别于造模前、给药第0天、7天、14天、21天测定各组小鼠体重。
2.3小鼠空腹血糖测定
分别于给药第0天、7天、14天、21天小鼠禁食不禁水后,测定小鼠空腹血糖。
2.4小鼠口服糖耐受量测定
各组小鼠于第21天禁食不禁水12h后灌胃相应药物,30min后按照2.0g/kg剂量灌胃25%的葡萄糖溶液,分别记录灌胃葡萄糖之后的0、30、60、120min血糖变化并按照公式计算曲线下面积(AUC):
其中,X1、X2、X3、X4分别代表0、30、60、120min的血糖值。
2.5脏器指数测定
各组小鼠摘除眼球取血后,颈椎脱曰处死小鼠后,迅速取出肝脏和肾脏,用生理盐水洗净,滤纸吸干水分,称重。按以下公式计算脏器指数:脏器指数=脏器重量(mg)/体重(g)。
2.6血清超氧化物歧化酶(SOD)、一氧化氮(NO)及肝脏丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、总胆固醇(TC)、甘油三酯(TG)测定
各组小鼠采血后,分离血清,依照试剂盒说明书测定SOD、NO;分离肝脏,匀浆后,依照试剂盒说明书测定MDA、GSH-Px、TC及TG。
2.7统计方法
试验数据采用SPSS17.0统计软件进行统计分析,统计量用均值±标准差表示,组间比较用单因素方差分析
3实验结果
3.1藏荆芥提取物对糖尿病模型小鼠体重影响
表1藏荆芥提取物对糖尿病模型小鼠体重的影响
注:与模型组比较,*P<0.05,**P<0.01。
各组小鼠在造模前,均精神状态良好,无异常行为,饮食、饮水正常,表1结果显示,造模前,各组小鼠体重正常,无显著性差异。造模后,除正常组外,其余各组小鼠均出现毛发无光泽,活动减少,出现饮食量增加、饮水量增加、尿量增加、体重增长缓慢等“三多一少”症状,即糖尿病临床所特有的症状,与空白组相比,其余各组小鼠体重均增长缓慢,均有极显著性差异(P<0.01)。灌胃给药7日后,藏荆芥各剂量组和阳性对照组小鼠症状得到明显改善,体重增加较为明显,藏荆芥除低剂量组外,中、高剂量组体重与模型组比较,均有显著性差异(P<0.05),给药14天和21天后,藏荆芥各剂量组小鼠体重仍呈现持续增长的趋势,除低剂量组外,中、高剂量组与模型组比较,均呈现显著性差异(P<0.05)。本实验结果说明,藏荆芥提取物可明显改善糖尿病模型小鼠“三多一少”症状,动物体重增加明显。
3.2藏荆芥提取物对糖尿病模型小鼠空腹血糖的影响
表2藏荆芥提取物对糖尿病模型小鼠空腹血糖的影响
注:与模型组比较,*P<0.05,**P<0.01。
表2结果表明,造模前,各组小鼠血糖均正常,无显著性差异。造模后,与空白组比较,给药前各组小鼠血糖均有显著性上升,呈现极显著性差异(P<0.01)。给药7天后,藏荆芥各组和阳性对照组小鼠血糖出现显著性下降,其中,与模型组比较,藏荆芥提取物低剂量组血糖有显著性差异(P<0.05),中、高剂量组血糖有极显著性降低(P<0.01)。给药14天和21天后,给药组小鼠血糖呈现稳定下降的趋势,与模型组比较,藏荆芥低、中、高三个剂量组均呈现显著性或极显著性下降(P<0.05或0.01)。该实验结果说明,藏荆芥提取物对糖尿病模型小鼠具有降低血糖作用,且呈现一定的剂量依赖性。
3.3藏荆芥提取物对糖尿病模型小鼠口服糖耐受量的影响
表3藏荆芥提取物对糖尿病模型小鼠口服糖耐受量的影响
注:与模型组比较,*P<0.05,**P<0.01。
表3结果显示,在灌胃葡萄糖溶液前,模型组血糖明显高于其他各组,与各给药组、空白组血糖比较,均有显著或极显著性差异(P<0.05或0.01)。灌胃葡萄糖溶液30min、60min、120min后,各组血糖水平均有明显上升,其中,30min时,各组血糖处于最高水平,60min和120时,血糖水平均有所下降。灌胃葡萄糖溶液30min后,各给药组与模型组比较,血糖水平均低于模型组,藏荆芥低、中、高剂量组与模型组比较,均显著或极显著低于模型组(P<0.05或0.01);灌胃葡萄糖溶液60min、120min后,藏荆芥各剂量组血糖水平均低于模型组,且均有显著或极显著性差异(P<0.05或0.01)。藏荆芥各剂量组曲线下面积均小于模型组,且都呈极显著性差异(P<0.01)。口服糖耐受量实验说明,藏荆芥提取物能提高糖尿病模型小鼠的糖耐受量。
3.4藏荆芥提取物对糖尿病模型小鼠脏器指数的影响
表4藏荆芥提取物对糖尿病模型小鼠脏器指数的影响
注:与模型组比较,*P<0.05,**P<0.01。
表4结果表明,糖尿病模型小鼠肝脏指数、肾脏指数均有升高,与空白组比较,有极显著性差异(P<0.01)。藏荆芥对糖尿病模型小鼠肝脏指数影响较小,与模型组比较,无显著性差异。肾脏指数说明,藏荆芥提取物可降低糖尿病模型小鼠肾脏指数,与模型组比较,各剂量组均有显著性差异(P<0.05),这可能与藏荆芥对糖尿病模型小鼠肾脏有保护作用有关。
3.5藏荆芥提取物对糖尿病模型小鼠血清SOD、MDA、NO、GSH-Px、TC、TG的影响
表5藏荆芥提取物对糖尿病模型小鼠血清SOD、NO、及肝脏MDA的影响
注:与模型组比较,*P<0.05,**P<0.01。
表6藏荆芥提取物对糖尿病模型小鼠肝脏GSH-Px、TC、TG的影响
注:与模型组比较,*P<0.05,**P<0.01。
表5结果显示,糖尿病模型小鼠SOD活性明显下降,而藏荆芥各剂量组及阳性对照组的SOD活性升高明显,与模型组比较,有显著或极显著性差异(P<0.05或0.01);NO指标结果表明,糖尿病模型小鼠给予藏荆芥及阳性对照药后,NO含量有了明显下降,其中,藏荆芥低、中、高剂量组与模型组比较,均有显著性差异(P<0.05);MDA测定结果说明,藏荆芥各剂量组及阳性对照组MDA含量均呈现下降趋势,与模型组比较,有显著或极显著性差异(P<0.05或0.01);藏荆芥和阳性对照药可明显增加GSH-Px活性,与模型组比较,藏荆芥低、中、高剂量组均有显著性差异(P<0.05);TC测定结果表明,藏荆芥各剂量及阳性对照药可降低TC含量,藏荆芥三个剂量组与模型组比较,对TC含量的降低明显,有极显著性差异;TG测定结果表明,藏荆芥三个剂量组及阳性对照组的TG含量均呈现降低趋势,藏荆芥低、中、高三个剂量组下降明显,与模型组比较,有显著性差异(P<0.05)。
四氧嘧啶是自由基激活剂,其可使胰岛中超氧阴离子及过氧化氢等自由基浓度增高,从而对胰岛β细胞产生毒性损伤,致细胞损伤坏死,引起胰岛素的合成分泌障碍,进而引发糖尿病。本发明所制备的藏荆芥提取物可降低四氧嘧啶诱导糖尿病模型小鼠血糖水平,提高糖耐受量,改善“三多一少”症状,恢复体重正常增长,说明藏荆芥提取物具有明确的预防和治疗糖尿病作用。有研究表明,糖尿患者体内的高血糖水平与自由基所引发的机体氧化应激存在一定的关系。随着病程的发展,糖尿病患者由于其自身的内源性抗氧化系统功能紊乱,造成机体清除自由基能力的减弱。因此,降低脂质过氧化反应及提高抗氧化剂稳定性是预防和治疗糖尿病的重要机制之一。本发明所制备的藏荆芥提取物能降低糖尿病模型小鼠血清NO及肝脏MDA含量,提高血清SOD及肝脏GSH-Px活性,说明藏荆芥提取物具有增强机体内源抗氧化系统活性,减轻体内氧化应激反应,缓解糖尿病发展的作用;此外,藏荆芥提取物还可降低糖尿病模型小鼠肝脏中TC、TG含量,说明藏荆芥提取物可改善四氧嘧啶性糖尿病模型小鼠脂质代谢紊乱状况。
综合上述药效学实验结果,本发明的藏荆芥提取物具有降血糖作用,可用于糖尿病的预防与治疗。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (2)
1.一种具有降血糖作用的藏荆芥提取物,其特征在于以藏荆芥为原料,经粉碎后过20目筛,按照体积质量比加入5-10倍量(V/m)50%-90%乙醇,回流提取1-3次,每次提取1-3小时,滤过,滤液合并,在55-75℃条件下减压回收乙醇,得浸膏;将浸膏中加入3-5倍量(m/m)热水,搅拌分散均匀,用乙酸乙酯或正丁醇进行萃取,回收溶剂,得到乙酸乙酯萃取物或正丁醇萃取物,即得到藏荆芥提取物。
2.权利要求1所述藏荆芥提取物在制备预防和治疗糖尿病的药物中的应用。
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