CN101002793A - 白芨多糖作为预防和治疗消化性溃疡药物的应用 - Google Patents
白芨多糖作为预防和治疗消化性溃疡药物的应用 Download PDFInfo
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Abstract
本发明属于生物制药技术领域,具体涉及白芨多糖提取物作为预防和治疗消化性溃疡药物的应用。本发明涉及的白芨多糖是由天然药物白芨块茎,通过离子交换层析与凝胶过滤层析等步骤获得的。动物试验表明,白芨多糖分别对无水乙醇和乙酰水杨酸致小鼠胃溃疡有较好的抑制作用和显著的促进愈合作用。可制备成预防和治疗消化性溃疡的药物。
Description
一、技术领域
本发明属于生物制药技术领域,具体涉及白芨多糖提取物作为预防和治疗消化性溃疡药物的应用。
二、背景技术
消化性溃疡,是消化系统的多发病,常见病,是指在消化道内接触胃液部分的内膜组织所发生的局限性缺损,包括胃溃疡和十二指肠溃疡。现代医学认为胃溃疡是攻击因子与胃粘膜防御因子之间的平衡失调所致。【胡伏莲,消化性溃疡发病机制的现代理念.[J].中华消化杂志,2005, 25(3):189-190】攻击因子主要指胃酸、胃蛋白酶,幽门螺杆菌(HP)感染,药物等,防御因子主要包括胃粘膜血流量、碳酸氢盐和粘液的分泌、细胞膜的完整性、细胞再生以及前列腺素生成、消化道激素分泌等。
根据作用机制的不同,现在胃溃疡的药物治疗有如下几类:
1.H2受体拮抗剂:用于临床已30年,促进溃疡病愈合疗效显著,但并未能防止复发,不能根治溃疡病。
2.质子泵抑制剂(PPI):抑酸效果比前者更强,能使溃疡愈合更快,但停药后也仍不免复发,不能使溃疡得到根治。且PPI目前价格较贵,多不作为一线药物应用。
3.胃粘膜保护剂:如枸橼酸铋钾,在胃酸条件下产生胶体,形成弥散性的保护层覆盖于溃疡面上,促进溃疡粘膜再生和溃疡愈合,不引起酸反弹,并通过铋离子对HP进行杀灭。
4.中和胃酸药:如碳酸氢钠,氢氧化铝等。治标不治本,仅用于症状轻、间歇发作的患者作为临时缓解症状用。
5.抗HP药物,如抗生素,是治疗HP阳性胃溃疡患者的必要药物之一。
6.其他,如M1受体阻断剂,前列腺素和中药等。
传统中药治疗溃疡副作用较低,在增强胃黏膜防御能力,提高胃溃疡愈合质量,减少复发等发面比较有优势,但是中药的成分复杂,药效分子和具体的作用机制不明确,限制了其发展。
白芨[Bletilla striata(Thunb.)Reichb.f.],又名双肾草、紫兰、凉姜,是兰科白芨属,多年生草本植物。入药的白芨块茎具备收敛止血、清热利湿、消肿生肌的功效,用于治疗外伤和消化道出血,消除疮疡肿毒,愈合溃疡等症【凌一揆中药学,北京:科学技术文献出版社.2003第5版.】其中,有关中药白芨对胃粘膜的保护和促进溃疡愈合方面的研究已经比较多。但是还没有白芨的提取物纯品的相关研究结果公开。
现代科学对白芨成分的分析始于二十世纪三十年代。研究证明,白芨的成分主要是葡萄甘露聚糖。近年,本实验室通过离子交换层析与凝胶过滤层析得到白芨多糖提取物纯品,并利用现代色谱和光谱技术进行分析,首次报道了一种具有确定组成和结构的新的葡萄甘露聚糖【孙剑涛,张峻峰,王春明,罗熠专利号:CN200410065732.5】。研究表明,自行纯化的白芨多糖(BSP),可以促进血管内皮生长因子(VEGF)表达,促进血管内皮细胞增殖,促进一氧化氮(NO)与肿瘤坏死因子(TNF-α)分泌。【Chunming Wang,Jiantao Sun,Junfeng Zhang,et al.A polysaccharide isolated from the medicinal herb Bletilla striata inducesendotheliai cells proliferation and vascular endothelial growth factorexpression in vitro.Biotechnology Letters,2006,28:539-543】白芨多糖(BSP)的以上生物学功能,都是与白芨促进愈合的传统治疗作用相符的。
三、发明内容
本发明所要解决的技术问题是白芨多糖提取物纯品作为预防和治疗消化性溃疡药物的应用。
本发明涉及的白芨多糖是由天然药物白芨块茎,通过离子交换层析与凝胶过滤层析等步骤获得的。具体见发明专利“一种颅内动脉瘤液体栓塞剂及其制备方法”【孙剑涛,张峻峰,王春明,罗熠,专利号:CN200410065732.5】。
本发明涉及的白芨多糖抗消化性溃疡作用是通过建立小鼠实验性胃溃疡模型,分别研究其对无水乙醇和乙酰水杨酸致小鼠胃溃疡的预防和愈合的作用。本发明所述的白芨多糖提取物纯品可制备成抗消化性溃疡的药物,在制备预防和治疗消化性溃疡药物中应用。
本发明的有益效果:在现有的治疗消化性溃疡药物中,化学合成药往往不能根治溃疡,价格高而且副作用大。传统中药治疗溃疡副作用较低,在增强胃黏膜防御能力,提高胃溃疡愈合质量,减少复发等发面比较有优势,但是中药的成分复杂,药效分子和具体的作用机制不明确,疗效不显著,限制了其发展。本发明从中国传统中药白芨中提取了多糖纯品,用于预防和治疗消化性溃疡,具有分子结构确定、疗效高、副作用小、成本低、具有预防和治疗双重功能等优点。
四、具体实施方式
1、白芨多糖对无水乙醇致小鼠胃溃疡的预防的影响
取ICR小鼠40只,体重18~22g,随机分为四组,雌雄各半,分别为生理盐水对照组,白芨多糖高、低剂量组(100、50g/kg),枸掾酸铋钾组(100mg/kg)。各组分别按20ml/kg体积灌胃给药,每天1次,连续6天,于末次给药前24h禁食不禁水,末次给药后1h,每只小鼠灌胃无水乙醇150ul,建立小鼠胃溃疡模型。
1h后脱颈处死各组小鼠,结扎贲门,幽门,注入1%甲醛2ml,取胃浸泡在1%甲醛中半小时。沿胃大弯剪开,展开平铺于玻璃板上,观察记录,数码相机拍照,电脑进行指数计算,并计算胃溃疡抑制率。
计算溃疡指数和溃疡抑制率(愈合率):采用Guth标准【Guth PH,Aures D,Paulsen G.Topical aspirin plus HCl gastric lesions in therat.Cytoprotective effect of prostagiandin,cimetidine,and probanthine.Gastroenterology 1979;76:88-93】并做修改,具体方法如下:按溃疡或糜烂面积大小给予计分,①:每3个点状溃疡(粘膜缺损小于1mm或出血性糜烂小点,称点状溃疡)计1分②条状出血:电脑标尺测量溃疡面的最大长径和垂直于最大长径的最大宽径(换算成真实长度),二者的乘积即为此溃疡面的记分。1mm宽及以下者每mm长为1分;1-2mm宽者每mm长为2分;2-3mm宽者每mm长为3分。所有计分累积即为小鼠的溃疡指数。溃疡抑制率=(平均模型组溃疡指数-平均实验组溃疡指数)/平均模型组溃疡指数。结果统计见表一:白芨多糖对无水乙醇致小鼠胃溃疡有较好的抑制作用。
表一 白芨多糖对小鼠无水乙醇型胃溃疡的抑制作用
组别 | 动物数量(只) | 剂量(mg/kg) | 溃疡指数 | 抑制率 |
生理盐水组白芨多糖高剂量组白芨多糖低剂量组枸掾酸铋钾组 | 10101010 | 20ml/kg10050100 | 89.74±12.1543.62±10.6260.11±11.0721.35±3.16 | 051.3933.0276.21 |
(和生理盐水组相比,p<0.05。)
2、白芨多糖对无水乙醇致小鼠胃溃疡的愈合的影响
取ICR小鼠40只,体重18~22g,随机分为四组,雌雄各半,分别为生理盐水对照组,BSP高、低剂量组(100、50g/kg),枸掾酸铋钾组(100mg/kg)。各组禁食不禁水24h后,每只小鼠灌胃无水乙醇150ul,1h后,各组分别按20ml/kg体积灌胃给药,每天1次,连续6天,其中分别于第3天,第5天给药前13h禁食不禁水,给药前1h每只小鼠灌胃无水乙醇150ul,以使得胃溃疡持续发生,症状明显以便计算指数。
末次给药6h后脱颈处死各组小鼠,结扎贲门,幽门,注入1%甲醛2ml,取胃浸泡在1%甲醛中半小时。沿胃大弯剪开,展开平铺于玻璃板上,观察记录,数码相机拍照,电脑进行指数计算,并计算胃溃疡愈合率。
计算溃疡指数和溃疡抑制率(愈合率)同实施例2。结果统计见表二:白芨多糖对无水乙醇致小鼠胃溃疡有显著的促愈合作用。
表二 白芨多糖对小鼠无水乙醇型胃溃疡的愈合作用
组别 | 动物数量(只) | 剂量(mg/kg) | 溃疡指数 | 愈合率 |
生理盐水组白芨多糖高剂量组白芨多糖低剂量组枸掾酸铋钾组 | 10101010 | 20ml/kg10050100 | 52.77±15.429.74±2.7613.92±5.4316.31±5.08 | 081.5473.6269.09 |
(和生理盐水组相比,p<0.05。)
3、白芨多糖对乙酰水杨酸致小鼠胃溃疡的预防的影响
取ICR小鼠40只,体重18~22g,随机分为四组,雌雄各半,分别为生理盐水对照组,白芨多糖高、低剂量组(100、50g/kg),枸掾酸铋钾组(100mg/kg)。各组分别按20ml/kg体积灌胃给药,每天1次,连续6天,于末次给药前24h禁食不禁水,末次给药后1h,每只小鼠按150mg/kg乙酰水杨酸的剂量灌胃,建立乙酸水杨酸致小鼠胃溃疡模型。
4h后脱颈处死各组小鼠,结扎贲门,幽门,注入1%甲醛2ml,取胃浸泡在1%甲醛中半小时。沿胃大弯剪开,展开平铺于玻璃板上,观察记录,数码相机拍照,电脑进行指数计算,并计算胃溃疡抑制率。计算溃疡指数和溃疡抑制率(愈合率)同实施例2。结果统计见表三:白芨多糖对乙酰水杨酸致小鼠胃溃疡有较好的抑制作用。
表三 白芨多糖对小鼠乙酸水杨酸型胃溃疡的抑制作用
组别 | 动物数量(只) | 剂量(mg/kg) | 溃疡指数 | 抑制率 |
生理盐水组白芨多糖高剂量组白芨多糖低剂量组枸掾酸铋钾组 | 10101010 | 20ml/kg10050100 | 80.73±17.4847.71±7.2552.69±7.7328.72±4.24 | 040.9034.7364.42 |
(和生理盐水组相比,p<0.05。)
4、白芨多糖对乙酰水杨酸致小鼠胃溃疡的愈合的影响
取ICR小鼠40只,体重18~22g,随机分为四组,雌雄各半,分别为生理盐水对照组,白芨高、低剂量组(100、50g/kg),枸掾酸铋钾组(100mg/kg)。各组禁食不禁水24h后,每只小鼠按150mg/kg乙酰水杨酸的剂量灌胃,4h后,各组分别按20ml/kg体积灌胃给药,每天1次,连续6天,其中分别于第3天,第5天给药前16h禁食不禁水,给药前4h每只小鼠按150mg/kg乙酰水杨酸的剂量灌胃,以使得胃溃疡持续发生。
末次给药6h后脱颈处死各组小鼠,结扎贲门,幽门,注入1%甲醛2ml,取胃浸泡在1%甲醛中半小时。沿胃大弯剪开,展开平铺于玻璃板上,观察记录,数码相机拍照,电脑进行指数计算,并计算胃溃疡愈合率。计算溃疡指数和溃疡抑制率(愈合率)同实施例2。结果统计见表四:白芨多糖对乙酰水杨酸致小鼠胃溃疡有显著的促愈合作用。
表四 白芨多糖对小鼠乙酸水杨酸型胃溃疡的愈合作用
组别 | 动物数量(只) | 剂量(mg/kg) | 溃疡指数 | 愈合率 |
生理盐水组白芨多糖高剂量组白芨多糖低剂量组枸掾酸铋钾组 | 10101010 | 20ml/kg10050100 | 59.57±8.2610.03±4.0421.38±3.5614.57±5.27 | 083.1764.1176.74 |
(和生理盐水组相比,p<0.05。)
Claims (1)
1、白芨多糖提取物纯品在制备预防和治疗消化性溃疡药物中的应用。
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CN101429254B (zh) * | 2008-12-05 | 2011-05-04 | 云南植物药业有限公司 | 一种白及多糖及其制备方法和新用途 |
CN107158406A (zh) * | 2017-06-06 | 2017-09-15 | 南京大学 | 葡甘聚糖糖基化的SiO2纳米颗粒及其制备方法和应用 |
CN109010349A (zh) * | 2018-10-29 | 2018-12-18 | 湖北中医药大学 | 白芨寡糖在改善肠道微生态中的应用 |
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Cited By (4)
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CN101429254B (zh) * | 2008-12-05 | 2011-05-04 | 云南植物药业有限公司 | 一种白及多糖及其制备方法和新用途 |
CN107158406A (zh) * | 2017-06-06 | 2017-09-15 | 南京大学 | 葡甘聚糖糖基化的SiO2纳米颗粒及其制备方法和应用 |
CN107158406B (zh) * | 2017-06-06 | 2020-07-14 | 宁国市吴越医药科技有限公司 | 葡甘聚糖糖基化的SiO2纳米颗粒及其制备方法和应用 |
CN109010349A (zh) * | 2018-10-29 | 2018-12-18 | 湖北中医药大学 | 白芨寡糖在改善肠道微生态中的应用 |
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