CN113018372A - 一种灵芝中药组合物及其制备方法 - Google Patents
一种灵芝中药组合物及其制备方法 Download PDFInfo
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- CN113018372A CN113018372A CN202110173180.3A CN202110173180A CN113018372A CN 113018372 A CN113018372 A CN 113018372A CN 202110173180 A CN202110173180 A CN 202110173180A CN 113018372 A CN113018372 A CN 113018372A
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Abstract
本发明涉及一种灵芝中药组合物及其制备方法,该方法以灵芝为主药,玉竹、川贝、罗汉果、金银花、桔梗、甘草、鱼腥草为辅药,经混合、粉碎、醇提、水提、提取物浓缩干燥、合并提取物等步骤制备获得灵芝中药组合物。本发明制备的灵芝中药组合物经小鼠活体实验验证,能够显著改善通过香烟烟雾诱导的肺损伤,减轻炎症细胞浸润肺组织,降低肺脏指数,抑制肺部胶原蛋白的增长,预防肺脏纤维化的形成,降低肺部其他并发症的发生。
Description
技术领域
本发明属于中药组合物制备技术领域,具体涉及一种灵芝中药组合物及其制备方法。
技术背景
灵芝,担子菌纲多孔菌科灵芝属真菌,是我国中医药宝库中的珍品,素有仙草之誉,《神农本草经》和《本草纲目》都对灵芝的功能有详细的记载,服用灵芝能够健脾、养胃、增元气、排毒养颜、抗癌止瘤、强身健体、调理血压,适用于多种人群。灵芝作为药材被历版《中华人民共和国药典》所收载。
玉竹,百合科多年生植物的干燥根茎,有养阴润燥,生津止渴功效,可用于治疗肺胃阴伤,燥热咳嗽,咽干口渴,内热消渴等病症。现代药理研究证实玉竹具有调节机体免疫功能,抑制结核杆菌生长,降血糖,降血脂,延长耐缺氧时间,强心,抗氧化,延缓衰老等功效。
川贝,百合科植物的干燥鳞茎,清热润肺,利咽开音,滑肠通便。主治肺热燥咳,干咳少痰,阴虚劳嗽,痰中带血,瘰疬,乳痈,肺痈。现代药理学研究证实川贝有祛痰镇咳功效,能抑制大肠杆菌及金黄色葡萄杆菌的生长繁殖,同时川贝中的川贝碱可起降压作用。
罗汉果,葫芦科植物罗汉果的干燥果实,清热润肺,利咽开音,滑肠通便。用于治疗肺热燥咳,咽痛失音,肠燥便秘。现在药理学研究证实罗汉果在止咳方面有疗效,又可用于脑水肿,大面积烧伤和烫伤的水肿,防治急性肾功能衰竭病和降低眼球内压,治疗急性青光眼。
金银花,忍冬科植物忍冬的干燥花蕾或初开的花,清热解毒,疏散风热。中医研究表明金银花可用于治疗臃肿疔疮,喉瘰,丹毒,热毒血痢,风热感冒,温病发热。现代药理学研究表明,金银花中含有绿原酸,木犀草素苷等活性成分,对上呼吸道感染致病病毒感染有较强的抑制作用,同时可以增强机体免疫力,抗肿瘤,消炎,解热,止血等。
桔梗,桔梗科植物桔梗的干燥根,宣肺,利咽,祛痰,排脓。用于咳嗽痰多,胸闷不畅,咽痛音哑,肺痈吐脓。现代药理研究表明,桔梗具有祛痰、镇咳、降血糖、抗炎、抑制胃液分泌和抗溃疡等作用。
甘草,豆科植物甘草的干燥根和根茎,具有补脾益气,清热解毒,祛痰止咳,缓急止痛,调和诸药的功效。临床上用于治疗脾胃虚弱,倦怠乏力,心悸气短,咳嗽痰多,脘腹,四肢挛急疼痛,痈肿疮毒等症状,同时甘草可缓解药物毒性。
鱼腥草,三白草科植物蕺菜的新鲜全草或干燥地上部分,清热解毒,消痈排脓,利尿通淋。用于肺痈吐脓,痰多喘咳,热痢,热淋,痈肿疮毒。现代药理学表明,具有抗菌、抗病毒、利尿、提高免疫力等作用。
本发明以灵芝为主药,玉竹、川贝、罗汉果、金银花、桔梗、甘草、鱼腥草为辅药,开发一种具有润肺消炎功效的灵芝中药组合物。
发明内容
本发以灵芝为主药,玉竹、川贝、罗汉果、金银花、桔梗、甘草、鱼腥草为辅药,经混合、粉碎、醇提、水提、提取物浓缩干燥、合并提取物等步骤制备获得灵芝中药组合物。通过小鼠活体实验验证,灵芝中药组合物具有润肺消炎的功效。
一种灵芝中药组合物及其制备方法具体步骤如下:
(1)按照重量份称取灵芝2~8份、玉竹2~8份、川贝3~9份、罗汉果5~8份、金银花4~7份、桔梗1~4份、甘草1~4份、鱼腥草 5~8份。混合后粉碎,得混合中药粉末。
(2)在混合中药粉末中按照1g∶20mL~80mL料液比加入95%乙醇,采用冷浸法,常温下搅拌20h,过滤,滤液经减压浓缩和冷冻干燥处理,得醇提物。
(3)将滤渣晾干,按照1g∶20mL~80mL料液比加入蒸馏水,采用水煎法,在80℃~90℃水浴中搅拌提取3h,过滤,滤液经减压浓缩和冷冻干燥处理,得水提物。
(4)将醇提物和水提物混合,得灵芝中药组合物。
(5)小鼠活体实验验证灵芝中药组合物的润肺消炎功效。
步骤(2)和步骤(3)中的减压浓缩条件为真空度0.09~0.10MPa、水浴温度60℃~70℃。
步骤(2)和步骤(3)中的冷冻干燥条件为真空度0.10MPa、温度-20℃~-30℃。
本发明的有益技术效果体现在以下几个方面:
1、本发明在中药理论基础上,将灵芝与其他中草药合理配伍,优化配伍比例,开发的灵芝中药组合物能够显著改善通过香烟烟雾诱导的肺损伤,减轻炎症细胞浸润肺组织,降低肺脏指数,抑制肺部胶原蛋白的增长,预防肺脏纤维化的形成,降低肺部其他并发症的发生。
2、本发明灵芝中药组合物是通过醇提和水提方法进行完全提取,按照提取率比例进行还原混合获得的,工艺简单,适用于大规模工业化生产。
3、本发明以中药配伍理论为基础,精心挑选中草药材,按照一定的重量比进行配方组合,运用恰当合理的制备方法,获取其中的有效成分,制成中药组合物,为研制具有润肺消炎作用的药物提供了理论基础和技术支撑。
附图说明
图1灵芝中药组合物制备工艺流程图;
图2病理切片结果图。
具体实施方式
下面结合具体的实施例,对本发明做进一步说明。本发明的特点和优点将随着描述而更清楚,但示例性的实施例仅仅用来说明本发明,并不对本发明的范围构成任何限制。
实施例1
1、称取灵芝2g、玉竹2g、川贝3g、罗汉果7g、金银花4g、桔梗3g、甘草4g、鱼腥草6g,混合后粉碎,得混合中药粉末。
2、在混合中药粉末中按照1g∶70mL料液比加入95%乙醇,采用冷浸法,常温下搅拌20h,过滤,滤液经减压浓缩和冷冻干燥处理,得醇提物。
3、将滤渣晾干,按照1g∶70mL料液比加入蒸馏水,采用水煎法,在85℃水浴中搅拌提取3h,过滤,滤液经减压浓缩和冷冻干燥处理,得水提物。
4、将醇提物和水提物混合,得灵芝中药组合物。31g中药原料可制备获得14.6g灵芝中药组合物。
灵芝中药组合物得率表
实施例2
1、以SPF级雄性昆明小鼠为实验动物,测定实施例1中制备的灵芝中药组合物润肺消炎功效。将40只雄鼠随机分为正常对照组1、正常对照组2、模型组、中药组合物组,共4组,每组10只。
2、按照以下实验方案测定实施例1中制备的灵芝中药组合物润肺消炎功效,实验开始28天后处死小鼠。
实验方案
正常对照组1:10只雄性昆明小鼠,每日暴露在动物房正常饲养环境中,按照0.2ml/20mg灌胃量,每日固定时间给小鼠灌胃生理盐水。
正常对照组2:10只雄性昆明小鼠,每日暴露在动物房正常饲养环境中,按照0.2ml/20mg灌胃量,每日固定时间给小鼠灌胃0.1% DMSO溶液。
模型组:10只雄性昆明小鼠,第1、2周暴露在动物房正常饲养环境中,第3、4周进行烟熏处理,按照0.2ml/20mg灌胃量,每日固定时间给小鼠灌胃0.1%DMSO溶液。
中药组合物组:10只雄性昆明小鼠,第1、2周暴露在动物房正常饲养环境中,第3、4周进行烟熏处理,中药组合物灌胃剂量为 750mg/kg/d,溶于0.1%DMSO溶液,按照0.2ml/20mg灌胃量,每日固定时间给小鼠灌胃。
烟熏处理方法为:将小鼠置于80×70×50cm纸箱内,点燃4支香烟置于箱内自然燃烧,密闭处理1h,每天熏烟1次。
3、测定指标及方法
小鼠行为活动观察:每天在灌胃和造模前后观察小鼠是否有死亡症状出现。
肺脏指数测定:肺脏指数=肺脏质量(mg)/体质量(g)。
组织病理学观察:取肺左叶处小块组织,放在4%多聚甲醛里固定,常规脱水、石蜡包埋、HE染色,制成切片后使用普通光学显微镜观察肺组织损伤程度。对组织切片分析平均内衬间隔(MLT)、肺泡破坏指数(DI)、平均肺泡隔厚度(MAST)三个指标。
羟脯氨酸(HYP)测定:用试剂盒(购自南京建成生物工程研究所)检测肺组织中羟脯氨酸含量,测定按照试剂盒方法进行。
统计与分析:采用SPSS 17.0软件,结果用均值±标准差表示,以P<0.05来表示差异存在显著性意义。
4、测定结果
小鼠行为活动观察结果:灌胃及造模后均未观察到小鼠死亡。
肺脏指数:动物造模后,肺脏受到了损伤,肺脏指数会随之发生改变。与正常组比较,模型组肺脏指数增大,表示肺脏可能发生了水肿、充血或肿大。中药组合物组小鼠肺脏指数指标有所降低,中药组合物组与模型组对比存在显著性差异。
肺脏指数测定结果
注:*P<0.05,与模型组相比,存在显著性差异,具有统计学意义。
组织病理学:小鼠烟熏处理后,肺脏出现明显的炎症反应,肺泡间隔大量断裂,肺泡腔明显增大。通过观察肺脏病理组织切片,比较模型组与正常对照组及中药组合物组的平均内衬间隔(MLT)、肺泡破坏指数(DI)、平均肺泡隔厚度(MAST)。结果显示,模型组MLT 及DI值均较正常对照组及中药组合物组高,且存在显著性差异,MAST 值模型组较正常对照组及中药组合物组低,且存在显著性差异。
肺脏平均内衬间隔(MLT)、肺泡破坏指数(DI)、平均肺泡隔厚度(MAST)测定结果
注:*P<0.05,与模型组相比,存在显著性差异,具有统计学意义。
病理切片比较:通过病理切片观察,发现正常对照组几乎没有炎症反应。模型组有严重的炎症反应,肺泡壁内被炎性细胞浸润,肺组织基本实化,肺泡结构严重破坏。中药组合物组的炎症反应明显减轻,部分肺泡壁有炎性细胞浸润,肺泡壁较完好,肺泡结构较正常。病理切片结果图请见说明书附图2。
羟脯氨酸(HYP)含量:机体胶原蛋白的主要成分之一为羟脯氨酸,它占胶原蛋白氨基酸的13%。肺脏组织中HYP含量是反映肺脏纤维化程度的直接指标。肺脏纤维化时,组织内胶原纤维含量增加,可通过检测肺脏组织HYP含量,来判断肺损伤的程度。模型组与正常对照组相比,羟脯氨酸含量明显增加,存在极显著性差异。中药组合物组与模型组相比,羟脯氨酸含量明显降低,在一定程度上有效缓解了肺损伤,存在显著性差异。
肺脏羟脯氨酸(HYP)含量测定结果
注:*P<0.05,**P<0.01,与模型组相比,存在着显著性和极显著性差异。
以上所述仅是本发明的优选实施方式,应当指出,本发明的内容,对于本技术领域的普通技术人员来说,在不脱离本发明技术原理的前提下,还可以做出若干改进和变型,这些改进和变型也应视为本发明的保护范围。
Claims (3)
1.一种灵芝中药组合物及其制备方法,其特征在于包括以下步骤:
(1)按照重量份称取灵芝2~8份、玉竹2~8份、川贝3~9份、罗汉果5~8份、金银花4~7份、桔梗1~4份、甘草1~4份、鱼腥草5~8份。混合后粉碎,得混合中药粉末。
(2)在混合中药粉末中按照1g∶20mL~80mL料液比加入95%乙醇,采用冷浸法,常温下搅拌20h,过滤,滤液经减压浓缩和冷冻干燥处理,得醇提物。
(3)将滤渣晾干,按照1g∶20mL~80mL料液比加入蒸馏水,采用水煎法,在80℃~90℃水浴中搅拌提取3h,过滤,滤液经减压浓缩和冷冻干燥处理,得水提物。
(4)将醇提物和水提物混合,得灵芝中药组合物。
(5)小鼠活体实验验证灵芝中药组合物的润肺消炎功效。
2.根据权利要求1所述的一种灵芝中药组合物及其制备方法,其特征在于:步骤(2)和步骤(3)中的减压浓缩条件为真空度0.09~0.10MPa、水浴温度60℃~70℃。
3.根据权利要求1所述的一种灵芝中药组合物及其制备方法,其特征在于:步骤(2)和步骤(3)中的冷冻干燥条件为真空度0.10MPa、温度-20℃~-30℃。
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