CN112168860A - 一种治疗无复流现象的中药组合物及其制备方法 - Google Patents
一种治疗无复流现象的中药组合物及其制备方法 Download PDFInfo
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Abstract
本发明属于中药领域,具体涉及一种治疗无复流现象的中药组合物及其制备方法,该组合物由以下原料药组成:红景天、广枣、人参。本发明的组合物可对经皮冠状动脉介入治疗后无复流症进行改善治疗,制成颗粒剂,使用便利,刺激性小,患者的顺应性好,尤其适用于婴幼儿。
Description
技术领域
本发明属于中药领域,具体涉及一种治疗无复流现象的中药组合物及其制备方法。
背景技术
经皮冠状动脉介入治疗(PCI)是治疗冠心病的一种有效手段,而无复流现象(no-reflow phenomenon,NR)是PCI术后常见的并发症。其定义为PCI治疗后,已解除心外膜冠状动脉机械性梗阻,但血流仍持续减低或相应区域心肌仍然灌注不足,表现为造影剂排空延迟并伴随心肌缺血表现。一旦发生无复流现象,患者即刻可出现胸痛、心律失常、造影剂滞留、ST段的抬高,血流动力学异常,严重的可以出现低血压、心源性休克、甚至发生灾难性血管崩溃,立即死亡。无复流总体发生率约2%,在大隐静脉桥(SVG)行PCI中为10%~15%,而在急性心肌梗死(AMI)直接PCI中无复流发生率可高达30%。无复流患者的住院病死率和心肌梗死再发率较对照组增加5~10倍,是心肌缺血、心室恶性重构的预测因子。功能性无复流发生时,是暂时性微循环阻塞,微循环结构尚未破坏,具有可逆性,处理及时可以改善。而解剖型无复流会导致微血管结构受到破坏,一旦发生很难逆转。无复流的发生削弱了PCI的疗效,影响临床预后。
无复流现象属于冠状动脉微循环障碍,主要发病机制为心肌微血管闭塞。心肌微循环,指由心肌微动脉、毛细血管及心肌微静脉构成的微循环系统,因此应属中医学络脉理论范畴。中医认为无复流现象的病机属心络受阻、血瘀络阻。心络类似冠脉微循环,气血、脏腑、经络功能障碍可致心络瘀阻、血行不畅,甚或形成微小癥积(血栓),可类同冠脉微循环的完整性受损及灌注不足。PCI可造成局部的损伤、病变处微血栓、粥样硬化斑块碎片等微栓子脱落可以引起远端微血管栓塞、血流恢复产生微血管再灌注损伤等原因均可致冠脉循环的障碍,这与中医的微小癥积、血瘀内结、络脉瘀阻认识接近。当气血推动无力,不能充实于心络,心络失养而受损,血瘀内结阻于心络时,冠脉微循环障碍发生,故出现无复流现象。
无复流的发生机制是多种因素导致的微循环解剖结构损害或功能障碍、微血管血流受阻。治疗上,以抗凝药、抗血小板聚集药物、血管扩张剂等类药物改善。但上述药物具有增加出血风险、存在使用局限性、疗效不确切等问题和争议,表明针对无复流现象的防治尚有较大的探索空间。而中药具有多靶点、多层次干预的特色和优势,可同时干预PCI术后的多个病理环节,起到整体调节的作用。
发明内容
本发明的目的就是针对现有技术存在的缺陷,利用中医药理论,针对无复流的发生机制,采用活血化瘀、益气化痰的治法,进行辨证配伍组方,以改善冠脉血流速度,提供一种预防或治疗无复流现象的中药组合物及制备方法和用途。
本发明的第一个目的在于提供一种用于预防或治疗无复流现象的新的中药组合物,该组合物由以下原料药组成:红景天、广枣、人参。
上述中药组合物由以下重量份的原料药组成:红景天1-30份、广枣1-30份、人参1-30份。
优选地,上述中药组合物由以下重量份的原料药组成:红景天5-14份、广枣2-8份、人参5-20份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天6份、广枣3份、人参6份。
上述新的中药组合物,还包括黄连或者麦冬。
上述中药组合物由以下重量份的原料组成:红景天1-30份、广枣1-30份、人参1-30份,黄连1-30份或者麦冬1-30份。
优选地,上述中药组合物由以下重量份的原料药组成:红景天5-14份、广枣2-8份、人参5-20份、黄连2-15份或者麦冬10-25份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份、黄连10份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天6份、广枣3份、人参6份、黄连4份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份、黄连10份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份、麦冬24份。
在一个具体的实施方案中,本发明的中药组合物由以下重量份的原料药组成:红景天6份、广枣3份、人参6份、麦冬12份。
在一个具体的实施方案中,本发明的中药组合物中所述人参为园参或林下参或野山参或生晒参或红参等炮制品。
本发明提供的中药组合物,其中红景天益气补血、通脉养心,为君药;广枣行气活血,化瘀通络,为臣药;红参行佐使之功,佐以黄连或麦冬清热生津之品,调和红参燥热之性。诸药合用,共奏益气活血、复脉通络之功效。
红景天
红景天(Rhodiola crenulata(Hook.f.et Thoms.)H.Ohba)属传统藏药。中药化学成分研究表明红景天属植物其主要药理活性成分为红景天苷(Salidroside)及其苷元酪醇(Ptyrosol)、酪萨维(Rosavin或Rosavidine)、二苯甲基六氢吡啶(Pyridrde)、红景天素即草质素-7-氧-(3-氧-β-D吡喃葡萄糖基)-α-L-吡喃鼠李糖苷(Rhodiosin)和草质素-7-氧-α-L-吡喃鼠李糖苷(Rhodionin)等。
黄酮类化合物是红景天主要生物活性成分。研究发现红景天总黄酮质量浓度为0.2mg·mL-1时,对羟自由基、超氧阴离子自由基、DPPH自由基的清除作用均高于50%,且随浓度的增高其清除自由基作用增强。表明红景天黄酮在体外具有良好的抗氧化活性。红景天总黄酮可以抑制TGF-β1(5μg·L-1)诱导的大鼠心肌纤维化细胞(CFB)的增殖,其机制可能为降低细胞中谷胱甘肽过氧化物酶GSH-Px和谷胱甘肽GSH含量,降低Ⅰ型胶原蛋白ColⅠ和Ⅲ型胶原蛋白ColⅢ表达,而改善改善心肌纤维化。在红景天对大鼠力竭游泳能力和心肌线粒体的抗氧化能力的影响实验中发现,服用红景天煎剂可以通过提高线粒体SOD和GSH-Px活性,显著降低MDA含量,从而抑制线粒体过氧化脂质的形成,消除体内自由基对线粒体膜和肌浆网膜造成损伤,从而增强机体抗氧化能力。
红景天苷为红景天主要成分,其在心血管方面生物活性逐渐被研究人员所发现和关注。颜天华等在实验中,用垂体后叶素诱发小鼠急性心肌缺血,发现红景天苷能够抑制心电图T波抬高和J点升高,表明红景天苷能减轻垂体后叶素引起的心肌缺血再灌注损伤。通过对小鼠的缺氧-复氧实验,发现红景天苷能显著改善心肌细胞的存活率,同时降低血清乳酸脱氢酶(LDH)肌酸激酶(CK)的含量。在探索红景天有效成分抗缺血性心脏疾病的作用及机制中发现,红景天主要通过抗氧化、抗凋亡和促进缺血区血管新生,保护缺血区心肌,减小梗死面积。红景天苷对氧糖剥离心肌细胞的保护作用,主要通过保护线粒体功能,抑制线粒体凋亡途径来实现。
广枣
广枣(Choerospondias axillaris(Roxb.)Burtt et Hill)属传统蒙药,为心脑血管疾病治疗的主药。中药化学成分研究表明广枣含有有机酸、黄酮、酚酸、鞣制、香豆素、木脂素及多糖等成分,其药理活性主要表现为对心脑血管疾病的保护及治疗作用。目前研究主要集中在广枣总黄酮和广枣总有机酸两大类成分上。
广枣富含有机酸类成分,含量在5.22%-8.13%。同时该类成分生物活性显著,尤其在缺血性心脏病方面具有较好的作用。上世纪80年代,王乃利、姚新生等报道以抗ADP诱导血小板聚集为指标,从广枣的乙醇提取物中跟踪分离出原儿茶酸、没食子酸、鞣花酸等活血有效成分。汤喜兰等报道,以柠檬酸、苹果酸、酒石酸、琥珀酸等模拟广枣总有机酸发现其对心肌缺血再灌注损伤都具有保护作用,确认有机酸类成分为广枣治疗心肌缺血再灌注损伤的主要活性物质基础之一。
广枣富含的总黄酮,可明显抑制离体大鼠心肌组织中的腺苷酸环化酶(adenylatecyclase,AC)活性,使cAMP含量明显下降,进而改善缺血性心律失常。通过观察广枣总黄酮对培养大鼠心肌细胞的作用,表明广枣总黄酮在整体或细胞水平的抗心律失常作用主要是对心肌细胞的直接作用,其抗心律失常的机制可能与广枣总黄酮拮抗Ca2+内流和β受体阻滞作用有关。广枣总黄酮显著降低麻醉犬冠脉阻力,增加冠脉血流量,改善冠脉循环作用,增加心肌的供血和供氧,提高心肌对缺氧的耐受力,保护心肌缺血。广枣60%乙醇洗脱物对急性心肌缺血有明显的保护作用,可使结扎冠状动脉前降支造成急性心肌缺血大鼠的心电图明显改善,心肌梗死范围明显缩小,心肌缺血损伤程度减轻,呈现一定的量效关系。
人参
人参(Panax ginseng C.A.Mey)为五加科植物人参的干燥根和根茎。
人参含有皂苷、多糖、多肽、酶、微量元素、挥发油以及氨基酸等多种天然活性成分。皂苷类为其主要成分,其中人参皂苷Rb1、Rb2、Rc、Rd、Re、Rg1、Rg2是水参、生晒参和红参的共有成分,其含量占人参总皂苷的90%以上。
人参经炮制后,由白变红,产生红参中特有成分,其具有显著而独特的生理活性。鲜参中的皂苷类成分会发生糖苷键和酯键的断裂,生成脱羧、脱糖降解产物,而大分子量、高极性皂苷转化为小分子量、低极性皂苷,或C-20位天然构型发生了转变,生成某些特有皂苷,从而使红参生物活性发生改变,人参皂苷Rh1、Rh2、Rg3、Rg5、Rg6、Rs4-Rs 7、Rk1-Rk3等是红参的特有成分。红参还增加了人参炔二醇、人参炔三醇和麦芽酚,因此增加了红参的活性。
有文献研究了红参中特有成分的药理作用,其中人参皂苷Rh2对高脂饮食大鼠心肌缺血再灌注(I/R)损伤的保护作用研究中,模型组和实验组与假手术组比较,大鼠外周血EPCs、大鼠心肌IL-6水平和血清SOD与MDA水平等相关指标均显著回调。在人参皂苷Rh2对高脂膳食大鼠心肌缺血再灌注(IR)时内皮祖细胞数影响的实验中,研究人员发现,人参皂苷Rh2可增加高脂膳食大鼠心肌缺血再灌注后外周血内皮祖细胞数量,从而减轻心肌缺血再灌注损伤。红参中另一主要活性成分Rg3,同样被证明可改善大鼠心肌缺血/再灌注损伤所导致的心功能损害,其作用机制与抗细胞凋亡及抗炎性反应作用有关。
其中人参皂苷Rh2对高脂饮食大鼠心肌缺血再灌注(I/R)损伤有保护作用。人参皂苷Rg3具有舒张血管作用,逆转人胆管癌多药耐药细胞株QBC939/ADM的耐药性。人参皂苷Rg5可有效抑制胃癌细胞BGC-823细胞的侵袭、迁移。人参皂苷Rk1还能够抑制肝脏组织中GSH的耗竭和升高超氧化物歧化酶(SOD)水平,降低脂质过氧化产物MDA的水平。
黄连
黄连(Coptis chinensis Franch)为毛茛科黄连属植物黄连的干燥根茎。
黄连的主要生物活性组分为生物碱,该类成分包括:小檗碱,并含黄连碱、甲基黄连碱、掌叶防己碱、小檗红碱、表小檗碱、黄连碱、巴马汀、药根碱等,其中以小檗碱的含量最高。
动物试验表明,小檗碱有抗菌、抗病毒及原虫、利胆、抗腹泻、抗炎和抗脑缺血、抗微生物、降压、抗心肌缺血及心肌梗死、抗心律失常、抑制中枢神经系统、止腹泻、抗溃疡、利胆、降血糖、抑制DNA的合成、抑制血小板聚集等作用。
麦冬
麦冬为百合科植物麦冬(Ophiopogon japonicus(L.f)Ker-Gawl)的干燥块根。
麦冬含多种甾体皂苷:麦冬皂苷A、B、C、D,苷元均为假叶树皂苷元,另含麦冬皂苷B′、C′、D′,苷元均为薯蓣皂苷元;尚含多种黄酮类化合物:如麦冬甲基黄烷酮A、B,麦冬黄烷酮A、麦冬黄酮A、B,甲基麦冬黄酮A,B。
研究表明麦冬总皂苷及总多糖可显著增加小鼠心肌营养血流量。麦冬抗心肌缺血的作用机制可能与防止心肌细胞脂质过氧化及改善脂肪酸代谢有关,并呈现一定的量效关系。
麦冬水提物能够降低D-半乳糖所致的衰老大鼠的血液黏度。麦冬总多糖在大鼠颈总动脉结扎脑缺血实验中,结渣前给与400mg/kg和200mg/kg的麦冬总多糖,上述剂量均可使结扎后脑内乳酸含量显著降低,从而逆转缺血后酸中毒造成的各种损害。
在一个具体的实施方案中,本发明的中药组合物,其中还包含可药用辅料,所述的药用辅料是本领域技术人员公知的。
本发明的中药组合物可以按本领域技术人员公知的技术制成各种常用剂型,比如片剂、颗粒剂、胶囊剂、丸剂、散剂、口服液体制剂,优选颗粒剂。
本发明的另一目的,是制备本发明的中药组合物的方法。
本发明中药组合物所含的有效成分可以这样制备:将上述原料药干燥研成粉末,混匀,得到有效成分。
本发明中药组合物所含的有效成分还可以这样制备:
方法一:将上述原料药水提,得所述中药组合物的有效成分;
方法二:将上述原料药醇提,得所述中药组合物的有效成分;
方法三:将上述原料药水提醇沉,得所述中药组合物的有效成分。
本发明中药组合物所含的有效成分的制备步骤中,
水提条件为:加所述原料药总重量的6-12倍量水提取2-3次,每次1-3小时;
醇提条件为:加所述原料药总重量的4-10倍量40-80%乙醇提取2-3次,每次1-3小时;
水提醇沉条件为:加所述原料药总重量的6-12倍量水提取2-3次,每次1-3小时,滤过,滤液浓缩至60-70℃时相对密度为1.15-1.20的清膏,加乙醇使含醇量达40-70%,静置。
本发明中药组合物所含的有效成分更优选下列步骤制备:
方法一:取所述原料药,混合后加水提取2-3次,每次加水量相当于药材总重量的6-12倍,每次提取时间为1-3小时,合并提取液,滤过,滤液浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分;
方法二:取所述原料药,混合后用40-80%乙醇回流提取2-3次,每次乙醇用量为该两味药材总量的4-10倍,提取时间为1-3小时,合并提取液,回收乙醇并浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分;
方法三:取所述原料药,混合后加水提取2-3次,每次加水量相当于药材总重量的6-12倍,每次提取时间为1-3小时,合并提取液,滤过,滤液浓缩至60-70℃时相对密度为1.15-1.20的清膏,加入乙醇,使乙醇含量为40-70%,静置12-24小时,滤过,滤液浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分。
制得的有效成分可以直接入药服用或可药用辅料按本领域技术人员公知的技术制备成所需制剂。
上述制备方法仅对本发明所提制法进行列举,但不应将此理解为本发明制备方法仅仅限于上述所列举方法。
本发明的第三个目的,是提供上述中药组合物用于制备预防或治疗无复流现象的药物的应用。
优选地,上述中药组合物用于制备预防或治疗经皮冠状动脉介入治疗无复流现象的药物的应用。
本发明中,如无特别说明,其中:
术语“水提”是指原料药水煮后取汁的过程。
术语“醇提”是指将原料药中有效成分溶解在乙醇溶液中,以提取出有效成分的过程。
术语“水提醇沉”是指原料药经水提后的水提液经过加乙醇达到不同含醇量,某些药物成分在醇中溶解度降低而析出沉淀,固液分离后使水提液得以精制的过程。
术语“有效成分”,是指原料药经提取之后起治疗作用的提取物的干燥粉末。
与现有技术相比,本发明具有以下有益效果:
1、本发明遵循“气行则血行”的中医理论,采用上述药味随证配伍,以益气活血为治法,达到复脉通络的作用,可对经皮冠状动脉介入治疗后无复流症进行改善治疗。
2、采用本发明的中药组合物制成颗粒剂,使用便利,刺激性小,患者的顺应性好,尤其适用于婴幼儿。
3、本发明的中药组合物,制备方法简单制备成本低,安全无毒副作用。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述,但本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限定本发明的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
实施例1-18本发明中药组合物的制备
实施例1
取红景天8g、广枣6g、红参10g,混合后加水提取3次,每次加水量相当于药材总重量的6倍,每次提取时间为2小时,合并提取液,滤过,滤液浓缩至60℃时相对密度为1.30的清膏,干燥,粉碎成细粉,得有效成分。
实施例2
取红景天12g、广枣5g、园参18g,混合后用70%乙醇回流提取2次,每次乙醇用量为药材总量的6倍,提取时间为1.5小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例3
取红景天6g、广枣3g、红参6g,混合后加水提取2次,每次加水量相当于药材总重量的12倍,每次提取时间为1小时,合并提取液,滤过,滤液浓缩至70℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例4
取红景天8g、广枣3g、野山参6g,混合后用80%乙醇回流提取2次,每次乙醇用量为药材总量的6倍,提取时间为1.5小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例5
取红景天5g、广枣2g、红参5g,混合后用60%乙醇回流提取3次,每次乙醇用量为药材总量的8倍,提取时间为2小时,合并提取液,过滤,滤液浓缩至60℃时相对密度约为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例6
取红景天14g、广枣8g、红参20g,混合后用70%乙醇回流提取3次,每次乙醇用量为药材总量的6倍,提取时间为3小时,合并提取液,过滤,滤液浓缩至70℃时相对密度约为1.30的清膏,干燥,粉碎成细粉,得有效成分。
实施例7
取红景天12g、广枣5g、红参18g,黄连10g,混合后用70%乙醇回流提取2次,每次乙醇用量为药材总量的6倍,提取时间为1.5小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例8
取红景天6g、广枣3g、园参6g,黄连4g,混合后加水提取3次,每次加水量相当于药材总重量的12倍,每次提取时间为3小时,合并提取液,滤过,滤液浓缩至65℃时相对密度为1.15的清膏,加入乙醇,使乙醇含量为70%,静置24小时,滤过,滤液浓缩至60℃时相对密度为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例9
取红景天8g、广枣4g、红参8g,麦冬12g,混合后用60%乙醇回流提取2次,每次乙醇用量为药材总量的8倍,提取时间为1小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例10
取红景天6g、广枣3g、园参6g,麦冬12g,混合后用80%乙醇回流提取2次,每次乙醇用量为药材总量的10倍,提取时间为2小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例11
取红景天12g、广枣5g、红参18g,麦冬24g,混合后用80%乙醇回流提取2次,每次乙醇用量为药材总量的10倍,提取时间为3小时,合并提取液,过滤,滤液浓缩至60℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例12
取红景天18g、广枣5g、红参18g,麦冬24g,混合后用60%乙醇回流提取2次,每次乙醇用量为药材总量的6倍,提取时间为3小时,合并提取液,过滤,滤液浓缩至70℃时相对密度约为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例13
取红景天6g、广枣3g、园参6g,麦冬8g,混合后用60%乙醇回流提取2次,每次乙醇用量为药材总量的6倍,提取时间为1.5小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例14
取红景天8g、广枣3g、园参6g,麦冬8g,混合后用40%乙醇回流提取2次,每次乙醇用量为药材总量的4倍,提取时间为1小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
实施例15
取红景天5g、广枣2g、红参5g,黄连2g,混合后加水提取2次,每次加水量相当于药材总重量的12倍,每次提取时间为1小时,合并提取液,滤过,滤液浓缩至60℃时相对密度为1.15的清膏,加入乙醇,使乙醇含量为40%,静置24小时,滤过,滤液浓缩至70℃时相对密度为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例16
取红景天5g、广枣2g、园参5g,麦冬10g,混合后加水提取3次,每次加水量相当于药材总重量的6倍,每次提取时间为3小时,合并提取液,滤过,滤液浓缩至70℃时相对密度为1.20的清膏,加入乙醇,使乙醇含量为70%,静置12小时,滤过,滤液浓缩至60℃时相对密度为1.30的清膏,干燥,粉碎成细粉,得有效成分。
实施例17
取红景天14g、广枣8g、园参20g,黄连15g,混合后加水提取2次,每次加水量相当于药材总重量的6倍,每次提取时间为2小时,合并提取液,滤过,滤液浓缩至60℃时相对密度为1.20的清膏,加入乙醇,使乙醇含量为70%,静置24小时,滤过,滤液浓缩至70℃时相对密度为1.20的清膏,干燥,粉碎成细粉,得有效成分。
实施例18
取红景天14g、广枣8g、园参20g,麦冬25g,混合后加水提取3次,每次加水量相当于药材总重量的10倍,每次提取时间为1小时,合并提取液,滤过,滤液浓缩至60℃时相对密度为1.15的清膏,加入乙醇,使乙醇含量为60%,静置24小时,滤过,滤液浓缩至65℃时相对密度为1.25的清膏,干燥,粉碎成细粉,得有效成分。
对比例1
取红景天50g、红参20g,混合后加水提取3次,每次加水量相当于药材总重量的12倍,每次提取时间为3小时,合并提取液,滤过,滤液浓缩至60℃时相对密度为1.15的清膏,加入乙醇,使乙醇含量为70%,静置24小时,滤过,滤液浓缩至65℃时相对密度为1.25的清膏,干燥,粉碎成细粉,得有效成分。
对比例2
取红景天50g、广枣20g,混合后用60%乙醇回流提取2次,每次乙醇用量为药材总量的8倍,提取时间为1小时,合并提取液,过滤,滤液浓缩至65℃时相对密度约为1.25的清膏,干燥,粉碎成细粉,得有效成分。
试验例:本发明所述组合物对大鼠心肌梗死模型的治疗作用
1、实验动物、药物与仪器
1.1实验动物
Wistar大鼠,SPF级(无特定病原体级实验动物):体重210-250g,雌雄各半,动物年龄10-11周,购自斯贝福(北京)生物技术有限公司;许可证编号:11401500036047。
动物房:试验期间动物饲养于北京盈科瑞药物安全有效性研究有限公司,实验设施许可证:SYXK(京)2017-0026;设施管理遵循中华人民共和国国家标准GB14925-2001《实验动物环境及设施》。
喂养条件:采用人工光照12小时明暗周期,环境温度维持在20~24℃,湿度在40%~70%,每小时换气15次;动物饲养于聚碳酸酯小鼠饲养笼,每笼饲养同组的5只同性别大鼠;每2天更换一次清洁动物笼和垫料。
饲料:大小鼠生长维持饲料,购自斯贝福(北京)生物技术有限公司
饮用水:试验动物饮用水,动物可自由摄取,每日更换新的水瓶和新鲜水。
1.2药物
应用本发明实施例、对比例的处方进行实验研究。按照实施例2、实施例7、实施例11、对比例1-2的处方和制备方法将原料药得到的有效成分配置成浓缩液。
阳性对照品:地奥心血康胶囊(成都地奥制药集团有限公司);盐酸普萘洛尔片(天津力生制药股份有限公司)。
本实验中人类体重按照70kg计算,大鼠体重按照200g计算,按照体表面积则算后各组大鼠给予对应受试物剂量见下表。
表1:阳性对照品及受试物的等效剂量换算
受试物名称 | 临床剂量/天 | 折算后大鼠给药剂量/天 |
地奥心血康胶囊组 | 0.6g/人 | 0.054g/kg |
盐酸普洛奈尔片组 | 0.24g/人 | 0.0216g/kg |
本发明低剂量组 | 7.5g生药/人 | 0.675g生药/kg |
本发明中剂量组 | 17.5g生药/人 | 1.575g生药/kg |
本发明高剂量组 | 37.5g生药/人 | 3.375g生药/kg |
其中,中剂量临床剂量折算大鼠给药剂量换算方式如下:
17.5g生药/70kg*70kg*0.018/0.2kg=1.575g生药/kg
1.3仪器与试剂
仪器:电子天平:YP30002,上海越平仪器仪表有限公司;
分析天平:CPA225D,赛多利斯
医用冷藏箱:YC-300L中科美菱低温科技有限公司
超纯水系统:Milli-Q Reference A+法国Millipore SAS
病理取材台:OML-QCA湖北欧美莱医疗科技有限公司
试剂和药品:
2%红四氮唑溶液,生产厂家,上海研拓生物科技有限公司;
PBS溶液,生产厂家,上海宇玫博生物科技有限公司;
10%中性甲醛,生产厂家,湖北鑫润德化工有限公司;
0.9%氯化钠注射液,100ml/瓶,石家庄四药有限公司。
2、实验方法
取SD大鼠,雌雄各半,随机分组为15组,每组8只。
分为假手术组、模型对照组、阳性对照组1-2、对比例1-2组、实施例2、7、11的低中高剂量组,开始灌胃给药每天1次,假手术组和模型对照组给予蒸馏水,连续给药7天。
末次给药1h后,动物麻醉,对大鼠进行建模手术,手术方法如下:将大鼠取右侧卧位备皮后(上至上肢到脊柱的连线,下至肋骨最下端到脊柱的连线),动物腹腔注射10%水合氯醛溶液麻醉,于大鼠身下放置小电热毯,消毒铺单后,以心尖搏动最强的点为中心,作一横行切口,长度前后距胸骨和脊柱各lcm,,依次切开皮肤、深浅筋膜,钝性分离胸大肌和前锯肌,显露肋骨,充分止血后,由第4肋间入胸,钝性分离肋间肌(分离肋间肌的范围不可过大,不能超过皮肤切口的范围)。牵开肋骨,撕开靠中部心包,用镊子掀开左心耳,从左心耳下方2-3毫米处人针,缝扎的中点在左心耳和肺动脉圆锥的交界和心尖连线上,缝扎的方向应和左心耳的边缘平行,以无损伤缝合线进行缝扎,进针深度为1.5毫米左右,结扎的时候要控制打结的力度,尽量做到既能在打结的过程中可见到下方大面积心肌表面变得苍白,室壁运动明显减弱,但又不能力度过大,以防止将心肌扎断。结扎完成后要对心脏进行约20秒的观察,如果出现心律失常,尽快进行心脏按摩。假手术组仅暴露心脏不结扎。
根据上述方法结扎大鼠冠状动脉左前降支30min再灌注90min建立大鼠心肌IRI模型。再灌注后,动物开腹腔,腹主动脉取血处死动物,去心脏,使用TTC进行染色,拍照,测定心肌梗死面积,结果如下:
表2本发明对急性心肌缺血模型梗死面积的治疗作用
组别 | 梗死面积(%) |
假手术组 | - |
模型对照组 | 27.2±8.2<sup>&&</sup> |
阳性对照-地奥心血康胶囊组 | 20.6±7.2* |
阳性对照-盐酸普洛奈尔片组 | 12.6±5.3 |
对比例1 | 19.5±7.6 |
对比例2 | 20.6±3.4 |
实施例2-低剂量 | 21.9±2.9 |
实施例2-中剂量 | 20.4±5.8 |
实施例2-高剂量 | 18.5±6.5 |
实施例7-低剂量 | 20.5±3.9 |
实施例7-中剂量 | 18.7±5.7** |
实施例7-高剂量 | 17.2±6.8** |
实施例11-低剂量 | 18.2±3.4 |
实施例11-中剂量 | 17.5±7.1** |
实施例11-高剂量 | 16.1±8.7** |
“&”与假手术组(G1)比较p<0.05;“&&”与假手术组(G1)比较p<0.01
“*”与模型对照组(G2)比较p<0.05;“**与模型对照组比较p<0.01
从上表可以看出,与正常对照组相比,模型对照组在心肌梗死面积均具有统计学意义的差异。与模型对照组、对比例组相比,受试物组均具有抑制心肌梗死面积的作用,其中,实施例7和实施例11的中、高剂量组与模型组相比具有统计学意义的差异。
Claims (12)
1.一种治疗无复流现象的中药组合物,其特征在于,所述组合物由以下重量份的原料药组成:红景天1-30份、广枣1-30份、人参1-30份。
2.根据权利要求1所述的中药组合物,其特征在于,所述中药组合物由以下重量份的原料药组成:红景天5-14份、广枣2-8份、人参5-20份。
3.根据权利要求1或2所述的中药组合物,其特征在于,所述中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份。
4.根据权利要求1所述的中药组合物,其特征在于,所述中药组合物还包括黄连1-30份或者麦冬1-30份。
5.根据权利要求4所述的中药组合物,其特征在于,所述黄连2-15份,麦冬10-25份。
6.根据权利要求4或5所述的中药组合物,其特征在于,所述中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份、黄连10份。
7.根据权利要求4或5所述的中药组合物,其特征在于,所述中药组合物由以下重量份的原料药组成:红景天12份、广枣5份、人参18份、麦冬24份。
8.根据权利要求1-7任一项所述的中药组合物,其特征在于,所述人参为炮制品,优选为园参、林下参、野山参、生晒参或红参。
9.根据权利要求1-8任一项所述的中药组合物,其特征在于,所述中药组合物的剂型为片剂、颗粒剂、胶囊剂、丸剂、散剂或口服液体制剂。
10.一种制备权利要求1-9任一项所述中药组合物的方法,其特征在于,将所述原料药干燥研成粉末,混匀,得到有效成分;或将所述原料药水提或醇提或水提醇沉;任选的按技术人员公知的技术制成制剂。
11.根据权利要求10所述的制备方法,其特征在于,其中,
水提条件为:加所述原料药总重量的6-12倍量水提取2-3次,每次1-3小时;
醇提条件为:加所述原料药总重量的4-10倍量40-80%乙醇提取2-3次,每次1-3小时;
水提醇沉条件为:加所述原料药总重量的6-12倍量水提取2-3次,每次1-3小时,滤过,滤液浓缩至60-70℃时相对密度为1.15-1.20的清膏,加乙醇使含醇量达40-70%,静置。
12.根据权利要求10或11所述的任一种制备方法,其特征在于,其中:
方法一:取所述原料药,混合后加水提取2-3次,每次加水量相当于药材总重量的6-12倍,每次提取时间为1-3小时,合并提取液,滤过,滤液浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分;
方法二:取所述原料药,混合后用40-80%乙醇回流提取2-3次,每次乙醇用量为该两味药材总量的4-10倍,提取时间为1-3小时,合并提取液,回收乙醇并浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分;
方法三:取所述原料药,混合后加水提取2-3次,每次加水量相当于药材总重量的6-12倍,每次提取时间为1-3小时,合并提取液,滤过,滤液浓缩至60-70℃时相对密度为1.15-1.20的清膏,加入乙醇,使乙醇含量为40-70%,静置12-24小时,滤过,滤液浓缩至60-70℃时相对密度为1.20-1.30的清膏,干燥,粉碎成细粉,得有效成分。
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