CN116966252A - 一种治疗更年期动脉粥样硬化的中药组合物 - Google Patents
一种治疗更年期动脉粥样硬化的中药组合物 Download PDFInfo
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Abstract
本发明公开了一种治疗更年期动脉粥样硬化的中药组合物，涉及中药组合物技术领域，其技术方案要点是：所述组合物由穿山龙10‑20g，莪术5‑15g，赤芍5‑15g，黄连3‑7g组成。所述制备方法为：将穿山龙、莪术、赤芍和黄连各个药材加入水共同浸泡，其中水面高出药材顶面2cm；然后将各个药材和浸泡药材的水一起煎煮，过滤并收集滤液，在滤渣中加入与药平齐的水量二次煎煮，过滤并收集滤液，将两次收集到的滤液混合后，浓缩得到310ml的粘稠溶液，置于‑20℃冰箱中保存，得中药组合物。该中药组合物可疏风活血、祛瘀消癥，疏解血脉横逆风邪、祛除留滞血脉瘀滞、清化血脉癥瘕，可用于治疗更年期AS。
Description
技术领域
本发明涉及中药组合物技术领域，更具体地说，它涉及一种治疗更年期动脉粥样硬化的中药组合物。
背景技术
女性绝经后，动脉粥样硬化(AS)相关的心脑血管病发病率随年龄增长显著上升。女性绝经后的时间约占女性生命周期的二分之一，因此AS相关心脑血管病成为影响女性绝经后生命健康的重大问题。
一项大型临床研究从中国10个地区招募了302632名女性，进行为期9年随访。研究发现，与非绝经期女性相比，绝经后女性患致死性或非致死性冠心病的风险显著增加，绝经时间与致死性、非致死性心血管疾病皆呈负相关，即绝经年龄越小，冠心病风险越高，且随绝经时间的延长，冠心病风险持续增高。一项对1368名急性冠脉综合征患者斑块特征的研究显示，女性斑块破裂和易损斑块(如脂质斑块、薄纤维帽等)的发生随着年龄增长而增加。雌二醇是目前干预更年期综合征的重要手段，但研究却发现尽管雌二醇可维持血管内皮功能，但同时也可增加绝经期女性AS相关的心脑血管病发病率，并增加乳腺癌、子宫内膜癌、深静脉栓塞等发生风险。因此寻找具有雌激素样效应而无雌激素不良反应的药物，研究其类雌激素样机制，对防治心血管一类疾病至关重要。综上，更年期AS严重影响了女性生命健康，是全球范围内亟待解决的重大问题。
更年期女性，随着生理月经的减少或停止，隐曲气郁、血脉留滞留瘀、蕴而化热动风，易导致胸痹心痛、中风等，临床症状可见风逆血脉、瘀血留滞的胸痛、头晕、偏瘫以及蕴热化风所致的烘热、潮热、盗汗、烦躁等。血脉留滞、蕴热化风、横逆血脉经络，致气血运行失常；瘀血留滞，又可酿热化毒，损伤脏腑血脉。因此，“内风逆窜血脉、瘀毒互结”，致生血脉瘀滞、化为癥瘕，是更年期动脉硬化发生发展的主要病因病机。
发明内容
本发明的目的是提供一种治疗更年期动脉粥样硬化的中药组合物，可疏风活血、祛瘀消癥，疏解血脉横逆风邪、祛除留滞血脉瘀滞、清化血脉癥瘕，可用于治疗更年期AS。
本发明的上述技术目的是通过以下技术方案得以实现的：一种治疗更年期动脉粥样硬化的中药组合物，所述中药组合物由穿山龙10-20g，莪术5-15g，赤芍5-15g，黄连3-7g组成。
本发明进一步设置为：所述中药组合物中穿山龙、莪术、赤芍和黄连的重量比为3:2:2:1。
本发明进一步设置为：所述中药组合物的最佳配比为穿山龙15g，莪术10g，赤芍10g，黄连5g。
治疗更年期动脉粥样硬化的中药组合物的制备方法：将穿山龙、莪术、赤芍和黄连各个药材加入水共同浸泡，其中水面高出药材顶面2cm；然后将各个药材和浸泡药材的水一起煎煮，过滤并收集滤液，在滤渣中加入与药平齐的水量二次煎煮，过滤并收集滤液，将两次收集到的滤液混合后，浓缩得到310ml的粘稠溶液，置于-20℃冰箱中保存，得中药组合物。
治疗更年期动脉粥样硬化的中药组合物的应用：所述中药组合物用于治疗更年期动脉粥样硬化的药物的制备。
综上所述，本发明具有以下有益效果：该中药组合物包含穿山龙，莪术，赤芍，黄连。其中穿山龙擅祛风活血，尤善疏解血脉风邪而通血脉，为方中君药。莪术行气破血、散瘀消癥；赤芍清热凉血、活血祛瘀消癥。两者寒热并用，可消减血脉癥瘕，共为臣药。黄连一可引诸药入心经血脉，二可清化血脉蕴结热毒，为佐药。合用上述中药，可疏风活血、祛瘀消癥，疏解血脉横逆风邪、祛除留滞血脉瘀滞、清化血脉癥瘕，可用于治疗更年期AS。
附图说明
图1是本发明实施例中实验研究步骤示意图；
图2中A是本发明实验研究中小鼠主动脉全长斑油红O大体染色的代表图像；B是本发明实验研究中小鼠主动脉全长斑面积的相对定量分析；
图3中A是本发明实验研究中小鼠主动脉根部斑块油红O染色的代表图像(50X)；B是本发明实验研究中小鼠主动脉根部斑块面积的相对定量分析；
图4是本发明实验研究中小鼠主动脉根部HE染色图(200X)；
图5中A是本发明实验研究中小鼠主动脉根部Masson染色代表图像(40X)；B是本发明实验研究中小鼠主动脉根部胶原纤维含量的相对定量分析；
图6是本发明实验研究中小鼠血清激素水平(*P<0.05，**P<0.01)；
图7是本发明实验研究中小鼠血清脂质水平(*P<0.05，**P<0.01)；
图8是本发明实验研究中小鼠血清炎症因子水平(*P<0.05，**P<0.01)。
具体实施方式
以下结合附图1-8对本发明作进一步详细说明。
实施例：一种治疗更年期动脉粥样硬化的中药组合物，所述中药组合物由穿山龙10-20g，莪术5-15g，赤芍5-15g，黄连3-7g组成。所述中药组合物中穿山龙、莪术、赤芍和黄连的重量比为3:2:2:1。所述中药组合物的最佳配比为穿山龙15g，莪术10g，赤芍10g，黄连5g。
该中药组合物的制备方法如下：将穿山龙、莪术、赤芍和黄连各个药材加入水共同浸泡，其中水面高出药材顶面2cm；然后将各个药材和浸泡药材的水一起煎煮，过滤并收集滤液，在滤渣中加入与药平齐的水量二次煎煮，过滤并收集滤液，将两次收集到的滤液混合后，浓缩得到310ml的粘稠溶液，置于-20℃冰箱中保存，得中药组合物。所述中药组合物用于治疗更年期动脉粥样硬化的药物的制备。
下面通过实验来论证本发明所提供的中药组合物可用于制备治疗更年期动脉粥样硬化的药物。
实验动物：选择SPF级8周龄雌性ApoE-/-雌性小鼠[品系C57BL/6J]60只，体重18-22g，购自北京金牧阳实验动物养殖有限责任公司，许可证号:SCXK(京)2021-0013。饲养条件为2级，室温保持在(22士2)℃，相对湿度(55士5)％，光照时间7:00-19:00。
实验材料：
饲料：小鼠高脂饲料[77.8％基础饲料+10％猪油+10％蛋黄粉+2％胆固醇+0.2％胆盐]：由小黍有泰提供，货号D12108C。
药物：①阿托伐他汀钙片(商品名:立普妥):辉瑞制药有限公司，批号：FK8380，每片l0 mg；②中药组合物由穿山龙10-20g，莪术5-15g，赤芍5-15g，黄连3-7g组成，饮片购自北京同仁堂科技股份有限公司。将穿山龙、莪术、赤芍和黄连各个药材加入水共同浸泡，其中水面高出药材顶面2cm；然后将各个药材和浸泡药材的水一起煎煮，过滤并收集滤液，在滤渣中加入与药平齐的水量二次煎煮，过滤并收集滤液，将两次收集到的滤液混合后，浓缩得到310ml的粘稠溶液，置于-20℃冰箱中保存，得中药组合物，即实验中的高剂量浓度药液。将高剂量浓度药液稀释3倍后得到中剂量浓度药液，将高剂量浓度药液稀释9倍后得到低剂量浓度药液。
实验方法：
实验小鼠处理方法：
①ApoE-/-雌性小鼠高脂喂养8周后，行卵巢切除术。术前称小鼠体重，每10g重量腹腔注射0.1ml 5％的水合氯醛。注射完成后，腹位固定小鼠，用小鼠剃毛刀在其背部切口处进行剃毛。在背部躯干下1/3，离开脊柱两侧各1-2cm处作纵形切口，长约lcm，在分离肌层后，可见淡黄色的脂肪团，用小镊子拉出脂肪团，可找到一团细线状不规则呈肉色的组织即为卵巢，下接粗大管状组织为子宫角，此时在卵巢下端穿丝线结扎，减去卵巢及余线，将组织退回腹腔，缝合切口，放回笼中，随时观察小鼠手术切口清况、呼吸频率等。术后24小时存活小鼠给予高脂饲料喂养。
②假手术组ApoE-/-雌性小鼠做同样切口，只穿线不结扎，保留卵巢，切口缝合。
实验分组：将去势彻底的小鼠随机分为5组：模型组、阿托伐他汀组、中药低剂量组、中药中剂量组、中药高剂量组，每组10只。将假手术组10只作为对照组，阿托伐他汀组作为阳性药组。给予高脂饲料喂养并同时给予药物干预8周。每周对小鼠外观及体重进行检测。小鼠灌胃的药物剂量是根据成人每日用药临床推荐的常用剂量，按体重系数比折算而成，每周记录体重以调整药物剂量。
①假手术组：做双侧卵巢切口保留卵巢，高脂饲料喂养+蒸馏水0.3ml/只灌胃,每日一次；
②模型组：双侧卵巢切除，高脂饲料喂养+蒸馏水0.3ml/只灌胃,每日一次；
③阿托伐他汀组：双侧卵巢切除，高脂饲料喂养+阿托伐他汀钙片10mg/kg/d灌胃0.3ml/只，每日一次；
④中药低剂量组：双侧卵巢切除，高脂饲料喂养+1.73g/kg/d灌胃0.3ml/只，每日一次；
⑤中药中剂量组：双侧卵巢切除，高脂饲料喂养+5.2g/kg/d灌胃0.3ml/只，每日一次；
⑥中药高剂量组：双侧卵巢切除，高脂饲料喂养+15.6g/kg/d灌胃0.3ml/只，每日一次。
表1实验动物分组与给药
标本采集：给药后处死小鼠，处死前12h禁食，不禁水。称体重，小鼠摘除眼球取血，室温静置2h，3000rpm离心15min，分离血清。小鼠解剖后，剪下主动脉。每组取5只小鼠完整的主动脉用4％的组织固定液固定于15ml的离心管，用于油红O大体染色、HE和Masson染色；余小鼠主动脉组织迅速用液氮冻存，然后转至-80℃保存，用于相关基因和蛋白检测。
实验检测:
①血清检测：
脂质代谢相关指标的检测：测试盒微量法测定血清总胆固醇(TG)、甘油三酯(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平。
炎性因子相关指标的检测：Elisa试剂盒测定TNF-α、IL-1β、IL-6。
雌激素水平相关指标的检测：Elisa试剂盒测定雌二醇(E2)、孕酮(P4)的表达
②组织检测：
每组小鼠精细分离留下一条完整的全主动脉4％多聚甲醛固定，用于油红O染色，离体主动脉样品和冷冻主动脉根部切片(厚度为10μm)进行脂质沉积评估，观察动脉粥样硬化斑块面积及部位。
主动脉根部冰冻切片进行HE、Masson染色，分别观察主动脉根部及主动脉瓣处斑块内坏死核心数量、胶原含量、纤维含量。
数据整理与统计分析：所有实验结果均使用GraphPad Prism 8软件统计分析，结果以“平均值±标准差”表示。组间数据比较，符合正态分布且方差齐的，采用单因素方差(ANOVA)分析。若不符合正态分布，则需使用非参数检验。P<0.05为存在差异，具有统计学意义。
如图2所示:
与假手术组小鼠比较，模型组小鼠主动脉全长斑块形成增加(P<0.01)；
与模型组比较，给予阳性药阿托伐他汀(P<0.01)、中药组合物中剂量(P<0.05)和高剂量(P<0.01)可减少小鼠主动脉全长斑块面积；
给予中药组合物高剂量比低剂量的斑块面积减少明显(P<0.05)。
如图3所示:
与假手术组小鼠比较，模型组小鼠主动脉全长斑块形成增加(P<0.01)；
与模型组比较，给予阳性药阿托伐他汀(P<0.01)、中药组合物低剂量、中剂量、高剂量(P<0.01)均可减少小鼠主动脉根部斑块面积。
如图4、图5所示:
空白组小鼠主动脉结构致密，胶原纤维排布整齐。
与空白组相比,模型组血管基底部胶原疏松，存在断裂现象，面积明显减少(P＜0.01)。
与模型组比较,经中药治疗组小鼠斑块内部胶原纤维含量增加，基底部血管组织疏松、排列紊乱情况改善；高剂量组胶原纤维含量明显增加(P＜0.05)。
如图6中血清ELISA结果显示:
与假手术组相比，模型组的E2和P水平显著下降(P<0.01)；
与模型组比较，给予中药组合物中剂量组P升高(P<0.05)；高剂量组E2和P显著升高(P<0.01)。
与中药组合物低剂量组比较，高剂量组E2和P升高(P<0.05)。
如图7中血清生化结果显示:
与假手术组相比，模型组的TC、TG、LDL水平显著升高(P<0.01)，HDL水平显著下降(P<0.01)；
与模型组比较，给予阳性药阿托伐他汀组的炎症因子TC、TG、LDL下降(P<0.01)，HDL显著升高；低剂量组TC水平下降(P<0.01)；中剂量组TC、TG、LDL下降(P<0.05)；高剂量组TC、TG、LDL显著下降(P<0.01)，HDL显著升高(P<0.01)。
与中药组合物低剂量组比较，高剂量组TC、TG、LDL下降(P<0.05)、HDL显著升高(P<0.01)。
如图8中血清ELISA结果显示:
与假手术组相比，模型组的炎症因子TNF-α、IL-1β、IL-6水平显著升高(P<0.01)；
与模型组比较，给予阳性药阿托伐他汀组的炎症因子TNF-α、IL-1β、IL-6水平下降(P<0.001)，低剂量组IL-6水平下降(P<0.05)，中剂量组IL-1β(P<0.001)、IL-6(P<0.05)水平下降，高剂量组TNF-α(P<0.05)、IL-1β(P<0.001)、IL-6(P<0.001)水平下降；与中药低剂量组比较，中药高剂量组IL-1β(P<0.05)、IL-6(P<0.001)水平下降更为显著。
通过实验研究发现本发明所提供的中药组合物能够有效抑制去势APOE-/-小鼠炎症反应，降低去势APOE-/-小鼠主动脉斑块的形成。进而说明该中药组合物可用于治疗女性更年期AS。
本具体实施例仅仅是对本发明的解释，其并不是对本发明的限制，本领域技术人员在阅读完本说明书后可以根据需要对本实施例做出没有创造性贡献的修改，但只要在本发明的权利要求范围内都受到专利法的保护。
Claims (5)
1.一种治疗更年期动脉粥样硬化的中药组合物，其特征是：所述中药组合物由穿山龙10-20g，莪术5-15g，赤芍5-15g，黄连3-7g组成。
2.根据权利要求1所述的一种治疗更年期动脉粥样硬化的中药组合物，其特征是：所述中药组合物中穿山龙、莪术、赤芍和黄连的重量比为3:2:2:1。
3.根据权利要求1所述的一种治疗更年期动脉粥样硬化的中药组合物，其特征是：所述中药组合物的最佳配比为穿山龙15g，莪术10g，赤芍10g，黄连5g。
4.根据权利要求1所述的中药组合物的制备方法，其特征是：所述制备方法为：将穿山龙、莪术、赤芍和黄连各个药材加入水共同浸泡，其中水面高出药材顶面2cm；然后将各个药材和浸泡药材的水一起煎煮，过滤并收集滤液，在滤渣中加入与药平齐的水量二次煎煮，过滤并收集滤液，将两次收集到的滤液混合后，浓缩得到310ml的粘稠溶液，置于-20℃冰箱中保存，得中药组合物。
5.根据权利要求1所述的中药组合物的应用，其特征是：所述中药组合物用于治疗更年期动脉粥样硬化的药物的制备。
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