CN110090233B - 槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途 - Google Patents
槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途 Download PDFInfo
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Abstract
本发明涉及槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途。本发明的槐耳菌质提取物对浆细胞乳腺炎确实有效，比现有的治疗方法显著加速，例如传统的疗法大约需要6个月以上的时间的治疗，才能获得疗效，而本发明的槐耳菌质提取物在5周的时间内便使患者伤口愈合，分泌物显著减少。
Description
发明领域
本发明属于药物技术领域，具体涉及一种槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途。
技术背景
浆细胞乳腺炎(Plasma cell mastitis)是乳腺的一种慢性非细菌性炎症。准确的病理机制尚不清楚。由于临床表现具有不同的特点，常被冠以不同的名称，如“闭塞性乳腺炎”、“哺乳期乳腺炎”、“慢性乳腺炎”、“乳腺导管扩张症”、“粉刺样乳腺炎”等。浆细胞性乳腺炎多发生在中、老年女性，高峰年龄为50～60岁，也可发生于男性。在临床上常被误诊为一般细菌感染，乳腺结核，甚至误诊为乳腺癌误切乳房。
对于这种疾患的治疗，由于其主要病因不是感染，所以抗生素的治疗基本无益。手术治疗常常导致局部伤口反复破溃，反复发作。目前临床上这种疾患的最主要的治疗药物是皮质类固醇的应用。但是，众所周知，糖皮质激素的应用可导致股骨头坏死，感染等严重的毒副作用。显然，发展新的疗效确切、无毒副作用的制剂具有很重要的临床使用价值。
发明内容
目前，对于浆细胞异常导致的疾患没有确实有效的药物制剂可供临床应用，本申请的发明人意外地发现槐耳菌质提取物在预防和/或治疗浆细胞性乳腺炎方面有不错的疗效。因此，本发明的目的在于提供一种槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途。
本发明的上述目的是采用以下技术方案来实现的。
本发明提供一种槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途。
其中，所述槐耳菌质提取物为槐耳菌质水提物。
优选地，所述槐耳菌质提取物采用如下方法来制备：取槐耳菌质，加水煎煮3次，第一次加2-15倍量的水，煮沸2-10小时，过滤，滤液备用；第二、第三次分别加1-15倍量水，煮沸2-10小时，过滤，合并三次滤液，减压浓缩至相对密度为1.05～1.50(55℃)，即得。
优选地，所述槐耳菌质提取物采用如下方法来制备：取槐耳菌质，加水煎煮3次，第一次加7倍量的水，煮沸6小时，过滤，滤液备用；第二、第三次分别加5倍量水，煮沸6小时，过滤，合并三次滤液，减压浓缩至相对密度为1.35～1.40(55℃)，即得。
优选地，所述药物为颗粒，可以采用如下方法来制备：将上述制备的槐耳菌质提取物与适量的糊精、可溶性淀粉等辅料混匀，制粒，干燥，过筛，分装，即得。
在本发明中，“槐耳菌质”是指槐耳菌Trametes robiniophila Murr.在玉米蕊轴、麦麸等发酵基质上，于一定条件下经培养后所得的干燥菌质。可以使用本领域常规的方法获得，例如但不限于，中国专利申请96116920.6公开的“槐栓菌及其入药应用”。也可以通过商业渠道获得，例如但不限于，启东盖天力药业有限公司根据国家食品药品监督管理局批准的工艺生产，批准文号(国药准字Z20000108)。
在本发明中，针对的疾病为浆细胞性乳腺炎，对于其他的乳腺炎，例如“感染性乳腺炎”没有显现的效果。感染性的乳腺炎需要抗生素的的治疗才有效。
与现有的药物相比较，本发明至少具有以下有益效果：
在以前，浆细胞性乳腺炎常常会被误诊为“乳腺癌”，因此，会采用抗癌药物来进行治疗，但是事实上，抗癌的药物用于浆细胞性乳腺炎的治疗没有效果。过去曾经有报道用抗癌药氨甲喋呤治疗粒细胞性乳腺炎(Granulomatous mastitis)，氨甲喋呤在临床上具有众所周知的毒性作用，其带来的毒副作用远远大于其治疗乳腺炎带来的益处。即使氨甲喋呤用于乳腺炎的治疗，据国际文献报道(ANZ J.Surg.2003；73:247–249)，需要和皮质激素强的松合并应用才有效，治疗时间需要12个月，显然与本制剂相比(本制剂5周显著改善)疗效差的很远。
本发明的槐耳菌质提取物对浆细胞乳腺炎确实有效，比现有的治疗方法显著加速，例如传统的疗法大约需要6个月以上时间的治疗，才能获得疗效，而本发明的槐耳菌质提取物在5周的时间内便使患者伤口愈合，分泌物显著减少。
传统药物多有严重的毒副作用，如长期应用糖皮质激素导致的免疫低下的副作用；手术导致伤口迁延不愈等；抗生素对于不伴有感染的患者常常无效等。本发明的药物不仅快速，而且不具有严重的毒副作用。
附图说明
图1为槐耳菌质提取物对变性牛血清白蛋白诱导小鼠外周血液中游离浆细胞及其产生自身抗体IgA+-ASC细胞的影响。
具体实施方式
下面通过实施例详细说明本发明，应当理解，下述实施例仅用于说明本发明，而不以任何方式限制本发明的范围。
实施例1槐耳颗粒用于浆细胞性乳腺炎患者的治疗
槐耳菌质提取物的制备：取槐耳菌质(获自启东盖天力药业有限公司，批号：20161215)，加水煎煮3次，第一次加7倍量的水，煮沸6小时，过滤，滤液备用；第二、第三次分别加5倍量水，煮沸6小时，过滤，合并三次滤液，减压浓缩至相对密度为1.35～1.40(55℃)，即得。
槐耳颗粒的制备：
将上述制备的槐耳菌质提取物825g与254g糊精、127g可溶性淀粉等辅料混匀，制粒，干燥，过筛，制成1000g颗粒，分装，即得。
槐耳颗粒用于浆细胞性乳腺炎患者的治疗：
浆细胞性乳腺炎是一种由浆细胞异常导致的慢性疾患，既往用抗生素、皮质激素、甚至手术疗法均未见明显的疗效，治疗长达6个月以上，病情仍然迁延不愈。在本实施例中，采用前述制备的槐耳颗粒进行治疗，患者5周复查均呈现显著的改善。具体临床资料如下：
以上六位患者均患有浆细胞乳腺炎，并进行了手术。由于槐耳颗粒在临床应用过程中，具有调节免疫功能的作用，本次临床试验目的是观察槐耳颗粒是否对浆细胞乳腺炎有效，当患者来复查时，在六个患者中，有五个患者用药五周后，创口已经愈合。(而以往的患者需要半年以上才能愈合)，其中患者2合并细菌感染，在第六周的时候愈合。
实施例2槐耳菌质提取物对变性牛血清白蛋白诱导小鼠外周血液中游离浆细胞及其产生自身抗体IgA+-ASC细胞的影响
我们通过用变性的牛血清白蛋白，反复给小鼠灌胃，反复刺激小鼠的胃肠道的粘膜淋巴组织，导致肠道的B淋巴细胞激活，形成大量的浆细胞(Plasma cells)。此后给小鼠灌胃槐耳菌质提取物进行实验性治疗，发现槐耳菌质提取物可以显著减少浆细胞的形成，并抑制由浆细胞的持续活化而导致的自身抗体(如自身IgA1)的形成，具体研究过程如下。
1、材料与方法
主要试剂：
(1)淋巴细胞分离液(Ficoll-Paque TM PLUS，GE healthcare产品，Cat.No:17-1440-02,Lot No：10163635)；
(2)细胞蛋白转移抑制剂(BD Golg Stop TM Protein Transport Inhibitor，Cat.No:554715,Lot No:3165536.BD Biosciences,19975Terreyana Rd.,San Diego，CA92121，USA)；
(3)CD19-APC(BD Biosciences产品,Catalog No：340437。Lot No：3182916。19975Terreyana Rd.,San Diego，CA 92121，USA)；
(4)细胞固定与打孔液(Fixation and Permeabilization solution,Cat.No:554715。Lot No:3165536.BD Biosciences产品,19975Terreyana Rd.,San Diego，CA92121，USA)；
(5)自身IgA-FITC抗体(Gene Tech Company Limited，Catalog No.GF020429，LotNo：2013112901。
(6)改良型RPMI-1640培养液(Thermo Fisher Scientific(China)Co.,Ltd.产品，Catalog No：SH30809.01B，Lot No：NVM0346。北京市安定门东大街28号邮编：100007)；10％fetal bovine serum(FBS，GIBCO产品，Catalog No:16000-044。Lot No:1259720。GrandIsland，NY 14072，USA)；streptomycin与penicillin(HyClone Laboratories Inc.产品，Catalog No:SV30010，Lot No：J122331。925West 1800South，Logan，Utah，84321)。
(7)槐耳菌质提取物的制备：取槐耳菌质(获自启东盖天力药业有限公司，批号为20170718)，加水煎煮3次，第一次加7倍量的水，煮沸6小时，过滤，滤液备用；第二、第三次分别加5倍量水，煮沸6小时，过滤，合并三次滤液，减压浓缩至相对密度为1.35～1.40(55℃)，即得。
(8)香菇多糖、灵芝多糖、猴头多糖：陕西恒德堂植物原料提取厂提供。
主要实验仪器：流式细胞仪(Guava Easycyte System，Guava Technologies,Inc，25801Industrial Blvd.,Hayward,CA 94545，USA)
主要实验方法：选择BALB/C近交系小鼠，12周龄。体重18.0-27.5g。前5周隔日给予牛血清白蛋白(Bovine serum albumin，简称BSA，200mg/kg)酸化水溶液灌胃。从第6周起，给予模型小鼠BSA溶液尾静脉注射。剂量为20mg/kg(制备方法：将125mg BSA溶于50ml无菌生理盐水中)，每天1次,连续3天。从第8周起，给予模型小鼠葡萄球菌肠毒素(Staphylococcal Enterotoxin B，SEB。Sigma公司产品)SEB(0.5mg/kg,将2.5mg的SEB溶于40ml无菌生理盐水中，按体重进行尾静脉注射，其剂量为0.5mg/kg)。每周1次,连续3周。正常对照组尾静脉注射等容量的生理盐水。
从第9周开始到第12周结束的4周中，对上述造模成功的小鼠进行实验治疗。分别灌胃，低剂量槐耳菌质提取物组(12.09g生药/kg/day，连续灌喂28天)；中剂量槐耳菌质提取物组(24.18g生药/kg/day，连续灌喂28天)；及高剂量槐耳菌质提取物组(48.36g生药/kg/day，连续灌喂28天)。为了进行对比，还同步应用了以下药物：香菇多糖(24.18g生药/kg/day)，灵芝多糖(24.18g生药/kg/day)及猴头多糖(24.18g生药/kg/day)。
2、实验结果
实验结果如图1所示。结果显示，浆细胞异常增殖模型小鼠：与正常小鼠相比，灌胃变性牛血清白蛋白模型小鼠血液中CD19阳性细胞，即脱离正常粘膜部位，异常地进入外周血液循环中(mis-trafficking to systemic circulation)的B淋巴细胞(CD19+细胞)显著增多。同时，可分泌自身IgA抗体的B细胞(自身IgA+-ASC)也显著增多。模型组与正常对照组比较，均有显著性差异。经统计学检验，P＜0.001(one way ANOVA with T-test)。
低剂量(12.09g生药/kg/day)槐耳菌质提取物组：与模型组比较，外周血液循环中(mis-trafficking to systemic circulation)的B淋巴细胞(CD19+细胞)以及可分泌自身IgA抗体的细胞(自身IgA+-ASC)并未显著减少。P值均大于0.05。
中剂量(24.18g生药/kg/day)槐耳菌质提取物组：与模型组比较，外周血液循环中(mis-trafficking to systemic circulation)的B淋巴细胞(CD19+细胞)显著减少。可分泌自身IgA抗体的细胞(自身IgA+-ASC)显著降低。经统计学检验，P值小于0.01(one wayANOVA with T test)。
高剂量(48.36g生药/kg/day)槐耳菌质提取物组：与模型组比较，外周血液循环中(mis-trafficking to systemic circulation)的B淋巴细胞(CD19+细胞)以及可分泌自身IgA抗体的细胞(自身IgA+-ASC)均显著减少。经统计学检验，P＜0.001(one way ANOVAwith T test)。根据流式细胞的统计学分析，其作用强于低剂量与中剂量组。
采用香菇多糖(24.18g生药/kg/day)，灵芝多糖(24.18g生药/kg/day)及猴头多糖(24.18g生药/kg/day)与槐耳菌质提取物进行比较，与模型组比较，外周血液循环中(mis-trafficking to systemic circulation)的B淋巴细胞(CD19+细胞)以及可分泌自身IgA抗体的浆细胞(自身IgA+-ASC)并未显著减少。P值均大于0.05。
Claims (4)
1.槐耳菌质提取物在制备预防和/或治疗浆细胞性乳腺炎的药物中的用途，其中，所述槐耳菌质提取物为槐耳菌质水提物。
2.根据权利要求1所述的用途，其特征在于，所述槐耳菌质提取物采用如下方法来制备：取槐耳菌质，加水煎煮3次，第一次加2-15倍量的水，煮沸2-10小时，过滤，滤液备用；第二、第三次分别加1-15倍量水，煮沸2-10小时，过滤，合并三次滤液，减压浓缩至55℃下相对密度为1.05～1.50，即得。
3.根据权利要求1或2所述的用途，其特征在于，所述槐耳菌质提取物采用如下方法来制备：取槐耳菌质，加水煎煮3次，第一次加7倍量的水，煮沸6小时，过滤，滤液备用；第二、第三次分别加5倍量水，煮沸6小时，过滤，合并三次滤液，减压浓缩至55℃下相对密度为1.35～1.40，即得。
4.根据权利要求1或2所述的用途，其特征在于，所述药物为颗粒，采用如下方法来制备：将上述制备的槐耳菌质提取物与适量的糊精、可溶性淀粉混匀，制粒，干燥，过筛，即得。
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| CN101439147A (zh) * | 2007-11-19 | 2009-05-27 | 启东盖天力药业有限公司 | 槐杞黄颗粒及其制备方法和制药用途 |
| CN101933950A (zh) * | 2009-07-03 | 2011-01-05 | 启东盖天力药业有限公司 | 槐耳菌质水提物的用途 |
| CN104119426A (zh) * | 2013-04-24 | 2014-10-29 | 启东盖天力药业有限公司 | 一种槐耳多糖蛋白及其制备方法和用途 |
| CN106974943A (zh) * | 2017-01-22 | 2017-07-25 | 浙江中医药大学 | 一种槐耳醇提物的用途及制备方法 |
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| CN101439147A (zh) * | 2007-11-19 | 2009-05-27 | 启东盖天力药业有限公司 | 槐杞黄颗粒及其制备方法和制药用途 |
| CN101933950A (zh) * | 2009-07-03 | 2011-01-05 | 启东盖天力药业有限公司 | 槐耳菌质水提物的用途 |
| CN104119426A (zh) * | 2013-04-24 | 2014-10-29 | 启东盖天力药业有限公司 | 一种槐耳多糖蛋白及其制备方法和用途 |
| CN106974943A (zh) * | 2017-01-22 | 2017-07-25 | 浙江中医药大学 | 一种槐耳醇提物的用途及制备方法 |
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