CN116688053B - 一种用于改善肿瘤恶病质肌肉萎缩的中药组合物及制备方法和应用 - Google Patents
一种用于改善肿瘤恶病质肌肉萎缩的中药组合物及制备方法和应用 Download PDFInfo
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- CN116688053B CN116688053B CN202310803840.0A CN202310803840A CN116688053B CN 116688053 B CN116688053 B CN 116688053B CN 202310803840 A CN202310803840 A CN 202310803840A CN 116688053 B CN116688053 B CN 116688053B
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Abstract
本发明公开了一种用于改善肿瘤恶病质肌肉萎缩的中药组合物及制备方法和应用,该中药组合物主要由以下重量的原料药组成:人参7~12g、麦冬10~14g、五味子5~8g、刺五加7~11g、红景天8~12g、白术10~14g、陈皮8~12g、半夏10~14g、茯苓10~15g、红花10~15g、冬凌草10~15g、甘草5~8g、黄精10~15g、枸杞子10~14g。本发明中药组合物使用纯天然中药原料,来源广泛,无毒副作用,可有效改善肿瘤恶病质患者肌肉萎缩的情况,达到防治肺癌恶病质体重减轻,提高肿瘤患者的治疗效果和生存质量,延长患者生存时间的效果,是中药治疗肿瘤恶病质的一大创新。
Description
技术领域
本发明涉及一种用于改善肿瘤恶病质肌肉萎缩的中药组合物及制备方法和应用,属于中医药领域。
背景技术
肿瘤恶病质为持续性骨骼肌质量流失的多因素综合征,严重影响了患者的生活质量和肿瘤的治疗效果,缩短了患者的生存时间。肌肉代谢异常在肿瘤恶病质患者中较为常见,主要包括肌肉蛋白质分解增加,合成的肌肉蛋白减少,其中骨骼肌蛋白的严重消耗造成肌肉萎缩是恶性肿瘤患者出现恶病质的主要原因。然而相关的疗法在改善肿瘤恶病质肌肉萎缩症状方面收效甚微。
肿瘤恶病质在中医属“虚劳”的范畴,认为肿瘤恶病质属中医脾胃虚弱证,因脾为“后天之本”,“气血生化之源”,《素问》中有关于“脾主身之肌肉”的理论,中医认为脾脏为“仓禀之官”,脾在体合肉,主四肢。认为当脾胃运化功能正常,则人体肌肉发达,四肢得以濡养,故当健脾、补脾,同时助胃气以消化饮食,可很好改善肿瘤恶病质虚劳症状。临床上也有一些以健脾和胃法、脾肾双补法、补气养阴法等治疗肿瘤恶病质肌肉萎缩的中药组方,但其效果并没有达到预期。
发明内容
针对现有技术的不足,本发明的目的是提供一种用于改善肿瘤恶病质肌肉萎缩的中药组合物(中药方)及制备方法和应用,可以有效改善肿瘤恶病质患者肌肉萎缩的进程,防治肺癌恶病质体重减轻,提高治疗效果和生存质量,延长患者生存时间。
为了实现上述目的,本发明的技术方案之一是:
一种用于改善肿瘤恶病质肌肉萎缩的中药组合物,主要由以下重量的原料药组成:人参7~12g、麦冬10~14g、五味子5~8g、刺五加7~11g、红景天8~12g、白术10~14g、陈皮8~12g、半夏10~14g、茯苓10~15g、红花10~15g、冬凌草10~15g、甘草5~8g、黄精10~15g、枸杞子10~14g。
本发明的技术方案之一是:一种用于改善肿瘤恶病质肌肉萎缩的中药组合物的制备方法,制成水煎剂、片剂、丸剂、颗粒剂、膏剂。
进一步,水煎剂的制备方法为:按重量称取各原料药,混合,采用水煎法制备药液。
本发明的技术方案之一是:一种所述的中药组合物在制备改善肿瘤恶病质肌肉萎缩药物中的应用。
本发明中药组合物各原料药的功效:
人参,味甘、微苦,性温。归脾,肺经。功能主治:大补元气,复脉固脱,补脾益肺,生津,安神。用于体虚欲脱、肢冷脉微、脾虚食少、肺虚喘咳、津伤口渴。
麦冬,味微苦,性微寒。归心、肺、胃经。功能主治:润肺清心、泻热生津、化痰止呕、治嗽行水。用于治疗肺燥干咳、阴虚痨嗽、喉痹等病症。
五味子,味酸、甘,性温。归肺、心、肾经。功能主治:敛肺,滋肾,生津,收汗,涩精。用于肺虚喘咳,口干作渴,自汗,盗汗,劳伤羸瘦,梦遗滑精,久泻久痢。
刺五加,味辛、微苦,性温。归脾、肾、心经。功能主治:益气健脾、补肾安神。用于脾肺气虚,体虚乏力,食欲不振肺肾两虚,久咳虚喘,肾虚腰膝酸痛,失眠多梦。
红景天,味甘、涩,性寒。归肺经。功能主治:补气清肺、益智养心、收涩止血、散瘀消肿。用于体虚、气虚、乏力,也用于病后出现的畏寒、气短、没有力气的现象。
白术,味苦、甘,性温,归脾、胃经。功能主治:补气健脾、燥湿利水、止汗。用于脾气虚弱,食少便溏,痰饮水肿,表虚自汗,胎动不安。
陈皮,味苦、辛,性温。归肺、脾经。功能主治:燥湿化痰、理气健脾。可以用来治疗寒症、气滞、脾胃气滞、晚腹胀满、湿痰寒痰等。
半夏,味辛、性温。归脾、胃、肺经。功能主治:燥湿化痰,降逆止呕,消痞散结。用于湿痰寒痰,咳喘痰多,痰饮眩悸,风痰眩晕,痰厥头痛,呕吐反胃,胸脘痞闷,梅核气;外治痈肿痰核。
茯苓,味甘、淡,性平。归心、肺、脾、肾经。功能主治:利水渗湿、健脾宁心,用于水肿尿少、痰饮眩悸、脾虚食少、便溏泄泻、心神不安、惊悸失眠。
红花,味辛,性温。归心、肝经。功能主治:活血通经,散瘀止痛。临床多用于治疗经闭,痛经,恶露不行,癥瘕痞块,胸痹心痛,瘀滞腹痛,胸胁刺痛,跌扑损伤,疮疡肿痛。
冬凌草,味苦、甘,性微寒。归肺、胃、肝经。功能主治:清热、解毒、活血止痛,用于咽喉肿痛、扁桃体炎、蛇虫咬伤、风湿骨痛等。
甘草,味甜,性平。归脾、肺经,兼入胃、心经。功能主治:益气补中,祛痰止咳,解毒,缓急止痛,缓和药性。用于脾胃虚弱,倦怠乏力,心悸气短,咳嗽痰多,脘腹、四肢挛急疼痛,痈肿疮毒,缓解药物毒性、烈性。
黄精,性平、味甘。归脾、肺、肾经。功能主治:补气养阴,健脾,润肺,益肾。用于脾胃气虚,体倦乏力,胃阴不足,口干食少,肺虚燥咳,劳嗽咳血,精血不足,腰膝酸软,须发早白,内热消渴。
枸杞子,味甘,性平。归肝、肾经。功能主治:滋肾,润肺,补肝,明目。用于肝肾阴亏,腰膝酸软,头晕,目眩,目昏多泪,虚劳咳嗽,消渴,遗精。
本发明中药方中人参,大补元气,补益脾肾,黄精补益脾肾,益肾填精,两者共为君药;白术健脾益气,茯苓健脾渗湿,枸杞子、五味子补益肝肾,四药共助君药补益脾肾;刺五加补肝肾,祛风湿强筋骨,半夏燥湿化痰,降胃止呕,陈皮行气化痰,红花活血化瘀,红景天补气活血,冬凌草清热解毒化痰,六药佐君臣补肝肾、和脾胃,化痰活血,共为佐药;麦冬滋阴和胃,甘草健脾解毒,两者调和诸药为使。诸药协同,扶正祛邪,共凑健脾益肾,化痰活血之效;紧扣肿瘤恶病质脾肾两虚,气阴不足,痰瘀毒结之病机。
本发明中药组合物使用纯天然中药原料,来源广泛,无毒副作用,可有效改善肿瘤恶病质患者肌肉萎缩的情况,达到防治肺癌恶病质体重减轻,提高肿瘤患者的治疗效果和生存质量,延长患者生存时间的效果,是中药治疗肿瘤恶病质的一大创新。
附图说明
图1为中药方对腓肠肌肌纤维横截面积的影响(400×)。
图2为中药方对小鼠腓肠肌自噬体的影响(单箭头指示自噬体)。
图3为中药方对肺癌恶病质小鼠腓肠肌中自噬关键蛋白表达的影响。图中,A代表空白组;B代表模型组;C代表中药方组;D代表阳性药组。
具体实施方式
以下结合实施例对本发明的具体实施方式作进一步详细说明。
实施例1
一种用于改善肿瘤恶病质肌肉萎缩的中药组合物,主要由以下重量数的原料药组成:人参9g、麦冬12g、五味子6g、刺五加9g、红景天10g、白术12g、陈皮10g、半夏12g、茯苓15g、红花15g、冬凌草15g、甘草6g、黄精15g、枸杞子12g。
其制备方法为:按重量份数称取各原料药,混合,采用水煎法制备药液,浓缩后冷藏备用。
实施例2
一种用于改善肿瘤恶病质肌肉萎缩的中药组合物,主要由以下重量的原料药组成:人参7g、麦冬10g、五味子5g、刺五加7g、红景天8g、白术10g、陈皮8g、半夏10g、茯苓10g、红花10g、冬凌草10g、甘草5g、黄精10g、枸杞子10g。
其制备方法为:按重量份数称取各原料药,混合,采用水煎法制备药液,浓缩后冷藏备用。
实施例3
一种用于改善肿瘤恶病质肌肉萎缩的中药组合物,主要由以下重量的原料药组成:人参12g、麦冬14g、五味子8g、刺五加11g、红景天12g、白术14g、陈皮12g、半夏14g、茯苓12g、红花12g、冬凌草12g、甘草8g、黄精12g、枸杞子14g。
其制备方法为:按重量份数称取各原料药,混合,采用水煎法制备药液,浓缩后冷藏备用。
本发明基于“脾主肌肉”这一中医理论基础,采用健脾益肾,化痰活血的中药方治疗肿瘤恶病质肌肉萎缩,围绕中药方对恶病质小鼠体重、腓肠肌重量、腓肠肌横截面积以及自噬相关蛋白表达的影响,从小鼠动物实验探究中药方防治肺癌恶病质肌肉萎缩的作用机制和改善效果,为肿瘤恶病质的中医药防治提供临床用药配方。具体实验如下:
1材料与试剂
1.1细胞及动物培养
Lewis肺癌细胞,常规培养;采购6周龄雄性C57BL/6J小鼠,购于华兴实验动物养殖场,生产许可证号:SCXK(豫)2019-0002,实验动物使用许可证号:SYXK(豫)2020-0004,适应一周后开始构建模型。
1.2中药方制备及用药剂量
中药方制备:人参9g、麦冬12g、五味子6g、刺五加9g、红景天10g、白术12g、陈皮10g、半夏12g、茯苓15g、红花15g、冬凌草15g、甘草6g、黄精15g、枸杞子12g,称取药材,应用水煎法进行药物提取,所得提取物浓缩后,冷藏备用。所有药物采购于河南中医药大学第一附属医院,并在本实验室进行制备。
左旋肉碱(阳性药)是一种常应用于临床改善肿瘤恶病质的现代医学治疗常用药,可减缓肌肉耗损进程。
2方法
2.1制备Lewis肺癌恶病质肌肉萎缩小鼠模型
收取对数生长期Lewis细胞,调整细胞浓度为107个/mL。随机给8只小鼠编号作为空白组,其余32只小鼠于右前腋下制备肺癌恶病质模型,0.2mL/只。
2.2动物实验分组设计及给药
每周记录小鼠的瘤体体积和体重变化。造模第8天,32只小鼠随机分为模型组、中药方组、阳性药组,每组8只,各实验组给药途径:造模第8天,空白组和模型组用生理盐水、中药方组用中药方液、阳性药组用左旋肉碱(10mg/kg)分别灌胃给药,每天一次,给药3周。中药方按下面公式计算给药剂量,并与空白组对照比较。
小鼠给药剂量=人的临床常用量×动物等效剂量系数9.1(按体表面积),每只小鼠体重20g,每日灌胃体积0.2mL/次。
2.3动物取材
给药3周后,采用5%(w/v)的水合氯醛溶液对小鼠进行麻醉处理,在冰上取小鼠双后肢腓肠肌,在分析天平上准确称取腓肠肌的重量,每组选取6只小鼠的部分腓肠肌放入装有4%(w/v)多聚甲醛溶液的试管中在常温下保存,用于HE染色(苏木精-伊红染色法),其余腓肠肌装入冻存管中,在液氮中迅速冷却以后,转移到-80℃冰箱,用于后续的腓肠肌相关蛋白检测。
2.4腓肠肌HE染色和肌纤维横截面积检测
小鼠腓肠肌组织取材、脱水、石蜡包埋和切片处理后进行HE染色,步骤包括:脱蜡与水洗、苏木精染核、伊红染色细胞质、脱水透化、封固、镜下观察等。
小鼠腓肠肌HE切片置于倒置显微镜(Olympus,CX31)下拍照,使用Image pro plus6.0图像采集系统对肌纤维横截面进行圈定计算出400倍视野下单位切片的总面积,依据肌纤维的数量算出肌纤维横截面的平均值和标准差。
2.5透射电镜成像观察恶病质小鼠腓肠肌的肌纤维结构以及自噬体数量
将刚剥离的腓肠肌组织,称重后切取厚度在1mm以下的薄片,放入多聚甲醛-戊二醛混合固定液中,经过脱水、浸透、包埋聚合、切片及染色等步骤后,运用电子显微镜观察各组腓肠肌内肌纤维的结构以及自噬体数量的变化。
2.6Western blot(WB法)检测自噬相关蛋白
将取下的腓肠肌用胶原酶消化、过滤、离心、洗涤,再离心去上清得肌细胞,加入RIPA(组织快速裂解液):PMSF(苯甲基磺酰氟)=100:1的裂解液,收集组织蛋白后,使用BCA蛋白浓度测定试剂盒检测蛋白浓度。上样,转膜,孵育一抗(Beclin1、ULK1、LC3B/LC3A、P621:2000),二抗(山羊抗兔GAPDH 1:10000),化学发光显色。在凝胶成像系统中扫描条带。使用Image Lab软件进行数据整理。
2.7统计分析
采用IBM SPSS 22.0统计软件进行分析,样本量符合正态分布且方差齐的用单因素方差分析,然后再进行LSD检验的分析,检测P<0.05具有统计学意义。不符合正态分布者用非参数检验。数据结果用表示。
3结果
3.1小鼠一般情况观察
造模第7天,在小鼠右腋皮下可以触摸到明显肿块,肿瘤体积在0.698±0.351cm3,造模完成。与空白组比较,造模小鼠明显出现体表毛发暗淡,四肢无力,活动减少,体重下降,体现出肿瘤恶病质症状。
3.2中药方对肺癌恶病质小鼠体重的影响
给药3周后,与空白组比较,模型组小鼠体重、腓肠肌重量及肌纤维横截面积明显下降(P<0.05);与模型组比较,中药方组小鼠体重、腓肠肌重量及肌纤维横截面积明显升高,且具有统计学意义(P<0.05);中药方组与阳性药组比较,小鼠体重和腓肠肌重量没有明显差异(P>0.05),但肌纤维横截面积有差异(P<0.05),结果见表1和图1。
表1中药方对肺癌恶病质小鼠体重、腓肠肌重量及肌纤维横截面积的影响(n=8)
注:#与空白组比较,P<0.05;*与模型组比较,P<0.05;^与阳性药组比较,P<0.05。3.3中药方对小鼠腓肠肌纤维结构和自噬体的影响
给药3周后,与空白组比较,模型组腓肠肌肌纤维的排列散乱,细胞间界限增宽,明暗带分布不均匀,肌纤维横纹排列不整齐;与模型组相比,中药方组、阳性药组肌纤维的排列较为整齐,细胞间隙较紧密,明暗带分布较均匀,肌纤维横纹排列较为整齐。
与空白组比较,模型组腓肠肌自噬体数量明显增加(P<0.05);与模型组比较,中药方组腓肠肌自噬体数量明显减少(P<0.05),阳性药组腓肠肌中自噬体数量降低,但差异无统计学意义(P>0.05)(表2和图2)。
表2中药方对腓肠肌自噬体的影响(n=8)
注:#与空白组比较,P<0.05;*与模型组比较,P<0.05;^与阳性药组比较,P<0.05。3.4中药方对肺癌恶病质小鼠腓肠肌中自噬关键蛋白Beclin1、ULK1、LC3B/LC3A、P62表达的影响
给药3周后,与空白组比较,模型组小鼠腓肠肌中Beclin-1、ULK1、LC3B/LC3A、P62蛋白表达明显增加(P<0.05);与模型组比较,中药方组腓肠肌Beclin-1、ULK1、LC3B/LC3A、P62蛋白表达降低(P<0.05);与模型组比较,阳性药组腓肠肌Beclin-1、ULK1蛋白表达降低(P<0.05);各用药组之间蛋白表达未见明显差异(P>0.05),其中LC3B蛋白的表达超过LC3A的表达水平越高,两者比值大于1时,才可产生自噬流,比值越大,其自噬程度越明显,结果见表3和图3。
表3中药方对肺癌恶病质小鼠腓肠肌Beclin1、ULK1、LC3B/LC3A、P62蛋白表达的影响(n=8)
注:#与空白组比较,P<0.05;*与模型组比较,P<0.05;^与阳性药组比较,P<0.05。
4讨论
本实验复制了Lewis肺癌恶病质经典的小鼠模型,通过中药方和阳性药干预恶病质成瘤小鼠21天后发现:中药方用药组小鼠体重、肠肌重量及肌纤维横截面积明显增加,且自噬体数量显著减少;中药方组中与自噬相关的ULK1、Beclin-1、LC3B/LC3A、P62蛋白表达较模型组均明显下降,表明中药方对于小鼠自噬途径具有抑制作用,降低肌肉蛋白的降解,有效改善肺癌恶病质小鼠肌肉萎缩。且与阳性药左旋肉碱相似,可以增加小鼠腓肠肌重量。
以上表明,中药方突出中医药方剂和中药的优势,针对肿瘤恶病质的发病机制发挥作用,可通过抑制自噬途径相关蛋白的表达,改善肿瘤恶病质引起的体重减轻和肌肉萎缩的进程,从而改善肿瘤恶病质症状。
Claims (7)
1.一种用于改善肿瘤恶病质肌肉萎缩的中药组合物,其特征在于,由以下重量的原料药组成:人参7~12 g、麦冬10~14 g、五味子5~8 g、刺五加7~11 g、红景天8~12 g、白术10~14g、陈皮8~12 g、半夏10~14 g、茯苓10~15 g、红花10~15 g、冬凌草10~15 g、甘草5~8 g、黄精10~15 g、枸杞子10~14 g。
2.根据权利要求1所述用于改善肿瘤恶病质肌肉萎缩的中药组合物,其特征在于,由以下重量的原料药组成:人参9 g、麦冬12 g、五味子6 g、刺五加9 g、红景天10 g、白术12 g、陈皮10 g、半夏12 g、茯苓15 g、红花15 g、冬凌草15 g、甘草6 g、黄精15 g、枸杞子12 g。
3.根据权利要求1所述用于改善肿瘤恶病质肌肉萎缩的中药组合物,其特征在于,由以下重量的原料药组成:人参7 g、麦冬10g、五味子5 g、刺五加7 g、红景天8 g、白术10 g、陈皮8 g、半夏10 g、茯苓10 g、红花10 g、冬凌草10 g、甘草5 g、黄精10 g、枸杞子10 g。
4.根据权利要求1所述用于改善肿瘤恶病质肌肉萎缩的中药组合物,其特征在于,由以下重量的原料药组成:人参12 g、麦冬14 g、五味子8 g、刺五加11 g、红景天12 g、白术14g、陈皮12 g、半夏14 g、茯苓12 g、红花12 g、冬凌草12 g、甘草8 g、黄精12 g、枸杞子14 g。
5.一种如权利要求1-4任一项所述用于改善肿瘤恶病质肌肉萎缩的中药组合物的制备方法,其特征在于,所述中药组合物制成水煎剂、片剂、丸剂、颗粒剂、膏剂。
6.根据权利要求5所述用于改善肿瘤恶病质肌肉萎缩的中药组合物的制备方法,其特征在于,水煎剂的制备方法为:按重量称取各原料药,混合,采用水煎法制备药液。
7.一种如权利要求1-4任一项所述的中药组合物在制备改善肿瘤恶病质肌肉萎缩药物中的应用。
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