CN114886994A - 一种治疗肝癌的苗药及其制备方法 - Google Patents
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- CN114886994A CN114886994A CN202210369080.2A CN202210369080A CN114886994A CN 114886994 A CN114886994 A CN 114886994A CN 202210369080 A CN202210369080 A CN 202210369080A CN 114886994 A CN114886994 A CN 114886994A
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Abstract
本发明公开了一种治疗肝癌的组合物,本发明所述苗药由艾叶、嘎炯豆丢劳独、见血清制备而成。本发明具有祛风除湿,温经散寒的功效,以达到温经散寒、舒筋活络、凉血止血、清热解毒的疗效,适用于患有肝癌的人群,制成的苗药疗效显著,无毒副作用。
Description
发明领域:
本发明涉及一种治疗肝癌的苗药及其制备方法,属于药品、尤其是苗药技术的领域。
背景技术
肝癌是一种发生在肝脏的恶性肿瘤,与饮酒、病毒性肝炎、食用没变食物、遗传等有关,早期常无症状,晚期出现肝区疼痛、发热、乏力等,早期有治愈可能,中晚期治疗复杂,效果差异大。肝癌可分为原发性肝癌和继发性肝癌两大类,其中原发性肝癌是指肝细胞或肝内胆管上皮细胞发生的恶性肿瘤;继发性肝癌又称转移性肝癌,指身体其他器官的恶性肿瘤扩散或转移至肝脏。现目前的治疗方法有化学药物治疗、手术治疗、中药治疗、放化疗等。不管是药物治疗还是手术治疗都存在复发的风险,并产生癌症转移的可能、放化疗治疗的副作用大,不能长期使用。
苗医药被列为第二批国家级非物质文化遗产名录,苗族医对疾病的认识和立方遣药有其独特模式。苗族在长期生产和与疾病斗争的实践中,总结积累了丰富而宝贵的医药经验,逐渐形成了苗族医的“纲、经、症、疾”的理论模式,以及关于病因、疾病分类、诊断、治疗和预防等方面的具有浓郁民族特色的苗族医药理论。苗医认为,毒、亏、伤、积、菌、虫是导致人体生病的六种因素,简称六因。而六因归根结底都要用产生毒害力的方式才能导致人体生病,所以苗医素有“无毒不生病”之说。故针对肝癌,苗医采用清热解毒,消肿止痛的苗药,来除去体内癌细胞滋生的毒素,还能起到消肿止痛的作用,以达到治疗肝癌的目的。其治疗肝癌以提高机体抗病力,改善全身状况和症状,减轻化疗、放疗不良反应,来减少肝癌的复发几率。同时,副作用较少,安全性大,也无依赖性,能长期使用。
发明内容
本发明所要解决的技术问题在于提供一种治疗肝癌的苗药及其制备方法。本发明具有清热解毒,消肿止痛的功效,适用于患有肝癌的人群。
为解决以上技术问题,本发明采用以下技术方案实现:
一种治疗肝癌的苗药,其药用有效成分按照重量组分计算:由艾叶100-500份,嘎炯豆丢劳独100-500份,见血清1-50份,歪倒秀10-150份,莴冲岗10-150份,莴迷莎幼10-150份制备而成。
具体的说,所述的治疗肝癌的苗药,其药用有效成分按照重量组分计算:由艾叶200-400份,嘎炯豆丢劳独200-400份,见血清5-30份,歪倒秀30-100份,莴冲岗30-100份,莴迷莎幼30-100份制备而成。
更具体的说,所述的治疗肝癌的苗药,其药用有效成分按照重量组分计算:由艾叶250份,嘎炯豆丢劳独250份,见血清10份,歪倒秀50份,莴冲岗50份,莴迷莎幼50份制备而成。
前述治疗肝癌的苗药的制备方法为:按照比例称取方中所有药材,并按照常规的制备方法进行加工,制成相应的苗药制剂。
具体地说,前述治疗肝癌的苗药的制备方法为:按照比例称取方中所有药材,将其粉碎后混合均匀后,加水煎煮后得煎煮液,并按照常规的制备方法进行加工,制成相应的药物制剂。
所述药物制剂包括水煎剂、洗剂。
所述水煎液这样制备:按照比例称取方中所有药材,将其粉碎后混合均匀后,打包备用,使用前取该药包加1000-2000ml水浸泡10-30分钟后,先大火煎煮30-60分钟,再文火煎煮1-2小时,水煎液倒入浴盆中加入适量冷水,泡浴30-60分钟即得。
所述洗剂这样制备:按照比例称取方中所有药材,将其粉碎后混合均匀后,加10倍量的水浸泡30-60分钟,再煎煮2-3次,每次1-2小时,合并两次水煎液,过滤后每500ml灌装装袋,密封即得。
与现有技术相比,本发明具有以下有益效果:
本发明所述治疗肝癌的苗药由艾叶,嘎炯豆丢劳独,见血清,歪倒秀,莴冲岗,莴迷莎幼制备而成。各成分性质及功效如下:艾叶为菊科植物艾Artemisia argyi LevL etVant.的干燥叶;夏季花未开时采摘,除去杂质,晒干;味辛、苦,性温;有小毒;归肝、脾、肾经;具有温经止血,散寒止痛;外用祛湿止痒的功效;用于吐血,衄血,崩漏,月经过多,胎漏下血,少腹冷痛,经寒不调,宫冷***;外治皮肤瘙痒;醋艾炭温经止血,用于虚寒性出血。嘎炯豆丢劳独(又名八角枫)为八角枫科八角枫属植物华瓜木Alangium chinenese(Lour.)Harms[Stylidium chinense Lour.]和瓜木Alangium Platanifolium(Sieb.Et Zucc.)Harms[Marlea platanifolia Sieb.Et Zucc.]的根、须根或根皮;根全年可采,挖出后,除去泥沙,斩取侧根和须状根,晒干即得;夏、秋采叶及花,晒干备用或鲜用;性热,味麻、辣,属热药,入冷经,有大毒;具有祛风除湿,舒筋活络的功效。见血清为兰科植物脉羊耳兰Liparis nervosa(Thunb.)Lindl.的全草;性凉,味苦、涩,归肺、肾经;具有凉血止血,清热解毒之功效;常用于胃热吐血,肺热咯血,肠风下血,崩漏,手术出血,创伤出血,疮疡肿毒,毒蛇咬伤,跌打损伤。歪倒秀又称(柴胡)为伞形科植物柴胡Bupleurum chinense DC.或狭叶柴胡Bupleurum scorzonerifolium Willd.的干燥根;按性状不同,分别***,味淡、微甜,属两经药;主要功效是清热解毒、利湿消肿;对痈肿、咽痛、蛇伤等有较强的解毒消痈作用;一般用于治疗疮毒、咽喉肿痛、肠痈腹痛、毒蛇咬伤,也用于热淋涩痛、小便不利。莴迷莎幼又称半边莲为桔梗科植物半边莲Lobelia chinensis Lour.的干燥全草;夏季采收,除去泥沙,洗净,晒干;性热,味辣、苦;属热药,入冷经;具有清热解毒,利水消肿的功效;用于痈肿疗疮,蛇虫咬伤,臌胀水肿,湿热黄疽,湿疹湿疮。
方解:
母药:莴冲岗(白花蛇舌草),性平,味淡、微甜,属两经药,清热解毒,利湿消肿;莴迷莎幼(半边莲),性热,味辣、苦,属热药,入冷经,清热解毒,利水消肿。歪倒秀(柴胡),性热,味苦、涩,属热药,入冷经,疏肝理气。
子药:艾叶,性辛、苦、温,归肝、脾、肾经,温经止血,散寒止痛。嘎炯豆丢劳独(八角枫),性热,味麻、辣,属热药,入冷经,有大毒,祛风除湿,舒筋活络;见血清,性凉,味苦、涩,归肺、肾经,凉血止血,清热解毒;
本发明进行了大量的实验研究,以下为本发明实验研究的结果:
一、急性毒性试验
1、实验动物:SD大鼠,雌雄各半,体重180-220g,SPF级,购于贵州医科大学实验动物中心,实验动物使用许可证:SYXK(黔)2018-001。饲养条件:动物代谢笼分笼饲养,保持昼夜节律,室温20-22℃,相对湿度60%-70%,自由取食进水。
2、实验方法:
分组:取健康SD大鼠80只,适应性喂养5d,按随机数字表法随机分为8组,每组10只。完整皮肤40只,即完整实施例1组、完整实施例2组、完整实施例3组、完整空白组;破损皮肤40只,即分为破损实施例1组、破损实施例2组、破损实施例3组、破损空白组;给药:在给药前24h小时,腹腔注射10%水合氯醛麻醉(0.3mL/100g),用宠物剃毛器将动物背部脊柱两侧背毛剃除,再用8%硫化钠脱毛,范围约3cm*3cm;脱毛24h后检查脱毛皮肤是否完整。大鼠破损皮肤的急性毒性试验,处理方法按照上述方法脱毛,24h后用粗钝刀片轻瓜背部脱毛后的皮肤,不伤及皮下组织,以皮下渗血而不造成表皮破损为度。各给药组将本发明药物实施例1、2、3组药物均匀涂抹于大鼠皮肤脱毛区域,空白对照组给予相同体积的蒸馏水,给药24h后,去除药物,连续观察7d,包括动物的大小便及其颜色、呼吸状况、精神状态、皮肤异常反应,外观,行为活动、体重变化以及中毒反应的严重程度、起始时间等。对于死亡动物及濒死动物行大体解剖,其他动物在观察期结束后大体进行解剖,肉眼观察主要脏器颜色、体积、相对位置等情况。
4、结果:完整皮肤大鼠和破损皮肤大鼠,皮肤短期内接触本发明药物,其呼吸状况、精神状态、外观、行为活动均无异常,用药部位皮肤无异常反应,7d之内,无大鼠死亡。大鼠的体重增长正常,与空白对照组比较,给药组的体重增长无明显差异(P>0.05)。结果见表1,表2。
注:注:与空白对照组比较,*P<0.05
注:与空白对照组比较,*P<0.05
5、结论:由表1和表2可知,在完整皮肤的大鼠急性毒实验中,实施例1组、实施例2组、实施例3组与空白对照组比较,在用药后各时间点的体重增加无显著性差异(P>0.05);在破损皮肤的大鼠中,实施例1组、实施例2组、实施例3组与空白对照组比较,在用药后各时间点的体重增加无显著性差异(P>0.05);表明本发明药物组合在短时间内对大鼠不存在急性毒性,经皮肤是安全的。
二、皮肤刺激性
1、实验动物:豚鼠,雌雄各半,清洁级,由贵州医科大学实验动物中心提供,实验动物使用许可证:SYXK(黔)2018-001。
2、实验方法:取健康豚鼠40只,适应性喂养5d,按随机字表法分为4组,即实施例1组、实施例2组、实施例3组与空白对照组,每组10只。给药前24h将各组豚鼠背部左右两侧的毛对称剃除,每侧面积约3cm*3cm,用8%硫化钠溶液脱尽短毛,并用温水洗净。在豚鼠脱毛24h后,在各组动物左侧脱毛去,分别涂抹本发明药物实施例1、实施例2和实施例3,每只0.2mL,空白对照组涂抹等量蒸馏水。代谢笼分笼饲养。涂药4h后,用温水去除残留药物,观察1h、24h、48h、72h皮肤反应。按照皮肤刺激性反应评分标准计算各组、各时间点评分。红斑:0无红斑,1非常轻的红斑,2明显的红斑,3中度至重度的红斑,4中度红斑(鲜红色)至焦痂(深层损伤);水肿:0无水肿,1非常轻度水肿,勉强可见,2轻度水肿,边缘清晰,3中度水肿(皮肤***约1mm),4重度水肿(皮肤***大于1mm,并超过涂受试药物的区域)。
4、结果:各组皮肤刺激性实验结果如表3所示。
注:与空白对照组比较,*P<0.05
5、结论:上述试验均显示实施例各组在1h到72h之间对皮肤均无刺激作用,表明本发明苗药组合经皮肤给药没有刺激性作用。
三、实验例1:CCK-8法检测本发明药物对肝癌HepG2细胞增殖抑制作用的影响
1、实验细胞和材料:肝癌HepG2细胞购自于美国模式培养物集存库(ATCC);CCK-8试剂盒(日本同仁化学);DMEM培养基(Gibco),胎牛血清(杭州四季青),胰酶(碧云天),PBS(博士德)。
2、实验方法:将处于对数生长期的肝癌HepG2细胞消化后重悬,调整细胞密度为4×104/mL,接种于96孔培养板中,每孔100μL,置37℃、5%CO2培养箱中培养24h。移弃旧培养液,加入含不同浓度的本发明实施例1药物的完全培养基,浓度设为0、50、100、200、400、800μg/mL,每孔100μL,每组设5个复孔,放入培养箱继续培养,48后分别加入10μLCCK-8试剂,置培养箱孵育2h后取出测定450nm处吸光度值(OD值)。不加细胞和药物只含完全培养基的孔为空白对照组孔,只含细胞和培养基不加药物组为细胞对照组。计算细胞活力(%)=[OD(加药)-OD(空白)]/[OD(对照)-OD(空白)]×100。
4、实验结果:通过CCK-8检测不同处理组和不同时间点的肝癌HepG2细胞的存活和生长情况,结果如表4所示,本发明药物实施例1苗药组方各浓度药物处理HepG2细胞后的OD值明显低于对照组,说明本发明药物苗药组合可明显抑制肝癌细胞的增殖作用。
表4本发明药物实施例1对肝癌HepG2细胞的增殖作用
注:*代表与HepG2对照组相比P<0.05。
四、实验例2:划痕实验检测本发明药物对肝癌HepG2细胞迁移能力的影响
1、细胞培养和铺板:将处于对数生长期的肝癌HepG2细胞消化后重悬,调整细胞密度为2×105/mL,接种于6孔培养板中,每孔1mL,加培养基至2mL,置37℃、5%CO2培养箱中培养24h。
2、配制本发明药物实施例1培养基稀释液。待细胞铺满整个6孔板底面,用200μL枪头进行划痕,弃去6孔板旧培养基,PBS洗涤并弃净,加入相应的培养基和本发明药物,使终浓度为0、100μg/mL、400μg/mL。显微镜下拍照记录0h各组划痕宽度。
3、每隔24h更换新的培养基和相应浓度的药物,48h后显微镜下拍照各组细胞划痕距离。
5、结果:如图1所示。结果显示本发明药物实施例1苗药组肝癌细胞的迁移能力明显低于空白对照组(P<0.05),提示本发明药物组合可抑制肝癌细胞的迁移能力。
五、实验例3:Transwell实验检测本发明药物实施例1对肝癌细胞的侵袭能力的影响
1、基质胶铺板:将Matrigel以1:8浓度稀释,包被transwell小室底部膜的上室面,置37℃30min使Matrigel聚合成凝胶。
2、将处于对数生长期的肝癌HepG2细胞消化后重悬,用无血清培养基调整为单细胞悬液,以2×104/孔接种到Transwell上室,并加入本发明实施例1制得的苗药组合,使终浓度为0、100μg/mL、400μg/mL,下室加入600μL含20%血清的培养基,将培养板小室置入37℃、5%CO2培养箱中培养24h。
3、固定封片、染色和结果统计:取出Transwell小室,弃去孔中培养液,PBS洗涤2遍,甲醇固定30min,将小室适当风干;0.1%结晶紫染色30分钟,PBS冲洗干净,用棉签轻轻擦掉上层未迁移细胞。400倍显微镜下随机选取5个视野统计结果。
5、结果:如图2所示。结果显示本发明药物实施例1药物两个浓度组侵袭的细胞明显低于空白对照组(P<0.05),说明本发明药物实施例1苗药组合可有效抑制肝癌细胞的侵袭能力。
六、临床病例
病例1
王某,男,56岁,疲倦乏力,食欲减退,肝区不适、疼痛半年,经医院检查诊断为肝癌,住院治疗1个月,效果不理想,用本发明苗药组合物制成水煎液进行泡浴4个月,疼痛减轻,用药半年后,不良症状消失,一年后随访,人健在。
病例2
李某,女,64岁,右上腹疼痛并呈持续性钝痛2月,身体消瘦,食欲减退,腹部胀满,经医院诊断为原发性肝癌伴腹水,肝癌,于贵阳市某三甲医院行手术治疗,半年后复查发现肝内肿瘤复发,化疗5个疗程后出院。于家中用本发明苗药组合制成水煎液泡浴6个月,复查肿瘤未再增大,食欲增加,症状明显改善,肝功能相关指标明显好转,甲胎蛋白值明显下降,生活质量明显提高。
病例3
许某,男,58岁,自述腕腹胀痛,疲乏无力,实验室检查甲胎蛋白升高,肝功能异常,经B超检查发现肝右叶异常,CT检查,确诊为肝癌,住院治疗3月余,效果不佳。家中用本发明苗药组合制成水煎液泡浴4个月,腕腹胀痛感减轻,甲胎蛋白值明显降低,肝功能明显改善。
与现有技术相比,本发明选用艾叶,嘎炯豆丢劳独,见血清,歪倒秀,莴冲岗,莴迷莎幼组方,具有清热解毒,消肿止痛的功效,适应肝癌人群的有益效果。
附图说明
图1本发明药物对肝癌细胞迁移能力的影响;
图2本发明药物对肝癌细胞侵袭能力的影响。
具体实施方式
下面结合实施例对本发明作进一步的说明,但并不作为对本发明限制的依据。
实施例1。
处方:由艾叶500g,嘎炯豆丢劳独500g,见血清50g,歪倒秀150g,莴冲岗150g,莴迷莎幼150g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,打包备用,使用前取该药包加1000ml水浸泡10分钟后,先大火煎煮30分钟,再文火煎煮1小时,水煎液倒入浴盆中加入适量冷水,泡浴30分钟即得水煎液。
用法用量:每7天泡浴一次为一疗程,一次30分钟。
实施例2。
处方:艾叶100g,嘎炯豆丢劳独100g,见血清1g,歪倒秀10g,莴冲岗10g,莴迷莎幼10g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,加10倍量的水浸泡30分钟,再煎煮2次,每次1小时,合并两次水煎液,过滤后每500ml灌装装袋,密封即得洗剂。
用法用量:每次取两袋洗剂,加适量温水泡浴30分钟,每7天一次为一疗程。
实施例3。
处方:艾叶250g,嘎炯豆丢劳独250g,见血清10g,歪倒秀50g,莴冲岗50g,莴迷莎幼50g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,打包备用,使用前取该药包加1500ml水浸泡30分钟后,先大火煎煮50分钟,再文火煎煮1.5小时,水煎液倒入浴盆中加入适量冷水,泡浴45分钟即得水煎液。
用法用量:每7天泡浴一次为一疗程,一次30分钟。
实施例4。
处方:艾叶300g,嘎炯豆丢劳独200g,见血清20g,歪倒秀10g,莴冲岗50g,莴迷莎幼110g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,加8倍量的水浸泡60分钟,再煎煮3次,每次2小时,合并两次水煎液,过滤后每500ml灌装装袋,密封即得洗剂。
用法用量:每次取两袋洗剂,加适量温水泡浴30分钟,每7天一次为一疗程。
实施例5。
处方:艾叶100-500g,嘎炯豆丢劳独100-500g,见血清1-50g,歪倒秀10-150g,莴冲岗10-150g,莴迷莎幼10-150g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,打包备用,使用前取该药包加2000ml水浸泡20分钟后,先大火煎煮60分钟,再文火煎煮2小时,水煎液倒入浴盆中加入适量冷水,泡浴60分钟即得水煎液。
用法用量:每7天泡浴一次为一疗程,一次30分钟。
实施例6。
处方:艾叶100-500g,嘎炯豆丢劳独100-500g,见血清1-50g,歪倒秀10-150g,莴冲岗10-150g,莴迷莎幼10-150g制成。
工艺:按照比例称取方中所有药材,将其粉碎后混合均匀后,加12倍量的水浸泡45分钟,再煎煮3次,每次1.5小时,合并两次水煎液,过滤后每500ml灌装装袋,密封即得洗剂。
用法用量:每次取两袋洗剂,加适量温水泡浴30分钟,每7天一次为一疗程。
Claims (8)
1.一种治疗肝癌的苗药,其特征在于:其药用有效成分按照重量组分计算:由艾叶100-500份,嘎炯豆丢劳独100-500份,见血清1-50份,歪倒秀10-150份,莴冲岗10-150份,莴迷莎幼10-150份制成。
2.如权利要求1所述的治疗肝癌的苗药,其特征在于:其药用有效成分按照重量组分计算:由艾叶200-400份,嘎炯豆丢劳独200-400份,见血清5-30份,歪倒秀30-100份,莴冲岗30-100份,莴迷莎幼30-100份制成。
3.如权利要求1或2所述的治疗肝癌的苗药,其特征在于:其药用有效成分按照重量组分计算:由艾叶250份,嘎炯豆丢劳独250份,见血清10份,歪倒秀50份,莴冲岗50份,莴迷莎幼50份制成。
4.如权利要求1-3中所述治疗肝癌的苗药的制备方法,其特征在于:按照比例称取方中所有药材,并按照常规的制备方法进行加工,制成相应的苗药制剂。
5.如权利要求4所述治疗肝癌的苗药的制备方法,其特征在于:按照比例称取方中所有药材,将其粉碎后混合均匀后,加水煎煮后得煎煮液,并按照常规的制备方法进行加工,制成相应的药物制剂。
6.如权利要求5所述治疗肝癌的苗药的制备方法,其特征在于:所述药物制剂包括水煎液、洗剂。
7.如权利要求6所述治疗肝癌的苗药的制备方法,其特征在于:所述水煎液这样制备:按照比例称取方中所有药材,将其粉碎后混合均匀后,打包备用,使用前取该药包加1000-2000ml水浸泡10-30分钟后,先大火煎煮30-60分钟,再文火煎煮1-2小时,水煎液倒入浴盆中加入适量冷水,泡浴30-60分钟即得。
8.如权利要求6所述治疗膀胱癌苗药的制备方法,其特征在于:所述洗剂这样制备:按照比例称取方中所有药材,将其粉碎后混合均匀后,加10倍量的水浸泡30-60分钟,再煎煮2-3次,每次1-2小时,合并两次水煎液,过滤后每500ml灌装装袋,密封即得。
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