CN113456748A - 抗热应激中药及其制备方法 - Google Patents
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Abstract
本发明公开了抗热应激中药,该抗热应激中药主要是在藿香、石膏、金银花、菊花及薄荷5味具有清热解暑的药材组成的中药复方的基础上,还同时加入麦冬、酸枣仁用于镇静、宁心、生津,再佐以黄柏、凤尾草用于抗炎、抗菌,还加入山楂用于调节消化,制成抗热应激中药汤剂。本发明还提供了该抗热应激中药的制备方法,并通过实验确定该抗热应激中药的抗热应激机理。该抗热应激中药能有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响,其作用机理是通过调节机体发热机制、显著改善血液细胞状况、维持体内血液指标的稳定来增强机体抗热应激能力,本发明为后期抗热应激中药研究及在动物养殖上的应用提供参考。
Description
技术领域
本发明涉及中药配方领域,特别是涉及抗热应激中药配方及其制备方法。
背景技术
热应激是指当受到超过自身调节能力的高温刺激时人和动物所做出的非特异性应答反应的总和。由于我国南方夏季气温较高,持续时间长,以及畜牧业集约化养殖方式的快速发展,高温引起的热应激会引起动物体内生理生化过程发生改变,进而对畜禽的健康和生产性能产生不良影响,引起动物新陈代谢紊乱,生产性能下降,免疫力减弱,甚至死亡。给养殖业带来巨大损失,因此,近年来找寻安全、有效的方法来提高畜禽抗热应激能力在养殖业倍受关注。
目前养殖场比较常用的一些抗热应激措施是物理降温缓解热应激带来的不良影响,如防止太阳福射、加强通风、喷淋风机降温、湿帘风机降温、冷风机和屋顶喷淋降温法等。虽然降低环境温度是缓解热应激最有效的方法,但在实际生产中其作用效果是有限的或由于成本太高难以应用。
目前抗热应激的添加剂比较常见的是电解质类、维生素类、镇静药类、抗生素类、有机酸类、中草药粉散剂类等。而这些添加剂抗热应激效果都不够理想,镇静药类及抗生素类添加剂还会产生毒副作用及药物残留,而目前大多数抗热应激中草药添加剂是以中草药简单粉碎后以粉末方式添加,导致中药生物利用度降低而影响抗热应激效果。
抗热应激中药分为清热类、抗惊厥类、调节代谢类、调整中枢神经功能、增强免疫力类等,现有的技术中,还缺乏针对养殖动物的适合的抗热应激中药制剂,及其抗热应激中药的作用机理。
发明内容
本发明的目的在于克服现有技术中的上述缺陷,提供了抗热应激中药及其制备方法,并探讨在热应激条件下抗热应激中药对小鼠存活率、生长性能、血常规和抗氧化能力的影响,结果表明,该抗热应激中药能有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响,并明确了该抗热应激中药的作用机理,为后期抗热应激中药研究及在动物养殖上的应用提供参考。
为实现上述目的,本发明提供了抗热应激中药,主要由下述重量份数的材料制备而成:凤尾草3~5份、藿香3~5份、麦冬3~5份、山楂3~5份、金银花3~5份、酸枣仁3~5份、石膏3~5份、菊花3~5份、黄柏3~5份、及薄荷3~5份。
作为本发明的进一步改进,所述的抗热应激中药,主要由下述重量份数的材料制备而成:凤尾草3份、藿香3份、麦冬3份、山楂3份、金银花3份、酸枣仁3份、石膏3份、菊花3份、黄柏3份、及薄荷3份。
上述抗热应激中药的制备方法,首先,按重量配比取用于制备抗热应激中药的所有材料,先用清水漂洗至少1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡至少50min,然后用大火煮开后,再用小火煮至少50min,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮30min以上,再次过滤取滤液,合并两次过滤取得的滤液,浓缩制成生药含量为0.2~0.5g/mL的抗热应激中药,放入3~6℃冰箱中冷藏,备用。
作为本发明的进一步改进,所述抗热应激中药的制备方法,首先,按重量配比取用于制备抗热应激中药的所有材料,先用清水漂洗1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡50min,然后用大火煮开后,再用小火煮50min,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮30min,再次过滤取滤液,合并两次过滤中取得的滤液,浓缩制成生药含量为0.2g/mL的抗热应激中药,放入4℃冰箱中冷藏,备用。
与现有技术相比,本发明提供的抗热应激中药的有益效果在于:在藿香、石膏、金银花、菊花及薄荷5味具有清热解暑的药材组成的中药复方的基础上,还同时加入麦冬、酸枣仁用于镇静、宁心、生津,再佐以黄柏、凤尾草用于抗菌、抗炎,还加入山楂用于调节消化,将这些中药在中兽医理论指导下进行组合后,经煎煮提取有效成分,制成抗热应激中药,提高中药生物利用度,可极大发挥其抗热应激的作用;通过抗热应激实验,可以确定本发明抗热应激中药机理,是通过调节机体发热机制、改善血液功能、维持体内血液指标的稳定来增强机体抗热应激能力,各成分具有相互协同促进的作用,有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响。
本发明提供的抗热应激中药的制备方法简单,制成的抗热应激中药制剂易于吸收,生物利用度高。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
本发明提供了抗热应激中药,主要由下述重量份数的材料制备而成:凤尾草3份、藿香3份、麦冬3份、山楂3份、金银花3份、酸枣仁3份、石膏3份、菊花3份、黄柏3份、及薄荷3份。
发明人在研究过程中发现,清热解毒的中草药可以提高动物的生长性能、调节机体免疫和增强抗氧化的功能,缓解热应激对动物的影响,其中,以藿香、黄柏、石膏可有效减轻热应激对小鼠肠道结构和黏膜的损害;山楂、黄柏、藿香和凤尾草均具有抗炎的作用。通过进一步实验并结合现代药理研究发现,凤尾草味淡、微苦,性凉,具有清热利湿、解毒止痢,凉血止血的作用,有显著的抗菌抗炎效果;麦冬味甘、微苦,性微寒,归胃、肺、心经,有养阴润肺、益胃生津、清心除烦,能有效缓解口干渴的症状,可改善胃黏膜的血液循环,可抗心肌损伤和抗炎,提高受损心肌细胞的活力和搏动频率;金银花可显著降低热应激时动物的直肠温度,还具有抗炎的作用;菊花既能清肝火,又能散风热,具有增加心肌收缩力和抗炎的作用;薄荷能降低温热环境下动物的体温和呼吸频率,也有抗炎的作用;酸枣仁性平,味甘、酸,归心、肝、胆经,具有补肝、宁心、敛汗、生津的作用。
在本发明实施例1中,在藿香、石膏、金银花、菊花及薄荷5味具有清热解暑的药材组成的中药复方的基础上,还同时加入麦冬、酸枣仁用于镇静、宁心、生津,再佐以黄柏、凤尾草用于抗菌、抗炎,还加入山楂用于调节消化,制成抗热应激中药。该抗热应激中药各成分具有相互协同促进的作用,有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响。
本发明实施例1的抗热应激中药的制备方法,首先,按重量配比取用于制备抗热应激中药的所有材料,合计30份,即为生药重量,先用清水漂洗1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡50min,然后用大火煮开后,再用小火煮50min,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮30min,再次过滤取滤液,合并两次过滤取得的滤液,浓缩制成生药含量为0.2g/mL的抗热应激中药,放入4℃冰箱中冷藏,备用。
其中,所述生药含量为:生药重量/浓缩制成的抗热应激中药的体积。
实施例2
本发明实施例2与实施例1的主要区别在于:所述抗热应激中药,主要由下述重量份数的材料制备而成:凤尾草5份、藿香5份、麦冬5份、山楂5份、金银花5份、酸枣仁5份、石膏5份、菊花5份、黄柏5份、及薄荷5份。
所述抗热应激中药的制备方法,首先,按重量配比取用于制备抗热应激中药的所有材料,合计50份,即为生药重量,先用清水漂洗2遍,将清洗好的材料放入砂锅中用蒸馏水浸泡至少1h,然后用大火煮开后,再用小火煮1h,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮40min,再次过滤取滤液,合并两次过滤取得的滤液,浓缩制成生药含量为0.5g/mL的抗热应激中药,放入3℃冰箱中冷藏,备用。
本发明实施例2的抗热应激中药与实施例1相比,虽然配方中各材料的配比不同、制备方法的条件也不同,但是该抗热应激中药各成分具有相互协同促进的作用,也能有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响。
实施例3
本发明实施例3与实施例2的主要区别在于:所述抗热应激中药,主要由下述重量份数的材料制备而成:凤尾草4份、藿香4份、麦冬4份、山楂4份、金银花4份、酸枣仁4份、石膏4份、菊花4份、黄柏4份、及薄荷4份。
所述抗热应激中药的制备方法,首先,按重量配比取用于制备抗热应激中药的所有材料,合计40份,即为生药重量,先用清水漂洗1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡1h,然后用大火煮开后,再用小火煮1h,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮40min,再次过滤取滤液,合并两次过滤取得的滤液,浓缩制成生药含量为0.33g/mL的抗热应激中药,放入6℃冰箱中冷藏,备用。
本发明实施例3的抗热应激中药与实施例2相比,虽然配方中各材料的配比不同、制备方法的条件也不同,但是该抗热应激中药各成分具有相互协同促进的作用,也能有效提高动物在高温条件下的存活率,降低热应激对动物生长的不利影响。
为了明确实施例1~3任一抗热应激中药的抗热应激机理、及抗热应激的效果,以实施例1~3中任一实施例的抗热应激中药作为复方组用于小鼠进行抗热应激实验。
所述抗热应激中药的抗热应激实验方法,包括下述步骤:
S1.实验动物分组:选择体重为23~30g,雌雄各一半的小鼠做为实验动物,将小鼠分为对照组、热应激实验组、及复方组,每组有10只小鼠,雌雄各一半,称取并记录各组小鼠的体重,即为初始体重;
S2.适应性处理:在每天固定时间,给对照组和热应激实验组分别按体重灌服0.5mL/20g生理盐水,给复方组按体重灌服0.2mL/20g的抗热应激中药,灌服前禁食禁水2h,按上述操作连续灌服5天后,称取并记录各组小鼠的体重,即为热应激前体重;
S3.热应激处理:将步骤S2中除对照组外,其余各组置于40℃的恒温箱内热应激1h,连续热应激2天,热应激期间持续给药,各组小鼠最后一次热应激前禁食12h,热应激结束后,统计小鼠存活率,称取并记录各组存活小鼠的体重,即为热应激后体重;
S4.指标测试,热应激结束后,统计小鼠存活率,称取并记录各组存活小鼠的体重,即为热应激后体重;然后脱颈椎处死小鼠,分别对各组小鼠采集血液样品测定血常规,采集肝脏和肾脏并测定各自的丙二醛MDA含量和谷胱甘肽过氧化物酶GSH-Px活性。
其中,血常规测定方法为:在试验结束时,小鼠摘眼球采血,收集血液于抗凝管中,用全自动血液细胞分析仪,测定和计算小鼠全血中白细胞计数WBC、红细胞计数RBC、血红蛋白HGB、平均红细胞体积MCV、平均血红蛋白含量MCH、红细胞压积HCT、淋巴细胞计数Lymph、单核细胞计数Mon、及中性粒细胞数目Gran这9项指标;
小鼠肝脏和肾脏MDA含量和GSH-Px活性测定方法为:小鼠处死后,剖取肝脏和肾脏,用预冷的4℃生理盐水洗净表面残血,滤纸吸干表面水分放在-80℃保存备用;用丙二醛测试盒,其批号为20201104,购自南京建成生物研究所,测定丙二醛MDA含量;用谷胱甘肽过氧化物酶测试盒,其批号为20201014,购自南京建成生物研究所,测定谷胱甘肽过氧化物酶GSH-Px活性。
采集的数据采用spass软件进行单因素方差分析,用LSD法进行多重比较,P<0.05表示差异显著。
各组小鼠体重的测定结果见表1:
表1小鼠体重的测定结果
注:*表示热应激实验组内热应激前后体重有显著性差异P<0.05,无标注表示差异不显著P>0.05,各组间无显著性差异P>0.05。
由表1可知,与初始体重相比,只有对照组小鼠体重增加,其他两组小鼠经热应激后体重均下降,说明热应激能导致小鼠体重减轻;对热应激前后进行比较,热应激实验组小鼠热应激前后体重差异显著P<0.05;复方组小鼠热应激后体重有所下降,但热应激前后体重差异不显著P>0.05;热应激后,复方组与对照组差异不显著P>0.05。
高温会使心肌细胞受到损伤,造成心衰,会使动物胃肠道缺血、淤血,使胃黏膜受到损伤,还会破坏气管粘膜的完整性,有研究表明,高温条件下肉牛直肠温度上升,呼吸频率加快,引起脱水、内脏器官损伤,最终导致大量死亡;结合表1结果证明,热应激组小鼠热应激后体重显著下降P<0.05,是与热应激导致脱水有关;复方组热应激前后体重差异不显著,说明试验中复方组的抗热应激中药对小鼠热应激有保护作用,能减轻热应激引起的脱水。
小鼠存活率的结果见表2
表2小鼠存活率结果
由表2可知,热应激实验组死亡率最高,为50%,复方组能显著降低热应激引起的死亡。
表1及表2的结果表明本发明实施例复方组的抗热应激中药,能提高小鼠对高温环境的适应能力,在高温条件下有较高的存活率,降低热应激对小鼠生长的不利影响。
小鼠血常规的测定结果见表3。
表3小鼠血常规测定结果
注:同行肩标小写字母不同表示差异显著P<0.05,字母相同或无标记表示差异不显著P>0.05。
由表3数据表明,热应激实验组小鼠平均红细胞体积MCV、平均血红蛋白含量MCH、白细胞计数WBC、单核细胞数量Mon均显著上升,与对照组、复方组均有显著性差异P<0.05;
复方组的平均红细胞体积MCV、平均血红蛋白含量MCH、白细胞计数WBC、单核细胞数量Mon、中性粒细胞数目Gran与热应激实验组相比显著下降P<0.05,与对照组无显著性差异P>0.05。
血液是动物机体重要的组成部分,有交换、运输和防御的功能,是体内各组织器官之间以及体内组织器官与外界环境的桥梁,热应激期间机体维持体内血液指标的稳定是耐热的基础。
白细胞在机体内具有重要的防御作用,其含量的升高多是由于机体体内发生的感染、发炎、组织坏死、物理性刺激等而引起,白细胞的升高是炎症反应的指标之一,而机体在热应激下,外周血白细胞含量升高,通过代偿机制提高血液白细胞数来抵抗热应激的负面效应;有报道指出,人体在发热后表现出明显的缺水状况,红细胞体积变大,环境高温时动物可通过加快呼吸频率进行辅助散热,呼吸频率的加快迫使红细胞数和血红蛋白含量增加以供给机体耗氧量的需求。
热应激组小鼠平均红细胞体积MCV、平均血红蛋白含量MCH、白细胞计数WBC、单核细胞数量Mon均显著高于对照组,是由于热应激会加速动物脱水,使血液浓缩,另外热应激加速呼吸,从而使上述血液指标显著增加。
复方组的平均红细胞体积MCV、平均血红蛋白含量MCH、白细胞计数WBC、单核细胞数量Mon、中性粒细胞数目Gran均显著低于热应激组,说明复方组的抗热应激中药对小鼠热应激炎症反应及脱水有显著的缓解效果。
小鼠肝脏和肾脏的丙二醛MDA和谷胱甘肽过氧化物酶GSH-Px活性测定结果见表4:
表4小鼠肝脏和肾脏MDA含量和GSG-Px活性的测定结果
由表4可知,三个组的肝脏、肾脏的MDA、GSH-Px活性均无显著性差异即P>0.05。
MDA为自由基与脂质发生过氧化物反应的氧化终产物,被认为是氧化应激的标志物,可以反映细胞脂质过氧化程度,MDA含量越少,说明机体抗氧化能力越强;GSH-Px是机体内重要的过氧化物酶,具有清除体内氢过氧化物和保护细胞结构和功能完整的作用。本试验中,三个组的肝脏、肾脏的MDA、GSH-Px活性均无显著性差异,即P>0.05,因此,复方组对小鼠抗氧化能力无明显作用,说明复方组的抗热应激中药不是通过提高GSH-Px活力、降低MDA含量发挥抗热应激作用。
综上所述,本发明的抗热应激中药能减少热应激小鼠体重下降和降低热应激小鼠的死亡率,其作用机理是通过调节机体发热机制、显著改善热应激小鼠的血液细胞状况,有效改善血液功能、维持体内血液指标的稳定来增强机体抗热应激能力,达到抗热应激的效果,且抗热应激效果显著。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (4)
1.抗热应激中药,其特征在于,主要由下述重量份数的材料制备而成:凤尾草3~5份、藿香3~5份、麦冬3~5份、山楂3~5份、金银花3~5份、酸枣仁3~5份、石膏3~5份、菊花3~5份、黄柏3~5份、及薄荷3~5份。
2.根据权利要求1所述的抗热应激中药,其特征在于,主要由下述重量份数的材料制备而成:凤尾草3份、藿香3份、麦冬3份、山楂3份、金银花3份、酸枣仁3份、石膏3份、菊花3份、黄柏3份、及薄荷3份。
3.权利要求1所述的抗热应激中药的制备方法,其特征在于,首先,按重量配比取用于制备抗热应激中药的所有材料,先用清水漂洗至少1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡至少50min,然后用大火煮开后,再用小火煮至少50min,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮30min以上,再次过滤取滤液,合并两次过滤取得的滤液,浓缩制成生药含量为0.2~0.5g/mL的抗热应激中药,放入3~6℃冰箱中冷藏,备用。
4.根据权利要求3所述的抗热应激中药的制备方法,其特征在于,首先,按重量配比取用于制备抗热应激中药的所有材料,先用清水漂洗1遍,将清洗好的材料放入砂锅中用蒸馏水浸泡50min,然后用大火煮开后,再用小火煮50min,过滤,取滤液,滤渣再加蒸馏水,使蒸馏水没过滤渣,然后,煎煮30min,再次过滤取滤液,合并两次过滤中取得的滤液,浓缩制成生药含量为0.2g/mL的抗热应激中药,放入4℃冰箱中冷藏,备用。
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