高温条件下猪散热调节特性及中草药复合制剂的作用
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摘要
夏季高温高湿天气易诱发动物的热应激。猪汗腺不发达,发生热应激时,调动外周血液循环、强化肺的呼吸与肾的排泄功能以散热,散热受阻,影响血液及血管生理,易造成组织器官的损伤,危及健康,给养猪业带来巨大损失。石膏、藿香、苍术、黄柏等具有清热祛湿功效,促进散热功效。本研究通过人工模拟高温高湿环境,诱发猪产生热应激,研究上述四味中药复合制剂对高温条件下猪的散热调控作用,旨在为畜舍环境控制、营养卫生调控技术提供依据。
本研究包括以下五个试验部分：
1、高温条件下猪生理生化指标的变化及中草药复合制剂的作用
在人工气候仓模拟夏季高温高湿环境,对中国小型实验猪(CEMP,农大Ⅰ系)进行为期10d的高温刺激(40℃,5h/d)。此外,用不同配比的藿香、苍术、黄柏和石膏提取物,制备成复合制剂,研究高温对猪生产性能和血清生化指标的影响及复合制剂的作用。试验选用2月龄实验猪48头,分为常温组、高温应激组(以下简称高温组)、高温中草药复合制剂组(以下简称复合制剂组)三个处理,每组16头。于试验的第1、3、6、10d,每组中选4头猪,用数字体温表测定直肠温,称重,静脉采血后分离血清,测定血清皮质醇、热休克蛋白70含量。结果显示高温组的猪体温升高并伴有热性喘息、下痢稀水、倦怠、少食等症状；同时猪日采食量和日增重显著下降(P<0.05),血清皮质醇、热休克蛋白70含量显著升高(P<0.05)。相对于高温组,复合制剂组猪的日增重和日采食量显著增高,且血清皮质醇、热休克蛋白70含量均显著降低(P<0.05)。以上结果表明,复合制剂可以显著缓解高温应激、改善猪的生产性能。
2、高温条件下猪皮肤微循环特征及中草药复合制剂的作用
本部分研究了复合制剂对高温条件下猪移动血细胞浓度、血管活性因子一氧化氮、内皮素-1含量的影响。试验处理同试验1,在试验的第1、3、6、10d,从每组中选4头猪,应用激光多普勒血流仪(Perflux 4001 master)对猪体表固定点进行测定。空腹前腔静脉采血,每头猪10mL,测定血清中NO和ET-1含量的变化。结果显示：高温组猪的移动血细胞浓度第1、3、6、10d均显著高于常温组(P<0.05),NO含量呈逐渐下降趋势,在第10d显著低于常温组(P<0.05)；ET-1含量第10d显著高于常温组(P<0.05)。复合制剂组第10d的NO含量显著提高、移动血细胞浓度显著降低(P<0.05),但ET-1含量无显著变化(P>0.05)。高温条件下,移动血细胞浓度趋于增加,至10d时血流速度减慢,NO和ET-1释放失衡,血管舒张作用减弱,收缩加强,不利于机体散热。第10d的试验结果表明,复合制剂可以通过对移动血细胞浓度、NO的调控,促进机体散热。
3、高温条件下猪全血黏度、红细胞变形指数的变化及中草药复合制剂的作用
本部分研究了高温环境下猪全血黏度、红细胞变形指数和红细胞压积的动态变化及复合制剂的影响。试验处理同试验1,在试验的第1、3、6、10d,从每组中取4头猪,空腹前腔静脉采血,每头10mL,分析血液流变学参数。每头猪5mL新鲜血样放入加入肝素钠(0.2mg/mL)的塑料试管中,其中1mL通过Wintrobe测试分析细胞压积,其余的用MVIS-02035血液流变自动分析系统在200、1的剪切率下分析全血黏度等血液流变学参数。结果显示：高温组的猪全血黏度高切变率值在第10d显著高于常温组(P<0.05),红细胞变形指数在第6、10d显著低于常温组(P<0.05),红细胞压积在第6、10d均显著高于常温组(P<0.05)。添加复合制剂后,全血黏度、红细胞变形指数和红细胞压积均得到了明显的恢复,尤其在高温的第10d治疗效果最明显(P<0.05)。说明持续高温刺激增加血液黏度减少血液灌注量,红细胞疲劳影响养分供给能力,复合制剂可以降低热应激产生的血管生理损伤。
4、高温条件下猪肺、肾组织的损伤及中草药复合制剂的作用
本部分详细研究了复合制剂对猪的肺与肾组织损伤的修复作用,并对猪肺、肾脏进行了病理学分析。试验处理同试验1,在试验的第1、3、6、10d,从每组中取4头猪,麻醉宰杀,取肺、肾组织,于4%中性甲醛溶液中固定。石蜡包埋,5μm连续切片,H-E染色,镜检。通过病理学分析发现,高温刺激对仔猪肺、肾脏组织均造成了不同程度的损伤。说明高温持续下去,可能导致微循环负荷加重,从而影响微循环对组织的灌流,最终导致组织细胞缺血缺氧及营养失调、组织损伤,以及严重的代谢紊乱。添加复合制剂后,对猪的肺与肾组织损伤具有良好的修复作用。
5、高温条件下猪下丘脑AVP、cAMP含量的变化及中草药复合制剂的作用
本部分详细研究了复合制剂对猪下丘脑精氨酸加压素和环磷酸腺苷水平的影响,通过二者含量的动态变化分析复合制剂对体温的双向调节作用,探讨复合制剂促进机体散热、发挥解热作用的机理。试验处理同试验1,在试验的第1、3、6、10d,从每组中选4头猪,麻醉宰杀,快速断头取脑,以灰结节及视交叉之间的中心点为中心确定下丘脑的位置,取出,测定下丘脑组织AVP和cAMP的含量。结果显示：高温组猪的精氨酸加压素在试验的第3、6、10d显著降低(P<0.05),复合制剂组在试验的第6、10d显著高于高温组(P<0.05)；高温组环磷酸腺苷含量在试验的第10d显著升高(P<0.05),而复合制剂组在试验的第10d显著低于高温组(P<0.05)。持续高温刺激下cAMP浓度增加,维持高体温以散热,复合制剂有缓解作用。
When exposed to a hot and damp environment, animals are susceptible to heat stress. The sweat glands of pigs are under development, when heat stress happens, pigs radiate heat through starting perpheral blood circulation, stengthening lung'srespiration and kidney exretion.When the animal can not radiate heat well under heat stress, further changes in the blood and other organs will be happened. Furthermore, heat stress results in the tissue injuries, impacts on the absorption of nutrition and leads to a decrease in growth and production performance. Some studies have shown that the extracted complex, prepared mainly from gypsum, wrinkled giant hyssop, cortex phellodendron, etc., is able to promote the heat radiation and remove the rheumatism. In the present study, the simulated high-temperature environment were used to study the effects of four kinds of gypsum and complex preparation on heat dissipation in heat-stress pigs, aim to provide the basis for envionment control and nutritional hygiene manipulation techniques.
The current study includes the following five experimental parts.
1、Variations of physical and biochemical index and effects of complex preparation in heat-stress piglets
Chinese experimental piglets (CEMP) were housed in the artificial climate climber with simulating high-stress environment for 10 days (40℃,5h/d). Variation of physiological and biochemical index affected by complex preparation were used to study the effects of heat stess on production performance and serum biochemical targets in pigs. In this paper,48 2-month piglets were randomly divided into three groups:normal temperature group (NTG), heat treatment group (HTG), Chinese herbal medicine extract compound agents group (CHG) under heat stress. On day 1,3,6 and 10, we randomly selected 4 pigs from each group, then weighed and collected the fresh blood from the vein, and separated serum and measured the blood levels of heat-stress protein 70. The result showed that temperature of the HTG was higher and appeared the symptoms of pant, dysentery, tiredness and losing appetite, etc. Furthermore comparing with the HTG, the daily feed intake and the weight gain heighten(P<0.05) and the serum cortisol and heat-stress protein 70 decreased (P<0.05). The complex preparation could improve growth performance of heat-stress piglets significantly.
2、Characteristics of skin microcirculation and effects of complex preparation in heat-stress piglets
This part reported the effects of complex preparation on blood cells concentration and NO and ET-1 contents of the blood vessel. On day 1,3,6 and 10, we randomly selected 4 pigs from each group, and used Laser Doppler Flowmetry to determine the surface capillary blood flow of pigs. Ten ml fresh blood was collected from the front cavity vein of each pig and the changes of NO and ET-1 contents were determined. The result showed that the blood cell concentration of the HTG was higher than that of the NTG on d 1,3,6, and 10(P<0.05); the NO content of the HTG gradually dropped, and was significantly lower than that of the NTG on d 10(P<0.05). In the ETG, the blood cells concentration decreased and NO content increased significantly on d 10(P<0.05), but no significantly influence on the ET-1 content was observed (P>0.05). The result showed that heat stress caused an increase in the blood cells concentration, a decrease in the speed of blood flow on the day of 10, an imbalance in the NO and ET-1 contents and the blood condensation, thus influencing the heat dissipation. The complex preparation could regulate the blood cells concentration, thus improving the rate of heat dissipation in piglets exposed to high-ambient temperature.
3、Variations of whole blood viscosity and red blood cell (RBC) deformation index and effects of complex preparation in heat-stress piglets
This part studied the effects of complex preparation on whole blood viscosity (WBV), RBC deformation and hematocrit (HCT). On day 1,3,6, and 10, we randomly selected 4 pigs from each group, collected 10 mL fresh blood from the front cavity vein of each pig, and analyzed the hemorheological parameters. Five mL fresh blood sample of each pig was put into plastic test tubes containing sodium heparin(0.2 mg/mL), of which 1.0 mL of blood was used for testing the packed cell volume(PCV) by the Wintrobe method; the rest of the blood was used to determine the whole blood viscosity(WBV) at the shear rates of 200 s-1 and 1 s-1, and other hemorheological parameters. The result showed that compared with the NTG, the WBV shear rate value of the HTG was significantly higher on d 10(P<0.05), the RBC deformation index was significantly lower on d 6 and 10, and the HCT was significantly higher on d 6 and 10. After the administration of the Chinese herbal medicine extract complex preparation, the RBC deformation index of the therapy groups showed substantial restoration to their normal state, especially on d 10 (P<0.05). This showed that continuous heat stress can increase the blood viscosity to reduce the amount of blood perfusion and RBC fatigue affect nutient supply capicity. Further, the Chinese herbal medicine extract complex preparation can alleviate the heat-stress induced pathological injuries.
4、Damages of lung and kidney tissues and effects of complex preparation in heat-stress piglets
This part studied the recovery of complex preparation on lung and kidney tissues injuries, and performed pathological analysis on the lungs and kidneys. On d 1,3,6, and 10, 4 pigs in each experimental group were killed, and their lung and kidney tissues were extracted. First, the lung and kidney tissues were fixed in 4%neutral formalin; thereafter, these tissues were finally embedded in paraffin wax, cut into 5μm successive sections, stained with hematoxylin-eosin(H-E) dye, and observed under the microscope. After performing the pathological analysis, heat stress caused various injuries to lung and kidney tissues in piglets. The result showed the persistent heat stress resulted in the heavy load to circulation system, and influenced the blood supply to heat-exchange surfaces, leading to cell tissues to lack of oxygen and blood, the imbalance of nutrition, tissues injuries and a series of serious metabolic disorder. The Chinese herbal medicine extract complex preparation can alleviate the heat-stress induced pathological injuries of lungs and kidneys.
5、Variation of the contents of AVP and cAMP in hypothalamus and effects of complex preparation in heat-stress piglets
This part studied the effects of complex preparation on AVP and cAMP contents inhypothalamus, analyzed the function of regulating the temperature through the dynamic changes of the contents of AVP and cAMP, and explored the mechanism that the anti-heat-stress of Chinese herbal medicine. On day 1,3,6, and 10, four pigs in each experimental group were selected and determined the changes of the temperature by digital thermometer. These pigs were anaesthetized and then killed, and rapidly extracted the brains from their heads. Taking tuber cinereum and optic chiasma as the center, determined the place of hypothalamus and extracted them, and determined the contents of AVP and cAMP. The result showed the temperature of the HTG went up, and the content of AVP was significantly lower on d 3,6 and 10(P<0.05); the content of AVP of the ETG was significantly higher than that of the HTG(P<0.05). The content of cAMP of the HTG was significantly higher on d 10 (P<0.05), while the content of cAMP of the ETG was significantly lower than that of the HTG on d 10 (P<0.05).The results showed that continuous high temprature can improve the concentration of cAMP and maintain a high body temprature to lose heat. The complex preparation can alleviated the effects.
本研究包括以下五个试验部分：
1、高温条件下猪生理生化指标的变化及中草药复合制剂的作用
在人工气候仓模拟夏季高温高湿环境,对中国小型实验猪(CEMP,农大Ⅰ系)进行为期10d的高温刺激(40℃,5h/d)。此外,用不同配比的藿香、苍术、黄柏和石膏提取物,制备成复合制剂,研究高温对猪生产性能和血清生化指标的影响及复合制剂的作用。试验选用2月龄实验猪48头,分为常温组、高温应激组(以下简称高温组)、高温中草药复合制剂组(以下简称复合制剂组)三个处理,每组16头。于试验的第1、3、6、10d,每组中选4头猪,用数字体温表测定直肠温,称重,静脉采血后分离血清,测定血清皮质醇、热休克蛋白70含量。结果显示高温组的猪体温升高并伴有热性喘息、下痢稀水、倦怠、少食等症状；同时猪日采食量和日增重显著下降(P<0.05),血清皮质醇、热休克蛋白70含量显著升高(P<0.05)。相对于高温组,复合制剂组猪的日增重和日采食量显著增高,且血清皮质醇、热休克蛋白70含量均显著降低(P<0.05)。以上结果表明,复合制剂可以显著缓解高温应激、改善猪的生产性能。
2、高温条件下猪皮肤微循环特征及中草药复合制剂的作用
本部分研究了复合制剂对高温条件下猪移动血细胞浓度、血管活性因子一氧化氮、内皮素-1含量的影响。试验处理同试验1,在试验的第1、3、6、10d,从每组中选4头猪,应用激光多普勒血流仪(Perflux 4001 master)对猪体表固定点进行测定。空腹前腔静脉采血,每头猪10mL,测定血清中NO和ET-1含量的变化。结果显示：高温组猪的移动血细胞浓度第1、3、6、10d均显著高于常温组(P<0.05),NO含量呈逐渐下降趋势,在第10d显著低于常温组(P<0.05)；ET-1含量第10d显著高于常温组(P<0.05)。复合制剂组第10d的NO含量显著提高、移动血细胞浓度显著降低(P<0.05),但ET-1含量无显著变化(P>0.05)。高温条件下,移动血细胞浓度趋于增加,至10d时血流速度减慢,NO和ET-1释放失衡,血管舒张作用减弱,收缩加强,不利于机体散热。第10d的试验结果表明,复合制剂可以通过对移动血细胞浓度、NO的调控,促进机体散热。
3、高温条件下猪全血黏度、红细胞变形指数的变化及中草药复合制剂的作用
本部分研究了高温环境下猪全血黏度、红细胞变形指数和红细胞压积的动态变化及复合制剂的影响。试验处理同试验1,在试验的第1、3、6、10d,从每组中取4头猪,空腹前腔静脉采血,每头10mL,分析血液流变学参数。每头猪5mL新鲜血样放入加入肝素钠(0.2mg/mL)的塑料试管中,其中1mL通过Wintrobe测试分析细胞压积,其余的用MVIS-02035血液流变自动分析系统在200、1的剪切率下分析全血黏度等血液流变学参数。结果显示：高温组的猪全血黏度高切变率值在第10d显著高于常温组(P<0.05),红细胞变形指数在第6、10d显著低于常温组(P<0.05),红细胞压积在第6、10d均显著高于常温组(P<0.05)。添加复合制剂后,全血黏度、红细胞变形指数和红细胞压积均得到了明显的恢复,尤其在高温的第10d治疗效果最明显(P<0.05)。说明持续高温刺激增加血液黏度减少血液灌注量,红细胞疲劳影响养分供给能力,复合制剂可以降低热应激产生的血管生理损伤。
4、高温条件下猪肺、肾组织的损伤及中草药复合制剂的作用
本部分详细研究了复合制剂对猪的肺与肾组织损伤的修复作用,并对猪肺、肾脏进行了病理学分析。试验处理同试验1,在试验的第1、3、6、10d,从每组中取4头猪,麻醉宰杀,取肺、肾组织,于4%中性甲醛溶液中固定。石蜡包埋,5μm连续切片,H-E染色,镜检。通过病理学分析发现,高温刺激对仔猪肺、肾脏组织均造成了不同程度的损伤。说明高温持续下去,可能导致微循环负荷加重,从而影响微循环对组织的灌流,最终导致组织细胞缺血缺氧及营养失调、组织损伤,以及严重的代谢紊乱。添加复合制剂后,对猪的肺与肾组织损伤具有良好的修复作用。
5、高温条件下猪下丘脑AVP、cAMP含量的变化及中草药复合制剂的作用
本部分详细研究了复合制剂对猪下丘脑精氨酸加压素和环磷酸腺苷水平的影响,通过二者含量的动态变化分析复合制剂对体温的双向调节作用,探讨复合制剂促进机体散热、发挥解热作用的机理。试验处理同试验1,在试验的第1、3、6、10d,从每组中选4头猪,麻醉宰杀,快速断头取脑,以灰结节及视交叉之间的中心点为中心确定下丘脑的位置,取出,测定下丘脑组织AVP和cAMP的含量。结果显示：高温组猪的精氨酸加压素在试验的第3、6、10d显著降低(P<0.05),复合制剂组在试验的第6、10d显著高于高温组(P<0.05)；高温组环磷酸腺苷含量在试验的第10d显著升高(P<0.05),而复合制剂组在试验的第10d显著低于高温组(P<0.05)。持续高温刺激下cAMP浓度增加,维持高体温以散热,复合制剂有缓解作用。
When exposed to a hot and damp environment, animals are susceptible to heat stress. The sweat glands of pigs are under development, when heat stress happens, pigs radiate heat through starting perpheral blood circulation, stengthening lung'srespiration and kidney exretion.When the animal can not radiate heat well under heat stress, further changes in the blood and other organs will be happened. Furthermore, heat stress results in the tissue injuries, impacts on the absorption of nutrition and leads to a decrease in growth and production performance. Some studies have shown that the extracted complex, prepared mainly from gypsum, wrinkled giant hyssop, cortex phellodendron, etc., is able to promote the heat radiation and remove the rheumatism. In the present study, the simulated high-temperature environment were used to study the effects of four kinds of gypsum and complex preparation on heat dissipation in heat-stress pigs, aim to provide the basis for envionment control and nutritional hygiene manipulation techniques.
The current study includes the following five experimental parts.
1、Variations of physical and biochemical index and effects of complex preparation in heat-stress piglets
Chinese experimental piglets (CEMP) were housed in the artificial climate climber with simulating high-stress environment for 10 days (40℃,5h/d). Variation of physiological and biochemical index affected by complex preparation were used to study the effects of heat stess on production performance and serum biochemical targets in pigs. In this paper,48 2-month piglets were randomly divided into three groups:normal temperature group (NTG), heat treatment group (HTG), Chinese herbal medicine extract compound agents group (CHG) under heat stress. On day 1,3,6 and 10, we randomly selected 4 pigs from each group, then weighed and collected the fresh blood from the vein, and separated serum and measured the blood levels of heat-stress protein 70. The result showed that temperature of the HTG was higher and appeared the symptoms of pant, dysentery, tiredness and losing appetite, etc. Furthermore comparing with the HTG, the daily feed intake and the weight gain heighten(P<0.05) and the serum cortisol and heat-stress protein 70 decreased (P<0.05). The complex preparation could improve growth performance of heat-stress piglets significantly.
2、Characteristics of skin microcirculation and effects of complex preparation in heat-stress piglets
This part reported the effects of complex preparation on blood cells concentration and NO and ET-1 contents of the blood vessel. On day 1,3,6 and 10, we randomly selected 4 pigs from each group, and used Laser Doppler Flowmetry to determine the surface capillary blood flow of pigs. Ten ml fresh blood was collected from the front cavity vein of each pig and the changes of NO and ET-1 contents were determined. The result showed that the blood cell concentration of the HTG was higher than that of the NTG on d 1,3,6, and 10(P<0.05); the NO content of the HTG gradually dropped, and was significantly lower than that of the NTG on d 10(P<0.05). In the ETG, the blood cells concentration decreased and NO content increased significantly on d 10(P<0.05), but no significantly influence on the ET-1 content was observed (P>0.05). The result showed that heat stress caused an increase in the blood cells concentration, a decrease in the speed of blood flow on the day of 10, an imbalance in the NO and ET-1 contents and the blood condensation, thus influencing the heat dissipation. The complex preparation could regulate the blood cells concentration, thus improving the rate of heat dissipation in piglets exposed to high-ambient temperature.
3、Variations of whole blood viscosity and red blood cell (RBC) deformation index and effects of complex preparation in heat-stress piglets
This part studied the effects of complex preparation on whole blood viscosity (WBV), RBC deformation and hematocrit (HCT). On day 1,3,6, and 10, we randomly selected 4 pigs from each group, collected 10 mL fresh blood from the front cavity vein of each pig, and analyzed the hemorheological parameters. Five mL fresh blood sample of each pig was put into plastic test tubes containing sodium heparin(0.2 mg/mL), of which 1.0 mL of blood was used for testing the packed cell volume(PCV) by the Wintrobe method; the rest of the blood was used to determine the whole blood viscosity(WBV) at the shear rates of 200 s-1 and 1 s-1, and other hemorheological parameters. The result showed that compared with the NTG, the WBV shear rate value of the HTG was significantly higher on d 10(P<0.05), the RBC deformation index was significantly lower on d 6 and 10, and the HCT was significantly higher on d 6 and 10. After the administration of the Chinese herbal medicine extract complex preparation, the RBC deformation index of the therapy groups showed substantial restoration to their normal state, especially on d 10 (P<0.05). This showed that continuous heat stress can increase the blood viscosity to reduce the amount of blood perfusion and RBC fatigue affect nutient supply capicity. Further, the Chinese herbal medicine extract complex preparation can alleviate the heat-stress induced pathological injuries.
4、Damages of lung and kidney tissues and effects of complex preparation in heat-stress piglets
This part studied the recovery of complex preparation on lung and kidney tissues injuries, and performed pathological analysis on the lungs and kidneys. On d 1,3,6, and 10, 4 pigs in each experimental group were killed, and their lung and kidney tissues were extracted. First, the lung and kidney tissues were fixed in 4%neutral formalin; thereafter, these tissues were finally embedded in paraffin wax, cut into 5μm successive sections, stained with hematoxylin-eosin(H-E) dye, and observed under the microscope. After performing the pathological analysis, heat stress caused various injuries to lung and kidney tissues in piglets. The result showed the persistent heat stress resulted in the heavy load to circulation system, and influenced the blood supply to heat-exchange surfaces, leading to cell tissues to lack of oxygen and blood, the imbalance of nutrition, tissues injuries and a series of serious metabolic disorder. The Chinese herbal medicine extract complex preparation can alleviate the heat-stress induced pathological injuries of lungs and kidneys.
5、Variation of the contents of AVP and cAMP in hypothalamus and effects of complex preparation in heat-stress piglets
This part studied the effects of complex preparation on AVP and cAMP contents inhypothalamus, analyzed the function of regulating the temperature through the dynamic changes of the contents of AVP and cAMP, and explored the mechanism that the anti-heat-stress of Chinese herbal medicine. On day 1,3,6, and 10, four pigs in each experimental group were selected and determined the changes of the temperature by digital thermometer. These pigs were anaesthetized and then killed, and rapidly extracted the brains from their heads. Taking tuber cinereum and optic chiasma as the center, determined the place of hypothalamus and extracted them, and determined the contents of AVP and cAMP. The result showed the temperature of the HTG went up, and the content of AVP was significantly lower on d 3,6 and 10(P<0.05); the content of AVP of the ETG was significantly higher than that of the HTG(P<0.05). The content of cAMP of the HTG was significantly higher on d 10 (P<0.05), while the content of cAMP of the ETG was significantly lower than that of the HTG on d 10 (P<0.05).The results showed that continuous high temprature can improve the concentration of cAMP and maintain a high body temprature to lose heat. The complex preparation can alleviated the effects.
引文
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