富硒女贞子缓解山羊热应激效果的研究

发布时间：2018-08-30 19:55

【摘要】：中兽医病因病理及辩证诊治的基础理论认为,热应激是由于高热邪气所致的阳盛阴虚病证,而中药女贞子具有滋阴补肾之功效；本课题组前期研究结果显示富硒女贞子能有效降低急性热应激小鼠的死亡率,缓解热应激造成小鼠的肝脏损伤程度,既提高热应激小鼠血清总蛋白、球蛋白和白蛋白的含量及免疫功能。因此我们对富硒女贞子进行了抗山羊热应激的研究,通过富硒女贞子安全性试验和抗热应激疗效试验,观察山羊生产性能、血液学、血清生化指标及对山羊外周血淋巴细胞增殖活性、细胞因子分泌、抗氧化活性的影响,来探索其抗山羊热应激的效果及作用机制。试验一富硒女贞子对山羊的安全性试验旨在研究富硒女贞子对山羊的用药安全性。选取30头成年产奶山羊随机均分为5组,其中C组饲喂基础日粮,CLL组基础日粮中添加女贞于30 g/头/d,LSLL组、MSLL组、HSLL基础日粮中分别添加富硒女贞子10g/头/d、30 g/头/d、50 g/头/d,给药组于试验开始的第5d给药,连续5d。经产奶量、血液中WBC、RBC、HGB含量、血清中TP、ALB、 ALT、AST、CK、LDH的水平测定。结果显示：富硒女贞子和普通女贞子均能提高山羊的产奶量,在试验1～7、22-38 d产奶量增长率由大到小的排序为：LSLL组MSLL组HSLL组CLL组C组；CLL组能显著提高山羊RBC(P0.05),MSLL组、CLL组均能显著提高山羊HGB含量(P0.05)；LSLL组、MSLL组、HSLL组和CLL组均能显著提高山羊血清中TP含量(P0.05),LSLL组、MSLL组和CLL组均能显著提高山羊血清中ALB含量(P0.05),LSLL组能显著降低血清中AST活性(P0.05)；添加富硒女贞子对山羊WBC含量、ALT、CK、LDH活性均无显著影响(P0.05)。说明日粮中添加富硒女贞子在提高山羊产奶量的同时,亦提高了血液RBC、HGB和血清TP、ALB的含量,降低了血清AST活性,而对山羊WBC、ALT、CK、LDH舌性影响不明显,提示基础日粮中按≤50 g/头/d剂量添加富硒女贞子,连用5 d,对山羊是安全的。试验二富硒女贞子对山羊热应激的影响为观察富硒女贞子对热应激山羊生产性能、血液学和血清生化指标的影响。选取30头泌乳期萨能山羊随机均分为5组,其中HS组饲喂基础日粮,CLL组基础日粮中添加女贞子10g/头/d,LSLL组、MSLL组、HSLL组基础日粮中分别添加富硒女贞子5 g/头/d、10 g/头/d、20 g/头/d。试验期为2014年7月10日至8月27日,给药组于每旬前3天混饲给药,给药4批次。经产奶量、乳成分、血液中WBC、RBC、HGB含量、血清中TP、ALB、ALT、AST、LDH、IL-2、IFN-γ、IL-6、SOD、GSH-Px、ACTH、COR的水平测定。结果显示：日粮中添加富硒女贞子能提高热应激山羊的产奶量,产奶量下降率由小到大的排序为：MSLL组LSLL组HSLL组CLL组HS组,其中MSLL组较非热应激期增加了2.89%,极显著高于HS组的下降10.07%(P0.01)；显著升高热应激山羊乳脂肪含量(P0.05)、提高了乳蛋白含量；明显提高了血清中TP、ALB含量；28 d时HSLL组的HSP70表达量、LSLL组的LDH活性显著低于HS组,LSLL组的IL-2、IFN-γ含量、SOD活性显著高于HS组(P0.05),14 d时MSLL组、HSLL组、CLL组的COR浓度均显著低于HS组(P0.05),对乳糖、总乳固体、非脂乳固体、WBC、RBC、HGB、ALT、AST、 IL-6的影响不明显(P0.05)。表明富硒女贞子对热应激山羊具有通过调节内分泌-免疫、改善机体代谢而提高产奶量的作用,尤以5～10g/头/d富硒女贞子的作用效果为佳。试验三富硒女贞子对山羊外周血淋巴细胞增殖活性、细胞因子分泌及抗氧化活性的影响分析富硒女贞子对山羊外周血淋巴细胞增殖活性、细胞因子分泌及抗氧化活性的影响。采用MTT法测定富硒女贞子多糖、蛋白质、齐墩果酸对山羊外周血淋巴细胞增殖活性的影响,并且对其培养上清细胞因子IL-2、IL-4、IL-6、IFN-γ水平和SOD活性进行检测。结果显示：富硒女贞子多糖、蛋白质和齐墩果酸体外均具有促进ConA刺激山羊外周血淋巴细胞分裂增殖的作用,且富硒女贞子蛋白质组分、齐墩果酸组分的作用效果明显优于普通女贞子组,各相应浓度比较差异显著比例分别达到63.64%、40.00%1富硒和普通女贞子均极显著提高外周血淋巴细胞中SOD的活性(P0.01),且富硒女贞子的作用效果明显优于普通女贞子；富硒女贞子可刺激淋巴细胞产生IL-2,但与对照组比较差异不显著(P0.05)。结果表明富硒女贞子多糖、蛋白质和齐墩果酸均具有促进山羊外周血淋巴细胞增殖活性的作用,同时明显提高其抗氧化能力。结论：富硒女贞子,按≤50g/头/d,连用5 d,对山羊具有较好的安全性：能缓解山羊热应激影响导致的产奶量下降(提高了2.89%),以5～10g/头/d富硒女贞子的作用效果为佳,且作用效果优于普通女贞子。其作用机制是通过HPA轴来调低热应激性升高的ACTH、COR,调升热应激降低的IL-2、IL-6、IFN-γ水平,使得机体维持良好的内环境稳态,提高其免疫功能,同时提高血清TP、ALB含量、GSH-Px、SOD活性,降低LDH活性、HSP70表达量,从而使机体降低应激性代偿,维持更接近正常期的产奶量。
[Abstract]:According to the basic theory of etiology, pathology and dialectical diagnosis and treatment of Chinese veterinary medicine, heat stress is due to yang-Sheng-yin deficiency caused by high-heat pathogenic qi, while Ligustrum lucidum has the effect of nourishing yin and tonifying kidney. The degree of visceral injury can not only increase the content of serum total protein, globulin and albumin, but also improve the immune function of heat-stressed mice.Therefore, we studied the anti-heat stress effect of Se-enriched Ligustrum lucidum on goats. To explore the effect and mechanism of anti-heat stress on peripheral blood lymphocyte proliferation, cytokine secretion and antioxidant activity. Experiment 1 The safety test of Se-enriched Ligustrum lucidum on goats was designed to study the medication safety of Se-enriched Ligustrum lucidum on goats. The basic diet of CLL group was fed with 30 g/head/d, LSLL group, MSLL group, HSLL base diet was fed with 10 g/head/d, 30 g/head/d, 50 g/head/d of Ligustrum lucidum enriched with selenium, and the treatment group was fed on the 5th day of the experiment. The milk yield, WBC, RBC, HGB content in blood, TP, ALB, ALT, AST, CK, LDH levels in serum were measured. The results showed that both Se-enriched Ligustrum lucidum and common Ligustrum lucidum could increase the milk yield of goats. The order of increase rate of milk yield from large to small was as follows: LSLL group MSLL group CLL group C group; CLL group RBC (P 0.05), MSLL group, CLL group could significantly increase the HGB content of goats (P 0.05); LL group can significantly increase the content of TP in goat serum (P 0.05), LSLL group, MSLL group and CLL group can significantly increase the content of ALB in goat serum (P 0.05), LSLL group can significantly reduce the activity of AST in serum (P 0.05); adding Se-enriched Ligustrum lucidum has no significant effect on WBC content, ALT, CK, LDH activity in goats (P 0.05). The content of RBC, HGB, TP and ALB in blood and the activity of AST in serum were increased, but the tongue of WBC, ALT, CK and LDH in goats were not affected obviously. It was suggested that adding Se-enriched Ligustrum lucidum to basal diet for 5 days was safe for goats. Thirty lactating Saanen goats were randomly divided into 5 groups, including HS group fed basal diet, CLL group fed basal diet supplemented with 10g Ligustrum lucidum per day, LSLL group, MSLL group, HSLL group fed basal diet supplemented with 5g Ligustrum lucidum enriched with selenium respectively. D, 10 g / head / d, 20 g / head / d. The trial period was from July 10 to August 27, 2014. The administration group was administered in four batches three days before every ten days. The milk yield, milk composition, WBC, RBC, HGB content in blood, serum TP, ALB, ALT, AST, LDH, IL-2, IFN-gamma, IL-6, SOD, GSH-Px, ACTH, COR levels were determined. The order of decreasing rate of milk yield and milk yield of high heat stress goats from small to large was: HS group of LSLL group of MSLL group of HSLL group, HS group of CLL group, MSLL group increased by 2.89% compared with non-heat stress group, significantly higher than HS group decreased by 10.07% (P 0.01); significantly increased milk fat content of heat stress goats (P 0.05), increased milk protein content; The expression of HSP70 in HSLL group and LDH activity in LSLL group were significantly lower than those in HS group at 28 d, while the content of IL-2, IFN-gamma and SOD activity in LSLL group were significantly higher than those in HS group (P 0.05). The COR concentration in MSLL group, HSLL group and CLL group was significantly lower than that in HS group at 14 d (P 0.05), and the effects on lactose, total milk solids, non-fat milk solids, WBC, RBC, HGB, ALT, AST, IL-6 were not clear. The results showed that the Se-enriched Ligustrum lucidum could increase milk yield by regulating Endocrine-Immunity and improving metabolism of the body. The effect of Se-enriched Ligustrum lucidum 5-10g/head/d was the best. The effects of Se-enriched Ligustrum lucidum on the proliferation, cytokine secretion and antioxidant activity of peripheral blood lymphocytes of goats were analyzed.The effects of Se-enriched Ligustrum lucidum polysaccharide, protein and oleanolic acid on the proliferation activity of peripheral blood lymphocytes of goats were determined by MTT method. The results showed that Se-enriched Ligustrum lucidum polysaccharides, proteins and oleanolic acid could promote the proliferation of peripheral blood lymphocytes stimulated by ConA in vitro, and the effects of Se-enriched Ligustrum lucidum protein components and oleanolic acid components were significantly better than those of ordinary Ligustrum lucidum group, and the corresponding concentrations were significantly different in proportion. Selenium-enriched Ligustrum lucidum L. and common Ligustrum lucidum L. both significantly increased SOD activity in peripheral blood lymphocytes (P 0.01), and the effect of Se-enriched Ligustrum lucidum L. was better than that of common Ligustrum lucidum L. Se-enriched Ligustrum L. could stimulate lymphocytes to produce IL-2, but there was no significant difference between the two groups (P 0.05). Conclusion: Selenium-enriched Ligustrum lucidum has better safety for goats when it is used for 5 days at a dose of <50g/head/d. It can alleviate the decrease of milk yield caused by heat stress in goats (2.89%) and enrich them at a dose of 5-10g/head/d. Ligustrum selenium has a better effect than Ligustrum lucidum. Its mechanism is to lower the levels of ACTH, COR, IL-2, IL-6 and IFN-gamma in the elevated heat stress through HPA axis, so that the body can maintain a good homeostasis of the internal environment, improve its immune function, and increase the content of TP, ALB, GSH-Px, SOD activity in serum. Reduce the activity of LDH and the expression of HSP70, so that the body can reduce stress compensation and maintain milk production closer to normal period.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S853.7

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