牡蛎粗多糖对热应激雄性动物血液生化指标及生精功能的影响
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摘要
本试验的目的是通过建立热应激模式,研究牡蛎多糖对动物血液生化指标以及对睾丸组织形态学变化的影响,进行了如下试验。
1、热应激模式的建立
将21只小鼠随机分为3组,即空白对照组、热应激A、热应激B组,每组7个小鼠。热应激A组处理分为两个步骤,即热应激Ⅰ阶段(38℃应激45min)和热应激Ⅱ阶段(42℃应激20min);热应激B组处理为一个步骤,即热应激Ⅰ阶段(39℃应激65min);应激6d后检测小鼠抗氧化指标(SOD、MDA、GSH-PX)和其相关激素(ACTH、COR、T_3、T_4),选择理想的小鼠模型。结果:热应激A组小鼠血清的SOD、MDA、GSH-PX、ACTH、COR、T_3、T_4,热应激B组的SOD、MDA、COR、T_3与空白对照组对比差异极显著(P<0.01)或显著(P<0.05);热应激B组的GSH-Px、ACTH、T_4与空白对照组对比差异不显著(P>0.05)。结论:热应激A组模式即38℃应激45min+42℃再应激20min,作为热应激模型。
2、牡蛎粗多糖对热应激雄性小鼠生化指标的影响
利用建立好的热应激模型,将6周龄成年雄性小鼠75只随机为5组,即设立空白对照组、热应激组、牡蛎多糖高、中、低剂量热应激组,每组3个重复,每个重复5只,空白对照组、热应激对照组饲喂基础日粮,牡蛎多糖高、中、低剂量热应激组饲喂基础日粮中分别添加1.8%、1.2%和0.6%牡蛎粗多糖,试验周期为28d。研究牡蛎粗多糖对热应激状态下雄性小鼠血液中Na~+、K~+、Cl~-、MDA、SOD、GSH-P_X、AKP、总蛋白、白蛋白和球蛋白及肝脏组织匀浆中ALT、AST等生化指标。结果:除AKP指标外,热应激对照组与空白对照组间差异显著(P<0.05)或极显著(P<0.01);牡蛎粗多糖添加组与热应激组比较,能改善小鼠血液中Na~+、K+、Cl-、MDA、SOD、GSH-P_X、AKP、总蛋白、白蛋白和球蛋白及肝脏组织匀浆中ALT、AST等生化指标,其中牡蛎粗多糖添加组能够极显著的提高小鼠SOD活性(P<0.01),高、中添加组显著(P<0.05)或极显著(P<0.01)提高Na~+、K~+、TP、ALB的浓度,显著降低血清Cl~-、ALT浓度(P<0.05);牡蛎粗多糖高、中、低剂量组组间比较发现,高、中剂量组与低剂量组比较,极显著提高SOD活性(P<0.01),显著提高TP含量(P<0.05),显著降低Cl-浓度(P<0.05),显著降低MDA活性(P<0.05),其他剂量组间差异不显著(P>0.05)。结论:热应激情况下,牡蛎粗多糖对调节血清电解质平衡及改善雄性小鼠的生化指标有一定的作用,不同添加剂量组间以高、中剂量组的作用较为显著。
3、热应激状态下牡蛎粗多糖对小鼠睾丸组织形态的影响
测定上述试验睾丸等组织的脏器指数,并观察热应激对小鼠睾丸组织形态的影响。结果:牡蛎粗多糖高、中、低剂量热应激组的各脏器指数较热应激组均有所升高,其中高、中组的肝脏指数与热应激组比较差异显著(P<0.05),但各组间差异不显著(P>0.05);显微镜下观察发现,热应激组睾丸生精细胞结构发生改变,管壁变薄、细胞排列不整齐、腔内精子数量较空白对照组明显减少,高、中剂量组睾丸生精细胞结构基本正常,管壁变薄、细胞排列较整齐、腔内精子较热应激组明显增多;低剂量组睾丸生精细胞排列较不整齐、腔内精子数量比高、中剂量组少,比热应激组多。结论:牡蛎粗多糖对改善热应激情况下,小鼠的睾丸组织形态有较明显的保护作用。
4、牡蛎粗多糖对夏季高温对公猪精液品质的影响
夏季高温情况下,选择某猪场8头种公猪,随机分为2组,即牡蛎粗多糖试验组、对照组,对试验期间的精液量、精子活力进行统计,并采集试验前后精液,检测精子密度、精子畸形率、精液抗氧化MDA、SOD、GSH-P_X指标。结果:牡蛎粗多糖试验组的射精量、精子活力比对照组都有一定水平的提高,但差异不显著(P>0.05);牡蛎粗多糖试验组的精子密度增加0.32亿个/mL,差异不显著(P>0.05),精子畸形率减少了4.26%,差异显著(P<0.05);试验组精液SOD、GSH-P_X显著提高(P<0.05);MDA含量极显著的降低(P<0.01)。结论:在夏季高温情况下,牡蛎粗多糖对精液抗氧化指标和种公猪的生精功能有较明显的改善作用。
The purpose of this paper was to establish the heat stress model, researching the influence of the animal blood biochemical indicators and the morphological changes of testicular tissue with oyster polysaccharide, and made the following experiment.
1. Establish the model of heat stress
Twenty-one male mice were randomLy divided into 3 groups, blank control group, heat stress group A and heat stress group B, each group 7 mice. the processing of group A was divided into two steps, stageⅠ(38℃45min) and stageⅡ(42℃stress 20min); the processing of group B with the step of stageⅠ(39℃stress 65min). Mice antioxidant indexes (SOD, MDA, GSH - P_X) and relevant hormone (ACTH, COR, T_3, and T_4) was tested, selecting the better model. Results: the indexes of group A’s SOD, MDA, GSH - P_X, ACTH, COR, T_3, T_4 and group B’s SOD, MDA, COR, T_3, contrast with that of control group, the differences were extremely significant (P < 0.01) or significant (P < 0.05). And group B’s GSH-Px, ACTH, T4 compared with that of control group, no significant differences (P > 0.05). Conclusion: The model of group A with heat stress 38℃for 45min and 42℃for 20min is the better model.
2. The influence of biochemical indexes with feeding of oysters polysaccharide under heat stress in male mice
With the heat stress model of group A, 75 male mice were randomLy divided into 5 groups, blank control group, heat stress control group, high, middle and low doses of oyster polysaccharide, each group 3 repetition with 5 mice. Blank control and heat stress control group were fed with based diet, and adding 1.8%, 1.2% and 0.6% oyster polysaccharide in based diet were fed for high, middle and low group, researching on the level of Na~+, K~+, Cl~-, MDA, SOD, GSH-P_X, AKP, TP and ALB in male mice blood and AST, ALT biochemical indexes in liver tissue homogenate under the condition of heat stress. Results: in addition to the AKP, the differences between the heat stress control group and control group were significant (P < 0.05) or extremely significant (P < 0.01). Oyster polysaccharide adding group compared with heat stress group can improve the biochemical indexes of Na~+, K~+ Cl ~-, MDA ,SOD, GSH - P_X, AKP, total protein, albumin and globulin in mice blood and AST,ALT in liver tissue homogenate. Oysters polysaccharide adding group can significantly increase mice’SOD (P < 0.01), and high, medium adding group can significantly (P< 0.05)or very significantly(P< 0.01) improve the concentrations of Na~+, K~+, TP and ALB, significantly reduce serum Cl~-, ALT concentration (P < 0.05). Oyster polysaccharide high, medium group compared with low dose group, can very significantly improve SOD (P < 0.01), significantly improve the TP content (P < 0.05), significantly reduce Cl~- concentration (P < 0.05) and significantly reduce the MDA activity (P < 0.05) respectively. The differences between other doses were no significant (P > 0.05). Conclusions: under the thermal stress situation, oyster polysaccharide can adjust the balance of serum electrolyte and improve the biochemical indexes of male mice. Among the groups with different doses,the role of high, middle dose group was better.
3. Detection of the influence of oyster polysaccharide on the testicle organizational form in thermal stress condition
The internal organs indices,such as testicle ,above the experiment, was determined and the influence of hot stress on mouse testicle organization shape was observated. Results: the viscera indices of oysters polysaccharide high, medium and low doses group improve, compared with that of heat stress, and the differences of viscera indices between high, medium group and heat stress group were significant (P < 0.05), but no significant differences between each group (P > 0.05). In microscope, heat stress group’s testicular cell structure change, lumen become thin and cells do not arrange tidily, sperm count in the cavity is significantly reduced compared with that of control group. The testicular cell structure of high, middle dose group is basicly normal, wall thin and cells arrange tidily, lumen sperm compared with that of heat stress group are significantly increase. The testicular cells of low dose group arrange less tidily, lumen sperm counts less than that of high, medium dose group,but more than that of heat stress group. Conclusions: oysters polysaccharide can obviously improve the protection of mice testicle organization forms in thermal stress circumstance.
4 The effects of oysters polysaccharides on the boar semen quality under summer high temperature.
Under summer high temperatures, eight stock boars selected from a farm were divided into 2 groups randomLy, namely oyster polysaccharide test group and control group .During the experiments, semen volume, sperm motility were statistified, and semen collected before and after experiments, was testing sperm concentration, abnormal sperm and semen antioxidant MDA, SOD, GSH-P_X indexs. Results: Oyster Polysaccharide could improve the levels of experimental group’s ejaculate volume and sperm motility than the control group, but the disparation was not significant (P > 0.05); The sperm density of oyster polysaccharide testing group was increased by 0.32 million / mL, the difference was not significant (P > 0.05), teratospermia rate decreased by 4.26%, the difference was significant (P < 0.05); testing group semen SOD, GSH-P_X were increased significantly (P < 0.05);the content of MDA was significantly lowed (P < 0.01). Conclusion: Under summer high temperatures, the oyster polysaccharide has a more significant improvement on semen antioxidant index and spermatogenic function of boars.
1、热应激模式的建立
将21只小鼠随机分为3组,即空白对照组、热应激A、热应激B组,每组7个小鼠。热应激A组处理分为两个步骤,即热应激Ⅰ阶段(38℃应激45min)和热应激Ⅱ阶段(42℃应激20min);热应激B组处理为一个步骤,即热应激Ⅰ阶段(39℃应激65min);应激6d后检测小鼠抗氧化指标(SOD、MDA、GSH-PX)和其相关激素(ACTH、COR、T_3、T_4),选择理想的小鼠模型。结果:热应激A组小鼠血清的SOD、MDA、GSH-PX、ACTH、COR、T_3、T_4,热应激B组的SOD、MDA、COR、T_3与空白对照组对比差异极显著(P<0.01)或显著(P<0.05);热应激B组的GSH-Px、ACTH、T_4与空白对照组对比差异不显著(P>0.05)。结论:热应激A组模式即38℃应激45min+42℃再应激20min,作为热应激模型。
2、牡蛎粗多糖对热应激雄性小鼠生化指标的影响
利用建立好的热应激模型,将6周龄成年雄性小鼠75只随机为5组,即设立空白对照组、热应激组、牡蛎多糖高、中、低剂量热应激组,每组3个重复,每个重复5只,空白对照组、热应激对照组饲喂基础日粮,牡蛎多糖高、中、低剂量热应激组饲喂基础日粮中分别添加1.8%、1.2%和0.6%牡蛎粗多糖,试验周期为28d。研究牡蛎粗多糖对热应激状态下雄性小鼠血液中Na~+、K~+、Cl~-、MDA、SOD、GSH-P_X、AKP、总蛋白、白蛋白和球蛋白及肝脏组织匀浆中ALT、AST等生化指标。结果:除AKP指标外,热应激对照组与空白对照组间差异显著(P<0.05)或极显著(P<0.01);牡蛎粗多糖添加组与热应激组比较,能改善小鼠血液中Na~+、K+、Cl-、MDA、SOD、GSH-P_X、AKP、总蛋白、白蛋白和球蛋白及肝脏组织匀浆中ALT、AST等生化指标,其中牡蛎粗多糖添加组能够极显著的提高小鼠SOD活性(P<0.01),高、中添加组显著(P<0.05)或极显著(P<0.01)提高Na~+、K~+、TP、ALB的浓度,显著降低血清Cl~-、ALT浓度(P<0.05);牡蛎粗多糖高、中、低剂量组组间比较发现,高、中剂量组与低剂量组比较,极显著提高SOD活性(P<0.01),显著提高TP含量(P<0.05),显著降低Cl-浓度(P<0.05),显著降低MDA活性(P<0.05),其他剂量组间差异不显著(P>0.05)。结论:热应激情况下,牡蛎粗多糖对调节血清电解质平衡及改善雄性小鼠的生化指标有一定的作用,不同添加剂量组间以高、中剂量组的作用较为显著。
3、热应激状态下牡蛎粗多糖对小鼠睾丸组织形态的影响
测定上述试验睾丸等组织的脏器指数,并观察热应激对小鼠睾丸组织形态的影响。结果:牡蛎粗多糖高、中、低剂量热应激组的各脏器指数较热应激组均有所升高,其中高、中组的肝脏指数与热应激组比较差异显著(P<0.05),但各组间差异不显著(P>0.05);显微镜下观察发现,热应激组睾丸生精细胞结构发生改变,管壁变薄、细胞排列不整齐、腔内精子数量较空白对照组明显减少,高、中剂量组睾丸生精细胞结构基本正常,管壁变薄、细胞排列较整齐、腔内精子较热应激组明显增多;低剂量组睾丸生精细胞排列较不整齐、腔内精子数量比高、中剂量组少,比热应激组多。结论:牡蛎粗多糖对改善热应激情况下,小鼠的睾丸组织形态有较明显的保护作用。
4、牡蛎粗多糖对夏季高温对公猪精液品质的影响
夏季高温情况下,选择某猪场8头种公猪,随机分为2组,即牡蛎粗多糖试验组、对照组,对试验期间的精液量、精子活力进行统计,并采集试验前后精液,检测精子密度、精子畸形率、精液抗氧化MDA、SOD、GSH-P_X指标。结果:牡蛎粗多糖试验组的射精量、精子活力比对照组都有一定水平的提高,但差异不显著(P>0.05);牡蛎粗多糖试验组的精子密度增加0.32亿个/mL,差异不显著(P>0.05),精子畸形率减少了4.26%,差异显著(P<0.05);试验组精液SOD、GSH-P_X显著提高(P<0.05);MDA含量极显著的降低(P<0.01)。结论:在夏季高温情况下,牡蛎粗多糖对精液抗氧化指标和种公猪的生精功能有较明显的改善作用。
The purpose of this paper was to establish the heat stress model, researching the influence of the animal blood biochemical indicators and the morphological changes of testicular tissue with oyster polysaccharide, and made the following experiment.
1. Establish the model of heat stress
Twenty-one male mice were randomLy divided into 3 groups, blank control group, heat stress group A and heat stress group B, each group 7 mice. the processing of group A was divided into two steps, stageⅠ(38℃45min) and stageⅡ(42℃stress 20min); the processing of group B with the step of stageⅠ(39℃stress 65min). Mice antioxidant indexes (SOD, MDA, GSH - P_X) and relevant hormone (ACTH, COR, T_3, and T_4) was tested, selecting the better model. Results: the indexes of group A’s SOD, MDA, GSH - P_X, ACTH, COR, T_3, T_4 and group B’s SOD, MDA, COR, T_3, contrast with that of control group, the differences were extremely significant (P < 0.01) or significant (P < 0.05). And group B’s GSH-Px, ACTH, T4 compared with that of control group, no significant differences (P > 0.05). Conclusion: The model of group A with heat stress 38℃for 45min and 42℃for 20min is the better model.
2. The influence of biochemical indexes with feeding of oysters polysaccharide under heat stress in male mice
With the heat stress model of group A, 75 male mice were randomLy divided into 5 groups, blank control group, heat stress control group, high, middle and low doses of oyster polysaccharide, each group 3 repetition with 5 mice. Blank control and heat stress control group were fed with based diet, and adding 1.8%, 1.2% and 0.6% oyster polysaccharide in based diet were fed for high, middle and low group, researching on the level of Na~+, K~+, Cl~-, MDA, SOD, GSH-P_X, AKP, TP and ALB in male mice blood and AST, ALT biochemical indexes in liver tissue homogenate under the condition of heat stress. Results: in addition to the AKP, the differences between the heat stress control group and control group were significant (P < 0.05) or extremely significant (P < 0.01). Oyster polysaccharide adding group compared with heat stress group can improve the biochemical indexes of Na~+, K~+ Cl ~-, MDA ,SOD, GSH - P_X, AKP, total protein, albumin and globulin in mice blood and AST,ALT in liver tissue homogenate. Oysters polysaccharide adding group can significantly increase mice’SOD (P < 0.01), and high, medium adding group can significantly (P< 0.05)or very significantly(P< 0.01) improve the concentrations of Na~+, K~+, TP and ALB, significantly reduce serum Cl~-, ALT concentration (P < 0.05). Oyster polysaccharide high, medium group compared with low dose group, can very significantly improve SOD (P < 0.01), significantly improve the TP content (P < 0.05), significantly reduce Cl~- concentration (P < 0.05) and significantly reduce the MDA activity (P < 0.05) respectively. The differences between other doses were no significant (P > 0.05). Conclusions: under the thermal stress situation, oyster polysaccharide can adjust the balance of serum electrolyte and improve the biochemical indexes of male mice. Among the groups with different doses,the role of high, middle dose group was better.
3. Detection of the influence of oyster polysaccharide on the testicle organizational form in thermal stress condition
The internal organs indices,such as testicle ,above the experiment, was determined and the influence of hot stress on mouse testicle organization shape was observated. Results: the viscera indices of oysters polysaccharide high, medium and low doses group improve, compared with that of heat stress, and the differences of viscera indices between high, medium group and heat stress group were significant (P < 0.05), but no significant differences between each group (P > 0.05). In microscope, heat stress group’s testicular cell structure change, lumen become thin and cells do not arrange tidily, sperm count in the cavity is significantly reduced compared with that of control group. The testicular cell structure of high, middle dose group is basicly normal, wall thin and cells arrange tidily, lumen sperm compared with that of heat stress group are significantly increase. The testicular cells of low dose group arrange less tidily, lumen sperm counts less than that of high, medium dose group,but more than that of heat stress group. Conclusions: oysters polysaccharide can obviously improve the protection of mice testicle organization forms in thermal stress circumstance.
4 The effects of oysters polysaccharides on the boar semen quality under summer high temperature.
Under summer high temperatures, eight stock boars selected from a farm were divided into 2 groups randomLy, namely oyster polysaccharide test group and control group .During the experiments, semen volume, sperm motility were statistified, and semen collected before and after experiments, was testing sperm concentration, abnormal sperm and semen antioxidant MDA, SOD, GSH-P_X indexs. Results: Oyster Polysaccharide could improve the levels of experimental group’s ejaculate volume and sperm motility than the control group, but the disparation was not significant (P > 0.05); The sperm density of oyster polysaccharide testing group was increased by 0.32 million / mL, the difference was not significant (P > 0.05), teratospermia rate decreased by 4.26%, the difference was significant (P < 0.05); testing group semen SOD, GSH-P_X were increased significantly (P < 0.05);the content of MDA was significantly lowed (P < 0.01). Conclusion: Under summer high temperatures, the oyster polysaccharide has a more significant improvement on semen antioxidant index and spermatogenic function of boars.
引文
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