| Epinephelus coioides is high-grade seaculture fish in China,high temperature stress is the key environmental factors which influence the growth and survival.The digestive tract is inclined to bring one of the main target organ of grouper Heat stress,while Heat Shock Protein(Heat Shock Protein,HSP)playing an important role in cell stress damage repair and inhibit apoptosis,can effectively protect the body against stress injury.As HSPs' inducers,Teprenone Capsules has been reported in the related research and application of the protection of gastric mucosa damage in the human,rat and lifestock but not seen in the research of aquatic animal.This paper analyzed the effect of Teprenone to the gastrointestinal tissue of Epinephelus coioides by feeding mixture feedstuffs with different doses of Teprenone Capsules.The content of the research mainly included the following four parts:(1)The influence of Teprenone Capsules to Epinephelus coioidesWe observed the data and studied the effect of different doses of Teprenone Capsules(100 mg/Kg,200 mg/Kg,400 mg/Kg)to liver function index in Epinephelus coioides by feeding mixture feedstuffs.Results showed that compared with the control group(0 mg/Kg),the level of AST,GTP and TBIL in low-dose group(100 mg/Kg)and mid-dose(200 mg/Kg)were not significant different(p>0.05),while the level of AST and TBA of Epinephelus coioides in high-dose group(400 mg/Kg)significantly increased to 411.6±50.1(?mol/L)and 24.8±5.1(?mol/L).Results suggested that the low-dose(100 mg/Kg)or the mid-dose(200 mg/Kg)of Teprenone Capsules feed additives had no effect on Epinephelus coioides' liver function indicators.And high-dose(400 mg/Kg)of Teprenone Capsules damaged for Epinephelus coioide's liver function,proved to have certain toxicity.(2)The histologic analysis of heat shock induced injury protection of Teprenone Capsules to Epinephelus coioides' gastrointestinalThrough the digestive tract tissue biopsies,we observed the histology changes of intestinal in Epinephelus coioides after feeding with Teprenone Capsules and heat shock damaging.Results showed that the length,width of intestinal mucosal villus and the thickness of the intestinal muscle layer in the treatment groups obviously higher than the control group.After feeding Teprenone Capsules For 15 d,the length,width of grouper intestinal mucosal villus and the thickness of intestinal muscle layer of treatment groups were completely different;In low-dose group(100 mg/Kg),the growing stability of the length and the width of the intestinal mucosal villus was good;in the mid-dose(200 mg/Kg),the stability of thickness of intestinal muscle layer is good.Contrasted 48 h with 0h after stressed by Ascending 8 ? higher to temperature stressed 2 h,the length and width of the intestinal mucosa villus and the intestinal muscle layer thickness had no significant differences before and after stress in processing groups.Histological observation showed that Teprenone Capsules can not only promote the length and width of intestinal mucosal villus and the growth of intestinal muscle layer thickness,but also can maintain the growing stability of intestinal tissue.Contrast found that mid-dose group(200 mg/Kg)of Teprenone Capsules were best for stability effect of intestinal villi and muscle layer.(3)The heat shock induced effect of Teprenone Capsules to digestive tract HSPs in Epinephelus coioidesAfter bserving the results of ELISA,we found that the heat shock induced effect of Epinephelus coioides digestive tract HSPs was obvious after adding different doses of Teprenone Capsule.The results showed that at 0 h and 48 h after high temperature stressed,the induced expression of HSP70,HSP60 and HSP90 of treatment group were obviously higher than control group.The expression level of HSP70,HSP60 and HSP90 was about 160ng/L,25 ng/L and 14 ng/L,the heat shock levels of HSP70 was much higher than that of HSP60 and HSP90;the expression levels of HSPs in stomach were higher than that in the intestinal tract.Within 3~24 h after high temperature stressed,compared to the control group,the HSPs of the group added Teprenone Capsules did not differ significantly.Results show that for Teprenone Capsules can improve gastrointestinal HSP 70 expression of fish,the higher the initial expression level can improve the subsequent thermal stress resistance,characterized by reduced susceptibility of heat shock;And each treatment group after heat shock recovery HSPs remained high expression level.(4)The immune enzyme activity of the digestive tract related Teprenone Capsules to Epinephelus coioidesThrough the observation of enzyme immune,we found that adding Teprenone Capsules had a significant impact on immune enzyme activity of the Epinephelus coioides gut.The trend of MT was first falls then rises,6 h was the lowest,but there were no significant differences between the treatment group and the control group(0 mg/Kg),only the low-dose group(100 mg/Kg)declined slower.Cyt-C showed a trend of low after the first high level,and the peak is given priority to the 6 h,on the contrary of the change of MT level,it may be one of the reasons to the inhibition of MT reducing.HSP70,HSP60 and HSP90 main at 6 h reached the highest peak,peak of cytochrome C is given priority to with 6 h,because of Cyt-C has an indispensable role in respiratory electron transport chain,however,Cyt-C variation,relative to other proteins are more likely to lead to the electron transfer problems of the organism and apoptosis,HSPs to a certain extent,ease the apoptosis.Low dose for Teprenone Capsules fish body heat shock response can be improved after the digestive tract antioxidant ability,at the same time,relatively lower malondialdehyde and hydroxyl free radical level,there by lowering the body's stress damage. |
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