| Transportation of livestock and poultry is an essential part of animal husbandry.Physiological disorder of animal organism,disease and even death as well as the decline of meat quality and animal welfare can be resulted by crowding,jolting,vibration,noise,temperature change,fasting,water deprivation and other complex stressors in the process of transportation,which brings huge economic losses to animal husbandry.Heat shock proteins are involved in a variety of life activities.As molecular chaperones,they have the function of assembling,folding and transshipment as well as anti-stress and anti-apoptosis,which may make a difference in transport stress.In this study,12 healthy male goats of similar weight(13.89±2.96Kg)were selected and randomly divided into 3 groups: The control group(no transported),two-hour transported group and six-hour transported group.The treatment groups were transported on the road at 28?-32? with the speed of 35-45km/h.Microscopical and ultrapathological changes of heart,liver,kidney,lung,trachea,bronchus,spleen,lymph nodes and small intestine of goats were systematically analyzed at different transportation time,and the distribution and expression of HSP27,HSP70 and HSP90 in the main organs of goats before and after transportation were discussed with the methods of HE staining,transmission electron microscopy,real-time PCR,immunohistochemistry and western blot,which provided basic data for the study of pathogenesis and solution of transportation stress in goats.The following were the main results.(1)The results of goat heart experiments showed myocardial fibers underwent granular degeneration,swelling and rupture of mitochondria,edema,hyperemia and inflammatory cell infiltration of mesenchyme,and the lesions were more serious in the group transported for six hours.These three proteins were mainly expressed in the cytoplasm of cardiac myocytes,while HSP27 and HSP90 were also expressed in the nucleus.Compared with the control group,the mRNA levels of HSP27,HSP70 and HSP90 were all improved in the two treatment groups.The protein expressions of HSP27,HSP70 and HSP90 were all increased in the 2 h transported group,but onlyHSP70 and HSP90 were increased in the 6 h transported group.(2)Goat liver experimental results showed that varying degrees of injuries were happened in the liver,which were included expansion and congestion of central venous,sinus gaps were minished,obvious grain and blister denaturation were appeared in the liver cells,a small amount of nucleus chromatin were gathered in the liver cells,and distortion and swelling of the mitochondria accompanied by breaking and disappearance of the cristae and appearance of vacuoles and lipid droplets in the cytoplasm,which were more serious in the group transported for six hours.HSP27 was distributed in the cytoplasm of hepatocytes,vascular endothelial cells and bile duct epithelial cells near the central vein and portal area.HSP70 and HSP90 were mainly located in the cytoplasm of hepatocytes near the central vein and portal area,and the HSP90 was also expressed in the nucleus.The mRNA levels of HSP27 and HSP90 were all increased in the 2 h transported group when compared with the control group,but in the 6 h transported group only HSP70 mRNA level was improved.Moreover,the protein expressions of HSP27 and HSP70 were all increased in the two transported groups.(3)The results of goat heart experiments showed that the structures of renal tubules and collecting tubules were disordered,the boundaries between the lumen were not clear and exfoliated epithelial cells and grume could be seen in the lumen,the glomeruli were congested and swollen,the capsular space narrowed down,and the mitochondria was swollen with the breakage and reduction of the cristae.The hyperemia and bleeding as well as the damage of lumen and glomerular were more obvious after six-hour transportation.These three proteins in the control group were mainly expressed in the renal tubules and collecting tubules and the expression was stronger after transportation.Positive reaction was also observed in the renal corpuscles of the treatment group.Compared with the control group,the mRNA and protein levels of HSP27 and HSP90 were all improved in the 2 h transported group.On the contrary,the mRNA level of HSP70 was decreased but the protein expression was increased in the 2 h transported group.In the 6 h transported group,only HSP27 mRNA level was increased,and the protein expressions of HSP27,HSP70 and HSP90 were all increased.(4)The experimental results of lung,trachea and bronchus in goat showed that the trachea and bronchial mucosa were edematous and hyperemic,with inflammatory cell infiltration and the lodging,rupture and desquamating of cilia,a small number of mucosal epithelial cells underwent degeneration and necrosis,the superfine lesions of cilia and goblet cells were obvious,the mitochondria was swollen and the cristae was dissolved and decreased.The alveolar cavities were collapsed or fused and the alveolar septa was widened,congested and infiltrated with inflammatory cells.Part of the alveolar epithelial cells was denatured and the exfoliated cells and debris could be seen in the alveolar space.The ultramicropathy of alveolar type II epithelial cells was obvious,the cell membrane of alveolar type I epithelium was uneven and the cytoplasm was vacuolated.The damage was more serious after 6 hours of transportation.HSP27 and HSP70 were expressed in trachea and bronchial epithelium,glandular epithelium and blood vessel endothelium,and the number of positively expressed inflammatory cells increased in the treatment group and was generally expressed in alveolar cells and bronchioloepithelial cells,while HSP90 was weakly expressed in tracheal and bronchial epithelium,glandular epithelium,alveolar type II epithelium and bronchiole epithelium.The protein expression of HSP70 in trachea was significantly improved in the 2 h and 6 h transported groups when compared with the control group.The mRNA and protein level of HSP70 in the lung were all increased after 2 hours' transportation.(5)The results of spleen and lymph node experiment in goat showed acute pathological changes in both lymph nodes and spleen after transportation.The lymphatic nodules and splenic nodules showed obvious hyperplasia,and the nuclei in lymphocytes and reticuloendothelial cells were broken in the group transported for six hours.Besides,the cell ultrapathological changes were obvious,cell apoptosis or necrosis could be seen.There were some differences of the location of these three proteins in the lymph nodes and spleen between before and after transportation.HSP27 was mainly distributed in diffuse lymphatic tissue,while HSP70 and HSP90 were mainly distributed in lymphatic nodules and splenic nodules.Compared with thecontrol group,the mRNA levels of HSP27,HSP70 and HSP90 in the lymph nodes and the spleen were all increased in the 2 h transported group.In the 6 h transported group,only HSP70 mRNA level in the lymph nodes and HSP90 mRNA level in the spleen were improved.The protein expression of HSP90 in the two treatment groups were significantly increased,and the protein expressions of HSP27 in the spleen and HSP70 in the lymphonodi were increased after 2 hours' transportation.On the contrary,in the 6 h transported group the protein expression of HSP70 in the spleen was decreased.(6)The experimental results of duodenum,jejunum and ileum of goat showed that the intestinal villus mucosa epithelium and lamina propria appeared hyperemia,hemorrhage and infiltration with inflammatory cells,a small amount of epithelial cells appeared degeneration,the microvilli were prostrate,the nuclei showed pyknosis and the mitochondria were swollen and proliferating.These three proteins were mainly expressed in intestinal villi,HSP27 and HSP70 were mainly expressed in cytoplasm,and HSP90 was expressed in both cytoplasm and nucleus.Compared with the control group,the mRNA level of HSP27 and HSP70 were all increased in the duodenum after 6 hours' transportation,and only HSP27 mRNA level was improved in the 2 h transported group.The protein expression of duodenal HSP90 was significantly increased after 6 hours' transportation.In the jejunum,the mRNA level of HSP27,HSP70 and HSP90 were all increased in the 2 h transported group.In the 6 h transported group only HSP70 mRNA level was improved.In the ileum,only HSP70 mRNA level was increased in the 2 h transported group.On the contrary,the mRNA levels of HSP27 were all decreased both in the two treatment groups as well as the protein expression.The expression of HSP70 in ileum was significantly increased after 2 hours' transportation when compared with the control group and the 6 h transported group.In conclusion,transportation stress can caused pathological damage to the main parenchymal organs and cavities of goats,and the damage was aggravated with the extension of transportation time.After transportation,the locations of HSP27,HSP70 and HSP90 were all changed in different organs,and the expressions of mRNA andprotein were also changed.These results suggested that the damage of goat organs can be caused by transport stress. |
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