运输应激对山羊免疫器官的病理损伤及热休克蛋白表达的影响

Effects of transport stress on pathological injury and expression of heat shock proteins of the immune organs in goats

本试验旨在研究运输应激对山羊免疫器官的病理损伤及热休克蛋白表达的影响。选取12只体质量相近的健康公山羊为实验动物,随机分为3组,对照组、运输应激2 h组和运输应激6 h组,以速度35~45 km/h模拟公路运输。运输结束后,立即采集所有山羊淋巴结和脾脏,应用HE染色、醋酸铀枸橼酸铅染色、免疫组织化学和Western blot等方法,分析运输应激山羊淋巴结和脾脏的病理变化及HSP27、HSP70和HSP90等3种热休克蛋白的表达情况。光镜观察显示,运输应激2 h组山羊淋巴结和脾脏均出现急性病理变化,淋巴小结和脾小结增生明显,淋巴结的髓索变粗,髓窦中有淡红色红染物质,脾脏红髓的红细胞急剧增多;运输应激6 h组淋巴细胞胞核出现核碎片,脾脏红髓的红细胞比2 h组有所减少。电镜观察发现,淋巴结和脾脏的淋巴细胞病变明显,发生凋亡或坏死。免疫组化显示,在淋巴结和脾脏中,HSP27主要分布在弥散淋巴组织,HSP70和HSP90主要分布在淋巴小结或脾小结。Western blot结果显示,与对照组相比,运输应激2h组与6h组淋巴结和脾脏中HSP90表达量分别显著和极显著升高(P<0.05,P<0.01),运输应激2 h组脾脏HSP27和淋巴结HSP70的表达量都显著升高(P<0.05);与对照组相比,运输应激2 h组和6 h组脾脏HSP70表达量分别下降(P>0.05)和极显著下降(P<0.01)。运输应激对山羊淋巴结和脾脏造成一定的病理损伤,3种热休克蛋白在淋巴结和脾脏都有表达,运输前后和不同运输时间3种热休克蛋白表达量发生了变化。

The purpose of this study was to investigate the effect of transport stress on the pathological injury of immune organs and the expression of heat shock proteins in goats.T welve healthy male goats of similar weight were selected as experimental animals and were randomly divided into three groups:control group,two-hour transport stress group and six-hour transport stress group.The transport stress groups were transported on road with the speed of 35-45 km/h.After transportation,the lymph nodes and spleens of all goats were collected immediately.The pathological changes,and the distribution and expression of HSP27,HSP70 and HSP90 of lymph nodes and spleen under transport stress were analyzed with the methods of HE staining,uranium-citrate staining,immunohistochemical staining and Western blot(WB).Optical microscope observation showed that acute pathological changes were found in goat lymph nodes and spleens,lymph nodules and splenic nodules showed obvious hyperplasia,the medullary cords of the lymph nodes thicken,and red exudates was found in the medullary sinus,and red blood cells increased rapidly in the red pulp of the spleen.The nuclei of lymphocytes and reticuloendothelial cells were fragmented after six-hour transportation,and the number of erythrocytes in the red pulp of the spleen of six-hour transport group than that of two-hour transport group.Electron microscopic observation showed that lymphocyte lesions in lymph nodes and spleen were obvious,and apoptosis or necrosis occurred.Immunohistochemistry showed that in lymph nodes and spleen,HSP27 was mainly distributed in diffuse lymphatic tissue,while HSP70 and HSP90 were mainly distributed in lymphatic nodules and splenic nodules.WB results showed that,compared with the control group,the expression levels of HSP90 in lymph nodes and spleen of the two-and six hour transport group were significantly and extremely significantly increased,respectively(P<0.05 and P<0.01).The expression level of HSP27 in spleen and HSP70 in lymph nodes was significantly increased after twohour transportation(P<C0.05).Compared with the control group,the expression of HSP70 in the spleen of the two- and six hour transport groups was significantly decreased(P>0.05)and ex-tremely significantly decreased(P<0.01).In conclusion,transport stress caused certain damage to the lymph nodes and spleens of goats.Three heat shock proteins were positively expressed in lymph nodes and spleen.The expression levels of three heat shock proteins were changed before and after transportation and at different transportation time.