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目的:探讨吉林双阳梅花鹿鹿茸水溶性蛋白组分应用的安全性,为鹿茸蛋白的开发应用提供实验依据。方法:将新鲜双阳梅花鹿鹿茸进行高速组织匀浆,离心取上清,用饱和硫酸铵进行沉淀,PBS溶解沉淀,最后通过SephadexG25分子筛层析方法过滤除盐,初步获取鹿茸水溶性蛋白组分。将58只ICR小鼠随机分为生理盐水灌胃组、鹿茸蛋白溶液灌胃组、鹿茸蛋白溶液皮下注射组和鹿茸蛋白溶液腹腔注射组,按6.7 mg.kg-1给药,每天2次,第28天处死小鼠,取心、肝、肾和肺组织进行病理形态学观察。结果:鹿茸总蛋白溶液采用Sephadex G25层析过滤除盐后,平均浓度为0.685 g.L-1。病理形态学切片显示:鹿茸蛋白溶液灌胃组(50.0%)、皮下注射组(91.7%)和腹腔注射组(87.5%)小鼠可见散在的心肌细胞肌浆凝聚;蛋白溶液灌胃组(40.0%)、皮下注射组(64.3%)和腹腔注射组(77.8%)小鼠肝组织有炎细胞浸润和不同程度灶状坏死;3只皮下注射组小鼠肾小管上皮有不同程度的玻璃样变性;3只蛋白溶液灌胃组小鼠有肺炎病变;皮下注射组及腹腔注射组各有3只小鼠肺间隔增宽。结论:不同给药途径的鹿茸蛋白溶液对小鼠心肌组织、肝脏组织有一定程度的病理性损害,对肾、肺组织无明显影响。
Objective: To investigate the safety of the antler water-soluble protein component of sika deer antler in Shuangyang, Jilin Province, and to provide an experimental basis for the development and application of antler protein. Methods: The fresh Shuangyang sika deer antler was homogenized at high speed. The supernatant was collected by centrifugation, precipitated with saturated ammonium sulphate and dissolved in PBS. Finally, the desalted water was fractionated by SephadexG25 molecular sieve chromatography. 58 ICR mice were randomly divided into normal saline group, antler protein solution gavage group, antler protein solution hypodermic injection group and antler protein solution intraperitoneal injection group, administered with 6.7 mg.kg-1 twice a day, Mice were sacrificed on the 28th day, heart, liver, kidney and lung tissues were observed for pathomorphology. Results: After the antler total protein solution was desalted by Sephadex G25 chromatography, the average concentration was 0.685 g.L-1. Histopathological sections showed that sarcolemmal aggregates of scattered cardiomyocytes were observed in pilose antler protein solution-fed group (50.0%), subcutaneous injection group (91.7%) and intraperitoneal injection group (87.5% %, Subcutaneous injection group (64.3%) and intraperitoneal injection group (77.8%) mouse liver tissue infiltration of inflammatory cells and varying degrees of focal necrosis; 3 subcutaneous injection group of mouse renal tubular epithelial cells with varying degrees of glassy degeneration ; 3 protein solution gavage mice had pneumonia lesions; subcutaneous injection group and intraperitoneal injection group each 3 mice broadened pulmonary septum. Conclusion: The antler protein solution of different routes of administration has a certain degree of pathological damage to the myocardium and liver of mice, but has no obvious effect on the kidney and lung tissues.