Sprague-Dawley大鼠肝微粒体蛋白浓度检测方法学研究

Study on Methodology of Protein Concentration in Liver Microsomes of Sprague-Dawley Rats

目的建立Sprague-Dawley(SD)大鼠肝微粒体蛋白浓度测定的紫外分光光度检测法。方法SD大鼠尾静脉连续3d注射0.9%氯化钠注射液2ml,第4d解剖大鼠,取肝脏,应用差速离心法提取大鼠肝微粒体,用Lowry法测定肝微粒体蛋白浓度。结果牛血清蛋白标准曲线范围为0～500μg/ml,回归方程为:Y=0.0007X+0.0161(r=0.9982),低、中、高浓度的回收率分别为95.85%、102.69%、101.28%,日内和日间精密度分别为3.83%、1.73%、0.57%及10.25%、2.22%、0.98%。结论本实验建立的Lowry蛋白测定法准确可靠,重复性和稳定性均良好,可为进一步研究用药后肝微粒体酶活性的改变提供依据。

Objective To establish the UV determination methodology of protein concentration in liver microsomes of Sprague-Dawley （SD）rats. Methods After the SD rats were intravenous administered with 0.9% NaCl once daily for 3 d, and liver microsome was extracted by different velocity centrifugation. The Lowry method was used to measure the concentration of protein in liver microsome. Results There was a good linearity within the range of 0-500μg/ml between the concentration and absorbance. The regression equation was Y=0.O007X＋0.0161 （r=-0.9982）of bovine blood serum albumin standard curve. The recoverys were 95.85%, 102.69%, 101.28% （n= 3）, within-day RSD of 3.83%, 1.73%, and 0.57%, and the between-day RSD of 10.25%, 2.22%, and 0.98%. Conclusion The methods of measuring protein concentration was sensitive and accurate in SD rats by ultraviolet spectrophotomater, and it was credible to provide reference for measuring the activity of the hepatic microsomal enzyme.