一种新型优化的肺组织羟脯氨酸水平测定方法的建立  

Establishment of a new and optimized assay method of HYP in lung tissue

作　　者：彭方毅[1,2] 周欢[1] 姜海蓉[1] 袁兵占[1] 崔玉花[1] 

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]重庆市江津区中心医院,重庆402260

出　　处：《重庆医学》2014年第11期1354-1356,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(30571714;60873103);国家自然科学基金重点资助项目(30830090);重庆市科委自然科学基金资助项目(CSTC;2008BB5331;2009BB7231);教育部科技重点资助项目(210178)

摘　　要：目的建立大鼠肺组织羟脯氨酸（HYP）水平测定方法，以评价大鼠肺组织纤维化程度。方法比较不同酸水解时间、氧化时间、显色时间等对大鼠肺组织HYP水平测定结果的影响，据此建立大鼠肺组织HYP水平测定方法，并初步应用于博莱霉素诱导大鼠肺纤维化HYP水平的测定。结果通过优化得出HYP水平测定最佳条件为110℃7．50mol盐酸（HCL）水解16h，常温氧化10min，60℃显色25min。本方法灵敏度为0．067／μg／mL，回收率为88．85％～110．88％，平均CV为4．70％～6．60％。该法应用于博莱霉素诱导大鼠肺纤维化研究中，模型组HYP水平明显高于对照组。结论该法灵敏度与回收率均较高、重现性较好，可用作临床对肺纤维化程度判定的定量方法。Objective To establish the assay method of hydroxyproline（HYP） level in rat lung tissue for evaluating the lung fi- brosis degree. Methods The impact of different acid hydroiyzable time,oxidative time,developing time on the assay results of the HYP level in rat lung tissue was studied. On this basis, the assay method for determining the HYP level was established and prelim- inarily applied in the detection of the HYP level in rat lung tissue. Results The optimal condition for the HYP level detection in rat lung tissue was afollows：7.50 mol HCL was hydrolyzed for 16 h at 110℃ ,oxidized for 10 min at room temperature and devel- oped for 25rain at 60℃. The sensitivity of this method was 0. 067 μg/mL. The recovery rate and the average CV of this method were 88.85%-110.88% and 4.70%-6.60%, respectively. In the study of bleomycin induced rat lung fibrosis,the HYP level of the model group was obviously higher than that of the control group. Conclusion This method has high sensitivity,high recovery rate and good reproducibility, and may be used as a reliable quantitative method to judge the lung fibrosis level in clinic.

关 键 词：大鼠 肺 组织 羟脯氨酸 检测 方法 优化 

分 类 号：R285.5[医药卫生—中药学]