一种新型优化的肺组织羟脯氨酸水平测定方法的建立

摘要

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 hydrolyzable 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 as follows :7 .50 mol HCL was hydrolyzed for 16 h at 110 ℃ ,oxidized for 10 min at room temperature and devel-oped for 25min at 60℃ .The sensitivity of this method was 0 .067μg/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 .%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℃7．50 mol盐酸（HCL）水解16 h ，常温氧化10 min ，60℃显色25 min。本方法灵敏度为0．067μg/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。