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单相酶扩散法测定SLE患者血清DNA酶的浓度 |
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陈宜芳 曾凡钦 许德清 李伯有 林宝珠
【摘 要】 目的 建立DNase检测的方法，并探讨血清DNase浓度异常在SLE发病中的意义。方法 于国内首次用单相酶扩散法检测了63例SLE患者。结果 SLE患者血清中DNase浓度明显低于正常人，活动期患者明显低于缓解期患者，血清DNase的浓度下降与DNA含量升高呈负相关。用于DNase检测的琼脂板中最适DNA和EB浓度分别为30%和2.5%，该法检测DNase的灵敏度可达2 μg/L。结论 用单相酶扩散法检测血清DNase浓度简便、灵敏、可靠，血清DNase水平低下在SLE的发生发展中起着一定作用，测定患者DNase浓度在研究SLE复杂的免疫发病机制上具有重要意义。
【关键词】 单相酶扩散法 脱氧核糖核酸酶 红斑狼疮，系统性
Measurement of serum deoxyribonuclease in patients with SLE by single radial enzyme diffusion method
CHEN Yifang ZENG Fanqin XU Deqing,et al.
(Sun Yat-sen Memorial Hospital,Sun Yat-sen University of Medical Sciences,Guangzhou 510120)
【Abstract】 Objective To establish the method detecting DNase and measure serum DNase in SLE patients.Methods The concentration of DNase was measured by single radial enzyme diffusion method in 63 serum samples of SLE patients.Results The concentration of DNase in SLE patients was significantly lower than that in healthy individuals.Patients at active stage showed much lower DNase level than patients at inactive stage,and decrease of serun DNase was negatively correlated with the increase of serum DNA.The optimal concentration of DNA and EB in agar plate was 30% and 2.5%,and the sensitivity detecting DNase was up to 2 μg/L.Conclusion The single radial enzyme diffusion method for measuring DNase is simple,sensitive and reliable.The decrease of serum DNase may play an important role in the development of SLE.
【Key words】 Single radial enzyme diffusion method Deoxyribonuclease Lupus erythematosus,systemic
系统性红斑狼疮(SLE)患者血循环中DNA水平升高〔1～3〕，但其升高的原因迄今尚不清楚。正常机体清除DNA的途径主要是通过DNA酶(DNase)的降解作用，本文建立了DNase测定的单相酶扩散法，用于检测SLE患者血清的DNase浓度，并藉以探讨血清DNase的浓度异常与DNA含量升高的相互关系。
1 材料与方法
1.1 实验材料与病例选择
1.1.1 主要试剂：三羟甲基氨基甲烷(Tris)：Fluka公司产品，配成Tris-HCI缓冲液，即0.05 mol Tris，0.1 mol NaCl,用HCl调pH至7.2；牛胸腺DNA：Sigma公司产品，用去离子水配成6 g/L，室温搅拌2～3小时，储存于4 ℃；牛胰DNA酶I(DNase)：Sigma公司产品，用去离子水配制成1 g/L的储存液，－20 ℃保存；RNA酶A(RNa[1] [2] 下一页
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