Moesin抗体与动脉粥样硬化相关疾病的关系
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Discussion on the Relationship Between Momesin Antibody and Atherosclerosis-related Diseases
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    目的 通过重组表达的方法获得纯化的膜突蛋白(moesin), 利用纯化的重组moesin作为抗原建立间接酶联免疫吸附实验(ELISA),检测并探讨抗膜突蛋白抗体与动脉粥样硬化相关疾病的关系。方法 设计针对moesin的引物,通过PCR扩增,得到moesin的编码DNA,构建出能够表达moesin的重组质粒,用异丙基硫代半乳糖苷(IPTG)在适当温度和条件下诱导表达,通过SDS-PAGE电泳鉴定所表达的蛋白,并对重组蛋白进行纯化。利用纯化的重组moesin作建立间接ELISA,采用酶联免疫吸附试验检测120例动脉粥样硬化相关疾病(20例急性冠状动脉综合征、30例原发性高血压、20例颈动脉硬化、20例糖尿病及30例血脂异常症)患者血清抗膜突蛋白抗体,观察抗膜突蛋白抗体的阳性率。结果 表达产物经SDS-PAGE电泳分析,确定所得表达产物为moesin;抗原的最佳包被浓度为4 mg /L,抗体的最佳稀释倍数为1∶100,酶标二抗的最佳稀释倍数是1∶20 000。在120例动脉粥样硬化相关疾病中,抗膜突蛋白抗体的阳性检出率较高,达25%~70%,显著高于正常对照组的5%(P<0.01);急性冠状动脉综合征组阳性检出率高达70%,高于其它疾病组(P<0.05),其它各亚型疾病组之间差异无显著性。结论 成功地表达了重组人moesin;建立了最佳的重组moesin作为抗原检测抗膜突蛋白抗体的间接ELISA反应条件;抗膜突蛋白抗体在动脉粥样硬化相关疾病患者中有较高的检出率。

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    Aim Obtain purified moesin protein by recombinant,purified recombinant moesin protein was plated as coating antigen of indirect ELISA. And discussion on the relationship between momesin antibody and atherosclerosis-related diseases. Methods A pair of primers were designed according to the sequence of the moesin. The moesin gene was amplified by PCR. PCR products were then inserted into a prokaryotic expression plasmid, Using isopropyl thiogalactoside (IPTG) induced expression in the appropriate temperature and conditions. Recombinant moesin protein was analyzed by SDS-PAGE. Recombinant moesin protein will be purified. Purified recombinant moesin protein was applied to establish the indirect ELISA. Adsorption assay in 120 patients(20 patients with acute coronary syndrome,30 patients with primary hypertension,20 patients with carotid atherosclerosis,20 patients with diabetes and 30 patients with hyperlipidemia) with diseases related to atherosclerosis anti-moesin antibody by ELISA, compared the positive rate of anti-moesin antibody of blood. Results The expression product was recombinant moesin by SDS-PAGE2) The optimal antigen concentration was 4 mg/L, the optimal antibody dilution was 1∶100, the optimal HRP-IgG dilution was 1∶20 000. In 120 cases of diseases related to atherosclerosis, positive rate is higher protein antibody against membrane process, up to 25%~70%, significantly higher than that of normal control group 5% (P<0.01) acute coronary syndrome group, the positive rate of 70%, higher than the other groups (P<0.05), the difference between other subtypes of disease no significant (P>0.05). Conclusion Successfully expressed the recombinant human moesin protein Establish the best indirect ELISA of Purified recombinant moesin protein was plated as coating antigen detect moesin antibody Moesin antibodies in patients with atherosclerotic disease associated with high detection rate.

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毛 华, 杜 峰 ,胥 勇, 朱晋坤, 文 美, 刘廷筑. Moesin抗体与动脉粥样硬化相关疾病的关系[J].中国动脉硬化杂志,2013,21(06):549~553.

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  • 收稿日期:2013-02-27
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