Asprosin的表达纯化及其对小鼠在体心脏功能的作用

Expression and purification of asprosin and its effects on cardiac function in mice

目的利用大肠埃希菌原核表达并纯化asprosin蛋白,研究其对心功能改善的影响。方法从GenBank获取asprosin编码序列,根据大肠埃希菌密码子偏好性进行密码子优化,全基因合成,连接于表达载体,进行IPTG诱导表达并纯化。通过对冠脉左前降支结扎再放松,建立小鼠心功能损伤模型。将30只小鼠随机分成3组:假手术组(sham)、心功能损伤组(MI/R)和心功能损伤并注射重组蛋白组(MI/R+r Asp)。通过心动超声检测左室功能,对心功能损伤程度进行评价,进而研究Asprosin对心功能改善的影响。结果经过原核表达并纯化,目标蛋白纯度大于95%,内毒素含量小于<0.1 EU/μg蛋白,适于进行细胞及动物研究。成功建立了心功能损伤模型,与单纯损伤组相比,外源给予重组asprosin蛋白后,小鼠心脏心功能明显改善(P<0.05)。结论 Asprosin蛋白具有抑制心功能损伤,改善心功能的作用。

Objective To purify asprosin protein expressed in Escherichia coli expression system and to study its effect on cardiac function. Methods Coding sequence of asprosin was obtained from Gen Bank. Codon optimization was performed according to the codon preference of E. coli. After gene synthesized,recombinant plasmid was made. Asprosin was then induced and purified by Ni-affinity purification. The mouse model of impaired cardiac function was established by ligating and relaxing the left anterior descending coronary artery. 30 mice were randomly divided into 3 groups： sham operation group（ sham）,cardiac dysfunction group（ MI/R） and cardiac dysfunction plus injection of recombinant asposin protein group（ MI/R ＋ r Asp）. The left ventricular function was detected by echocardiography to determine the improving effect of recombinant asprosin protein on cardiac function. Results After prokaryotic expression and purification,the purity of the target protein was higher than 95%,and the endotoxin content was less than 0. 1 EU/μg protein,which was suitable for cell and animal studies. After the recombinant asprosin protein was given,the left ventricular function of the mice was improved significantly（ P〈0. 05）. Conclusions Asprosin acts as a myocardial protective molecule to improve cardiac function.