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Progress in the expression of P-glycoprotein in brain metastases of lung cancer andrelated TCM research

TMR Theory and Hypothesis 2018年第1期1卷 18-25页

作者： shuai zhui Ling-Yu Qi Xue Wang Hua-Yao Li Jia Li Jing Yang Shandong University of TCM Jinan 250000 China Qingdao University Qingdao 266071 China Department of Oncology Weifang Traditional Chinese HospitalWeifang 261031Shandong Province China

Brain metastases are common intracranial tumors, and their occurrence not only represents a high degree of malignancy, but also often is the major factor in treatment failure and poor prognosis. Primary site of brain metastases often occur in lung. P-glycoprotein is a member of the （ATP binding cassette） transporter superfamily,which is closely related to the development of lung metastases. It is the main reason for influencing the drug through the blood_brain barrier into the brain tissue, and it also is an important factor affecting the treatment of brain metastases. According to the theory of traditional chinese mddicine, the pathogenesis of brain metastases is due to phlegm, poison, stasis, virtual and so on. The principle of treatment is to promote blood circulation, remove phlegm turbidity. In recent years, the impact of Chinese herbal medicine on P-glycoprotein is increasing. This paper analyzes the mechanism and components of the relevant Chinese medicine on P-glycoprotein. It provides a reference for clinical rational drug use.

关键词： Lung cancer Brain metastasis P-glycoprotein Blood-brain barrier Chinese medicine treatment Research progress

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温度响应的黄曲霉毒素B_(1)纳米抗体重组表达、复性及生物活性

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食品科学 2023年第18期44卷 141-148页

作者：张乐平涂追李燕萍李小江帅文苑张航何庆华南昌大学食品科学与资源挖掘全国重点实验室江西南昌330047 南昌大学食品学院江西南昌330047 南昌大学中德联合研究院江西南昌330047

将不同长度类弹性蛋白多肽(elastin-like polypeptide,ELP)与纳米抗体融合表达,获得具有温度响应能力的抗黄曲霉毒素B_(1)(aflatoxin B_(1),AFB_(1))纳米抗体。采用递归定向连接克隆得到重组表达载体pET30a-G8-ELP20、pET30a-G8-ELP40、pET30a-G8-ELP60、pET30a-G8-ELP80,分别转化大肠杆菌BL21(DE3)菌株,表达后经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)分析显示4种融合蛋白均以不溶性包涵体形式存在于菌体沉淀中,对其进行变性处理后,分别采用稀释复性、可逆相变循环(inverse transition cycling,ITC)纯化、透析复性、柱上复性4种方式对包涵体蛋白进行复性。SDS-PAGE分析4种复性方式分别获得的4种融合蛋白,结果显示其纯度差异不显著,但ITC纯化蛋白得率最高,分别为83%、90.7%、89.5%、88.3%;间接竞争酶联免疫吸附实验测定复性后融合蛋白活性,结果表明由稀释复性法获得的G8-ELP80重组蛋白IC50最低(4.35 ng/mL);浊度分析法测得融合不同长度ELP标签纳米抗体的相变温度分别为45、38、32、28℃;圆二色谱分析显示4种融合蛋白二级结构均以β-折叠、β-转角为主,与预估结果相符。本研究将ELP标签与抗AFB1纳米抗体融合表达,实现了纳米抗体的温度刺激响应性,系统比较了不同复性方式对蛋白特性的影响,为后续AFB1检测分析提供了基础。

关键词：纳米抗体类弹性蛋白融合表达可逆相变循环包涵体复性

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