基于多肽阵列技术构建的细胞穿膜肽靶向抑制炎性痛大鼠NR2B磷酸化

发布时间：2018-05-06 20:42

本文选题：多肽阵列技术 + 炎性疼痛　；参考：《实用医学杂志》2017年19期

【摘要】：目的我们已经基于多肽阵列技术构建了可以靶向抑制Fyn与PSD95相互作用的穿膜短肽Fyn P,本实验旨在验证Tat-LK15运载此短肽抑制炎性痛大鼠NMDA受体3B亚基(NR2B)磷酸化进而治疗炎性疼痛的可行性。方法 Tat-LK15与Fyn P短肽连接形成稳定的细胞穿膜肽Tat-LK15/Fyn P,乱序Fyn P作为对照(Tat-LK15/m Fyn P);然后检测腹腔注射Tat-LK15/Fyn P对完全弗氏佐剂(CFA)炎性痛模型大鼠脊髓背角(SCDH)Fyn与PSD-95的相互作用的影响,并且测定SCDH磷酸化NR2B(p-NR2B)表达水平的变化,另外测定腹腔注射此复合物后炎性痛大鼠机械缩足反射阈值(MWT)及热缩足反射持续时间(TWL)的变化。结果 Tat-LK15/Fyn P注射后3 d,炎性痛模型大鼠SCDH Fyn与PSD-95的结合受到抑制,Fyn P及Tat-LK15/m Fyn P无此作用,炎性疼痛大鼠p-NR2B蛋白水平的表达下降51%(P0.01),Fyn P及Tat-LK15/m Fyn P注射后p-NR2B后的表达无明显变化。Tat-LK15/Fyn P腹腔注射后3、7、14及21 d炎性痛大鼠的MWT显著增高,TWL显著缩短(P0.01),Fyn P及Tat-LK15/m Fyn P无此治疗作用。结论 Tat-LK15可有效运载此短肽抑制慢性炎性痛大鼠Fyn与PSD95相互作用,从而抑制NR2B的磷酸化激活,进而对炎性疼痛大鼠具有治疗作用。
[Abstract]:Objective We have constructed a transmembrane short peptide (Fyn), which can specifically inhibit the interaction between Fyn and PSD95, based on polypeptide array technology. The aim of this study was to verify the effect of Tat-LK15 on the phosphorylation of NMDA receptor 3B subunit NR2B in inflammatory pain rats. The feasibility of treating inflammatory pain. Methods Tat-LK15 was linked with Fyn P short peptide to form a stable cellular transmembrane peptide Tat-LK15/Fyn P, and Fyn P was used as control. The effect of intraperitoneal injection of Tat-LK15/Fyn P on the interaction between PSD-95 and SCDHN Fyn in spinal dorsal horn of rats with complete Freund's adjuvant inflammatory pain was detected. The expression of SCDH phosphorylated NR2BU p-NR2B) was measured, and the changes of mechanical foot reflex threshold (mttt) and thermospasm reflex duration (TWL) after intraperitoneal injection of the compound were also measured in rats with inflammatory pain. Results 3 days after injection of Tat-LK15/Fyn P, the binding of SCDH Fyn to PSD-95 was inhibited in inflammatory pain model rats. Fyn P and Tat-LK15/m Fyn P had no such effect. The expression of p-NR2B protein decreased in inflammatory pain rats. The expression of Fyn P and Tat-LK15/m Fyn P were not changed after p-NR2B injection. The MWT of Tat-LK15 / Fyn P was significantly increased after intraperitoneal injection of Tat-LK15 / Fyn P. The MWT of Tat-LK15 / Fyn P group was significantly higher than that of Tat-LK15 / Fyn P rats and 21 days after intraperitoneal injection of Tat-LK15 / Fyn P, MWT was significantly shorter than that of P0.01Fyn P and Tat-LK15/m Fyn P in inflammatory pain rats. Conclusion Tat-LK15 can effectively transport this short peptide to inhibit the interaction between Fyn and PSD95 in chronic inflammatory pain rats, thus inhibit the activation of NR2B phosphorylation, and thus has therapeutic effect on inflammatory pain rats.
【作者单位】：解放军421医院麻醉科;广州军区广州总医院麻醉科;
【基金】：国家自然科学基金面上项目(编号:81371233) 广东省科技计划项目(编号:2014A020212554)
【分类号】：R402

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