左旋千金藤啶碱D1激动作用增加6-OHDA损毁大鼠的DARPP-32磷酸化效应

为了进一步阐明SPD对大鼠纹状体突触后D1受体的激动作用特性,本文应用反磷酸化在体内测定及放射配体结合方法,分别观察SPD对6-OHDA损毁大鼠纹状体DARPP-32体内磷酸化作用及突触后D1受体密度的影响.结果表明:皮下给予SPD(20,40 mg/kg,21 d),损毁侧纹状体DARPP-32体外32P]的掺入量较健侧下降50%(P<0.01).换言之,损毁侧纹状体内DARPP-32的磷酸化程度增加了.然而,SPD使损毁导致D1受体上调的作用减弱(Bmax 从385.0±26.1 fmol/mg 降至319.7±20.1 fmol/mg水平).因此,SPD激动D1受体,使6-OHDA损毁大鼠纹状体内DARPP-32磷酸化作用加强,而受体密度减少.这是SPD调节脑内D1受体信号转导功能的重要机制.

Increased phosphorylation of DARPP-32 by D1 agonistic action of l-stepholidine in the 6-OHDA-lesioned rat striatum

In order to explore the characteristics of l-stepholidine (SPD) activating the postsynaptic D(1) receptors, the effects of SPD on DARPP-32 phosphorylation in vivo with back-phosphorylation assay and on the postsynaptic D(1) receptor densities with radioligand assay were observed in the striatum of 6-OHDA-lesioned rat. The results showed that following subcutaneously administration of 20 or 40 mg/kg SPD for 21 d, (32)P phosphate incorporation into the DARPP-32 protein in the denervated striatum showed a 50% reduction (P<0.01) vs the intact striatum, indicating an increase of DARPP-32 phosphorylation in vivo in the denervated striatum. However,the D(1) receptor B(max) was decreased from 385.0+/-26.1 to 319.7+/-20.1 fmol/mg protein. It is suggested that D(1) agonist action of SPD decreases the D(1) receptor density but increases the phosphorylation of DARPP-32 in the striatum of 6-OHDA-lesioned rat, which may be responsible for the regulation of D(1) receptor signal transduction in brain neurons.