中国比较医学杂志Issue(7)：39-45,7.DOI:10.3969/j.issn.1671.7856.2015.007.009
SiRNA 干扰慢性炎性痛大鼠患侧背根神经节 MrgC表达及 PKCε磷酸化的研究
SiRNA interference on expression of MrgC and phosphorylation of PKCεin ipsilateral dorsal root ganglion of rats with chronic inflammatory pain
摘要
Abstract
Objective To observe the Interference effects of siRNA( small interference RNA) intrathecal injection on the expression of mRNA and protein of MrgC, on PWTs ( paw withdrawal thresholds) and the phosphorylation of PKCεSer729 in ipsilateral DRG( dorsal root ganglion) of rats with chronic inflammatory pain induced by CFA( complete freund’ s adjuvant).Methods 16 health adult male SD (Sprague-Dawley) rats were randomly divided into 2 groups: control siRNA group and MrgC siRNA group, 8 rats in each group.After success of intrathecal catheterization, corresponding siRNA was injected in rats for 4d, once a day, 5μg/d per rat.The model of chronic inflammatory pain was established by CFA (0.1ml per rat) subcutaneously injected into the right hind paw at 4th day post-administration, then two groups were administrated corresponding siRNA on alternate day and executed at the 11th day post-administration.The PWTs were measured at 5 time points of pre-intrathecal catheterization, pre-administration, 4th day post-administration(0h post-CFA injection), 5th day post-administration(24h post-CFA injection), 11th day post-administration(7 d post-CFA injection). The expression of MrgC mRNA in ipsilateral DRG was detected by fluorogenic quantitative PCR, and the expression of MrgC and the phosphorylation of PKCε Ser729 in ipsilateral DRG was detected by immumofluorescence method.Result Compared with 4th day post-administration, PWTs of both two groups at 5th day post-administration decreased significantly ( P<0.01 ) .While there was no significant difference of PWTs between two groups at every detective time point. Compared with control siRNA group, the expression of MrgC mRNA and the rate of MrgC positive cells in MrgC siRNA group both decreased significantly ( P<0.01,P <0.05), whereas the rate of p-PKCε Ser729 positive cells increased obviously ( P<0.05) at 11th day post-administration.Conclusion MrgC siRNA can effectively interfere the expression of mRNA and protein of MrgC in L4-L6 ipsilateral DRGs of rats with chronic inflammatory pain induced by CFA, and the siRNA interference on MrgC can obviously up-regulate the phosphorylation of PKCε Ser729, while it has no significant effect on PWTs of rats.关键词
小干扰RNA/MrgC/蛋白激酶Cε亚基Ser729磷酸化/慢性炎性痛/背根神经节Key words
SiRNA/MrgC/Phosphorylation of PKCεSer729/Chronic inflammatory pain/Dorsal root ganglion分类
医药卫生引用本文复制引用
林小溪,方芳,方剑乔,刘盈君..SiRNA 干扰慢性炎性痛大鼠患侧背根神经节 MrgC表达及 PKCε磷酸化的研究[J].中国比较医学杂志,2015,(7):39-45,7.基金项目
国家自然科学基金资助项目（81202755）；浙江省医药卫生科技计划（2015KYA172）。 （）
