摘要
目的:利用鞘内注射前强啡肽mRNA的反义寡聚核苷酸,观察由福尔马林引起的大鼠后脚炎性水肿程度的变化,来证明脊髓背角中强啡肽(dynorphin,Dyn)A的量可能影响外周炎症反应的大小。方法:在3组成年Wistar大鼠(每组5只)预先在鞘内分别注入对抗前强啡肽mRNA的反义寡聚核苷酸(AS-ODN)、生理盐水(NS)、和反序的反义寡聚核苷酸(RS-ODN);4h后,再给各组动物一侧后脚掌皮下注射福尔马林(0.5%,100μl),并分别在注射后的1h、2h、3h三个时间点,各以5只动物先检查后脚掌直径,以福尔马林注射前的直径为对照,算出直径增加的百分数,作为炎性水肿变化的指标,并在脚掌检测后立即处死动物作免疫组化检测,确定各时间点大鼠两侧脊髓背角浅层中DynA(1—8)免疫反应样物质的光密度值。结果:①3组动物在福尔马林注射后的3个时间点,注射侧后脚掌的炎性水肿均呈进行性的加重,但在鞘内注入AS-ODN的实验组,各个时间点的水肿程度均明显地小于注射了生理盐水和RS-ODN的两个对照组;而在两个对照组相应的各时间点之间无显著性差异。②在免疫组化检测中,可看到三组动物注射侧脊髓背角中都出现DynA(1-8)的光密度值随测定值时间推后而增加,实验组各时间点的测定值均明显地小于两个对照组相应时间点的测定值,但后两组的各相应时间点的测定值之间无显著性差异。结论:鞘内注射反义核苷酸后福尔马林引起的脊髓中强啡肽A的表达上调被阻止,并且平行地出现了局部炎性水肿发展的被阻止。
Objective: We utilized intrathecal (i.t.) application antisense oligedeoxynucleotide (AS-ODN) to induce prodynorphin (Pro-Dyn) mRNA and observed the degree of inflammatory edema after the formalin injection in rats to prove whether the quantity of dynorphin A in nerve centre could effect the degree of periphery inflammation. Method: The rats were pretreated with i.t. application of AS-ODN, normal saline (NS) or the reverse ODN (RSODN) separately. The rat's paw was injected with formalin and observed the incremental percentages of diameter difference(%) combined with the immunocytochemical evidences that intensified expression of dynorphin A(1-8)(Dyn A)(OD units) in spinal dorsal horn after the formalin injection in 1h,2h and 3h time slices.Resuit:①All the injected paws showed inflammatory edema with a progressive aggravation in three inspect times after the formalin injection, at the same time interval. The formalin induced incremental percentages of diameter differences of injected paws in i.t. AS-ODN group were smaller than that in i.t NS or i.t RS-ODN group but there were no differences in i.t NS group and in i.t RS-ODN group. ②The up-expression of Dyn A in ipsilateral dorsal horn to the formalin injection increased gradually follow the formalin injection in all groups of all inspect times, and there were markedly reducer in i.t. AS-ODN group than that in i.t NS or i.t RS-ODN group but there were no differences in i.t NS group and i.t RS-ODN group in all inspect times.Conclusion: Application of AS-ODN induced prodynorphin mRNA could suppress the increase expression of Dyn A in the spinal dorsal horn along with prevention on the local inflammatory edema.
出处
《中国康复医学杂志》
CAS
CSCD
北大核心
2006年第6期510-513,共4页
Chinese Journal of Rehabilitation Medicine
关键词
前强啡肽
反义寡聚核苷酸
强啡肽A
炎性水肿
脊髓背角
大鼠
prodynorphin
antisense oligodeoxynueleotide
dynorphin A
inflammatory edema
spinal dorsal horn
rat