摘要
目的探讨脊髓原钙粘蛋白20（PCDH20）在大鼠骨癌痛形成中的作用。方法SPF级雌性SD大鼠36只，体重180～200g，采用随机数字表法，将大鼠随机分为4组（n=9）：假手术组（S组）、骨癌痛组（BCP组）、慢病毒对照组（LC组）、PCDH20siRNA-LV组（P组）。LC组和P组分别于脊髓内注射对照慢病毒和PCDH20 siRNA慢病毒4μl。1周后建立骨癌痛模型，采用右侧胫骨上段骨髓腔内注射Walker256乳腺癌细胞的方法制备。于病毒注射前1d（T1）、骨癌痛模型建立前1d（T2）、模型建立后7、14和21d（T3、T4、T5）时测定机械缩足阈值（MWT）。骨癌痛模型建立后21dMWT测定结束后，取术侧胫骨，HE染色后光镜下观察胫骨癌细胞侵犯情况；取脊髓组织，采用免疫组化法和Western blot法测定脊髓PCDH20和突触后致密物蛋白质95（PSD95）蛋白的表达水平，采用RT-PCR法测定脊髓PCDH20和PSD95mRNA的表达水平。结果BCP组、LC组及P组肿瘤细胞突破骨髓腔，侵犯骨皮质，骨皮质结构严重破坏；与S组比较，BCP组、LC组和P组B～L时MWT明显降低，BCP组和LC组脊髓PCDH20和PSD95mRNA及其蛋白表达水平明显上调，P组脊髓PCDH20蛋白水平上调（P〈0．05）；与BCP组比较，LC组各时点MWT、脊髓PCDH20和PSD95mRNA及其蛋白表达水平比较差异无统计学意义（P〉0．05），P组Td和B时MWT明显升高，脊髓PCDH20和PSD95mRNA及其蛋白表达水平下调（P〈0．05）。结论PCDH20通过调控大鼠脊髓PSD95的表达，调节兴奋性突触的功能，从而参与骨癌痛的形成。
Objective To investigate the effects of protocadherin 20 （PCDH20） in the spinal cord in the development of bone cancer pain （BCP） in rats. Methods Thirty-six SPF female Sprague-Dawley rats, weighing 180-200 g, were randomly divided into 4 groups （n = 9 each）： sham operation group （group S）, BCP group, lentivirus control group （group LC） and PCDH20 siRNA lentivirus group （group P）. Control lentivirus and lenti- virus containing PCDH20 siRNA 4 μl were injected into the ipsilateral spinal cord in groups LC and P, respectively. One week later, BCP was induced by injection of Walker 256 breast cancer cells into the upper segment of bone marrow of right tibia. The mechanical paw withdrawal threshold （MWT） was measured at 1 day before injection of lentivirus （TE ）, 1 day before BCP （T2 ）, and 7, 14 and 21 days after BCP （T3-5 ） . Three rats in each group were sacrificed after measurement of the MWT at 21 day after BCP and the tibia on the operated side was obtained for examination of invasion of the cancer cells with light microscope. The spinal cord was removed for determination of the expression of PCDH20 and postsynaptic density 95 （PSD95） protein （by Western blot） and mRNA （by RTPCR）. Results In groups BCP, LC and P, the cancer cells grew out of the bone and destroyed the cortical bone seriously. Compared with group S, the MWT was significantly decreased at T3-5 in groups BCP, LC and P, the ekpression of PCDH20 and PSD95 protein and mRNA was up-regulated in groups BCP and LC, and the expression of PCDH20 was up-regulated in group P （P 〈 0.05）. Compared with BCP group, no significant change was found in the MWT and expression of PCDH20 and PSD95 protein and mRNA in group LC （ P 〉 0.05）, and the MWT was significantly increased at T4.s and the expression of PCDH20 and PSD95 protein and mRNA was down-regulated in group P （ P 〈 0.05）. Conclusion PCDH20 is involved in the development of BCP through regulating the expression of PSD95 in the spinal cor
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2012年第12期1445-1448,共4页
Chinese Journal of Anesthesiology
基金
基金项目：国家自然科学基金（81070890,30872441）
