光感基因技术对脊髓栓系神经源性膀胱组织及其受体P2X1、P2X3的调控

Application of channelrhodopsin-2(ChR2) technology on tethered cord syndrome and the regulation of receptors P2X1 and P2X3 expression in neurogenic bladder tissue

目的研究光感基因技术对脊髓栓系神经源性膀胱组织及其受体P2X1、P2X3的影响。方法将60只未成年SD雌性大鼠(尿流动力学检查无异常)随机分组,建立脊髓栓系神经源性膀胱动物模型,分为对照组(n=15)、脊髓栓系无光感基因组(n=21)、脊髓栓系有光感基因组(n=21)。HE染色观察膀胱组织的病理学改变,Real time PCR和Western blotting检测膀胱逼尿肌受体P2X1、P2X3在分子和蛋白水平的表达情况。结果在建模过程中,诱发电位监测发现术中有3只动物脊髓受损,其余动物建模成功。与脊髓栓系无光感基因组比较,在HE染色结果中脊髓栓系有光感基因组逼尿肌细胞形态大小相对均一,肌纤维排列较有序,趋近于对照组。Real time PCR和Western blotting检测结果显示,与对照组比较,脊髓栓系无光感基因组P2X1、P2X3 mRNA和相对蛋白表达量均升高,差异有统计学意义(P<0.05);而经蓝光照射一定时间后,脊髓栓系有光感基因组中两种受体mRNA和相对蛋白表达量均降低,与脊髓栓系无光感基因组比较,差异有统计学意义(P<0.05);与对照组比较,差异无统计学意义(P>0.05)。结论光感基因技术对脊髓栓系神经源性膀胱组织及其受体P2X1、P2X3有一定的调控作用。

Objective To study the effects of activation of channelrhodopsin-2(ChR2)on tethered cord syndrome and the regulation of P2X1 and P2X3 expression in neurogenic bladder tissue.Methods Sixty female SD female rats(no abnormalities in urodynamics)were used to establish a tethered neurogenic bladder animal model,and then randomly divided into control group(n=15),tethered cord without ChR2 group(n=21),and tethered cord with ChR2 group(n=21).HE staining was used to observe the pathological changes of bladder tissue.Real time PCR and Western blotting were used to detect the expression of bladder detrusor receptors P2X1 and P2X3 at the mRNA and protein levels,respectively.Results During the modeling process,evoked potential monitoring showed that three animals had spinal cord injury during operation.The remaining animals were successfully established as tethered cord syndrome model.Compared with the tethered cord without without ChR2 group,the photoreceptor detrusor cell morphology of the tethered cord with ChR2 group was relatively uniform in the HE staining results,and the muscle fibers were arranged in an orderly manner,which was close to the control group.The results of Real time PCR and Western blotting showed that the expressions of P2X1 and P2X3 at the molecular and protein levels were higher in the tethered cord without ChR2 group than the healthy control group(P<0.05).After a certain time by blue light stimulation,the expressions of P2X1 and P2X3 levels in the tethered cord with ChR2 group were decreased(P<0.05).Conclusion The ChR2 technology has certain significance for the treatment of tethered cord syndrome with neurogenic bladder.