ABSTRACT
Objective:To explore the possible mechanism of electroacupuncture to improve detrusor hyperreflex after suprasacral spinal cord injury. Methods:A total of 60 female Sprague-Dawley rats were included. According to the random number table, twelve were selected as the blank group, twelve as the sham operation group, and the remaining 36 were made neurogenic bladder models using modified T10 spinal cord transection. After that, twelve of them were randomly selected as the model group and twelve were as the electroacupuncture group from the model rats that met the requirements. On the 19th day after modelling, Ciliao (BL32), Zhongji (RN3) and Sanyinjiao (SP6) were taken for electroacupuncture. After seven days of continuous treatment, urodynamic testing was performed, content of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in detrusor was determined by ELISA, and the level of phosphorylation of myosin light chain kinase (p-MLCK) of detrusor was determined by Western blotting. Results:Compared with the blank group and the sham operation group, the maximum bladder capacity and bladder compliance significantly reduced (P < 0.01), and the base pressure and leakage point pressure of bladder significantly increased (P < 0.01); the content of cAMP and PKA in detrusor reduced (P < 0.01), p-MLCK in detrusor reduced (P < 0.05) in the model group. Compared with the model group, the maximum bladder capacity and bladder compliance increased (P < 0.01), the base pressure of the bladder and the pressure at the leak point decreased (P < 0.05); the contents of cAMP and PKA protein in detrusor increased (P < 0.05), the p-MLCK in detrusor increased (P < 0.05) in the electroacupuncture group. Conclusion:Electroacupuncture at Ciliao, Zhongji and Sanyinjiao points could improve the bladder function of rats with detrusor hyperreflex after complete spinal cord injury, and its mechanism may be related to up-regulating the expression of cAMP and PKA, phosphorylating and inactivating p-MLCK, which promote relaxation of detrusor.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the inhibitory effect of adenovirus-mediated interleukin-24 (AdVIL-24) in conjunction with ionizing radiation on the growth of CNE-2Z human NPC cells in vitro and in vivo and underlying mechanisms.</p><p><b>METHODS</b>The CNE-2Z cells were transfected with AdVIL-24 alone or combined with radiotherapy. The transfection efficacy of AdVIL-24 in CNE-2Z cells was determined by RT-PCR and Western blot. Cell growth was assessed by MTT assay, apoptosis was detected by flow cytometry through double staining of cells with propidium iodide (PI) and the expressions of P21, P27, cyclin E, CDK2, Bax, Bcl-2 and caspase-3 were detected with semi-quantitative RT-PCR and western blot, respectively. Anti-tumor effect of AdVIL-24 was observed using CNE-2Z human nasopharyngeal carcinoma transplanted tumor in athymic nude mouse model. The volume and weight of the xenografted tumors were measured and the expressions of P21, P27, cyclin E, CDK2, Bax, Bcl-2, caspase-3, CD34 and VEGF and the microvessel density in xenografted tumors were determined by immunohistochemistry analysis.</p><p><b>RESULTS</b>AdVIL-24 plus radiotherapy induced cell growth inhibition (P < 0.05, Q3d, 4d = 0.916, 1.050) , cell cycle G1 phase arrest(50.37%, P < 0.05, Q = 1.042) and apoptosis (48.82%, P < 0.05, Q = 1.042) , substantial up regulations of P21, P27, the ratio of Bax/Bcl-2 and cleaved caspase-3 and down regulations of cyclin E and CDK2 (P < 0.05, QP21 = 0.959, QP27 = 0.956, Qcyclin E = 1.078, QCDK2 = 1.046, QBax/Bcl-2 = 0.995) in vitro. In the xenografted tumors, AdVIL-24 plus radiotherapy induced cell growth inhibition (P < 0.05, Qvolume14 = 1.053, Qweight = 1.004) , apoptosis (P < 0.05, Q = 0.974) , substantial upregulation of P21, P27, the ratio of Bax/Bcl-2 and cleaved caspase-3 and downregulation of cyclin E and CDK2 (P < 0.05; QP21 = 0.920, QP27 = 0.937, QcyclinE = 1.060, QCDK2 = 1.019, QBax/Bcl-2 = 0.982, Qcleaved-Caspase-3 = 0.927) , decreased the tumor vessel CD34 expression and microvessel density. AdVIL-24 potentially blocked the radiation-induced enhancement of VEGF.</p><p><b>CONCLUSION</b>AdVIL-24 gene therapy combined with radiotherapy may be a novel and effective treatment strategy for human NPC.</p>