The Expression Of TRAP220,PMLC And MLCK In Brain Tissue Of Pitients With Refractory Epilepsy And Clinical Significance

[Purpose] TRAP220 (thyroid hormone receptor- associated protein) functions as a coactivator for nuclear receptors and stimulates transcription by recruiting the TRAP mediator complex to hormone responsive promoter regions. Thus, TRAP220 enhances the function of thyroid/steroid hormone receptors such as thyroid hormone and oestrogen receptors. This study investigated the expression of TRAP220 mRNA and protein level in epileptic brains comparing with human control.[Methods] We examined the expression of TRAP220 mRNA and protein levels in temporal lobes from patients with chronic pharmacoresistant epilepsy who have undergone surgery.[Results] Expression of TRAP220 mRNA and protein was shown to be decreased significantly in the temporal cortex of the patients with epilepsy. cDNA Microarray result showed that the cy5/cy3 value of TRAP220 was 0.181. FQ-PCR result showed mean value of correspondence gene copy value of TRAP220/18s in epileptic group is 10.56×10-5, and 53.49×10-5 in the control group. Western blot gray scale was0.5963±0.218 3(±s) in intractable epilepsy and 0.9933±0.130 2(±s) in control (p<0.05);Immunohistochemistry OD value 0.3068±0.1081(±s)in intractable epilepsy and 0.6825±0.1576(±s) in control (p<0.05).[Conclusions] Our work showed that a decrease in TRAP220 mRNA and protein levels may be involved in the pathophysiology of epilepsy and may be associated with impairment of the brain protective effect and congtive ability. [Purpose] To learn Phosphorylation of Myosin light chain(P-MLC) and Myosin Light Chain Kinase(MLCK) protein expressional levels in temporal lobe and hippocampus tissue of patients with refractory temporal lobe epilepsy and the role it plays in mossy sprouting.[Method] Immunohistochemistry and immunofluorescence were used to check P-MLC and MLCK protein levels in refractory temporal lobe epilepsy, comparing to the control group.[Result] Immunohistochemistry results showed the protein level of P-MLC up-regulated obviously in refractory epileptic temporal lobe, the optical density value is 0.3461±0.1270 in refractory epilepsy and 0.1904±0.0267 in the control group(P<0.05). In epileptic hippocampus tissue, the optical density value is 0.4134±0.0227 and 0.0955±0.0605 in the control group (P<0.05). The similar result also got in immunofluorescence, the fluorescence intensity value in epileptic temporal lobe is 32.408±1.5244 and 11.254±1.4060 (P<0.05) in the control group (P<0.05); the fluorescence intensity value in epileptic hippocampi is 29.428±1.5704 and 9.53±0.8856 (P<0.05) in the control group (P<0.05). Immunohistochemistry and immunofluorescence results showed that there is no obvious changes of MLCK protein level between the epileptic group and the control group.[Conclusion] The function of P-MLC and MLCK are associated with mossy fiber sprouting closely, P-MLC protein level up regulated in refractory temporal epilepsy, but there is no changes of MLCK. It is indicated that the dynamic balance between MLCK and dephosphorylating enzyme was broken is likely the reason of P-MLC up-regulation, and P-MLC up-regulation play an important role in mossy fiber sprouting process.