TMEM16A And TMEM16B Expression On DRG Neurons In CCI Model Rat

Objective:To compare different concentration NFA intervention after CCI model rat,CCI model rat and normal rat of Thermal Withdrawal Latency(TWL)changes; TMEM16 A and TMEM16 B levels of RNA and protein expression changes in DRG neurons.Aimed to explore mediation role of Ca CCs in Neuropathic pain.Methods:After making the CCI model,hot plate test was used to detect thermal withdrawal latency(TWL). Respectively 1 d before surgery,postoperative 1,3,5,7,9,12, 14 d different concentrations of the NFA intervention neuropathy rational pain rat CCI model,CCI pyrocondensation model rats and normal rats foot reflex latency detection. Immunofluorescence assay was used to show the distribution of TMEM16 A and TMEM16 B on the DRG neurons;RT-PCR and Western Blot techniques were used to detect the expression of m RNA and protein of TMEM16 A and TMEM16 B on DRG neurons of CCI model after intervented by different concentrations of NFA.Results:(1)TWL of CCI group was significantly shorter than normal,after giving 10μM,50μM and 300μM NFA,the TWL of each groups were longer than the CCI group(P<0.01,n=6);the TWL of 50μM NFA intervented group was significantly longer than the 10μM NFA interved group(P<0.05,n=6);but there were no statistical differences between the 300μM and 50μM NFA interved groups.(2) Immunofluorescence showed TMEM16 A were mainly in the DRG neurons.(3) m RNA level of TMEM6 A in CCI group was significantly higher than normal group. As the increasing of concentrations of NFA,m RNA levels of TMEM16 A on DRG neurons decreased gradually compared with CCI group,the difference has statistically significane(P<0.01,n=6);m RNA level of TMEM16 B in CCI group was significantly decreased than normal group. As the increasing of concentrations of NFA, m RNA levels of TMEM16 B on DRG neurons higher gradually compared with CCI group,the difference has statistically significane(P<0.01, n=6).(4)Expression of TMEM16 A in CCI group was significantly higher than the normal group; as the increasing of concentrations of NFA,expression of TMEM16 A protein on DRG neurons gradually reduced compared with CCI group,the difference has statistically significance(P<0.01,n=6).Expression of TMEM16 B in CCI group was significantly reduced than the normal group; as the increasing of concentrations of NFA, expression of TMEM16 B protein on DRG neurons gradually higher compared with CCI group, the difference has statistically significance(P<0.01, n=6).Conclusion: Niflumic acid can change the expression of TMEM16 A and TMEM16 B on DRG neurons of neuropathic pain rats.which prompt that TMEM16 A and TMEM16 B may be involved in the occurrence or the regulation of neuropathic pain.