| Inflammatory pain is a kind of pathological pain,refers to the noxious stimulation after removal of persistent pain also pathological phenomenon,which is one of the most common diseases in the clinical type of pain,affecting people's quality of life seriously.Due to inflammatory pain itself repeatedly in the course,the treatment is very difficult.So the mechanism of the peripheral and central nervous system is the focus to research.At present,the clinical treatment of inflammatory pain by peripheral sensitization drugs inhibit:the first is the NSAIDs,during the inflammation,through the inhibition of cyclooxygenase activity,reduce prostacyclin production,relieve inflammatory to loss the pain.But,which can cause gastrointestinal discomfort?renal dysfunction and other side effects;Another is opiates,generated by itself is widely distributed in the peripheral tissues of immune cells,will increase in the number of parts of opioid receptors on primary neurons of the inflammatory state,system or local given opioid compounds some can reduce inflammatory pain,but its reward effect on the nervous system or strengthening effect caused by addiction;Finally the cannabinoids in models of inflammation,local administration of cannabinoid receptor agonists can activate CB1 receptor activation produces analgesia passive excitatory adenosine cyclase by primary afferent fibers.But it will cause Neurotoxicity.In recent years,drugs for central sensitization treatment such as norepinephrine reuptake inhibitors,serotonin and other antiepileptic drugs in five are exposed to the inadequacy of their treatment of inflammatory pain,therefore seek more safe,effective and convenient treatment is imperative,clinical practice and experimental studies demonstrated that acupuncture for inflammatory pain can play a good analgesic effect,but the exact mechanism is not clear.Generally,inflammatory pain and the relationship between ion channels and receptors,neurotransmitter are important ideas to study on acupuncture treatment of inflammatory pain.At the same time,new research confirms that nociceptive calcium and voltage-gated calcium channels in neurons of the free state played an important role in the peripheral and Naka Min in the process,and CaBPs by the special EF type specific binding with free calcium ion concentration and internal induced changes in cell signaling,which play the effect of pain relief.In order to play in the process of EA for this study by complete Freund's adjuvant(CFA)model rats as the research object,the integrated use of behavior,morphology,in-depth study of EA on inflammatory pain in a CFA-rat model of spinal ganglia and spinal dorsal horn in the CaBPs affect the molecular biology and physiology of EA by combining the expression of proteins to change the internal calcium concentration of neurons and cells in calcium regulated signal transduction pathways,regulate neurotransmitters such as CGRP,the number of release and postsynaptic NMDA receptor Glu model,and ultimately improve the central sensitization of spinal dorsal horn,providing theoretical basis for revealing the mechanism of acupuncture analgesia.Purpuse:To discuss the analgesic effect of EA on Inflammatory pain of rats' ST3 6 and neuronal CaBPs expression in spinal cord and DRGs,explain the mechanism of the analgesic,and provide experimental basis for clinical application.Methods:64 male SD rats,were randomly divided into Control.Nacl.CFA.CFA&EA groups,16 rats in each group.The Nacl group left subcutaneous were injected with 50ul 0.9%Nacl;Group CFA and CFA&EA were injected with 50ul CFA in left of the hind paws to induce localized inflammatory pain,48h.CFA&EA were followed by an optimal EA treatment protocol(20/100Hz,1-3 mA,30 min)at 11:00 and 12:00 a.m post-CFA in conscious isoflurane anesthesia aerosol rats.The EA was conducted at acupuncture point ST36.The pain-threshold changes were measured by paw withdrawal latency(PWL)to a noxious thermal stimulus and assessed by mechanical stimulation and thermal stimulation.The changes of pain threshold after modeling and after acupuncture in three stages were plantar rats,Rats were killed immediately,with 4%formaldehyde perfusion or were directly sacrificed respectively by Fluorescence Immunohistochemical method and Western Blot method.The expression of CaBPs in the DRG and dorsal horn of spinal cord in rats was detected.Results:1 Behavioral changes of rats in each group after EAHeat pain threshold:CFA and Nacl group were performed either with the group or internal groups are basically the same in the three phase experiments;CFA group and CFA&EA group after EA compared with CFA group and Nacl group are basically the same,the decline in the CFA&EA group(P<0.05);in the CFA group increased in comparison after EA and CFA&EA group(P<0.05).The mechanical threshold:CFA group and Nacl group,in the three phase experiments were performed either with the group or internal groups have differences but without statistical significance.Before electric acupuncture,CFA group and CFA&EA group compared with CFA group and Nacl group,the pain threshold decreased(P<0.05);after EA,the CFA&EA group compared to the CFA group value increased(P<0.05)2 Expression changes of PV,CB and NECAB1/2 positive neurons in the spinal ganglia after EAPV,CB and NECAB1/2 positive neurons in the Control group,Nacl group,the change trend of the expression of CFA and CFA&EA groups are basically the same,Control group and Nacl group had no obvious difference,CFA group compared with Control group and Nacl group compared with the expression(P<0.05),CFA&EA group and CFA group compared with the expression(P<0.05);at the same time,the number of CGRP positive cells were quantitatively found,CFA group compared to Control group and Nacl group had mild upregulation of CFA&EA,The expression of EA in group was lower than that in group CFA,and the number of CGRP positive cells in the four groups was better than that in group(Control 30.1±3.4,Nacl 28.3±9.4,CFA 45.0±6.7,CFA&EA 33.5 ±8.7).On PV,CB and NEC AB 1/2 positive neurons were counted,there are about 70?80%in DRG neurons are small neurons associated with pain related,and co expression and have different degrees of pain and classical neurotransmitter CGRP in each group of the(PV:1.1±3.76%,CB:1.2±10.5%),NECAB12 and CGRP co expression(NECAB1:28± 6.17%,NECAB2:16.2±3.94%).3 Changes of PV,CB and NECAB1/2 positive expression in spinal cord dorsal horn before and after EAIn the Control group,Nacl group,CFA group and CFA&EA group,the four groups in the spinal dorsal horn of PV,CB,NECAB1/2 positive expression trend of material are basically the same,and there are a lot of distribution in the pain associated with the superficial layers of the spinal dorsal horn,and Four kinds of basic protein and CGRP positive fiber co expression.Based on a large and equal magnification in the dorsal horn of CaBPs peripheral neurons across the ganglion over the projection of CGRP positive fibers were found in statistics in Nacl group,Control group,CFA group and CFA&EA group in the basic trend showing consistency(Control 49.5±5.8,Nacl 50.4±4.4,CFA 75.9±7.7,CFA&EA 48.7±6.7),Control group and Nacl group did not change significantly,CFA group compared with Control group and Nacl group compared with the expression(P<0.05),CFA&EA were down regulated compared with the CFA group(P<0.05).The results of WB showed that PV,CB and NECAB1/2 content in the dorsal horn of the spinal cord in the Control group,Nacl group,consistent changes of CFA group and CFA&EA group shows same trends,Control group and Nacl group showed no significant change;in the model,compared with CFA group and CFA&EA group in Control group and Nacl group protein content decreased(P<0.05);CFA&EA group of EA intervention in protein content increased(P<0.05).Conclusion:This study discusses the analgesia mechanism of acupuncture for CFA-induced inflammatory pain through behavioral science,chemistry and molecular biology.In the process of the EA can effectively up-regulate the protein expressions of PV,CB and NECAB1\2,enhancing the numbers of these cells in DRGs and spinal dorsal horn in CFA rats and make the neuropeptide expressions with quantity increase.Four kinds of CaBPs is mainly done by intracellular calcium ion buffer power for the analgesic action of inflammatory pain,But the relationship between CaBPs and neuropeptides further studies are needed in the process of acupuncture analgesia... |
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