| Purpose:Using streptozocin one-time injection inducediabetes peripheral neuropathy model rats,through observation of the“Adjust Zang DredgeMeridian”electric acupuncture's effect on diabetic peripheral neuropathy model rats' thermal sensitivity,nerveconduction velocity,the ultrastructure of pathomorphology,nerve cells apoptosis ratio and GRP78 ?Caspase-12 protein,to analyze the correlation between those changes and the level of endoplasmic reticulum stress(ERS),and thus to indicate the mechanism that “Adjust Zang Dredge Meridian” electric acupuncture can protect rats' nerve from apoptosis.Methods:86 rats were used in the experiment,5 healthy rats of which as normal control group.The diabetic rat-model was built through one-time injection of STZ,and then divided into 3 groups randomly,i.e.,model control group,electroacupuncture intervention group and medicine intervention group.In the same circumstance,maintenance feeding was given to the normal control groupand onerat was grabbed randomly each time as normal control observation.Electroacupunctureintervention group received the intervention of “ Adjust Zang DredgeMeridian”electroacupuncture.Medicineintervention group received mecobalamine intervention.After models are built successfully,electroacupuncture and drug interventionare carried out.Monitor rats' general conditions,weight and blood glucose changes.Thermal sensitivity was detected before building,when model succeed,after 4 weeks and 8 weeks of intervention.Nerve conduction velocity was detected before intervention,after 12 weeks of intervention.We detected ultrastructure of sciatic nerve pathological morphology with Electron Microscopyafter 12 weeks of intervention.Detected adopting nerve cells apoptosis ratio with TUNEL staining techniques,and detected GRP78?Caspase-12 protein level of DPN rats with immunofluorescence technique and Western Blot technique.Results:1.Modeling of diabetic rats and changes of related indicators1.1 General condition of experimental rats: the DM modeling rate was 97.5%.Rats in normal control group were generally in good health condition,including color and gloss,reaction speed,drinking and feeding.Rats in normal control group showed poor color and gloss,slow reaction and obvious polydipsia,polyphagia,polyuria characters.Rats in intervention groups showed fair color and gloss,relatively slow reaction andpolydipsia,polyphagia,polyuria characters.1.2 Weight changes:Rats in normal control group showed steady weight growth,while 3modeling groups showed significant weight loss after model success.After 12 weeks of intervention,weight loss in experimental control group was significant losscompared to that of when model success(P<0.01).Rats of electroacupuncture intervention group and medicine intervention group showed significant weight loss after modeling.In the progression of disease course,we detected an upward weight change.After 4 weeks of intervention,that change was detected in eletroacupuncture intervention group,and 8weeks in medicine intervention group(P<0.05).After 4 weeks of intervention,rats weight in electroacupuncture intervention group was significantly higher than that of medicine intervention group(P<0.05),but there was no significant differences between them after8 weeks and 12 weeks of intervention(P>0.05).1.3 Glysemia: when model succeed,rats' random glysemialevel in experimental control group,electroacupuncture intervention group and medicine intervention group were significantly higher than that in both normal control group and before modeling(P<0.01).In the progression of disease course,we detected a upward trend in glysemia level in those 3groups.After 12 weeks intervention,there was no significant differences detected in those 3groups(P>0.05).1.4 Thermal sensitivity results: In the progression of disease course,a significant fall of thermal sensitivitywas detected.Comparison among groups:no significant difference of thermal sensitivity was detected before and after modeling,after 4W and 12 W of intervention,thermal sensitivity of intervention groups were higher than that of model group(P<0.01).After 4W of intervention,thermal sensitivity of electroacupuncture intervention group and medicine intervention showed no apparent diffrence.After 12 W of intervention,thermal sensitivity in medicine intervention group was significantly lower comparedto that of electroacupuncture intervention group(P<0.01).In-groupcomparison:Nosignificant difference was detected before and after modeling.After 4W and 12 W of intervention,thermal sensitivity in model control group and intervention groupswere showed significantfall than that before modeling and model success(P<0.05).2.Resultof tibial nerve conduction velocity test: after modeling,there was nosignificantdifference of Sensory nerve conduction velocity(SCV)and Motor nerve conduction velocity(MCV)detected between all the groups.Comparison among groups after 12 weeks intervenion:both electroacupunctureintervention and medicineintervention could significantly improve SCV ? MCV(P<0.01).Compared with medicine intervention group,electroacupuncture intervention groupfunctioned better,but not significant(P>0.05).In-group comparison: compared with before the intervention,SCV?MCV level weresignificantly lower(P<0.01).3.After 12 w of intervention,sciatic nerves were taken and detected in 1500 time stransmission electronmicro scope.Structural integrity myelin sheath of myelinated nerve fiber of normal group rats sciatic nervepackage around axons,electron density uniform inside the nervous axon;sciatic nerve of model control group demyelinating changed,layer seriously separated.Thecorrespondent change in eletroacupuncture intervention group and medicine intervention group were reduced,with layer osteoporosis happening in certain nerve fiber.4.After 12 w of intervention,we detected a large number of TUNEL positive cellsaround sciatic nerve of model group.The amount of TUNEL positive cells of electroacupuncture intervention group and medicineintervention groupwerein between normal control group and model control group.Compared with the normal control group,the proportion of apoptosis cells were significantly increased in model control group,the eletroacupuncture intervention group and medicineintervention group(P<0.01).Compared to model control group,the proportion of apoptosis cells significantly decreased in eletroacupuncture intervention group and medicineintervention group(P<0.01).The proportion of apoptosis cellsshowed no significant difference between the twointervention groups(P>0.05).5.After 12 w of intervention,immunefluorescence tests revealed:compared with the normal control group,GRP78 and Caspase-12 fluorescence of sciatic nerve in model control group significantlyenhanced.Compared with themodel control group,GRP78 and Caspase-12 fluorescenceof sciatic nerve in eletroacupuncture intervention group and medicine intervention group were significantly decreased,with no obvious differences between the two interventiongroups.After 12 w of intervention,the expression of protein GRP78 and Caspase-12 of sciatic nerve in model control group wasobviously enhancedcompared to the normal control group(P<0.05).Compared with themodel control group,the expression ofprotein GRP78 and Caspase-12 of sciatic nerve in the twointervention groups were significantly decreased(P<0.05),no significant difference between the two intervention groups.Conclusion:1.STZ one-time injection method was employed in the experiment to ensure the high success modeling rate.Diabetic modeling rats showed similar symptoms with human being,and showed significant decrease in weight and thermal sensitivity.“Adjust Zang Dredge Meridian” electric acupuncture has asignificant effect on increasing the weight and thermal sensitivity of DM ratsand DPN rats,while has no significant effect on blood glucose falling.2.“Adjust Zang Dredge Meridian” electric acupuncture has asignificant effect on improving tibial nerve conduction velocity.3.“Adjust Zang Dredge Meridian” electric acupuncturesignificantly alleviates the pathological injury and facilitates the remediation of sciatic nerve,thus to play a role of protection and intervention.4.“Adjust Zang Dredge Meridian” electric acupuncture has an significant effect on decreasing nerve cells apoptosis ratio of DPN rats and restraining ERS,thus to protect.5.“Adjust Zang Dredge Meridian” electric acupuncture has an significant effect on reducing GRP78 ? Caspase-12 protein,which are the iconic protein of sciatic nerve ERS,through which way to restrain ERS and protect. |
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