Yi Sugar, Protein Kinase C On Stz Rats And The Influence Of Sodium Channel Experiment

Diabetic peripheral neuritis belongs to a complication of diabetes mellitus,are chronic neuropathy, the most common complication of diabetes. Because thedisease with hands and feet numbness and pain as the main performance, so alsoknown as the diabetic peripheral neuritis. Peripheral neuropathy in diabetescan occur in the early time. Its pathogenesis is characterized with sensorydisorder symmetries appear, generally lower limb lesions probability more thanthe upper extremities, early manifestations of abnormal sensation, and pain withpain as the main performance of the late. Main show is numb, crawling, ant, hotor cold limbs, sense of electric shock, appear even gloves or socks kind feeling,often accompanied by pain and temperature sensation diminish or disappear.Another kind of expression is the symmetry of the pain and paresthesia, painwith cold pain or burning, stabbing pain is the most common, the degree of painsevere even unbearable, and pain at night will increase.Protein kinase C is a group of phospholipid dependent on Ca2+activatedprotein serine/threonine kinase. In a variety of immune receptor mediatedsignal transduction can be two triglyceride (DAG) and calcium activated. PKCis an important second messenger, the extracellular information transfer intocells, thereby regulating cell growth, contraction, secretion, conduction,membrane permeability, extracellular matrix and gene expression and so on. Inthe diabetic state, these functions are abnormal.Sensory neurons in the voltage-gated sodium channel (VGSCs) in a varietyof peripheral nerve injury caused by chronic neuropathic pain diseases playsan important role, such as trauma, nerve compression, diabetic neuropathy, virusor chemical factors caused by peripheral nerve injury.These peripheral nerveinjury induced neuropathic pain showed spontaneous pain, hyperalgesia, burning pain and allodynia. Some research indicated that, VGSCs blocker to treat theneuropathic pain, such as anticonvulsants, antiarrhythmic agents and localanesthetics.In recent years, there is a great deal of the peripheral nerve injury inducedby spinal dorsal root ganglion (DRG) sodium channel change study, found thatDRG VGSCs is activated, VGSCs type, quantity, distribution andelectrophysiological properties are changed, resulting in DRG increasedneuronal excitability, discharge frequency increases, the generation of ectopicdischarges wait, these are closely related with neuropathic pain.This paper adopts to rats by intraperitoneal injection of STZ modes ofreplication of rat model of diabetes mellitus, again through the Chinese medicineYi Tang Kang oral gavage intervention, on the diabetic model rat nociceptivebehavior in rats, to explore Yi Tang Kang on diabetic rat model of peripheralnerve protective effect; Using immunohistochemistry and RT-PCR method oftraditional Chinese medicine, sugar Kang Gan prognosis of diabetic model ratdorsal root ganglion tissue in PKC, Nav1.7, Nav1.8, Nav1.9protein and mRNAexpression levels were detected, of PKC in diabetes pathogenesis changes inexpression levels and traditional Chinese medicine Yi Tang Kang treats thediabetes peripheral nerve pathological change part mechanism of action.Purpose：Through the experiment study the influence of Yi Tang Kang on STZrat protein kinase C（PKC）and sodium channels Nav1.7，Nav1.8，Nav1.9，and exploresthe therapy effect of Yi Tang Kang on the diabetic peripheral neuropathy andpathological pain。Material and method：Weight of200～250g Wister rats are randomly dividedinto blank control group，the placebo group and Yi Tang Kang group，10ratsare in each group。Blank control group is feeded physiological saline once daily，the placebo group is feeded5ml placebo once daily after STZ mode，Yi Tang Kang group is feeded5ml Yi Tang Kang once daily after STZ modeling，each group isfeeded six weeks. Hot plate experiment detect rats pain behavioral changes, tosure STZ rats have diabetes peripheral neuropathy, produces pathological pain.Using immunohistochemistry and RT-PCR detection methods to detect PKC and Nav1.7,Nav1.8, Nav1.9changes.Results：1. Results of pain behavior experimentThe placebo group3weeks after modeling, began to appear the time of lickingfeet extend, and Yi Tang Kang group did not appear this phenomenon untill4weeks,and in the next weeks of treatment, the time of licking feet gradually declined.Placebo group compared with the Yi Tang Kang group during the3-6weeks, thetime of licking feet was significantly different（P<0.01）.The rate of increased pain threshold of placebo group was significantlyhigher than the rate of Yi Tang Kang group.2. Results of PKC experiment:Immunohistochemical results:, PKC expression was significantlyupregulation in dorsal root ganglia(DRG) of placebo group rats, compared withthe placebo group, protein levels of PKC in Yi Tang Kang group rats weresignificantly downregulation.RT-PCR results: PKC mRNA expression was significantly upregulation in DRGof placebo group rats, compared with the placebo group, the levels of PKC mRNAin Yi Tang Kang group rats were significantly downregulation.3. Results of sodium channels experiment:Immunohistochemical results: Nav1.7, Nav1.8protein expression in DRG ofplacebo group rats was significantly higher than that in the bland control grouprats, and Yi Tang Kang group Nav1.7, Nav1.8protein expression levels aresignificantly lower than placebo group.Nav1.9protein expression in DRG of eachgroup rats was no significant difference.RT-PCR results: Nav1.7, Nav1.8mRNA expression in DRG of placebo group rats was significantly higher than that in the bland control group rats, and Yi TangKang group Nav1.7, Nav1.8mRNA expression levels are significantly lower thanplacebo group.Nav1.9mRNA expression in DRG of each group rats was no significantdifference.Conclusion：1.STZ rats pain threshold increased after three weeks of modeling, that indicateSTZ rats suffer diabetic peripheral neuropathy (DPN) after three weeks ofmodeling. Yi Tang Kang have the effect on lowering pain threshold of STZ rats,that reduce the symptoms of diabetic neuropathic pain.2.PKC in DRG of DPN rats was highly expressed, Yi Tang Kang can downregulatethe expression of PKC, and inhibit PKC activation, that is its role in thetreatment of DPN pathological pain.3.Nav1.7, Nav1.8in DRG of DPN rats was highly expressed, Yi Tang Kang candownregulate the expression of Nav1.7, Nav1.8, may have blocking effect on Nav1.7,Nav1.8channels, which may be one of the mechanisms of Yi Tang Kang improvesymptoms of diabetic neuropathic pain.