A Study Of The Functions Of PKC And MAPK Signal Transduction Pathways In Regulating The Gastric Motility Of SD Rats Through Electro-acupuncturing At The "zusinli"

In its physiologic condition,the mechanism of intracellular signaltransduction for digestive smooth cell contraction comprises ofcalcium-mediated and non-calcium-mediated. Transduction of calcium-mediatedsmooth cell information, which plays an important role in the modulationmechanism,，It is reported calcium and inositol thiphosphate are obviouslyincreased in anrum smooth cell of rabbit by electro-acupuncturing at“Zusanli”[1-2],Electro-acupuncturing presents regulation of gastric movement bycalcium-mediated intracellular information transduction.,is relatively clear, yetnot the only factor influencing smooth cell contraction.。It testifies G proteincoupled calcium sensitivity modulation mechanism, by means of disproportionof regulatory protein light chain for myosin between calcium andphosphorylation.， Studies in recent years show that in the thin filament-related proteins regulation mechanism, a new type of protein kinase C (PKC) canregulate the important stages in the MAPK signal transduction pathwaysthrough the PKC isozyme, i.e. ERK (extracellular-signal regulated proteinkinase)’s activity[3]stimulates PKC—MAPK to make the actin thinfilament-related protein, which is in the form of calmodulin-binding protein(CaD) and resembling troponin (CaP) phosphorylation, slide and the smoothmuscle cells contract[3-5]. As the upstream kinase of MAPK, PKC can regulatethe gastric movement by stimulating P44/42MAPK pathways[6-9]。Some otherexperiments have reported that acupuncturing at stomach meridian can makegastric mucosa PKC activity increase[68],The experiment proves that it is thesame with the non-calcium-mediated mechanism of regulating gastricmovement by electro-acupuncturing，Experiments in our lab also show that theenhanced gastric movement brought about by electro-acupuncturing at the“Zusanli” of the rats is likely to be related to the positive regulation of thehomeobox holax genes in the retinoic acid pathways and the negative regulationof the ptgs2genes in the phospholipase C pathways ultimately[82]。The subjectusing the mature technology of the functional classification of gene chiptechnology, The changes in the gastricoelectric activities of SD rats afterelectro-acupuncturing are observed. Gastric biopsy samples (gastric antrum,0.8-1cm from the pylorus) are selected and the PKC and MAPK signaltransduction pathways are tested through gene-chip detection and western blottest to find out the expressions of correlated protein CaD and CaD, to observethe contraction and relaxation activities of the role of acupuncture serum onisolated rat primary gastric smooth muscle cells,, and further validateacupuncture regulate gastrointestinal function of PKC and MAPK cell signalingpathway. demonstration of this regulatory network "acupuncture- gastrointestinal hormone-G protein-PKC and MAPK pathway (thin filamentregulation mechanism)-gastrointestinal motility”. to provide scientific andtheoretical basis for further explore the acupuncture mechanism.Methods：1、The SD rats are divided into three groups at random, namely “Zusanli”,non-acupoint and normal control groups. The changes in the gastricoelectricactivities of SD rats after electro-acupuncturing are observed。2、isolated from the normal rat’s gastric antrum smooth muscle cells, at thesame time, to obtain Each group of rat’s blood by carotid artery and isolated theblood serum，then， under a microscope to record the different treatmentfactors of gastric smooth muscle cells in different serum morphologicalchanges。3、Gastric biopsy samples (gastric antrum,0.8-1cm from the pylorus) areselected and the PKC and MAPK signal transduction pathways are testedthrough gene-chip detection and western blot test to find out the expressions ofcorrelated protein CaD and CaD。Results：1、Electro-acupuncturing at “Zusinli” to Bring a Two-way Regulation tothe Gastric Smooth Muscle Movement。2、Electro-acupuncturing at “Zusanli” can regulate part of the genericexpression in the PKC and MAPK signal transduction pathways。3、Electro-acupuncturing at “Zusanli” can regulate part of the correlatedprotein CaD&CaP expression in the PKC and MAPK signal transductionpathways。4、 The needling “Zusanli”， could promote contraction of normalrat’sisolated gastric antrum smooth muscle cells with the length of cell shortened。Conclusions：1、the gastric movement enhanced by acupuncturing is related to theup-regulation of MAPK6(ERK3), and the down-regulation of IL1R2， thegastric movement decreased by acupuncturing is related to the up-regulation ofFOS、CXCL9(MIG) and NOS2A (iNOS), and the down-regulation of MMP9。2、the gastric movement enhanced by acupuncturing is related to thedown-regulation of CaD and CaP， the gastric movement decreased byacupuncturing is related to the up-regulation of CaD and CaP。3、PKC and MAPK intracellular signal transduction pathways possible inregulating the gastric activities of SD rats through electro-acupuncturing at the“Zusinli”. is relatively clear, yet not the only way influencing acupunctureeffeet。...