| Objective: osteoarthritis (OA) is a multiple disease in the elderly,it’s pathological exchange includes articular cartilage degeneration,destructionand osteophytes formation. Traditional view is that the over expression ofproinflammatory factors IL-1, IL-6, TNF-α and protease classes MMP-1,MMP-3, MMP-9play important roles in OA. However, related gene knockoutmice animal experiments did not provide strong evidence, in the clinicaltreatment, application of anti-inflammatory drugs and protease inhibitors didnot achieve an ideal result. Therefore, the pathogenesis of OA need to be furtherclarified, in recent years, researchers showed that Wnt/β-catenin signal pathwayplays an important role in the pathogenesis of OA. The animal experimentshowed: specific increase the expression of β-catenin in knee joint cartilagecells of adult mice, the mice will appear OA-like change,which revealed thatthe up-regulation of Wnt/β-catenin signaling pathway lead to the occurrence ofOA. OA treatment can be divided into operation therapy and non operationtreatment, the electro-acupuncture therapy is clinically used as a non operationtreatment. The traditional acupuncture and electrical stimulation are combinedin electro-acupuncture.As the acupuncture stimulate the acupoints of OApatients, which leads to the release of analgesic substance, and electricalstimulation cause muscle contraction, which improve local blood circulation, these all can help damage repair. Therefore, this article trys to explore themechanism of electro-acupuncture treatment of rabbit knee OA model throughobserving the gross morphology changes of articular cartilage, the pathologicalchanges of cartilage cells and the expression of β-catenin in the cartilage cells.Methods:(1) The object and intervention measures: purchasing36NewZealand rabbits with health experiment from the animal center of LuzhouMedical College, randomly selected12rabbits as the normal control group,without any treatment, model group and EA group with12rats in each groupaccording to the Videman method, use plaster cast their left knees straightly for6weeks to build OA model.6weeks after modeling, Electro-acupuncturetreatment group accept acupuncture treatment, selection of left lower limb ofrabbits Zusanli, Liangqiu, Neixiyan,Waixiyan, the same time every day for25minutes, the treatment was continued for15days, the remaining two groups atthe same time fixed for25minutes every day.15days after the air embolismmethod executed three animal groups, open the left knee joint for grossobservation, then use the sharp blade to cut cartilage specimens of femoralcondylar subchondral bone of the medial zone, which is about0.5cm*0.5cm*0.5cm full cartilage tissue, making HE staining, immunohistochemistry andlight microscopy observing. Score according to the modified Mankin ’s code ofpoints, the score and the testing data were statistically analyzed.(2)Observationindex: visual observation three groups of rabbit knee cartilage grossmorphological changes, and record their modified Mankin ’s score; Cut a little rabbit knee medial femoral condyle cartilage in10%neutral FormaldehydeSolution fixed, paraffin embedded, sliced, and HE staining, observe the changeof content the morphology of cartilage cells,and record their cartilage cellsmodified Mankin ’s score. analysis of immunohistochemical expression:Using Rabbit anti-β-catenin kit for detection the β-catenin expression of leftknee joint medial condyle cartilage, and in the light microscope usingmorphometry to calculated β-catenin cell fraction.(3)Statistical methods:statistical analysis was performed using SPSS19.0for windows software.±s isused to express the result, the statistical difference (P <0.05), and significantdifference (P <0.01). Results:(1) cartilage gross observation: there were nonjoint cavity effusion in normal group, cartilage surface smooth appearance, ablue and white, transparent, deep color and bright, no defect and new biological,no edema and hyperaemia, according to the modified Mankin ’s cartilagestructure scores are0points.12rabbits of the model group cartilage gloss andtransparency decreased, color yellowish, partial cartilage surface with smallcracks and abnormal protuberance, articular surface is rough, most obviouseffusion in articular cavity. The structure of cartilage modified Mankin ’s results:3get2points,6is3points and3is4points. Electro-acupuncture treatmentgroup only4visible intra-articular effusion, and the amount less than the modelgroup, part of the cartilage surface a little ulcer, smooth cartilage surface gloss,compared with the normal control group, model group than the poor butimproved the structure of cartilage, Mankin ’s score results:3get1points,5is2 points,4is3points. Each score according to the modified Mankin ’scomparison of cartilage structure, the difference was statistically significant(P<0.01).(2) cartilage HE stain observing: normal group cartilage cell layersthick, cartilage cells arranged in neat rows, full and uniform dyeing.Chondrocytes and tidal line integrity modified Mankin ’s scores were0points.The model group staining showed articular cartilage was thinner, reducing thenumber of visible cracks, most of the surface, a crack depth of the subchondralbone, cartilage cells group together, irregular shape, tidal line disturbance.Cartilage cells modified Mankin ’s score5get2points,7get3points, tidal linescore4get1point,8get2points,both two scores were higher than normalgroup increased (P<0.01). Electro-acupuncture group staining showed that thecartilage layer is lower than that of the normal group was thinner than those inthe model group, but the thickness of the cartilage surface smooth, comparedwith the model group, and only a few small cracks formed, but the fissureconfined in cartilage, chondrocytes are plump, regular shape, small part ofcartilage slice I see a few cartilage cell aggregation, tide line seem moredisorder than normal group, cartilage cells modified Mankin ’s score6get1points,6get2points, scoring only2points1tide line, the rest are1points.Two scores were decreased than that in model group (P<0.01).(3)immunohistochemistry β-catenin testing results: under the microscope,brown particles within the chondrocyte is the expression of β-catenin. Thenormal group had β-catenin weakly positive expression, this experiment were extremely rare brown granules; model group had β-catenin expression stronglypositive, full-thickness cartilage visible dark brown particles.Electro-acupuncture treatment group than those in the model group decreasedthe expression of β-catenin, only in cartilage tissue surface, middle found a fewbrown granule. Conclusion:(1) the expression of β-catenin in articularcartilage is positively correlated with the damage of the cartilage, β-catenin ofseverely damaged cartilage is higher than that in the mild damaged cartilage.(2)electro-acupuncture therapy can reduce the degree of damage of articularcartilage, the mechanism may be that electro-acupuncture could decrease theexpression of β-catenin in cartilage cells, and indirectly inhibit theinflammatory gene transcription of downstream. |
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