| Objective: Osteoarthritis (OA) is a degenerative articularcartilage lesions as well as the main form of concomitant bone spurs, andchanges in the consolidated synovial lesions of chronic disease, inaddition to the stroke at the top of the second high morbidity of thedisease, and prevalence increases with increasing as the population ages,the incidence of obesity. Chondrocyte senescence and apoptosis has beenconfirmed that the formation and development is closely related to theformation and development of osteoarthritis. Silent information regulator2homolog protein1(SIRTl) was proved to be closely associated with cellaging. SIRTl gene has a certain degree of expression in humanchondrocytes and cartilage tissues, and the expression of cartilage cellsand osteoarthritis patients significantly lower than the normal expressionof articular cartilage cells. P53tumor suppressor gene(P53), also namedas gene housekeeper，it’s coding protein is a kind of transcription factors,control of cell cycle promoter, determine the cells to start dividing andcontrol cell apoptosis, the P53expression is closely related to theoccurrence and development of osteoarthritis. SIRT1may not only toregulate apoptosis through interaction with p53, but can also be regulated by Nuclear factor-κB inhibitory effect of peroxisome proliferatoractivated receptors to prolong life, through the regulation of forkhead boxclass O transcription factor to extend the life of cells and the interactionof other ways to control cell apoptosis and senescence.Electro-acupuncture as product of traditional Chinese medicine therapycombined with modern treatments of physical factors, the efficacy ofosteoarthritis has been an increasing number of clinically confirmed. Inthis experiment using a knee extensor braking method making animalmodel of osteoarthritis, by observing electro-acupuncture treatmentgroup’s morphological changes of rabbit knee cartilage after HE staining,and comparing SIRT1and p53expression values which measured byimmunohistochemical method with normal control group and modelgroup. Explore the possible mechanisms of electro-acupuncture treatmentin knee osteoarthritis from cell and molecular level. Methods:(1) Theobject and intervention measures: purchasing39New Zealand rabbitswith health experiment from the animal center of Luzhou MedicalCollege, randomly selected13rabbits as normal control group, no modelof knee osteoarthritis. The rest of the26rabbits using knee extensor tubetype plaster braking method to build osteoarthritis model, randomlydivided into the model group and the electro-acupuncture group13each.The model group and the electro-acupuncture group were taken straighttube type gypsum fixed his left knee. fixed for6weeks to build the knee osteoarthritis model. After6weeks, using the random number tablemethod including both model group and treatment group select each arabbit to death by randomly, the corresponding index changes, the leftknee joint was observed to determine whether the knee osteoarthritismodel successfully. Electro-acupuncture group received acupunctureintervention.At the end of this experiment, all of the rabbits using earmarginal vein injection of air embolism method to be put to death. cut therabbit knees, take part of the medial femoral condyle cartilage, observechange of the related parameters of each knee cartilage.(2) Outcome:①The naked eye observation method was used to observe the normalcontrol group, model group and electro-acupuncture group treatment ofrabbit knee internal gross morphological changes;②Cut a little rabbitknee medial femoral condyle cartilage using HE staining, observe thechange of content the morphology of cartilage cells, cartilage collagenand proteoglycan matrix of the light microscope, the cartilage’smorphology and optical microscope observation results using improvedMankin ’s score standard score;③Analysis of the quantitativeimmunohistochemistry: the expression of Rabbit SIRTl antibody kit andP53antibody kit for detection of rabbit knee medial femoral condylecartilage SIRTl and P53.(3) Statistical methods: statistical analysis wasperformed using SPSS19.0for windows software. x±s is used toexpress the result, the statistical difference (P <0.05), and significant difference (P <0.01). Results:(1) Articular cartilage gross observation:normal control group, no obvious knee effusion, the medial femoralcondyle cartilage smooth appearance, blue and white. Bright and noobvious crack; model group, most of the left knee effusion obviously,medial femoral condyle cartilage is dark yellow, part is beige losing theirluster, the surface is rough and crack, and the crack is visible, erosion andulcer formation, partial weight-bearing joints, cartilage full-thicknesscartilage defect is visible, bone exposure, osteophyte formation of newbiological; Electro-acupuncture treatment group showed most animalmedial condyle cartilage surface is not flat, slightly yellowish, scatteredpunctate hyperemia, occasionally erosion, Knee effusion and synovialhyperplasia appeared within the knee, it’s degree was lighter than in themodel group. The electro-acupuncture treatment group and model groupof knee surface and synovial according to histology Peng TaiPing methodcompares the rank sum of the two groups, naked eye view has astatistically significant difference (P <0.05), while no significantdifference under light microscope. According to the modified Mankin ’sscores compared electro-acupuncture treatment group and model groupcartilage structure, with significant statistical difference (P <0.01).(2) HEstaining pathological observation: normal control group, HE stainingshowed that the cartilage surface is smooth, the cartilage layer of uniformthickness, matrix with uniform; cartilage cells arranged in neat rows, visible to the cartilage cell size, tidal line complete; model group, HEstaining showed that the cartilage surface ulceration, deletion, thecartilage thickness was significantly thinner, staining obvious decrease,reducing the number of cartilage cells, can see irregular large somatacartilage cells clustering, tide line is disorder; Electro-acupuncture group,articular cartilage surface comparison rules, occasionally small fracture,cartilage cells arranged more neatly, cell number increased, part ofcartilage cell rupture. Cartilage thickness was significantly thicker,staining the mild decline, showed mild disorder of tidal line. Thepathological section HE staining were scored according to the modifiedMankin ’s comparison, comparison between the model group and theelectro-acupuncture group with significant statistical difference (P <0.01).The normal control group, electro-acupuncture treatment group andmodel group’s cartilage, chondrocytes, integrity and HE staining tide linewere scored according to the modified Mankin ’s has statisticallysignificant differences (P <0.01).(3) Immunohistochemical staining:normal control group under light microscope, the expression of SIRT1see lots of dense brown coloring articular cartilage cells, the number ofcartilage cells was great. And p53expression is although cartilage cellsbut only a small amount of cell brown coloring. Expression of SIRT1inmodel group under light microscope to see the articular chondrocyteslittle fawn colored, a small number of. Expression of cartilage cells and p53although less but most cells brown hyperchromatic, cell coloruniformity. Electro-acupuncture group under light microscope, theexpression of SIRT1see a lot of brown colored, slightly less quantity thanin the normal control group in articular cartilage, dark staining. Theexpression of p53chondrocytes showed a few nuclear brown staining,compared with normal control group, the number of slightly more inarticular cartilage. The normal control group, electro-acupuncturetreatment group and model group’s SIRT1and P53from the value ofgroups has statistically significant differences (P <0.01). Usingcorrelation analysis the normal control group, electro-acupuncturetreatment group and model group’s SIRT1and P53, groups werestatistically significant (P <0.01), We can believed both in normalpopulation and osteoarthritis the expression of SIRT1was negativelycorrelated P53. Conclusion:(1) Using tube type plaster fixation method,keep the rabbit knee in extension position six weeks, can successful setup the knee osteoarthritis model.(2) SIRT1plays a very important role inthe pathogenesis and development of osteoarthritis, Probably with itsvariety of ways to controlled cartilage degeneration and destructionrelated. such as regulation of p53gene expression.(3)Electrio-acupuncture can effectively improve the symptoms of kneeosteoarthritis, its mechanism may be related to many ways to worktogether,such as activation of cartilage SIRTl expression, reduce the content of p53, etc. to improve cartilage cells, promote cell survival andinhibit the apoptosis of cartilage cells. |
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