| Objective: Knee Osteoarthritis(KOA)is a common joint disease and occurred in the elderly patients,both men and women can be diseased.There are more than half people who over the age of 60 can be found the imaging KOA performance,the occurrence and development of KOA is a long-term chronic process.The late stage of KOA with knee pain symptoms,joint dysfunction was obviously worsened and resulted in a serious decline in the quality of life.As the proportion of older people in the community increase,the prevalence of the KOA increases significantly.In the background of the research of knee osteoarthritis developing and the research of traditional Chinese medicine has been vigorously developed,this study was designed the treatment of knee osteoarthritis with the ZhiLongHuoXueTongYu(ZLHXTY)capsule in the rats.By comparing the degree of damage,histomorphological change,the content of SOD and MDA,the expression of collagen? and MMP-13 in cartilage in each experimental groups,to explore the prevention and treatment of osteoarthritis by ZLHXTY capsule and its possible mechanism,through animal experiments certification that the ZLHXTY capsule can lubrication of knee cartilage,remission or treatment of knee osteoarthritis by invigorating liver and kidney,clearing and activating the channels and collaterals,promoting muscles and bones.Methods: The detailed experimental steps are as follows: Grouping: male SD rats as experimental subjects.Randomly divided into 4 groups as A: blank group,B: model group,C: ZHHYTY capsule treatment of low-dose group(0.05g/kg/d)and D: ZHHYTY capsule treatment of high-dose group(0.1g/kg/d).Each of group has 10 rats.The model establishment: A modified Hulth's operation was used to establish the KOA model in SD rate,KOA model was success established after 6 weeks of post-operation,and then each group was separately gavage for 8 weeks by drinking water,low-dose and high-dose ZLHXTY capsule.Then the knee synovial extract liquid was used to determine the content of SOD and MDA,The cartilage of the medial femoral condyle of the knee was excised quickly and the histomorphology were observed by HE-staining and scored by Mankin's score.The expression of MMP-13 and collagen ?was detected by immunohistochemistry and scored by the score method of positive staining cell.SPSS statistical software was used to analyze the data to compare the effects of ZHHYTY capsule on these indexes and to analyze and test the therapeutic effect of ZHHYTY Capsule on knee osteoarthritis.Results: General situation: All rats survived,no infection,death cases,the typical late-stage KOA model was established in the model group after 14 weeks of post-operation.(1)Results of synovial SOD(U/ml)content in each group A: 166.32 ± 19.86;B: 40.74 ± 12.06;C: 105.45 ± 18.88;D: 120.96 ± 8.49.Comparing with the other three groups,the activity of SOD in group B decreased obviously(p <0.01).And the results of SOD in group D was more obvious in the drug treatment groups(p <0.05 p = 0.032).(2)Results of synovial MDA(nmol/mgprot)content in each group,A: 15.79 ± 3.43;B: 51.82 ± 10.15;C: 34.65 ± 6.63;D: 23.66 ± 6.62.The content of MDA in group A was the lowest in each group,and comparison with A the result of MDA in group D was more obvious(p<0.05 p=0.032).The difference between the remaining groups was more significant(p <0.01).(3)HE-staining: The cartilage damage was the most serious in the model group,Cartilage thickness becomes decreased significantly and stained fading obviously,the cartilage surface depression,formation the cartilage defect area;The chondrocytes arranged in disorder,the surface cells clustered,the deep chondrocytes showed obvious nuclear compression and cytoplasmic hypertrophy.The chondrocytes have a tendency to reduce.Some areas showed cell-staining missing;tide line disappeared,cartilage fibrosis significantly.The ZLHXTY capsule treatment groups: Cartilage thickness to varying degrees of decreased and the staining to varying degrees of fading,cartilage surface rough edgy,not smooth enough,some cartilage irregular cracks.The surface chondrocytes increased,as diffused,in some areas chondrocytes formed clusters.Middle and deep chondrocytes showed a tendency of cytoplasmic hypertrophy,some of the cells showed obvious nuclear compressed,the Cytoplasm transparent swollen.The tidal lines are mostly complete.In the low-dose drug treatment group cartilage fibrosis can be more obvious.The degree of cartilage repair in group D was better than that in group C.(4)Mankin's score of cartilage: group A: 0.30 ± 0.48;B: 10.30 ± 1.89;C: 6.20 ± 1.03;D: 5.6 ± 1.26.In the drug treatment groups,the score of group C was higher than that of group D,but the statistically comparison have no significant(p> 0.05 p= 0.298).There was significant difference between the other groups(p<0.01).(5)Immunohistochemistry: In the group A,the type ? collagen was strongly brown positive expression,while MMP-13 protein was not expressed or was extremely weakly positive.In the group B,during the cartilage damage was more obvious,MMP-13 began to be strongly positive expression,and type ? collagen expression was significantly weaker than that in the group A,showed a weaker positive expression.In the ZLHXTY capsule treatment groups,MMP-13 and type ? collagen were all negatively correlated with each other,that is,the more severe the cartilage damage,the stronger the expression of MMP-13,and the weaker the positive expression of Collagen?.The reaction of MMP-13 staining deepened more clearly around the chondrocytes in group B and in the cytoplasm in group C;The reaction of Collagen ? staining deepened obviously in some of the cytoplasm and surrounding chondrocytes in group D.(6)The type ? collagen expression score: Group A: 11.40 ± 1.25;B: 3.40 ± 1.17;C: 5.70 ± 1.49;D: 6.30 ± 2.06.The results showed that there was no significant difference between the two ZLHXTY capsule treatment groups(p> 0.05 p=0.388).The differences between remaining groups were statistically significant(p<0.01).(7)MMP-13 expression score: group A: 0.2 ± 0.42;B: 8.40 ± 1.90;C: 6.70 ± 1.70;D: 5.30 ± 1.64.The difference between the two ZLHXTY capsule groups was statistically significant(p<0.05 p = 0.048).The difference between the group C and the group B was significant(p<0.05 p = 0.018).The comparisons between the remaining groups were statistically significant difference(p<0.01).Results: 1.The ZLHXTY capsule can increase SOD activity in SD rat,increase the body's antioxidant capacity,reduce oxygen free radicals in the synovial,cartilage damage,inhibition the synovitis,reduce inflammatory factor exudation,Improve cartilage matrix metabolism and promote cartilage repair.2.The ZLHXTY capsule can inhibit the expression of MMP-13 in cartilage,promote the synthesis of Collagen ?,strengthen the repair of cartilage matrix and enhance the cartilage matrix resistance to enzymatic hydrolysis,thus slowing the progression of OA disease. |
PDF Full Text Request