| Objective: To observe the mechanism of Duhuo Jisheng decoction in the treatment of knee osteoarthritis(OA)in rats through animal experiments,and to provide a theoretical basis for the clinical application of Duhuo Jisheng decoction in the treatment of OA.Methods:1.Grouping: Forty male SD rats were randomly divided into 4 groups: Blank group,Model group,Duhuo Jisheng decoction group(DHJS group),glucosamine sulfate group(GS group),10 rats in each group.2.Modeling: rats in the model group,GS group,and DHJS group were fasted for 12 hours,anesthetized by intraperitoneal injection of 0.3% pentobarbital sodium(30mg/kg),and0.2ml 4% papinase was injected into the knee joint cavity.After the start of the experiment,No The procedures described above were performed at 1,4,and 7 days.The blank group was treated with the same method as the model group,GS group,and DHJS group,and the equal volume of 0.9% sodium chloride injection was injected into the knee joint cavity.3.Administration: DHJS group was given Duhuo Jisheng decoction(2ml/100g)by gavage twice a day,GS group was given glucosamine sulfate(0.17 g/kg)as positive control,model group and blank group were given the same amount of 0.9% sodium chloride injection by gavage twice a day.The rats were treated with intragastric administration for8 weeks.4.Sampling: After gavage,the rats were fasted and water deprived for 12 hours,and then anesthetized by intraperitoneal injection of 0.3% pentobarbital sodium.After successful anesthesia,samples were collected.5.Observation indicators: the general condition,behavioral score,serum interleukin-6(IL-6)level,smad2,mmp-13 and TGF-β1 protein content in synovial fluid,imaging findings of knee joint,pathological morphology of knee cartilage and Mankin score of rats in each group were observed.6.Statistical analysis of experimental data.Results:1.General conditions: on the 8th day of modeling,the rats in the model group,DHJS group and GS group showed decreased spirit,reduced food intake,reduced daily activity,insensitive response to external stimuli,obvious swelling of bilateral knee joints,and subcutaneous nodules could be touched in some rats.After 8 weeks of TCM intervention in DHJS group,the above symptoms were improved.2.IL-6 level: the serum IL-6 level of SD rats in each group was detected,and it was found that: compared with the blank group,the serum IL-6 level of SD rats in the model group was significantly increased(P < 0.01);Compared with the model group,the serum level of IL-6 in the DHJS group and the GS group decreased significantly(P < 0.01).3.X-ray findings of bilateral knee joints: the blank group had good knee joint space,clear articular surface,and no obvious osteophytes.In the model group,the knee joint space was narrowed,the articular surface was blurred,and the boundary was unclear.The above manifestations of rats in DHJS and GS groups were improved.4.HE staining of knee cartilage: in the blank group,the cartilage surface was smooth,the tide line was clear,and the boundary between cartilage layer and subchondral bone was clear.In the model group,the cartilage surface structure was destroyed,the cartilage layer was thinner,the number of chondrocytes was reduced,the distribution of chondrocytes was disordered,and the boundary between cartilage layer and subchondral bone was not clear.Compared with the model group,the DHJS and GS groups showed improvement in the above performance.Compared with the blank group,the Mankin score of the model group was significantly increased(P < 0.01).Compared with the model group,the Mankin scores of the DHJS group and the GS group decreased significantly(P < 0.01).The results showed that after gavage of DHJS,the degree of articular cartilage injury in the knee of SD rats in the DHJS group was alleviated.5.Western Blot results of rat cartilage tissue: Duhuo-Jisheng decoction can increase the expression levels of TGFβ1 and Smad2 protein in cartilage tissue,reduce the expression of MMP-13 in cartilage tissue,protect and repair chondrocytes,and delay the progress of KOA.6.The behavioral scores of rats in each group were found(FIG.5-2): compared with the blank group,the behavioral scores of the model group were significantly increased,and there was significant difference(P < 0.01);Compared with the model group,the behavioral scores of the DHJS group and GS group were significantly decreased(PDHJS Groupvs Model Group < 0.01,PGS Groupvs Model Group < 0.05).The results showed that after gavage of DHJS,the symptoms of knee osteoarthritis in SD rats of DHJS group were relieved.Conclusions:Duhuo Zizhi Decoction may affect the regulation of TGF-βsignaling pathway,down-regulate the expression of MMP13 in cartilage,promote the expression of TGF-β1and Smad2 in cartilage,reduce the expression level of inflammatory factors in joint fluid,inhibit the degradation of cartilage matrix,play the role of anti-inflammatory,analgesic and joint protection,and finally play the role of prevention and treatment of KOA. |
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