Therapeutic Effect and Mechanism of Jipei Dilong Ointment on Acute Soft Tissue Injury in Rats

[Objectives]This study was conducted to observe the therapeutic effect of Jipei Dilong Ointment on rats with acute soft tissue injury caused by heavy objects and to explore its action mechanism.[Methods]Thirty six rats were randomly divided into six groups(control group,model group,high-dose Jipei Dilong Ointment group(JP-H),medium-dose Jipei Dilong Ointment group(JP-M),low-dose Jipei Dilong Ointment group(JP-L)and diclofenac group).Except for the Control group,other groups were subjected to modeling of acute soft tissue injury by the weight impact method.All administration was performed once a day for nine consecutive days.The local appearance score and activity disorder score were determined after soft tissue injury in rats.HE staining was used to detect the pathological changes of injured soft tissues in rats.RT-PCR was used to detect the relative mRNA expressions of Bax,Bcl-2,MMP-9 and TIMP-1 in injured soft tissues of rats.Western Blot was used to detect the protein expressions of MMP-9,TIMP-1,TLR4,MyD88 and NF-κB p65 in injured soft tissues of rats.Results were statistically analyzed.[Results]Compared with the model group,Jipei Dilong Ointment could significantly improve the appearance symptoms such as swelling and ecchymosis in the injured area and the movement function of the affected limb(P<0.05).It could also improve the infiltration of inflammatory cells and widening of the intermuscular space caused by injury.Among them,the JP-H group and the diclofenac group had more significant curative effects.After 9 d of administration,each administration group could significantly up-regulate the ratio of Bcl-2/Bax mRNA expression level(P<0.05 or P<0.01),and the ratio of MMP-9/TIMP-1 mRNA expression level showed a downward trend(P>0.05).The expression level of NF-κB p65 protein in each administration group was significantly decreased(P<0.01).The protein expression levels of TLR4 and MyD88 and the ratio of MMP-9/TIMP-1 protein expression level in each administration group decreased to varying degrees.Among them,the JP-H group and diclofenac group significantly decreased(P<0.05).[Conclusions]Jipei Dilong Ointment has the functions of relieving pain,swelling and inflammation.It could improve the local appearance,functional activity and tissue morphology of affected limbs in rats,and has a therapeutic effect on acute soft tissue injury in rats.Its mechanism of action might be related to the inhibition of TLR4/MyD88/NF-κB p65 signaling pathway and the regulation of Bcl-2/Bax and MMP-9/TIMP-1 balance.

Protective Effect and Molecular Mechanism of Shui People's Classic Prescription Jipei Dilong Ointment on Osteoarthritis in Rats

[Objectives]To explore the protective effect and molecular mechanism of Shui People s Classic Prescription Jipei Dilong Ointment on knee osteoarthritis in rats.[Methods]72 SPF male SD rats were divided into control group,model group,Jipei Dilong Ointment high,medium and low dose groups,and positive drug Diclofenac group,with 12 rats in each group.Except the control group,all other groups were injected with 0.2 mL of 2%papain and 0.3%L-cysteine mixture into knee joint cavity to establish osteoarthritis model,while the control group was injected with the same amount of normal saline.The Lequesne MG score was used to determine the success of the model.After successful modeling,external administration was given for 4 weeks.The histopathological changes in articular cartilage and synovium were observed by HE staining;the levels of TNF-α,IL-1βand COX-2 in serum were detected by ELISA,and the relative expression of MMP-9 and TIMP-1 mRNA in cartilage was detected by qRT-PCR;the relative expression of MMP-9,TIMP-1,TLR4,MyD88 and NF-κB in rat cartilage was detected by Western-blot.[Results]Compared with the model group,Jipei Dilong Ointment could significantly reduce the Lequesne MG score and Mankin s score of arthritic rats(P<0.01);significantly improve the pathological changes in articular cartilage and synovium,reduce tissue edema,necrosis,inflammatory cell infiltration and fibrous tissue proliferation;reduce the expression of MMP-9 mRNA(P<0.01)in different degrees and increase the expression of TIMP-1 mRNA(P<0.01);reduce the relative expression of MMP-9,TLR4,MyD88 and NF-κB protein in different degrees(P<0.01),and significantly increase the relative expression of TIMP-1 protein in cartilage(P<0.01).[Conclusions]Jipei Dilong Ointment can improve the joint injury of osteoarthritis rats,and the mechanism may be related to the regulation of TLR4/MyD88/NF-κB signaling pathway and the ratio of MMP-9/TIMP-1.