The Study Of Kidney-tonifying And Blood-activating Herbal Act On Knee Osteoarthritis By Modulation Of Immune Microenvironment

Objective1 Animal experimentTo assess the effect of kidney-tonifying and blood-activating herbal on knee osteoarthritis(OA)and to investigate its underlying mechanism of immune microenvironment modulation.2 Clinical trialTo evaluate the efficacy and safety of Long-bie Capsule in treating patients with knee osteoarthritis.Methods1 Animal experiment24 C57/BL6 mice were randomly assigned to Groups A,B and C.To establish the OA Model,Groups B and C were intraarticular injected with type II collagenase.Group A and were intragastric administrated with normal saline at a dosage of 0.2ml·10g-1 once per day.Group C was intragastric administrated with Long-bie Capsule solution at a equivalent dosage,0.2ml·10g-1 per day.The treatment lasted for 4 weeks.Consequently,serum and knee joints were collected.The expression of serum IL-6 was detected by ELISA.The expression of TLR4,MyD88,IRF-7 and MMP-13 was detected by Western blotting.We observed the structural changes and collagen degration by HE staining and Masson staining,respectively.The cartilage destruction was assessed by Safarin 0-fast green staining and OARSI histologic scale.2 Clinical trial48 patients with knee osteoarthritis were randomly assigned to experimental group and control group.The experimental group was treated with Long-bie Capsule and Celecoxib.The control group was treated with Long-bie Capsule placebo and Celecoxib.The treatment lasted for 4 weeks.We compared the WOMAC,SF-36,efficacy of TCM syndrome and adverse events between the two groups.Results1 Animal experiment1.1 There were significant differencs between the three groups in expression of IL-6(P<0.01).Expression of IL-6 in Group B was higher than Groups A and C(P<0.05).No significant differences were observed between Group A and C(P>0.05).1.2 The expression of IRF-7 was different between the three groups(P<0.05).Expression of IRF-7 in Group B was higher than Groups A and C(P<0.05).Expression of IRF-7 between Groups A and C was no significant different(P>0.05).1.3 There were significant differencs between the three groups in expression of TLR4(P<0.01).Both Groups B and C expressed TLR4 higher than Group A(P<0.05).However,there was no significant difference between Groups B and C(P>0.05).1.4 The expression of MyD88 was different between the three groups(P<0.05).Group B expressed MyD88 higher than Groups A and C(P<0.05).There was no difference between Groups A and C(P>0.05).1.5 MMP-13 expression was different in the three groups(P<0.05).Group B was higher than Groups A and C(P<0.05).There was no significant difference between Group A and C(P>0.05).1.6 In the femoral side,the OARSI scale was different between groups(P<0.01).Group B was higher than Groups A and C(P<0.05).Group C was higher than Group A(P<0.05).In the tibial side,the OARS I scale was different between groups(P<0.01).Both Group B and C were higher than Groups A(P<0.05)and there was no difference between Groups B and C(P>0.05).2 Clinical trial2.1 WOMAC pain scores reduced in both groups after 4-week treatment(P<0.01).The scores in experimental group was lower than in control group(P<0.05).2.2 WOMAC function scores reduced in both groups(P<0.01).The function scores in experimental group was lower than in control group(P<0.05).2.3 WOMAC stiffness scores had no significant changes after 4-week treatment(P>0.05).There were no differences between two groups(P>0.05).2.4 SF-36 PCS improved in both groups after treatment(P<0.05).The experimental group was higher than control group(P<0.05).2.5 SF-36 PCS improved in both groups after treatment,but there was no statistically significance(P>0.05).There were no differences between groups(P>0.05).2.6 The TCM syndrome scores decreased in both groups(P<0.01).The experimental group has lower score compared with control group(P<0.01).2.7 The total effective rates were 91.67%in experimental group and 83.33%in control group.The effective rate was higher in experimental group(P<0.05).Conclusion1 Animal experimentThe expression of TLR4,IRF-7,MyD88,IL-6 and MMP-13 increases in OA mice.The kidney-tonifying and blood-activating herbal may suppress the expression of IRF-7 by TLR4/MyD88 signaling pathway,resulting in less secretion of IL-6 and MMP-13,which alleviates inflammation and delays cartilage destruction.2 Clinical triralThe clinical trial indicates that Long-bie capsule may relieve pain and improve physical function when combination use with Celecoxib.Its efficacy is superior to using Celecoxib alone.Long-bie Capsule is a safe and effective drug for knee osteoarthritis.