| Objective:In this study,three kinds of moxibustion duration were used to treat experimental rabbit knee osteoarthritis.The amount of cartilage matrix proteoglycan and CHI3L1 gene expression in rabbit knee osteoarthritis were measured.The effects of different moxibustion duration treating experimental rabbit knee osteoarthritis and its regulatory effect on cartilage matrix proteoglycan and CHI3L1 gene were observed.This study aims to explore its possible therapeutic mechanism so as to provide experimental evidence for clinical moxibustion treatment of KOA.Methods:Fitfty 6 to 10 months of healthy Japanese white rabbits,weighted 2.5 ± 0.5kg,were randomly divided into 5 groups,ie.,10 minutes treatment group(10min group),20 minutes treatment group(20min group),40 minutes treatment group(40min group),control group and model group,with 10 rabbits in each.Experimenatl rabbit KOA model was induced by intra-articular injections with papain solution at the right knee of the rabbits in all treatment groups(10min group,20 min group,40 min group)and the model group.After the success of modeling,all treatment groups were treated by different duration of moxibustion treatment.Different duration of mild moxibustion was performed for acupuncture points “Xiyan”according to the three treatment groups(10 min group,20 min group,40 min group).Such a moxibustion treatment was performed once a day for 4 periods in total each of which lasts 5 days.A break for 2 days was required between 2 consecutive periods.The whole knee joints were taken out after we sacrificed all rabbits.The cartilage,synovial tissue and surrounding tissues of the right knee were collected.Pathological morphological analysis was performed by HE staining,safrania O / fast green staining and alcian blue staining.RT-PCR was used to detect the expression of CHI3L1 gene.The experimental data were analyzed by SPSS22.0.Results:1.HE staining:The degree of lesion(mainly degree of inflammation)from weak to strong:Control group <40min group <20min group ≤ 10 min group <model group(1)Control group: Morphology of the synovial is clear,no histopathological changes.(2)Compared with control group,40 min group showed a small amount of inflammatory infiltration.(3)Compared with 40 min group,20 min group showed severer inflammation.(4)Basically,10 min group and 20 min group share a same degree of inflammation which means no significant difference between 2 groups.But for 10 min group,there were more inflammatory cells scattered or focal infiltration in synovial area,hence strictly speaking,10 min group showed slightly heavier inflammation than 20 min group.(5)Model group showed the most serious lesion in all groups.A large number of neutrophils and pus infiltration were found.Also,local tissue necrosis,small amount of nuclear debris,more lymphocytes and plasma cell infiltration were observed.2.Safrania O / fast green staining: The OD value of the control group was higher than that of the model group,10 min group and 20 min group(p<0.05).The OD value of the model group was less than 10 min group,20 min group and 40 min group(p<0.05)10min group and 20 min group were lower than 40 min group(p<0.05);10min group and 20 min group showed no significant difference(p> 0.05).3.Alcian blue staining: The OD value of the control group was higher than that of the model group,10 min group and 20 min group(p<0.05).The OD value of the model group was lower than 10 min group,20 min group and 40 min group(p <0.05)(P<0.05).10 min group and 20 min group were lower than 40 min group(p<0.05).There was no significant difference between the 10 min group and the 20 min group(p>0.05).4.The expression level of CHI3L1 gene in the control group was lower than model group and 40 min group(p <0.05).The gene expression of the model group was lower than 40 min group,but higher than that of the 10 min group and 20 min group(P<0.05).10 min group and 20 min group were lower than 40 min group(p<0.05).There was no significant difference in gene expression between 10 min group and 20 min group(p>0.05).5.The correlation analysis between cartilage matrix proteoglycan and CHI3L1 gene expression level: The cartilage matrix proteoglycan in model group was negatively correlated with CHI3L1 gene expression level,the correlation coefficient r=﹣0.758,p=0.011.The correlation analysis showed no statistically significant in moxibustion treatment groups.Conclusion:1.In this study,rabbits KOA model was established by intra-articular injections with papain solution.The method was reliable,reproducible and featured by easy control and high success rate.2.Moxibustion has a therapeutic protective effect on cartilage matrix damage during KOA,which can lessen synovial inflammation and can effectively slow down the progression of KOA.Also,40 min group can significantly improve the cartilage matrix proteoglycan amount,or even close to the control level,which means 40 min group has the best therapeutic effect in all groups.10 min group and 20 min group share the same effect.3.(1)In the course of KOA,the expression of CHI3L1 gene in cartilage matrix increased,which indicated that the amount of CHI3L1 in cartilage matrix increased;(2)shorter duration(10min and 20min)moxibustion treatment can inhibit the expression of CHI3L1 gene in cartilage matrix.(3)Longer duration(40min)moxibustion can improve the expression level of CHI3L1 gene in cartilage matrix.4.(1)The amount of proteoglycan in model group was negatively correlated with the expression of CHI3L1 gene in the cartilage matrix.That is to say,proteoglycan was decreased whereas the expression of CHI3L1 gene in the cartilage matrix was increased in the course of KOA.(2)Different duration of moxibustion can affect the amount of proteoglycan and the expression of CHI3L1 gene in cartilage matrix. |
PDF Full Text Request