Effects of Bushen Qiangjin Jiaonang(补肾强筋胶囊)on knee subchondral bone in knee osteoarthritis rats and its mechanism of action:an experimental study
Objective:To explore the effects of Bushen Qiangjin Jiaonang(补肾强筋胶囊,BSQJJN)on subchondral bone of knee joint in knee osteoarthritis(KOA)rats and its mechanism of action.Methods:Twenty-four 6-week-old specific pathogen-free(SPF)-grade male Sprague-Dawley(SD)rats were randomized into normal group,model group,and BSQJJN group,8 cases in each group.The rats in model group and BSQJJN group were subjected to anterior cruciate ligament transection(ACLT)on the right hind knees for inducing KOA.Four weeks after successful modeling,the rats in BSQJJN group were intervened by intragastric administration with BSQJJN solution,while the ones in normal group and model group with deionized water,once a day for consecutive 4 weeks.Four weeks after the drug intervention,all rats were sacrificed,and their right hind knee joints were harvested for observing the gross structure,meanwhile,the knee joint specimens were scanned by using a Micro-CT scanner for analyzing the bone volume fraction(BVF),trabecular number(Tb.N),trabecular thickness(Tb.Th)and trabecular separation(Tb.Sp)in knee tissues of rats in each group.Additionally,the knee joint specimen sections were stained with hematoxylin-eosin(HE)and safranin O-fast green(SO-FG)for observing the knee subchondral bone tissues.Furthermore,the expres-sions of hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor A(VEGFA)in subchondral bone of knee joints were detected by using immunohistochemical staining,and the expressions of CD31 and Endomucin were detected by immunofluorescence stai-ning.Results:①Effects of BSQJJN on gross structure of knee joint in KOA rats.The changes,manifesting as severely worn cartilage,marked knee joint space narrowing(JSN),seriously damaged intra-articular structure and noticeable adhered surrounding tissues,were observed in rats of model group;while,compared to model group,the changes were comparatively slight in rats of BSQJJN group,just mild cartilage wear and slight JSN were observed.②Effects of BSQJJN on subchondral bone of knee joint in KOA rats.The results of MicroCT examination showed that,in model group,the trabeculae in subchondral bone were sparsely arranged,with obvious osteophytes around;while,compared to model group,the arrangement of trabeculae in subchondral bone of knee joint was denser in BSQJJN group.Additionally,compared to the normal group,the BVF was smaller,the trabeculae was fewer,the Tb.Th and Tb.Sp were greater in subchondral bone tissues of rats from model group and BSQJJN group(P=0.001,P=0.012;P=0.013,P=0.034;P=0.000,P=0.001;P=0.000,P=0.005);further com-parison revealed that the BVF was larger,the trabeculae was more,the Tb.Th and Tb.Sp were smaller in BSQJJN group compared to model group(P=0.001;P=0.039;P=0.022;P=0.039).The results of HE and SO-FG staining showed that the rats in model group and BSQJJN group exhibited obviously degeneration in knee joint cartilages,with the loose trabeculae in the subchondral bone;whereas,the de-gree of subchondral bone calcification was lower in BSQJJN group compared to model group.③Effects of BSQJJN on the expression of type H blood vessel formation-related proteins in subchondral bone of knee joint in KOA rats.The expressions of HIF-1α,VEGFA,CD31 and Endomucin in subchondral bone of knee joints were higher in model group and BSQJJN group compared to normal group(P=0.010,P=0.002,P=0.041,P=0.017;P=0.045,P=0.025,P=0.032,P=0.011),and was highest in model group(P=0.026,P=0.006,P=0.036,P=0.029).Conclusion:The BSQJJN can inhibit the expression of type H blood vessel formation-related proteins,and intervene the formation of type H blood vessels in the subchondral bone of knee joints in KOA rats,thereby improving the sclerotin of subchondral bone.
osteoarthritis,kneerats,Sprague-DawleyBushen Qiangjin Jiaonangsubchondral bonetype H blood vessels
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