ABSTRACT
OBJECTIVE：To ex plore the protective effects of Longbie capsule contained serum （called“LBJN”for short ）on the apoptosis of chondrocytes induced by YAP inhibitor verteporfin and its mechanism. METHODS ：Primary human knee osteoarthritis（OA）chondrocytes were extracted by two-step enzymatic digestion ，and then identif ied by toluidine blue staining and type Ⅱ collagen immunofluorescence staining. The effects of 2，5 μmol/L verteporfin alone or combined with 5％LBJN on cell apoptosis were detected by flow cytometry. Solvent control （0.1％ DMSO）and 5％ LBJN were set. Western blot assay was adopted to detect the expression of apoptosis related proteins （YAP，Bcl-2，cleaved-caspase-3） after treated with 0.1％DMSO（solvent control ），2 μmol/L verteporfin，2 μmol/L verteporfin+5％LBJN 和 0（blank control ），2.5％ LBJN and 5％ LBJN for 48 h. The expression of autophagy related proteins （mTOR，Beclin-1，LC3A/B） after treated with 0 （blank control ），2.5％，5％ LBJN for 48 h were det ected by Western blot assay. RESULTS ：The isolated cells accorded with the characteristics of chondrocytes. Compared with 0.1％DMSO， the apoptosis rates of cells were increased significantly after treated with 2，5 μmol/L verteporfin（P＜0.05），and the effects of the two concentrations were similar （P＞0.05）. Compared with verteporfin alone ，2，5 μmol/L verteporfin combined with 5％LBJN could significantly decrease the apoptotic rate of cells （P＜0.05）. Compared with 0.1％DMSO，the protein expression of YAP and Bcl-2 were decreased significantly after treated with 2 μ mol/L verteporfin （P＜0.05）， while the protein expression of cleaved-caspase-3 were increased significantly （P＜0.05）. Compared with 2 μmol/L verteporfin，protein expression of YAP and Bcl-2 were increased significantly after treated with 2 μmol/L verteporfin+5％LBJN（P＜0.05），while the protein expression of cleaved-caspase-3 were decreased significantly （P＜0.05）. Compared with blank control ，the protein expression of YAP ，Bcl-2 and Beclin-1 were increased significantly after treated with 2.5％，5％LBJN（P＜0.05），while protein expression of cleaved-caspase- 3 and mTOR were decreased significantly （P＜0.05）. CONCLUSIONS ：LBJN can block the apoptosis of chondrocytes induced by YAP inhibitor verteporfin ，and its mechanism may be related to regulating the expression of apoptosis related proteins and enhancing autophagy of chondrocytes.
ABSTRACT
OBJECTIVE： To study the effects of ligustrazine on miR-20b/VEGF and BMP2/Smad1 pathways in subchondral bone of knee osteoarthritis （KOA） model rats， and to investigate the mechanism of ligustrazine for KOA prevention and treatment. METHODS： Totally 18 healthy male SD rats were randomly divided into normal control group， model group and ligustrazine group， with 6 rats in each group. The rats in the latter two groups were used to establish KOA model by intra-articular injection of 4％ papain solution. From the 2nd day after the last injection， ligustrazine group was given intragastrical administration of Ligustrazine suspension （100 mg/kg） 2 mL； normal control group and model group were given intragastrical administration of isometrical normal saline， once a day， for consecutive 6 weeks. After the last after medication， the situation of bilateral knee articular cartilage of rats were observed after exposure. The knee joints of rats were sectioned and stained with HE. The pathological change of articular cartilage were observed by microscope and scored by modified Mankin’s score. mRNA expression of VEGF， BMP2 and Smad1， and the expression of miR-20b were detected by RT-PCR； the protein expression of VEGF， BMP2 and Smad1 were detected by Western blot assay. RESULTS： Model group and ligustrazine group suffered from cartilage injury of knee joint at varying degrees. Compared with normal control group， Mankin’s scores of knee joint and cartilage tissue were increased significantly in model group （P＜0.01）； mRNA and protein expression of BMP and Smad1， the expression of miR-20b in subchondral bone of model group were decreased significantly， while mRNA and protein expression of VEGF were increased significantly （P＜0.01）. Compared with model group， Mankin’s score of cartilage tissue were decreased significantly in ligustrazine group （P＜0.01）； mRNA and protein expression of BMP and Smad1， the expression of miR-20b in subchondral bone were increased significantly， while mRNA and protein expression of VEGF were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Ligustrazine can repair damaged articular cartilage in KOA model rats， the mechanism of which may be associated with inhibiting the protein expression of VEGF and activating BMP-2/Smad1 signaling pathway via up-regulating the expression of miR-20b， and promoting the degradation of VEGF mRNA in subchondral bone.
ABSTRACT
OBJECTIVE:To systematically review the efficacy and safety of Bushen huoxue herb and celecoxib in the treat-ment of knee osteoarthritis, and provide evidence-based reference for clinical treatment. METHODS:Retrieved from CNKI, CBM,VIP and Wanfang Database,Medline,PubMed and Cochrane Library,randomized controlled trials (RCT) about Bushen huoxue herb(test group)and celecoxib(control group)in the treatment of knee osteoarthritis were collected. Meta-analysis was per-formed by using Rev Man 5.3 software after data extraction and quality evaluation. RESULTS:Totally 15 RCTs were included,in-volving 1 129 patients. Results of Meta-analysis showed,the total effective rate [RR=1.09,95%CI(1.04,1.14),P0.05;Lysholm:MD=2.32,95%CI(-1.95,6.58),P>0.05;WOMAC:MD=-2.87,95%CI(-6.38,0.64), P>0.05] and the incidence of adverse reactions in 2 groups[RR=0.49,95%CI(0.22,1.09),P=0.08]. CONCLUSIONS:Bushen huoxue herb shows better efficacy and analgesic effect than celecoxib in the treatment of knee osteoarthritis,and is similar to cele-coxib in terms of improving knee function score and safety.