BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene ma...BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.展开更多
目的:探究血清甘油三酯-葡萄糖(TyG)指数、摄食抑制因子-1(nesfatin-1)、视黄醇结合蛋白4(RBP4)联合预测糖尿病视网膜病变(DR)的价值,为DR早期预测提供支持。方法:回顾性分析。收集2022-02/2023-12我院接诊的2型糖尿病(T2DM)患者164例...目的:探究血清甘油三酯-葡萄糖(TyG)指数、摄食抑制因子-1(nesfatin-1)、视黄醇结合蛋白4(RBP4)联合预测糖尿病视网膜病变(DR)的价值,为DR早期预测提供支持。方法:回顾性分析。收集2022-02/2023-12我院接诊的2型糖尿病(T2DM)患者164例的临床资料,按照眼底检查结果分为DR组43例(其中增殖性DR 19例,非增殖性DR 24例),不合并DR的T2DM组121例。入院后记录患者基本资料,检查血清TyG指数、nesfatin-1、RBP4水平。结果:DR组病程长于T2DM组,空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、低密度脂蛋白及TyG指数、RBP4水平高于T2DM组,高密度脂蛋白、nesfatin-1水平低于T2DM组(均P<0.001)。多因素Logistic回归分析可知,T2DM病程(OR=1.338,95%CI:1.059-1.690)、糖化血红蛋白(OR=5.065,95%CI:1.659-15.470)、低密度脂蛋白(OR=12.715,95%CI:2.385-67.790)、TyG指数(OR=23.057,95%CI:2.936-181.073)、RBP4(OR=1.319,95%CI:1.028-1.692)是T2DM患者发生DR的危险因素,nesfatin-1(OR=0.007,95%CI:0.003-0.016)为保护因素。绘制ROC曲线显示,TyG指数、nesfatin-1、RBP4均对T2DM患者并发DR具有一定预测价值,曲线下面积(areas under curve,AUC)分别为0.804、0.878、0.738,各指标联合预测时AUC为0.946,预测敏感度为83.72%、特异度为92.56%。增殖性DR患者TyG指数、RBP4水平高于非增殖性DR患者,nesfatin-1水平低于非增殖性DR患者(均P<0.05)。Spearman相关性分析显示,TyG指数、RBP4水平与DR病情程度呈正相关,nesfatin-1水平与DR病情程度呈负相关(r_(s)=0.557、0.392、-0.359,均P<0.05)。Pearson相关分析显示,T2DM并发DR患者TyG指数与nesfatin-1水平呈负相关,与RBP4水平呈正相关,nesfatin-1与RBP4水平呈负相关(r=-0.486、0.538、-0.592,均P<0.05)。结论:血清TyG指数、nesfatin-1、RBP4水平与DR发病风险及病情程度有关,可作为DR早期预测的标志物,且联合预测效能更好。展开更多
文摘BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.
文摘目的:探究血清甘油三酯-葡萄糖(TyG)指数、摄食抑制因子-1(nesfatin-1)、视黄醇结合蛋白4(RBP4)联合预测糖尿病视网膜病变(DR)的价值,为DR早期预测提供支持。方法:回顾性分析。收集2022-02/2023-12我院接诊的2型糖尿病(T2DM)患者164例的临床资料,按照眼底检查结果分为DR组43例(其中增殖性DR 19例,非增殖性DR 24例),不合并DR的T2DM组121例。入院后记录患者基本资料,检查血清TyG指数、nesfatin-1、RBP4水平。结果:DR组病程长于T2DM组,空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、低密度脂蛋白及TyG指数、RBP4水平高于T2DM组,高密度脂蛋白、nesfatin-1水平低于T2DM组(均P<0.001)。多因素Logistic回归分析可知,T2DM病程(OR=1.338,95%CI:1.059-1.690)、糖化血红蛋白(OR=5.065,95%CI:1.659-15.470)、低密度脂蛋白(OR=12.715,95%CI:2.385-67.790)、TyG指数(OR=23.057,95%CI:2.936-181.073)、RBP4(OR=1.319,95%CI:1.028-1.692)是T2DM患者发生DR的危险因素,nesfatin-1(OR=0.007,95%CI:0.003-0.016)为保护因素。绘制ROC曲线显示,TyG指数、nesfatin-1、RBP4均对T2DM患者并发DR具有一定预测价值,曲线下面积(areas under curve,AUC)分别为0.804、0.878、0.738,各指标联合预测时AUC为0.946,预测敏感度为83.72%、特异度为92.56%。增殖性DR患者TyG指数、RBP4水平高于非增殖性DR患者,nesfatin-1水平低于非增殖性DR患者(均P<0.05)。Spearman相关性分析显示,TyG指数、RBP4水平与DR病情程度呈正相关,nesfatin-1水平与DR病情程度呈负相关(r_(s)=0.557、0.392、-0.359,均P<0.05)。Pearson相关分析显示,T2DM并发DR患者TyG指数与nesfatin-1水平呈负相关,与RBP4水平呈正相关,nesfatin-1与RBP4水平呈负相关(r=-0.486、0.538、-0.592,均P<0.05)。结论:血清TyG指数、nesfatin-1、RBP4水平与DR发病风险及病情程度有关,可作为DR早期预测的标志物,且联合预测效能更好。
