Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation...Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation-induced emission luminogens(AIEgens)can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout.First,we developed a series of luminogens(i.e.,tetraphenyl ethylene(TPE)-NH_(2),TPE-2NH_(2),TPE-4NH_(2),TPE-COOH,TPE-2COOH,TPE-4COOH,and TPE-Ketoalkyne),each of which was then evenly mixed with MSU crystals.Next,optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope(CLSM).This approach was used for imaging standard samples of MSU,hydroxyapatite(HAP)crystals,and mixed samples with 1:1 mass ratio of MSU/HAP.We also imaged samples from mouse models of acute gouty arthritis,HAP deposition disease,and comorbidities of interest.Subsequently,CLSM imaging results were compared with those of compensated polarized light microscopy,and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line.Finally,CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi.As a promising candidate for MSU crystal labeling,TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples,animal samples,and human samples,and could precisely distinguish gout from HAP deposition disease.This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.展开更多
Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each gr...Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each group. Group 1 received 0.9% saline 2. 5 ml/kg per day by gastrogavage (ig) for 10 days; Group 2, 3 and 4 received STDT 0.125 g/kg, 1.0 g/kg and 8.0 g/kg per day respectively by ig for 10 days; Group 5 received colchicine 4. 5 mg/kg per day by ig for 4 days; and Group 6 was untreated. MSU crystals 10 mg /500ul containing polymyxin B 10 u/ml was injected into the knee joints of Group 1-5 to make rabbit arthritis models. Leukocytes in synovial lavage fluids was then counted and differentiated; pathological injury of synovial membranes was observed under HE staining; interleukin-1 beta (IL-1B), tumor necrosis factor alpha (TNFa) and leukotriene B4 (LTB4) content in synovial lavage fluids were determined by ELISA. Results: MSU caused a rapid leukocyte infiltration and increased production of IL-1B, TNFa and LTB4 2 hrs after intra-articular injection. STDT inhibited neutrophil infiltration in synovial fluids dose-dependently, protected the synovial membrane against pathological injury and reduced the production of IL-1B, TNFa and LTB4; while colchicine did not decrease the level of TNFa, but significantly inhibited neutrophil infiltration in synovial fluid and reduced the production of IL-11B and LTB4. Conclusion: STDT exerts an anti-inflammatory effect in rabbit model of acute MSU arthritis, its mechanism being probably due to the decrease of XL-1B, TNFa and LTB4 synthesis.展开更多
目的通过观察三黄胶囊对尿酸钠(MSU)致急性痛风性关节炎大鼠血清及膝关节液中白介素-β(IL-β)和肿瘤坏死因子-α(TNF-α)或环氧酶2(COX-2)水平的影响,探讨三黄胶囊治疗急性痛风性关节炎的机制。方法将大鼠分为正常组,模型组,三黄胶囊组...目的通过观察三黄胶囊对尿酸钠(MSU)致急性痛风性关节炎大鼠血清及膝关节液中白介素-β(IL-β)和肿瘤坏死因子-α(TNF-α)或环氧酶2(COX-2)水平的影响,探讨三黄胶囊治疗急性痛风性关节炎的机制。方法将大鼠分为正常组,模型组,三黄胶囊组(6.45 g生药/kg),秋水仙碱组(0.325 mg/kg)。正常组和模型组灌胃生理盐水;各给药组按剂量灌胃给药,连续14 d。末次给药1 h后,正常组右侧踝关节腔注射100 m L无菌1%磷酸盐缓冲液(PBS),其余各组注射100 m L MSU溶液,于造模后24 h用酶联免疫吸附试验(ELISA)检测血清和关节滑液中IL-β、TNF-α或COX-2水平;光镜下观察其对大鼠踝关节滑膜组织病理变化的影响。结果与正常组比较,模型组MSU注射进大鼠踝关节腔24 h后,在大鼠血清中IL-β、TNF-α和关节滑液中IL-β、TNF-α及COX-2的含量均明显增加(P<0.05);与模型组比较,三黄胶囊组及秋水仙碱组均显著降低MSU致大鼠痛风性关节炎血清IL-β、TNF-α及关节滑液中IL-β、TNF-α和COX-2水平(P<0.05);造模组滑膜组织的炎性细胞浸润、纤维组织增生、滑膜细胞增生、水肿、充血的病理改变较中药组和西药组严重。结论 MSU可以增加血清IL-β、TNF-α及关节滑液中IL-β、TNF-α和COX-2的水平;三黄胶囊可能通过降低IL-β、TNF-α和COX-2的水平产生抗MSU痛风性关节炎的作用。展开更多
基金Thisworkwas supported by the Shanghai Science and Technology Committee(No.22dz1204700)the NationalKeyR&D Program of China(Nos.2020YFA0803800 and 2017YFE0132200)+2 种基金the National Natural Science Foundation of China(Nos.82072510,21907034,21788102,21525417,and 51620105009)the Natural Science Foundation of Guangdong Province(Nos.2019B030301003 and 2016A030312002)the Innovation and Technology Commission of Hong Kong(No.ITC-CNERC14S01).
文摘Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation-induced emission luminogens(AIEgens)can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout.First,we developed a series of luminogens(i.e.,tetraphenyl ethylene(TPE)-NH_(2),TPE-2NH_(2),TPE-4NH_(2),TPE-COOH,TPE-2COOH,TPE-4COOH,and TPE-Ketoalkyne),each of which was then evenly mixed with MSU crystals.Next,optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope(CLSM).This approach was used for imaging standard samples of MSU,hydroxyapatite(HAP)crystals,and mixed samples with 1:1 mass ratio of MSU/HAP.We also imaged samples from mouse models of acute gouty arthritis,HAP deposition disease,and comorbidities of interest.Subsequently,CLSM imaging results were compared with those of compensated polarized light microscopy,and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line.Finally,CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi.As a promising candidate for MSU crystal labeling,TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples,animal samples,and human samples,and could precisely distinguish gout from HAP deposition disease.This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.
基金This program was supported by the National New Drugs Foundation(No.98-35-N-13)
文摘Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each group. Group 1 received 0.9% saline 2. 5 ml/kg per day by gastrogavage (ig) for 10 days; Group 2, 3 and 4 received STDT 0.125 g/kg, 1.0 g/kg and 8.0 g/kg per day respectively by ig for 10 days; Group 5 received colchicine 4. 5 mg/kg per day by ig for 4 days; and Group 6 was untreated. MSU crystals 10 mg /500ul containing polymyxin B 10 u/ml was injected into the knee joints of Group 1-5 to make rabbit arthritis models. Leukocytes in synovial lavage fluids was then counted and differentiated; pathological injury of synovial membranes was observed under HE staining; interleukin-1 beta (IL-1B), tumor necrosis factor alpha (TNFa) and leukotriene B4 (LTB4) content in synovial lavage fluids were determined by ELISA. Results: MSU caused a rapid leukocyte infiltration and increased production of IL-1B, TNFa and LTB4 2 hrs after intra-articular injection. STDT inhibited neutrophil infiltration in synovial fluids dose-dependently, protected the synovial membrane against pathological injury and reduced the production of IL-1B, TNFa and LTB4; while colchicine did not decrease the level of TNFa, but significantly inhibited neutrophil infiltration in synovial fluid and reduced the production of IL-11B and LTB4. Conclusion: STDT exerts an anti-inflammatory effect in rabbit model of acute MSU arthritis, its mechanism being probably due to the decrease of XL-1B, TNFa and LTB4 synthesis.
文摘目的通过观察三黄胶囊对尿酸钠(MSU)致急性痛风性关节炎大鼠血清及膝关节液中白介素-β(IL-β)和肿瘤坏死因子-α(TNF-α)或环氧酶2(COX-2)水平的影响,探讨三黄胶囊治疗急性痛风性关节炎的机制。方法将大鼠分为正常组,模型组,三黄胶囊组(6.45 g生药/kg),秋水仙碱组(0.325 mg/kg)。正常组和模型组灌胃生理盐水;各给药组按剂量灌胃给药,连续14 d。末次给药1 h后,正常组右侧踝关节腔注射100 m L无菌1%磷酸盐缓冲液(PBS),其余各组注射100 m L MSU溶液,于造模后24 h用酶联免疫吸附试验(ELISA)检测血清和关节滑液中IL-β、TNF-α或COX-2水平;光镜下观察其对大鼠踝关节滑膜组织病理变化的影响。结果与正常组比较,模型组MSU注射进大鼠踝关节腔24 h后,在大鼠血清中IL-β、TNF-α和关节滑液中IL-β、TNF-α及COX-2的含量均明显增加(P<0.05);与模型组比较,三黄胶囊组及秋水仙碱组均显著降低MSU致大鼠痛风性关节炎血清IL-β、TNF-α及关节滑液中IL-β、TNF-α和COX-2水平(P<0.05);造模组滑膜组织的炎性细胞浸润、纤维组织增生、滑膜细胞增生、水肿、充血的病理改变较中药组和西药组严重。结论 MSU可以增加血清IL-β、TNF-α及关节滑液中IL-β、TNF-α和COX-2的水平;三黄胶囊可能通过降低IL-β、TNF-α和COX-2的水平产生抗MSU痛风性关节炎的作用。
