骨关节炎兔血清中软骨寡聚基质蛋白和基质金属蛋白酶-3水平

目的 探讨血清软骨寡聚基质蛋白(COMP)与基质金属蛋白酶(MMP)-3检测应用于临床评估骨关节炎(OA)软骨病理改变的可能性.方法 伸直位石膏管型制动16只兔右后膝关节制作OA模型.以造模时间不同分为造模2周、造模6周,左后膝关节未造模故为对照组.X线影像学与病理观察模型关节的变化;评估关节软骨降解程度(OA积分);ELISA检测兔血清COMP、MMP-3水平;分析血清COMP、MMP-3水平与OA积分间的相关性.结果 (1)造模2周影像学变化较造模前不明显;造模6周兔胫骨平台边缘不光滑,关节间隙变窄,表面有毛刺样增生,胫骨平台及股骨内髁外侧可见唇样增生.(2)OA关节病变的形态学观察:造模2周兔关节软骨表面粗糙,表层裂隙;软骨细胞弥漫增多,排列紊乱;OA积分为(4.000±2.204)分.造模6周兔关节软骨表层可见较多裂隙延伸向下深达辐射层;裂隙周围可见脱水固缩坏死的软骨细胞且排列紊乱,部分成簇增生,各层结构不易分辨,有血管翳通过;OA积分为(10.620±1.408)分,与造模2周比,P=0.000.(3)造模2周兔血清COMP[(3.64±0.18)μg/L]、MMP-3[(1.99±0.81)μg/L]水平高于造模前[COMP(3.35±0.20)μg/L,MMP-3(1.61±0.71)μg/L];造模6周兔血清COMP[(3.96±0.44)μg/L]、MMP-3[(3.44±0.91)μg/L]水平高于造模前和造模2周,差异有统计学意义(P值均＜0.05).血清COMP、MMP-3水平与OA积分呈线性相关关系(r值均＞0.710,P值均小于0.05).结论 OA血清中COMP和MMP-3水平对评估OA软骨降解程度具有重要意义.

Abstract：

Objectiye To study the levels of cartilage oligomeric matrixprotein (COMP) and matrix metalloproteinase-3 (MMP-3) in the serum fluid of osteoarthritic rabbit models and their relationships with the severity of pathological changes, so as to investigate their correlation with osteoarthritis(OA). Methods The osteoarthritic animal models were get from immobilizing the right knees of 18 rabbits in full extension using plaster cast. Knee joint pathological changes of 2,6 weeks were examined for pathological severity of OA; ELISA sandwich method was used to measure the levels of COMP and MMP-3 in serum before and after modeling( at 2, 6 weeks respectively); X ray of model keens was also obtained in different period.Correlation analysis was performed to demonstrate the relationship between the levels of COMP, MMP-3 in the serum and the pathological severity of OA. Results ( 1 ) Morphological observations: immobilizing the right knees of rabbits in full extension using plaster cast was a reliable methed for osteoarthritic animal models and the typical histopathologic character was seen; the severity of osteoarthritisgradually increased with time extended. (2) The levels of COMP[(3.64 ±0. 18)μg/L], MMP-3 [(1.99 ±0. 81 ) μg/L]in the serum of 2 weeks osteoarthritic animal models were higher than those before immobilizing with plaster cast [COMP(3.35 ±0. 20) μg/L,MMP-3( 1.61 ±0. 71 ) μg/L]. The levels of COMP[(3.96 ±0. 44) μg/L],MMP-3[(3.44 ±0. 91) μg/L] of 6 weeks were much higher,with a significant difference(P ＜0.05). The levels of COMP, MMP-3 in serum had a linear correlation with the pathological severity of OA (r ＞0. 710,and P ＜ 0. 05 ). Conclusion The levels of COMP and MMP-3 in serum can help to predict and evaluate the progression of OA.     

慢性氟中毒大鼠软骨的损害及COLIXA3蛋白、表达的实验研究

目的 探讨不同程度的氟中毒是否影响染氟大鼠软骨组织中COLIXA3蛋白的表达.方法 选用3～4周龄健康雄性Wistar大鼠40只,按体质量随机分为5组,每组8只,分笼饲养,期间分别自由饮用含氟化钠0(对照)、25、50、100、150mg/L的蒸馏水,饲喂6月后建立氟中毒大鼠模型.应用光学显微镜分析实验大鼠骨组织的病理形态学变化过程.并采用免疫组织化学技术检测大鼠股骨干骺端COLIXA3蛋白的表达情况.结果 大鼠骨组织HE染色显示,各染氟组股骨干骺端出现不同程度软骨骨化,骨密度增加,具有硬化性氟骨症病变.对照组软骨组织未见明显异常.大鼠软骨细胞COLIXA3免疫组织化学染色结果为阳性,胞浆内可见有棕黄色颗粒,25、50、100 mg/L组在软骨组织中COLIXA3蛋白表达(23.3±4.5、41.2±5.6、26.4±7.5)增强.其中50、100 mg/L组表达与对照组(6.1±3.5)相比明显增加,组间比较差异有统计学意义(P均<0.05).150 mg/L组COLIXA3蛋白(13.3±4.2)较前面3组表达减弱,仍高于对照组,组问比较差异无统计学意义(P>0.05).结论 动物模型中,大鼠病理学为单纯性骨硬化表现.低剂量氟促进,高剂量抑制大鼠软骨细胞的增生.随着染氟时间的延长,外环境中氟浓度过高时,对软骨细胞就表现为氟离子的直接毒性作用.氟化物影响染氟大鼠软骨组织中COLIXA3蛋白的表达,低剂量氟可以促进COLIXA3蛋白的表达,随剂量增加氟的促进作用减弱.

Abstract：

Objective To explore whether different degrees of fluorosis influence the expression of cartilage COLIXA3 protein in fluorosis model rats. Methods Forty male Wistar rats 3 to 4 weeks old were randomly divided into 5 groups according to body mass, and these rats were fed with distilled water containing sodium fluoride(NaF) of 0(control), 25, 50, 100 and 150 mg/L for 6 months, respectively, in order to establish the animal model of drinking water type fluorosis. Pathomorphologieal changes of the osseous tissues of rats were analyzed under light microscope and transmission electron microscope, and the expression of COLIXA3 protein of femur metaphysis was examined by immunohistochemistry. Results HE staining showed different degrees of femoral metaphyseal ossification of cartilage in each experimental group, bone density increased, with sclerotic lesions of skeletal fluorosis. The control group showed no abnormal cartilage. Electron microscopy showed that the experimental groups with varying degrees of cartilage cell swelling, cell matrix fades, 50 mg/L group .showed hyperplasia, and 100,150 mg/L groups were observed with organelles decreased, part of the disintegration of the cartilage cell lacunae, lmmunohistochemical staining of rat chondrocytes COLIXA3 was positive, cytoplasm with brown granules, cartilage COLIXA3 protein expression(23.3 ± 4.5, 41.2 ± 5.6, 26.4 ~ 7.5) in the 25, 50 and 100 mg/L groups enhanced. Compared to the control group (6.1 ± 3.5), the expression of 50 and 100 mg/L groups was significantly increased, and the differences were statistically significant(all P < 0.05). The expression(13.3 ± 4.2)of COLIXA3 protein in 150 mg/L group was decreased compared with the previous three, but is still higher than that of control, and the difference was not statistically significant(P > 0.05). Conclusions There has pathological changes of sclerosing skeletal fluorosis in animal model. Low-dose fluoride promotes while high-dose inhibits cartilage cell proliferation. When fluorine concentration in external environment is too high and with extended exposure to fluoride, direct toxic effects of fluoride on cartilage cells is observed. Fluorine affects and promotes the expression of COLIXA3 protein in cartilage. Low-dose fluoride can promote COLIXA3 protein expression, as the dose increases (over 100 mg/L), the effect decreases.     

敲除趋化因子受体3对延缓原发性胆汁性肝硬化模型病情进展的作用

目的 建立原发性胆汁性肝硬化(PBC)动物模型,研究趋化因子受体3(CXCR3)及其配体干扰素诱导蛋白10(CXCL10)在PBC发病中的作用.方法 6～8周雌性C57BL/6J小鼠72只,分为3组:C57BL/6J野生鼠PBC模型组、正常对照组、CXCR3基因敲除(CXCR3-/-)C57BL/6J小鼠PBC模型组,PBC模型组小鼠每次腹腔注射5 mg/kg的聚肌苷酸(聚)胞苷酸(Poly I:C),每周2次,共24周,第8、16、24周处死小鼠,收集小鼠肝脏、血清标本;酶联免疫吸附试验(ELISA)检测血清抗线粒体抗体(AMA)和CXCL10滴度;病理分析肝脏炎性细胞浸润程度.计量资料两样本间比较选用t检验.结果 第8、16、24周对照组血清中AMA-M2的平均吸光度(A)值分别为0.17±0.04、0.19±0.07、0.20±0.06;PBC野生鼠模型的AMA-M2的平均A值分别为0.70±0.41、0.48±0.35、0.69±0.44;CXCR3-/-小鼠模型的平均A值分别为0.56±0.25、0.46±0.35、0.85±0.34.第8、16、24周PBC野生鼠模型组碱性磷酸酶(ALP)平均值分别为(115±33)、(119±32)、(133±52)U/ml;PBC CXCR3-/-鼠模型组ALP平均值分别为(106±29)、(112±29)、(122±60)U/ml;对照组小鼠ALP平均值分别为(65±7)、(70±9)、(77±10)U/ml.与对照组相比,PBC模型组的AMA滴度和ALP水平明显增高(P＜0.05);CXCR3-/-小鼠和野生鼠PBC模型组之间的AMA滴度和ALP水平差异无统计学意义.PBC模型组小鼠第8周可见小胭管周围单个核细胞浸润,胆管上皮细胞变性坏死,基底膜不完整;第16周可见小叶间胆管增生,炎性细胞浸润;第24周可见小胆管损伤、闭塞,胆管内胆栓形成,周围炎性细胞浸润,肉芽肿形成,炎症从门静脉区发展到肝实质.CXCR3-/-鼠PBC模型组病变程度较同期的野生鼠PBC模型组轻,第8周可见部分小胆管周围少量炎性细胞浸润,第16周汇管区炎性细胞浸润较前增多,至第24周时无胆栓和纤维间隔形成;CXCR3-/-小鼠模型组血清CXCL10的平均滴度较对照组明显增高;ELISA检测PBC模型组血清中CXCL10的平均滴度较对照组明显升高(P＜0.05),随着病程的进展,CXCL10滴度呈逐渐上升的趋势.与野生鼠PBC模型相比,CXCR3-/-小鼠模型CXCL10的平均滴度更高,第24周时两者差异有统计学意义.结论 敲除CXCR3有助于减轻和延缓小鼠PBC模型的病情进展.

Abstract：

Objective To generate an autoimmune cholangitis animal model and investigate whether CXCR3 and its ligand were involved in the pathogenesis of primary biliary cirrhosis (PBC). Methods Female C57BL/6 wild-type (WT) mice and CXCR3 knockout (CXCR3-/-) mice were injected with 5 mg/kg of poly I:C intra-peritoneally twice a week for 24 consecutive weeks. Liver specimens were collected to evaluate the degree of cell infiltration. AMA was assayed by ELISA. Differences in pathology were compared between CXCR3-/- mice and wild-type. Student's test was used to assess the differences. Results Anti-mitochon-drial antibody (AMA) and alkaline phosphatase (ALP) level were elevated significantly in the sera of all poly Ⅰ:C injected mice compared with control mice. AMA titers in serum were increased in the poly I:C injected WT mice compared with that of the control mice at week 8, 16, and 24 respectively (0.70±0.41 vs 0.17±0.04,0.48±0.35 vs 0.19±0.07, 0.69±0.44 w 0.20±0.06, P＜0.01). There was no significant difference between AMA titers in the serum of WT PBC mice and CXCR3-/- PBC mice (0.70±0.41 vs 0.56±0.25, 0.48±0.35 vs 0.46±0.35, 0.69±0.44 vs 0.85±0.34). Serum alkaline phosphatase (ALP) levels were raised among the poly I:Cinjected WT mice compared with WT controls (115±33) vs (65±7) U/ml; (119±32) vs (70±9) U/ml; (133±52) vs (77±10) U/ml. The serum ALP levels in CXCR3-/- PBC mice were (106±29), (112±29)and (122±60) U/ml at week 8, 16 and 24 respectively. There was no significant difference in ALP level between WT and CXCR3-/- mice PBC model. Considerable numbers of infiltrating cells were detected at the portal areas 8weeks after poly I:C injection, which progressed up to 24 weeks. At week 24, the interlobular bile ducts were lost and bile-plug were evident. Compared to WT mice model, this results revealed that CXCR3-/- mice, had fewer foci of inflammation and significant reduction in total inflammatory cells in their livers than those of the WT mice at 8, 16 and 24 weeks after injection of poly I:C. At week 24, there were no cholestasis orgranulomas in CXCR3-/- mice of PBC models. Compared to the control model, CXCL10 serum level was increased in PBC model. With the disease progression, CXCL10 serum level was elevated gradually. In comparison with wild mice, CXCR3-/- mice model had a higher serum level of CXCL10. At week 24, there was significant difference of CXCL10 serum level between wild-type mice and CXCR3-/- mice model. Conclusion In conclusion, these findings indicate that, CXCR3-/- mice might have a delayed onset and ultimately could not recruit sufficient effector cells to the liver for inflammation development.     

Alleviation of ischemia/reperfusion injury in ob/ob mice by inhibiting UCP-2 expression in fatty liver

AIM： To investigate the protective effect of target suppression of uncoupling protein-2 （UCP-2） on ischemia/ reperfusion （I/R） injury in fatty liver in ob/ob mice. METHODS： Plasmids suppressing UCP-2 expression were constructed, and transfected into fatty liver cells cultured in vitro and the ob/ob mouse I/R injury model. Serum tumor necrosis factor （TNF）-α levels, UCP-2 mRNA expression, alanine aminotransferase （ALT） levels in ob/ob mice were tested, and the pathological changes in fatty liver were observed in experimental and control groups. RESULTS： In ob/ob mouse I/R models, serum TNF-α levels were significantly higher than in normal controls. After the plasmids were transfected into the cultured cells and animal models, expression of UCP-2 rnRNA was significantly reduced as compared with that in the control group （2^1.56± 0.15 vs 2^-0.45± 0.15, p 〈 0.05）. In ob/ob mouse models, in which expression of UCP-2 was suppressed, serum ALT levels were significantly lower than those of other groups, and pathological analysis revealed that injury of liver tissues was significantly alleviated. CONCLUSION： The target suppression of UCP-2 expression in fatty liver can alleviate the I/R injury in the ob/ob mice.     

Effect of osteoporosis and intervertebral disc degeneration on endplate cartilage injury in rats

Objective:To investigate the effect of osteoporosis and intervertebral disc degeneration on the endplate cartilage injury in rats.Methods:A total of 48 female Sprague Dawley rats(3 months)were randomly divided into Groups A,B,C and D with 12 rats in each group.Osteoporosis and intervertebral disc degeneration composite model,simple degeneration model and simple osteoporosis model were prepared in Groups A,B and C respectively.After modeling,four rats of each group at 12th.18th and 24th week were sacrificed,Intervertebral height of cervical vertebra C6/C7 was measured.Micro-CT was used to image the endplate of cephalic and caudal cartilage at C6/C7 intervertebral disc.Abraded area rate of C6 caudal and C7 cephalic cartilage endplate was calculated,and then C6/C7 intervertebral disc was routinely embedded and sectioned.stained with safranin O to observe histological changes microscopically.Results:At 12,18 and24 weeks,intervertebral disc height of C6/C7 were(0.58±0.09)mm,(0.53±0.04)mm and(0.04±0.06)mm in Group A rats,(0.55±0.05)mm,(0.52±0.07)mm and(0.07±0.05)mm in Group B rats.At 24th week.intervertebral disc height of Group A rats was significantly lower than that of Group B rats(P0.05);intervertebral disc height of Groups A and B rats at each time point were significantly lower than that of Groups C and D(P0.05).There was no significantly statistical difference of intervertebral disc height between Groups C and D(P0.05).At 12 and 18 weeks,the abraded rate of C6 caudal and C7 cephalic cartilage endplate in Group A rats were significantly higher than that in Groups B.C and D rats(P0.05);the abraded rate in Group B was significantly higher than that in Groups C and D(P0.05).Microscopic observation of CT showed that ventral defects in C6caudal or C7 cephalic cartilage endplate in Groups A and B appeared after 12 weeks of modeling;obvious cracks were found in front of the C6 and C7 vertebral body,and cartilage defect shown the trend of"repairing"at 18 and 24 weeks after modeling.Conclusions:Intervertebral disc degeneration and osteoporosis can cause damage to the cartilage endplate.Co-existence of these two factors can induce more serious damage to the endplate.which has possitive correlation with intervertebral disc degeneration.Osteoporosis plays a certain role in intervertebral disc degeneration process,and accelerates the degeneration of intervertebral disc in a specific time window.     

生长抑素和生长激素联用对兔重症急性胰腺炎肠黏膜屏障损伤的保护作用

Objective To investigate the protective effect of somatostatin (SS)combined with growth hormone (GH) in treatment of intestinal mucosal barrier injury in rabbits with severe acute pancreatitis (SAP), as well as its clinical significance. Methods Seventy-two rabbits were equally assigned into model group (SAP group), SS treated group (SS group) and SS combined with GH treated group (SS + GH group). SAP models were induced by retro-injection of 5% sodium taurocholate into the pancreatic duct. After modeling, all rabbits were given 5 % glucose saline daily.The rabbits in SS group and SS+GH group were continuously Given SS (3.5μg·kg-1·h-1)for 48 hours. Besides, the rabbits in SS+GH group were subcutaneously injected with 0.15 IU/kg of GH at the 1st and the 24th hours after modeling. The levels of serum amylase, serum tumor necrosis factor-α (TNF-α) and plasma diamine oxidase were measured at the 6th, 12th, 24th and 48th hours after modeling. The pathological changes of pancreatic tissue and ileal mucosa were observed. Survival rate was calculated. Data were analyzed using SPSS 16.0 software. The univariate analysis was used to compare the difference among groups. Results In SS+GH group, the levels of serum TNF-α and plasma diamine oxidase were (2. 43 ± 0. 14) pg/ml and (4. 61 ± 0. 45) U/L at the 24th hour respectively, and were (2.08±0.23) pg/ml and (3.75±0.47) U/L at the 48th hour, respectively,which were lower than those in SAP group and SS group [(2.80 0.30) pg/ml and (8.74 ± 1.77)U/L, respectively, at the 24th hour; (2. 45±0.12) pg/ml and (5. 02±0.95) U/L, respectively, at the 48th hour)]with significant difference (P＜0.05). The inflammation in pancreas and ileal mucosa was alleviated, and the integrity of bowel mucosa was improved. Survival rate of SS+GH group was significantly higher than SAP group and SS group. There was no significant difference in level of serum amylase between SS+GH group and SS group. Conclusion The combination of SS with GH may enhance the function of intestinal mucosa barrier and improve the prognosis of SAP in rabbits.     

生长抑素和生长激素联用对兔重症急性胰腺炎肠黏膜屏障损伤的保护作用

Objective To investigate the protective effect of somatostatin (SS)combined with growth hormone (GH) in treatment of intestinal mucosal barrier injury in rabbits with severe acute pancreatitis (SAP), as well as its clinical significance. Methods Seventy-two rabbits were equally assigned into model group (SAP group), SS treated group (SS group) and SS combined with GH treated group (SS + GH group). SAP models were induced by retro-injection of 5% sodium taurocholate into the pancreatic duct. After modeling, all rabbits were given 5 % glucose saline daily.The rabbits in SS group and SS+GH group were continuously Given SS (3.5μg·kg-1·h-1)for 48 hours. Besides, the rabbits in SS+GH group were subcutaneously injected with 0.15 IU/kg of GH at the 1st and the 24th hours after modeling. The levels of serum amylase, serum tumor necrosis factor-α (TNF-α) and plasma diamine oxidase were measured at the 6th, 12th, 24th and 48th hours after modeling. The pathological changes of pancreatic tissue and ileal mucosa were observed. Survival rate was calculated. Data were analyzed using SPSS 16.0 software. The univariate analysis was used to compare the difference among groups. Results In SS+GH group, the levels of serum TNF-α and plasma diamine oxidase were (2. 43 ± 0. 14) pg/ml and (4. 61 ± 0. 45) U/L at the 24th hour respectively, and were (2.08±0.23) pg/ml and (3.75±0.47) U/L at the 48th hour, respectively,which were lower than those in SAP group and SS group [(2.80 0.30) pg/ml and (8.74 ± 1.77)U/L, respectively, at the 24th hour; (2. 45±0.12) pg/ml and (5. 02±0.95) U/L, respectively, at the 48th hour)]with significant difference (P＜0.05). The inflammation in pancreas and ileal mucosa was alleviated, and the integrity of bowel mucosa was improved. Survival rate of SS+GH group was significantly higher than SAP group and SS group. There was no significant difference in level of serum amylase between SS+GH group and SS group. Conclusion The combination of SS with GH may enhance the function of intestinal mucosa barrier and improve the prognosis of SAP in rabbits.     

慢性肾脏病患者游离脂肪酸与细胞因子及颈动脉病变的关系

Objective To investigate the serum level of free fatty acid (FFA) and explore its relationship with cytokines and atherosclerosis (AS) in chronic kidney disease (CKD).Methods The serum level of FFA was determined with enzymatic colorimetry.IL-1 β, IL-6 and TNFα were determined with ELISA.High-sensitivity C-reactive protein (hsCRP) was measured with immunoturbidimetry.Prevalence of atherosclerosis was detected with carotid ultrasonography.We evaluated the relationship between serum levels of FFA and IL-1β,IL-6, TNFα, hsCRP as well as the renal function in 130 adult patients with CKD, stratified according to the GFR ( based on the National Kidney Foundation/Kidney Dialysis Outcomes Quality Initiatives) and in 58 hemodialytic (HD) patients.The relationship between FFA level and cardiac geometry incidence in CKD patients was analyzed with logistic regression model.Results The serum level of FFA was significantly higher in CKD patients as compared with that in the healthy controls [(492.63 ± 143.59)vs (302.65 ± 142.18) μ mol/L, P ＜ 0.01], even in the early stage of CKD.The level of FFA increased with the progression of renal dysfunction.In the non-dialytic CKD group, the level of FFA was negatively related to GFR and positively related to the proteinuria (P ＜ 0.05), while in the HD group, it was positively correlated with dialysis duration ( P ＜ 0.05 ).The serum levels of FFA were higher in CKD patients with carotid artery atherosclerosis than those in patients without ( P ＜ 0.05 or ＜ 0.01 ).However, in both groups with impairment of renal function, the levels of FFA were positively correlated with hsCRP, IL-1 β, IL-6,TNFα and TG( all P ＜ 0.05 ).A positive correlation between the level of FFA and the clinical manifestations such as carotid intimal medial thickness (IMT) and AS was also found.A negative correlation was found between the level of FFA and the serum level of albumin and GFR( P ＜ 0.05).Conclusion Serum levels of FFA are significantly higher either in non-dialytic CKD or in HD patients and it is related with hsCRP, IL-1 β, IL-6, TNFα as well as carotid artery atherosclerosis, indicating that FFA is an independent risk factor of AS in CKD.     

脱氧雪腐镰刀菌烯醇致新西兰家兔关节损伤的实验观察

目的 观察脱氧雪腐镰刀菌烯醇(DON)对新西兰家兔膝关节软骨和滑膜的损伤.方法 将15只成年雄性新西兰家兔随机分为3组,对照组正常饲养,高、低剂量组分别投予0.10、0.05 mg/kg剂量的DON,经耳缘静脉注射隔日染毒.20 d后处死家兔,取双侧膝关节,进行常规病理检查,测定关节冲洗液白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、一氧化氮(NO)水平.结果 光镜下,可见低、高剂量组家兔膝关节软骨细胞变性、坏死.滑膜组织炎细胞浸润.关节冲洗液IL-1β、TNF-α、NO,组间比较差异有统计学意义(F值分别为19.396、18.195、22.136,P＜0.05);与对照组[(0.113±0.043)、(0.138±0.087)μg/L、(23.56±9.35)μmol/L]比较,高剂量组[(0.451±0.091)、(0.575±0.122)μg/L,(70.27±11.53)μmol/L]和低剂量组[(0.295±0.107)、(0.387±0.131)μg/L,(45.32±12.24)μmol/L]明显增高(P＜0.05),且高剂量组高于低剂量组(P＜0.05).结论 DON可导致家兔关节软骨和滑膜损伤,其损伤类似于人骨关节炎病变.DON可能是骨关节炎的病因之一.     

布洛芬与氨基葡萄糖对膝骨关节炎患者滑膜细胞增殖及软骨寡聚基质蛋白表达影响的比较

目的 探讨布洛芬与氨基葡萄糖对膝骨关节炎(OA)滑膜细胞增殖及软骨寡聚基质蛋白(COMP)表达的影响.方法 布洛芬与氨基葡萄糖含药血清培养早期和晚期滑膜细胞,四唑氮化合物/黄嘌呤氧化酶(MTS/PMS)法测定吸光度(A)值,hCOMP定量试剂盒测定COMP含量.采用等方差假设双侧t检验进行统计学处理.结果 通过MTS/PMS法观察确定布洛芬与氨基葡萄糖含药血清培养滑膜细胞观察点在第5～7天;氨基葡萄糖含药血清培养滑膜细胞A值[晚期组(0.054±0.021),早期组(0.777±0.034)]低于正常血清对照组(P＜0.05).布洛芬[晚期组(35.4±1.9),早期组(46.0±2.2)]与氨基葡萄糖含药血清[晚期组(36.6±1.3),早期组(48.8±1.3)]对软骨寡聚基质蛋白的表达也有明显降低作用(P＜0.05).结论 氨基葡萄糖可以抑制体外培养早期和晚期膝OA患者滑膜细胞增殖,而布洛芬与氨基葡萄糖皆可抑制体外培养滑膜细胞分泌COMP.     

Correlation of sonographic severity with biochemical markers of synovium and cartilage in knee osteoarthritis patients

OBJECTIVE: Ultrasonography can be used to detect soft tissue abnormalities within the joints that cannot be assessed using conventional X-rays. This study investigated the relationship between soft tissue and/or bony abnormalities on ultrasonography and the biochemical markers of the synovium and cartilage in the knee of osteoarthritis (OA) patients. METHODS: The knees from 51 OA patients who fulfilled the ACR criteria were enrolled in this study. Knee ultrasonography was performed in the affected knee joints using a 12 MHz linear probe to assess the presence of effusion, synovial proliferation, capsular distention, the length of osteophytes and the cartilage thickness. At the same time, the serum hyaluronic acid (HA) and the cartilage oligomeric protein (COMP) levels were measured by ELISA, and RIA was used to determine the serum osteocalcin levels. RESULTS: The patients with a longer medial osteophyte showed higher serum HA and COMP levels than those with a shorter one. The serum HA levels were significantly higher in those patients with a larger amount of effusion and/or synovial proliferation, which indicated inflammatory changes, than in those without. In addition, the severity of the capsular distention also correlated well with the serum HA and COMP levels. However, the length of the lateral osteophytes and the thickness of the femoral cartilage showed no correlation with the serum HA or COMP levels. In addition, the serum osteocalcin levels did not show any association with the above ultrasonographic parameters. CONCLUSION: This study demonstrated that the serum HA and COMP levels were elevated in the more severe OA patients by knee ultrasonography than in the less severe patients. This suggests that the detailed pathological changes in the soft tissue and/or bone of the OA joints on ultrasonography are directly reflected by the biochemical markers measured in the peripheral blood.     

Serum cartilage oligomeric matrix protein (COMP) levels in knee osteoarthritis in a Brazilian population: clinical and radiological correlation

OBJECTIVES: In this study we present data on serum cartilage oligomeric matrix protein (COMP) levels in a Brazilian population with isolated knee osteoarthritis (OA) compared to healthy controls. Clinical and radiological correlations with COMP levels were also evaluated. METHODS: Two hundred and seventy-two patients seen at the Rheumatology Division of the Federal University of S?o Paulo (UNIFESP) with a symptom of 'pain in the knees' for at least 3 months were invited to participate in this study. History and clinical examination were performed in all patients. Eighty-six patients with clinical isolated knee OA according to the American College of Rheumatology (ACR) criteria and without other causes of pain in the knee were included. Fifty-eight healthy individuals were selected, matched for age and sex, and used as controls. OA evaluation included Lequesne and Western Ontario and MacMaster Universities osteoarthritis index (WOMAC) questionnaires, visual analogue scale (VAS) for pain and standard knee X-rays. Blood samples were taken from all participants and serum COMP levels were measured by enzyme-linked immunosorbent assay (ELISA). OA radiological analysis was performed using the Kellgren and Lawrence (K/L) grading scale. RESULTS: Patients with symptomatic knee OA presented significantly higher serum COMP levels compared to healthy controls and to those with non-symptomatic narrowing of the articular space (p<0.001). Patients with clinical evidence of knee OA and without radiological abnormalities (K/L grade 0 or 1) had intermediate serum COMP levels, significantly higher than those observed in healthy controls (p<0.03). CONCLUSIONS: We observed increased serum COMP levels in patients with symptomatic radiological knee OA. High serum COMP levels may also indicate cartilage damage in selected symptomatic patients without significant radiological abnormalities.     

血清软骨寡聚基质蛋白检测在关节炎患者中的意义研究

目的 探讨血清软骨寡聚基质蛋白(COMP)在各类关节炎患者中的差别及对类风湿关节炎(RA)软骨破坏放射学改变的早期预测价值.方法 采用酶联免疫吸附试验(ELISA)方法测定并对比分析154例各类关节炎患者血清COMP水平的差异,RA患者各项临床指标及2年后关节放射学改良SHARP(vdH-Sharp)指数评分与COMP行相关性分析.结果 与仅有滑膜损害的其他关节炎患者及正常人群比较,RA患者血清COMP显著升高(P＜0.05).骨关节炎(OA)、银屑病关节炎(PSA)患者与正常人群比较血清COMP亦有差异,而与其他滑膜关节炎患者比较差异不大.RA、OA、PSA患者间血清COMP值比较差异无统计学意义.RA患者2年随访发现COMP与患者关节X线的改良SHARP评分前后差值呈正相关(P＜0.01,r=0.848).但与初诊时的抗环瓜氨酸肽(CCP)抗体、类风湿因子(RF)、关节功能、改良SHARP评分均无相关性,与初诊时的红细胞沉降率(ESR)、C反应蛋白(CRP)、晨僵时间、关节肿胀、压痛指数相关(P＜0.05).结论 COMP在各类软骨受侵犯关节炎尤其是RA患者血清中异常增高,提示其可为RA软骨病变早期诊断及判断软骨病变进展、预后和治疗效果的一项理想指标.     

Increased pentosidine, an advanced glycation end product, in serum and synovial fluid from patients with knee osteoarthritis and its relation with cartilage oligomeric matrix protein

BACKGROUND: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders-for example, rheumatoid arthritis. OBJECTIVE: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)-a marker of articular cartilage destruction. METHODS: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. RESULTS: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. CONCLUSION: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker.     

Serum cartilage oligomeric matrix protein and other biomarker profiles in tibiofemoral and patellofemoral osteoarthritis of the knee

OBJECTIVES: There is some evidence that tibiofemoral osteoarthritis (TFJ OA) and patellofemoral osteoarthritis (PFJ OA) may have different risk factors. To investigate the possibility that these conditions are separate disease entities, we compared biomarker profiles of patients with each disease. METHODS: Serum samples were taken from 222 patients who had knee pain and X-ray signs of knee OA. Eighty-two had only medial TFJ OA and 38 only PFJ OA in one or both knees. The remaining patients had either mixed disease or equivocal radiographic evidence of OA. The following biomarkers were measured in serum samples from baseline and follow-up visits: cartilage oligomeric matrix protein (COMP), glycosaminoglycan, keratan sulphate epitope 5D4, YKL-40, osteocalcin, C-telopeptide of type I collagen, hyaluronan and C-reactive protein. RESULTS: The two subsets of OA (TFJ and PFJ) had similar radiographic disease severity and there were no significant differences in the presence and patterns of pain scores (visual analogue scale and Western Ontario and McMaster Universities Osteoarthritis Index). No difference was found for the biomarkers between the two groups, with one exception. Both baseline and area under the curve per month COMP concentrations were significantly higher in the TFJ than the PFJ group (P<0.01). CONCLUSIONS: The reduced serum COMP in PFJ disease compared with TFJ OA could be due to small articular cartilage volume in the latter or to a qualitative difference in cartilage metabolism.     

Cartilage oligomeric matrix protein (COMP) in rheumatoid arthritis and its correlation with sonographic knee cartilage thickness and disease activity

Clinical Rheumatology - The aim of the study is to investigate the correlation of serum cartilage oligomeric matrix protein (COMP) levels with articular cartilage damage based on sonographic knee...     

The heritable determinants of cartilage oligomeric matrix protein

OBJECTIVE: Cartilage oligomeric matrix protein (COMP) is a cartilage matrix macromolecule. The protein is detectable in serum and has been investigated as a biomarker of osteoarthritis (OA). An association between COMP and OA has been shown, yet the precise factors governing serum levels of COMP remain unclear. The aim of this study was to determine whether genetic factors influence serum levels of COMP. METHODS: A classic twin study was conducted using COMP levels in serum obtained from healthy female twin volunteers. COMP levels were determined by an inhibition enzyme-linked immunosorbent assay method. The heritability of COMP was determined by comparing correlation among 160 monozygotic and 349 dizygotic twin pairs. Data on potential confounding factors, including age, body mass index, and the presence of OA as assessed by hand, hip, and knee radiographs, were included in the analysis. RESULTS: Serum levels of COMP showed a correlation of 0.72 (95% confidence interval [95% CI] 0.65-0.80) among monozygotic twin pairs and 0.47 (95% CI 0.39-0.55) in dizygotic pairs. This equated to an estimated heritability for COMP of 40% (95% CI 20-60%). Although age and body mass index were found to be significantly associated with COMP in regression analysis, taking the effects of these factors into account did not influence the estimate of heritability. CONCLUSION: This study showed that heritable factors influence serum levels of the cartilage matrix biomarker COMP. Together with other published data, the results suggest that genetic factors operate at an early stage in the etiologic pathways that influence the development of radiographically discernible OA.