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1.
Summary: The expression and activity of NF-kB in the synovium of collagen-induced arthritis (CIA) rats was detected in order to investigate the possible therapeutic effects of triptolide on rheumatoid arthritis (RA). The experimental Wistar rat model of CIA was set up by intradermal injection of emulsion of bovine collagen 11 and the successful rate of setting-up models was evaluated by arthritis index (AI). Rats were grouped randomly into three groups: normal, model and treatment group. The expression of TNF-α and IL-6 in synovial fluid was detected by ELISA, and the expression and activity of NF kB in synovium by immunohistochemistry method and by electrophoretic mobility shift assay (EMSA) respectively. As compared with normal group, the expression of TNF a and IL-6 in synovia (P〈0. 05), and the expression and activity of NF-kB (P〈0.05) in synovium were increased in model group. There was statistical difference in above-mentioned indexes between model group and treatment group. Triptolide may play a protective role in IRA via downregulating the expression and activity of NF-kB in synovium.  相似文献   

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Background There are few reports of a biological role for glycosyltransferases in the infiltration of osteoarthritic synovitis. The aim of this research was to investigate the expression and cellular location of β-1,4-galactosyltransferase Ⅰ (β-1,4-GaIT-Ⅰ) in a surgically-induced rat model of knee osteoarthritis (OA), and explore the role of β-1,4-GalT-Ⅰ in the pathogenesis of OA.Methods Male Sprague-Dawley rats were randomly divided into three groups: OA group, sham group and normal group. The model of OA was established in the right knees of rats by anterior cruciate ligament transaction (ACLT) with partial medial meniscectomy. Fibroblast-like synoviocytes (FLSs) obtained from normal rat synovial tissue were cultured.The expression of β-1,4-GalT-Ⅰ mRNA in the synovial tissue, articular cartilage and FLSs treated with tumor necrosis factor-α (TNF-α) were assayed by real-time PCR. Western-blotting and immunohistochemisty were used to observe the expression of β-1,4-GalT-Ⅰ at the protein level. Double immunofluorescent staining was used to define the location of the β-1,4-GalT-Ⅰ with macrophage-like synoviocytes, FLSs, neutrophils, and TNF-α in the OA synovium. The alteration of TNF-α in FLSs which were treated with lipopolysaccharide (LPS) and β-1,4-GalT-Ⅰ-Ab were detected by enzyme-linked immunosorbent assay (ELISA).Results The mRNA and protein expression of β-1,4-GalT-Ⅰ increased in synovial tissue of the OA group compared with the normal and sham groups at two and four weeks after the surgery, however, no significant difference appeared in the articular cartilage. Immunohistochemistry also indicated that the β-1,4-GalT-Ⅰ expression in OA synovium at four weeks after surgery increased sharply compared with the control group. β-1,4-GalT-Ⅰ co-localized with macrophage-like synoviocytes, FLSa, neutrophils and TNF-α in rat OA synovitis. Moreover, in vitro β-1,4-GalT-Ⅰ mRNA in FLSs was affected in a dose- and time-dependent manner in response to TNF-α stimulation. ELISA revealed that the expression of TNF-α was attenuated in FLSs in vitro when treated with anti β-1,4-GalT-Ⅰ antibody.Conclusion β-1,4-GalT-Ⅰ may play an important role in the inflammation process of rat OA synovial tissue which would provide the foundation for further researching into the concrete mechanism of β-1,4-GalT-Ⅰ in OA synovitis.  相似文献   

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This study investigated the expression and immune effect of TLR4 in human trophoblast cells. The expression level of TLR4 mRNA in normal and LPS-stimulated human term trophoblast cells (1 mg/L LPS, 12 h) was detected by RT-PCR. In LPS-stimulated human term trophoblast cells of TLR4-blocked group and non-TLR4-blocked group, and normal term trophoblast cells of blank control group, apoptosis rate was measured by flow cytometry (FCM), and the level of TNF-α determined by using enzyme linked immunosorbent assay (ELISA) respectively. RT-PCR results showed that the expression level of TLR4 mRNA in LPS-stimulated human trophoblast cells was significantly higher than that in normal cells (P〈0.01). FCM revealed that there was significant difference in apoptosis rate of LPS-stimulated human term trophoblast cells between TLR4-blocked group and non-TLR4-blocked group (P〈0.05), or between TLR4 antibody-blocked group and blank control group. ELISA indicated that the level of TNF-α in LPS-stimulated human trophoblast cells also had statistical differences between TLR4 antibody-blocked group and non-TLR4 antibody-blocked group (P〈0.05). Our results suggest that TLR4 plays an important role in the immunological mechanism of apoptosis and secretion of TNF-α of human term trophoblast cells stimulated by LPS.  相似文献   

5.
Background Fascin, an actin binding protein, usually expressed at a low level in normal epithelium, but is significantly increased in transformed epithelial cells and several common carcinomas. In this study, we examined the expression of fascin by immunohistochemistry in sinonasal epithelium with chronic inflammation (control group), exophytic papilloma (EP), inverted papilloma (IP) with dysplasia and cancerated IP (including caminoma in situ and invasive squamous cell carcinoma, SCC), and furthermore investigated the relationship between fascin expression and formation of malignant IP. Methods Fascin expression was immunohistochemically detected using monoclonal antibody against fascin in 86 paraffin embedded tissues, including 10 cases of sinonasal mucosa with chronic inflammation, 10 of EP, 45 of IP with dysplasia (45 cases were divided into three groups: IP with mild dysplasia, IP with moderate dysplasia, and IP with severe dysplasia, 15 cases each), and 21 of cancerated IP. Results The level of fascin expression was significantly higher in the neoplastic tissue than that in control group. Fascin expression increased gradually with the progression from sinonasal epithelium with chronic inflammation, IP with mild dysplasia, IP with moderate dysplasia, IP with severe dysplasia, to cancerated IP, and significant difference of fascin expression was observed between any two groups of the five. Conclusion Precancerous lesions of IP exhibit elevated levels of fascin that may be associated with carcinogenesis of IP. Fascin may play a role in the formation of IP and EP.  相似文献   

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In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kunming mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.  相似文献   

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The action mechanism of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in the genesis, development and degeneration of haemangioma was investigated by detecting their expression in the tissue of haemangioma in different phases by using the immunohistochemistry. Fifty paraffin-embedded specimens of skin capillary haemangioma were collected, which were documented in the Department of Pathology, Renmin Hospital of Wuhan University from 2000 to 2006. All samples were stained by regular HE method, and proliferative cell nuclear antigen (PCNA) was tested by immunohistochemical S-P method. The samples were classified according to the Mulliken criteria and the expression pattern of PCNA. Immunohistochemical S-P method was ap- plied to detect the expression of MMP-2 and TIMP-2 in proliferative and degenerative phases of cutaneous capillary haemangioma, and in normal skin tissues. In combination with the detection of the expression of factor Ⅷ-related antigen, it was verified that in haemangioma tissues, the cells expressing MMP-2 and TIMP-2 were vascular endothelial cells. The MMP-2 and TIMP-2 expression was quantitatively analyzed by image analysis system (HPIAS-1000), and one-way ANOVA(107) and SNK(q) test were done to analyze average absorbance (A) and positive area rate of immunohistochemically positive particles by using SPSS11.5. The results showed: (1) Among 50 samples of haemangioma, there were 26 proliferative haemangiomas, and 24 degenerative haemangiomas, respectively; (2) The expression of MMP-2 was weak in normal vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression of MMP-2 in proliferative group was significantly higher than in degenerative group and control group (normal skin) (P〈0.05), but there was no statistically significant difference between the latter two groups; (3) TIMP-2 was highly expressed in normal tissues, degenerative vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression level of TIMP-2 in proliferative phase was significantly lower than in degenerative phase (P〈0.05), and the expression of TIMP-2 in proliferative phase was significantly different from that in degenerative phase and normal tissues (P〈0.05). It was concluded that in proliferative phase of haemangioma, MMP-2 may promote over-proliferation of endothelial cells of haemangioma, and in degenerative phase, TIMP-2 can inhibit the proliferation of endothelial cells of haemangioma. The two substances play important roles in the genesis, development and degeneration of haemangiomas.  相似文献   

10.
Summary: The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG (5.61±0.78 mmol/L), FINS (15.42±5.13 mU/L) and Ho- meostasis model assessment-IR (HOMA-IR) (1.21±0.52) in GDM group were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively) (P〈0.01). The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group (7.56±2.31 mU/L and 0.47±0.26 respectively) (P〈0.01). There was no significant difference in the InsR expression level among the three groups (P〉0.05). TP of InsR with insulin stimulation was significantly decreased in GDM group (0.20±0.05) as compared with normal pregnancy group (0.26±0.06) (P〈0.01). TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group (0.31±0.06) (P〈0.01). TP of InsR with insulin stimu- lation was negatively related with HOMA-IR in GDM group (r=-0.525, P〈0.01). There was no correlation between the protein expression of InsR and HOMA-IR in GDM group (r=-0.236, P〉0.05). It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.  相似文献   

11.
目的研究狗慢性膝关节炎症期滑膜组织中各种细胞因子(TNF-α,IL-1β,IL-6,IL-10)表达水平。方法Beagle狗17条,分为正常组(n=8)和慢性非感染性炎症组(n=9)。分别取其膝关节滑膜组织,采用ELISA方法检测滑膜组织中各种细胞因子的表达水平。结果与正常组相比,慢性非感染性炎症组滑膜组织内IL-1β、IL-6浓度显著增高(P〈0.05),TNF-α、IL-10浓度无明显变化(P〉0.05)。结论慢性非感染性炎症期,狗膝关节滑膜组织中IL-1β、IL-6表达显著增加。  相似文献   

12.
外治法对实验性兔膝骨性关节炎滑膜中bFGF表达的影响   总被引:2,自引:0,他引:2  
目的:了解外治法对碱性成纤维细胞生长因子(bFGF)在实验性兔膝骨性关节炎滑膜中的表达分布情况的影响,探讨中医外治法治疗骨性关节炎的机理。方法:32只实验动物随机分为正常组、模型组、扶他林组和外治法组,采用股静脉结扎方法造模成功后,治疗1个月。用免疫组化SABC方法对实验动物膝关节滑膜bFGF的表达分布进行观察。结果:造模组24只动物膝关节滑膜中bFGF免疫组化阳性率为92%(22/24),且外治法组表达强于扶他林组和模型组。正常组8只动物的膝关节滑膜中免疫组化为阴性。结论:外治法能够促进实验动物膝关节滑膜bFGF的表达,从而为外治法治疗骨性关节炎在细胞因子水平提供理论依据。  相似文献   

13.
目的:以腺病毒(Ad)为载体、p53为治疗基因,研究p53基因对人白介素(IL)-1β诱导兔膝关节炎的疗效和机制。方法:以Ad-LacZ为对照,将Ad-p53和Ad-LacZ分别注入6只人白介素1β(hIL-1β)诱导兔膝关节炎模型的膝关节。分析滑膜组织病理改变和关节炎的相关指标变化,检测滑膜组织细胞凋亡情况;培养人类风湿关节炎(RA)患者的滑膜成纤维细胞(FLS),用四氮唑蓝比色法(MTT)测定p53对滑膜细胞生长及增殖的影响,流式细胞仪检测细胞周期和凋亡情况,测定上清IL-6的水平。此外,逆转录多聚酶链反应(RT-PCR)和蛋白印迹(Western blot)方法评价腺病毒能否将治疗基因p53有效转移至滑膜组织。结果:Ad-p53感染滑膜成纤维细胞和滑膜组织后,治疗基因p53 mRNA和蛋白的表达明显上调。p53能显著减轻hIL-1β诱导兔膝关节炎的炎症,抑制滑膜细胞增生,减少炎症细胞浸润。p53不能诱导RA-FLS凋亡,但能显著下调促炎症细胞因子IL-6的产生。结论:抑癌基因p53很可能主要通过抑制促炎症因子的产生来减轻关节和滑膜炎症,是RA基因治疗的有效因子之一。  相似文献   

14.

目的  探讨白细胞介素-24(IL-24)基因在膝关节骨性关节炎(KOA)疾病中的作用及意义,为KOA的早期诊断及基因治疗提供一定的科学依据。方法  选择该医院骨科原发性KOA的滑膜标本,提取总RNA,用实时逆转录聚合酶链反应(Real-time RT-PCR)检测KOA滑膜组织中IL-24基因的表达。结果  IL-24基因表达在KOA滑膜组织中降低(P <0.01)。结论  IL-24在膝关节骨性关节炎滑膜中的表达降低,且与KOA病变关系密切,为将来诊断和基因治疗膝关节骨性关节炎疾病提供参考。

  相似文献   

15.
Preventionofadhesionofrabbitkneejointwithchitosan:anexperimentalstudyYeGenmao(叶根茂);HouChunlin(侯春林)(DepartmentofOrthopaedics,C...  相似文献   

16.
陈森洲  张惠勤  骆耐香  李莎莎  黄耀峰 《重庆医学》2011,40(35):3540-3542,3641
目的探讨昆明山海棠(THH)在胶原诱导性关节炎(CIA)大鼠模型中对外周血及滑膜组织低氧诱导因子-1α(HIF-1α)表达的影响。方法将Wistar大鼠分为对照组(n=10,不做任何处理)、CIA模型组(n=10)、治疗组[包括地塞米松组(n=10)、THH高剂量组(n=10)、THH中剂量组(n=10)及THH低剂量组(n=10)]。实验前CIA模型组及治疗组大鼠建模。X线摄片观察关节的形态改变,逆转录-聚合酶链反应(RT-PCR)以及免疫组化法分别检测HIF-1α在外周血及滑膜组织中的表达情况。结果对照组大鼠踝关节软组织结构完整,足趾关节间隙清楚,CIA模型组大鼠踝关节周围软组织肿胀,足趾关节间隙模糊、狭窄;各治疗组分别与CIA模型组及与对照组比较,大鼠在外周血及滑膜组织中HIF-1α表达差异均有统计学意义(P<0.05)。结论 THH可能通过调节外周血以及滑膜组织中HIF-1α的表达而对CIA大鼠具有治疗作用。  相似文献   

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目的: 探讨单纯疱疹病毒1(HSV-1)型载体介导白细胞介素1受体拮抗基因(IL-1Rα)局部治疗兔膝关节炎的作用。方法: 采用改良Hulth法构建兔负重膝关节炎模型;造模成功后利用基因重组技术构建IL-1Rα基因单纯疱疹病毒1型表达载体(pHSV-IL-1Rα-LacZ),行兔膝关节腔内注射4周;手术造模和基因治疗后,抽取关节腔灌注液,采用ELISA分析IL-1Rα和IL-1的表达;采用HE染色和甲苯胺蓝染色方法观察局部治疗效果,pHSV-LacZ空白载体设为对照组。结果: 手术造模治疗组注射pHSV-IL-1Rα-LacZ后能明显持续抑制IL-1表达水平,且对膝关节软骨炎进展显著性抑制,与对照组比较,差异具有统计学意义(P<0.05)。结论: pHSV-LacZ是关节腔内局部基因治疗兔膝关节炎模型的理想基因运输体。关节腔内持续表达的IL-1Rα可能通过拮抗IL-1的作用治疗兔膝关节炎。  相似文献   

18.
目的 观察通痹灵对胶原诱导性关节炎(CIA)大鼠关节滑膜组织炎症和血管内皮生长因子(VEGF)表达水平的影响,以探讨通痹灵治疗类风湿性关节炎(RA)的作用机理。方法 大鼠尾根部皮下注射Ⅱ型胶原蛋白建立CIA模型,分组给药治疗,给药37d后处死动物,对动物膝关节滑膜病理切片后分别常规HE染色和免疫组化染色。对HE染色切片用显微镜观察炎症细胞浸润,巨噬细胞增生和纤维增生情况,对免疫组化染色切片观察血管内皮生长因子(VEGF)阳性细胞情况以了解各组动物关节滑膜VEGF的表达水平。结果 HE染色切片显示通痹灵高剂量组滑膜炎症细胞浸润,巨噬样A型细胞增生和纤维组织增生的程度均比模型组低(P<0.05或P<0.01),并与正常组相仿(P>0.05)。免疫组化染色切片显示通通痹灵和雷公藤高剂量组均能降低滑膜VEGF表达水平(P<0.05)。结论 通痹灵有改善大鼠关节滑膜炎症,抑制滑膜VEGF表达作用,这可能是通痹灵治疗关节炎取得临床疗效的机理之一。  相似文献   

19.
目的运用不依赖加压固定的体外关节制动造模方式,制备皮肤软组织健康的兔膝骨关节炎模型。方法选用雄性新西兰大白兔18只,按照随机数字表法随机分为正常组、传统Videman法造模组(简称Videman组)、悬吊式外固定法造模组(简称悬吊组),各6只。正常组不予任何处理,Videman组单纯采用管型高分子石膏完全制动兔左膝关节于伸直位,悬吊组采用悬吊式外固定支具有限制动兔左膝关节于伸直位。6周后解除关节制动,观察兔皮肤软组织和左膝关节情况,用Bates-Jensen伤口评估工具(the Bates-Jensen wound assessment tool,BWAT)对皮肤软组织健康情况进行评估,用Lequesne MG评分对左膝关节功能进行评估;空气栓塞法处死动物,对左膝关节软骨行大体观察和HE染色病理学观察,并用Pelletier评分及Mankin评分对软骨损伤情况进行评估。结果(1)皮肤软组织健康情况比较,与正常组和悬吊组相比,Videman组BWAT评分升高(P<0.05);悬吊组与正常组BWAT评分比较,差异无统计学意义(P>0.05)。(2)左膝关节功能比较,与正常组相比,Videman组和悬吊组Lequesne MG评分均显著升高(P<0.01);Videman组和悬吊组Lequesne MG评分比较,差异无统计学意义(P>0.05)。(3)正常组左膝关节软骨大体观察和HE染色观察结果均正常,Videman组和悬吊组均见典型的软骨破坏,符合膝骨关节炎病理学改变。与正常组相比,Videman组和悬吊组Pelletier评分及Mankin评分均显著升高(P<0.01),达到轻中度骨关节炎的评分等级;Videman组和悬吊组Pelletier评分及Mankin评分比较,差异均无统计学意义(P>0.05)。结论悬吊式外固定支具有限制动膝关节于伸直位的造模方式可成功制备皮肤软组织健康的兔膝骨关节炎模型,其造模效果与传统Videman造模法效果一致,但对皮肤软组织的保护能力强于后者。  相似文献   

20.
目的:研究补肾和柔肝中药对C57黑鼠膝骨关节炎滑膜中软骨寡聚基质蛋白(COMP)基因表达的影响。方法:将7周龄C57黑鼠随机分为空白组、模型组、补肾组、柔肝组、芬必得组,雌雄各半。除空白组外,其余各组先进行4周跑步造模,再分别予生理盐水、补肾方(密骨胶囊)、柔肝方(养血软坚胶囊)、芬必得灌胃。给药4周后取材,应用Trizol法抽提总RNA,RT-PCR技术半定量检测膝关节滑膜中COMP基因表达,采用天能GIS凝胶图像处理系统分析各组表达灰度值。结果:模型组与空白组相比,滑膜中COMP基因表达明显升高(P〈0.01),各药物组与模型组比较,滑膜中COMP基因表达明显降低(P〈0.05);补肾组与柔肝组比较,滑膜中COMP基因表达明显升高(P〈0.01),两者有显著性差异。结论:补肾和柔肝中药均可以下调C57黑鼠膝骨关节炎滑膜中COMP基因的表达水平,可以延缓滑膜的增生和纤维化,减少滑膜分泌炎症介质和软骨降解酶,对骨关节炎有明显的防治作用,且柔肝中药明显好于补肾中药。  相似文献   

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