Connective Tissue Growth Factor Promotes Fibrosis Downstream of TGFβ and IL‐6 in Chronic Cardiac Allograft Rejection

Cardiac transplantation is an effective treatment for multiple types of heart failure refractive to therapy. Although immunosuppressive therapeutics have increased survival rates within the first year posttransplant, chronic rejection (CR) remains a significant barrier to long‐term graft survival. Indicators of CR include patchy interstitial fibrosis, vascular occlusion and progressive loss of graft function. Multiple factors have been implicated in the onset and progression of CR, including TGFβ, IL‐6 and connective tissue growth factor (CTGF). While associated with CR, the role of CTGF in CR and the factors necessary for CTGF induction in vivo are not understood. To this end, we utilized forced expression and neutralizing antibody approaches. Transduction of allografts with CTGF significantly increased fibrotic tissue development, though not to levels observed with TGFβ transduction. Further, intragraft CTGF expression was inhibited by IL‐6 neutralization whereas TGFβ expression remained unchanged, indicating that IL‐6 effects may potentiate TGFβ‐mediated induction of CTGF. Finally, neutralizing CTGF significantly reduced graft fibrosis without reducing TGFβ and IL‐6 expression levels. These findings indicate that CTGF functions as a downstream mediator of fibrosis in CR, and that CTGF neutralization may ameliorate fibrosis and hypertrophy associated with CR.     

Age‐related expression,enzymatic solubility and modification with advanced glycation end‐products of fibrillar collagens in mouse lung

Changes in the expression of fibrillar collagens and post-translational modifications with advanced glycation end-products (AGEs) are often associated with tissue aging. Less is known about age-related changes in mouse lung tissue. Therefore, we studied the expression level and AGE load of fibrillar collagens in lungs from young (≤ 6 months), adult (15 months) and old (≥ 25 months) mice. The mRNA expression level was reduced in adult and old mice compared with the young. Old mice also showed a reduced protein level, whereas the adults even had more collagen protein. Fractionating of the fibrillar collagens into enzyme-soluble and insoluble collagens revealed a reduced solubility of collagens in old age. The enzymatic solubility of fibrillar collagens correlated inversely with the AGE load in the insoluble collagen as detected by the AGE-related fluorescence. While the intensity of the AGE-related fluorescence was increased in fibrillar collagens in response to age, the fluorescing AGE variant argpyrimidine was less affected. In summary, aging causes a reduced expression, lower enzymatic solubility and increased AGE load of fibrillar collagens in mouse lung tissue, but not all changes occur gradually with age.     

大黄酸对免疫性肝纤维化大鼠形成的影响

目的 观察大黄酸抗猪血清诱导的大鼠肝纤维化的效应并探讨其作用机制.方法 用猪血清腹腔内注射复制免疫损伤性肝纤维化模型,造模同时给予大黄酸口服作为肝纤维化防治组;免疫攻击16周后处死动物.行肝组织病理分析,免疫组化法检测肝内结缔组织生长因子(CTGF)、TGF-猹1表达.结果 16周后模型组大鼠形成典型的肝纤维化,肝内CTGF与TGF-猹1表达均显著增强,以大黄酸防治的大鼠,肝纤维化程度明显减轻,CTGF和TGF-猹1表达的阳性指数下降.结论 大黄酸能有效地减轻猪血清诱导的肝纤维化大鼠的肝损伤及纤维化程度,其机制可能与抑制TGF-猹1和CTGF表达有关.     

CTGF，PDGF和VEGF在臀肌挛缩带中的表达及其对挛缩带形成的作用

目的：探索细胞生长因子CTGF,PDGF和VEGF在臀肌挛缩带中的表达情况及其对臀肌挛缩症（GMC）的发病作用.方法：收集臀肌挛缩症患者挛缩带和病变旁肌肉的病理标本,运用逆转录-聚合酶链式反应（RT-PCR）、免疫组化和Western Blot技术检测细胞生长因子CTGF,PDGF和VEGF在病理组织中的表达情况.结果：细胞生长因子CTGF,PDGF和VEGF表达定位于成纤维细胞和血管内皮细胞的胞质中,并且呈强阳性表达,其在mRNA水平分别上调了3.5倍、5.8和2.7倍（P〈0.05）.蛋白水平的表达情况与mRNA一致,分别上调3.9倍、6.2倍和2.9倍（P〈0.05）.结论：细胞生长因子CTGF,PDGF和VEGF的表达上调可能是引起挛缩带产生的重要机制.     

Effect of houttuynia cordata aetherolea on adiponectin and connective tissue growth factor in a rat model of diabetes mellitus

Objective

To determine the effect of Houttuynia cordata Aetherolea on connective tissue growth factor and adiponectin in a rat model of diabetes mellitus (DM).

Methods

DM was induced in rats using streptozotocin (STZ) and high glucose-lipid animal feed. Animals were then treated with Houttuynia cordata Aetherolea for 8 weeks. Changes in connective tissue growth factor and adiponectin levels in rats were observed.

Results

Connective tissue growth factor and adiponectin levels in rats with DM improved after Houttuynia cordata Aetherolea treatment.

Conclusion

Houttuynia cordata Aetherolea had a positive effect on rats with DM by reducing levels of connective tissue growth factor and increasing adiponectin levels.     

决明子对糖尿病大鼠肾脏纤维化的抑制作用

目的:探讨决明子对糖尿病大鼠肾脏纤维化的保护作用及其可能机制。方法:将70只SD大鼠随机分为模型组(60只)和正常对照组(10只)。禁食12 h后大鼠1次性左下腹腔注射55 mg.kg-1链脲佐菌素(STZ)制备糖尿病模型,对照组给予等容量生理盐水。造模成功的大鼠随机分为模型组(生理盐水,4 mL.kg-1)、卡托普利组(10 mg.kg-1)和决明子低、中、高剂量组(1,5,10 g.kg-1),连续灌胃给药8周。检测空腹血糖、24 h尿白蛋白量、血肌酐及肌酐清除率,采用HE染色观察肾脏组织病理学变化,采用RT-PCR检测肾脏组织中转化生长因子(TGF-β1)及结缔组织生长因子(CTGF)的mRNA表达,采用Western blot检测肾脏Smad3和Smad6的蛋白表达。结果:与对照组相比,糖尿病模型组大鼠空腹血糖、血肌酐、24 h尿白蛋白量均显著升高(P<0.05),肌酐清除率显著下降(P<0.05),肾脏组织中TGF-β1和CTGF的mRNA表达均明显升高(P<0.05),Smad3蛋白表达亦显著升高(P<0.05),而Smad6蛋白表达则显著下降(P<0.05);与糖尿病模型组相比,决明子治疗组中的上述各指标均显著改善(P<0.05),肾脏病理学变化也明显减轻。结论:决明子可显著减轻糖尿病大鼠肾脏纤维化程度,其机制可能与抑制肾脏组织中TGF-β1,CTGF和Smad3,并促进Smad6的表达有关。     

The Role of Increased Connective Tissue Growth Factor in the Pathogenesis of Oral Submucous Fibrosis and its Malignant Transformation—An Immunohistochemical Study

Connective tissue growth factor (CTGF), a matricellular protein of the CCN family of extracellular matrix-associated heparin-binding proteins, is highly expressed in various organ fibrosis and several malignant tumors. Although a few studies have been conducted using CTGF in oral submucous fibrosis (OSF) and oral squamous cell carcinoma, no study has demonstrated its relation with various stages of OSF and its malignant transformation. The present study investigated the possible role of CTGF in the pathogenesis of OSF and its malignant transformation by using immunohistochemistry. Ten formalin-fixed paraffin-embedded tissue blocks, each of Stage 1 OSF, Stage 2 OSF, Stage 3 OSF, Stage 4 OSF, well- differentiated squamous cell carcinoma (WDSCC) with OSF and WDSCC without OSF were stained for CTGF by immunohistochemistry. Ten cases of healthy buccal mucosa (NOM) were included as controls. The present study demonstrated a statistically significant expression of CTGF in the epithelium and connective tissue of OSF and WDSCC with and without OSF cases against its complete absence in NOM. We observed an upregulation of CTGF expression from NOM to various stages of OSF to WDSCC with or without OSF. A gradual upregulation of the CTGF expression in various stages of OSF to WDSCC (with and without OSF) against its complete absence in NOM suggests that CTGF plays an important role in the pathogenesis of OSF and its malignant transformation.     

Molecular analysis of neocortical layer structure in the ferret

Molecular markers that distinguish specific layers of rodent neocortex are increasingly employed to study cortical development and the physiology of cortical circuits. The extent to which these markers represent general features of neocortical cell type identity across mammals, however, is unknown. To assess the conservation of layer markers more broadly, we isolated orthologs for 15 layer‐enriched genes in the ferret, a carnivore with a large, gyrencephalic brain, and analyzed their patterns of neocortical gene expression. Our major findings are: 1) Many but not all layer markers tested show similar patterns of layer‐specific gene expression between mouse and ferret cortex, supporting the view that layer‐specific cell type identity is conserved at a molecular level across mammalian superorders; 2) Our panel of deep layer markers (ER81/ETV1, SULF2, PCP4, FEZF2/ZNF312, CACNA1H, KCNN2/SK2, SYT6, FOXP2, CTGF) provides molecular evidence that the specific stratifications of layers 5 and 6 into 5a, 5b, 6a, and 6b are also conserved between rodents and carnivores; 3) Variations in layer‐specific gene expression are more pronounced across areas of ferret cortex than between homologous areas of mouse and ferret cortex; 4) This variation of area gene expression was clearest with the superficial layer markers studied (SERPINE2, MDGA1, CUX1, UNC5D, RORB/NR1F2, EAG2/KCNH5). Most dramatically, the layer 4 markers RORB and EAG2 disclosed a molecular sublamination to ferret visual cortex and demonstrated a molecular dissociation among the so‐called agranular areas of the neocortex. Our findings establish molecular markers as a powerful complement to cytoarchitecture for neocortical layer and cell‐type comparisons across mammals. J. Comp. Neurol. 518:3272–3289, 2010. © 2010 Wiley‐Liss, Inc.     

阿霉素肾病大鼠肾脏CTGF的表达及意义

目的:观察阿霉素肾病模型大鼠肾脏结缔组织生长因子(connective tissue growing factor,CTGF)的表达情况,探讨其与肾小球硬化发生发展的关系.方法:16只雄性SD大鼠随机分为对照组和模型组(n=8),模型组大鼠采用尾静脉注射阿霉素的方法建立肾病模型.分别检测两组大鼠的24h尿蛋白定量及血生化、肾功能指标,免疫组化染色及荧光定量PCR法分别检测大鼠肾脏CTGF蛋白和mRNA的表达.结果:与对照组比较,模型组大鼠的Alb明显降低,24h尿蛋白定量、TC、BUN、Cr及肾脏CTGF蛋白和mRNA的表达明显升高(P＜0.01).结论:CTGF过度表达是肾小球硬化发生发展的重要因素,可能是通过促进细胞外基质积聚导致肾小球硬化.