Transcriptional regulation of connective tissue growth factor by sphingosine 1-phosphate in rat cultured mesangial cells

Connective tissue growth factor (CTGF) is induced by transforming growth factor-beta (TGF-beta) via Smad activation in mesangial cells. We recently reported that sphingosine 1-phosphate (S1P) induces CTGF expression in rat cultured mesangial cells. However, the mechanism by which S1P induces CTGF expression is unknown. The present study revealed that S1P-induced CTGF expression is mediated via pertussis toxin-insensitive pathways, which are involved in the activation of small GTPases of the Rho family and protein kinase C. We also showed by luciferase reporter assays and chromatin immunoprecipitation that S1P induces CTGF expression via Smad activation as TGF-beta does.     

LncRNA TUG1 acts as a competing endogenous RNA to mediate CTGF expression by sponging miR-133b in myocardial fibrosis after myocardial infarction

Myocardial fibrosis (MF) is one of the basic causes of many cardiovascular diseases. Noncoding RNAs (ncRNAs), including microRNA (miRNA) and long noncoding RNA (lncRNA), have been reported to play an indispensable role in MF. The current work is focused on investigating the biological role of lncRNA taurine upregulation gene 1 (TUG1) in activating cardiac myofibroblasts as well as the underlying mechanism. The outcome revealed that after myocardial infarction TUG1 expression increased and miR-133b expression decreased in the rat model of MF. The expression level of TUG1 increased following AngII treatment in cardiac myofibroblast. TUG1 knockdown inhibited the Ang-II induced cardiac myofibroblast activation and TUG1 overexpression increased proliferation and collagen generation of cardiac myofibroblasts. Bioinformatic prediction programs predicted that TUG1 had MRE directly combined with miR-133b seed sequence, luciferase activity, and RIP experiments indicated that TUG1, acted as a sponger and interacted with miR-133b in cardiac myofibroblasts. Furthermore, a target of miR-133b was CTGF and CTGF knockdown counteracted the promotion of MF by miR-133b knockdown. Collectively, our study suggested that TUG1 mediates CTGF expression by sponging miR-133b in the activation of cardiac myofibroblasts. The current work reveals a unique role of the TUG1/miR-133b/CTGF axis in MF, thus suggesting its immense therapeutic potential in the treatment of cardiac diseases.     

Angiogenesis is not impaired in connective tissue growth factor (CTGF) knock-out mice.

Connective tissue growth factor (CTGF) is a member of the CCN family of growth factors. CTGF is important in scarring, wound healing, and fibrosis. It has also been implicated to play a role in angiogenesis, in addition to vascular endothelial growth factor (VEGF). In the eye, angiogenesis and subsequent fibrosis are the main causes of blindness in conditions such as diabetic retinopathy. We have applied three different models of angiogenesis to homozygous CTGF(-/-) and heterozygous CTGF(+/-) mice to establish involvement of CTGF in neovascularization. CTGF(-/-) mice die around birth. Therefore, embryonic CTGF(-/-), CTGF(+/-), and CTGF(+/+) bone explants were used to study in vitro angiogenesis, and neonatal and mature CTGF(+/-) and CTGF(+/+) mice were used in models of oxygen-induced retinopathy and laser-induced choroidal neovascularization. Angiogenesis in vitro was independent of the CTGF genotype in both the presence and the absence of VEGF. Oxygen-induced vascular pathology in the retina, as determined semi-quantitatively, and laser-induced choroidal neovascularization, as determined quantitatively, were also not affected by the CTGF genotype. Our data show that downregulation of CTGF levels does not affect neovascularization, indicating distinct roles of VEGF and CTGF in angiogenesis and fibrosis in eye conditions.     

老年慢性心力衰竭患者血清sST2、CTGF、suPAR与心功能的关系及其联合检测对心血管事件的预测价值

摘要 目的：探讨老年慢性心力衰竭（CHF）患者血清可溶性肿瘤生成抑制因子2（sST2）、可溶性尿激酶纤溶酶原激活物受体（suPAR）、结缔组织生长因子（CTGF）与心功能的关系及其联合检测对心血管事件（CVE）的预测价值。方法：选取2019年1月～2022年1月我院收治的150例老年CHF患者（CHF组）,其中纽约心脏病协会（NYHA）分级Ⅱ级49例、Ⅲ级72例、Ⅳ级29例,根据随访6个月是否发生CVE分为CVE组（n=34）和非CVE组（n=116）,另选取同期在我院进行体检的110例健康体检志愿者作为对照组。通过超声心动图检查左室收缩末期内径（LVESD）、左室射血分数（LVEF）、左心室舒张末期内径（LVEDD）,酶联免疫吸附试验法检测血清sST2、CTGF、suPAR水平。多因素Logistic回归分析老年CHF患者发生CVE的影响因素,Pearson或Spearman相关性分析老年CHF患者血清sST2、CTGF、suPAR水平与超声心功能指标和NYHA分级的相关性,受试者工作特征（ROC）曲线分析血清sST2、CTGF、suPAR对老年CHF患者发生CVE的预测价值。结果：CHF组血清sST2、CTGF、suPAR水平和LVESD、LVEDD高于对照组,LVEF低于对照组（P＜0.05）。NYHA分级Ⅱ级、Ⅲ级、Ⅳ级老年CHF患者血清sST2、CTGF、suPAR水平均依次升高（P＜0.05）。 老年CHF患者血清sST2、CTGF、suPAR水平与LVEF呈负相关,与LVESD、LVEDD、NYHA分级呈正相关（P均＜0.05）。NYHA分级≥Ⅲ级（OR=2.318）、N末端B型钠尿肽前体（NT-proBNP）升高（OR=1.104）、sST2升高（OR=1.641）、CTGF升高（OR=1.644）、suPAR升高（OR=1.892）为老年CHF患者CVE发生的独立危险因素,LVEF升高（OR=0.839）为其独立保护因素,差异均有统计学意义（P＜0.05）。血清sST2、CTGF、suPAR单独与联合预测老年CHF患者发生CVE的曲线下面积分别为0.788、0.782、0.771、0.936。结论：老年CHF患者血清sST2、CTGF、suPAR水平升高与心功能异常和CVE发生密切相关,联合检测血清sST2、CTGF、suPAR水平对老年CHF患者发生CVE的预测价值较高。     

Thrombopoietic-mesenchymal interaction that may facilitate both endochondral ossification and platelet maturation via CCN2

CCN2 plays a central role in the development and growth of mesenchymal tissue and promotes the regeneration of bone and cartilage in vivo. Of note, abundant CCN2 is contained in platelets, which is thought to play an important role in the tissue regeneration process. In this study, we initially pursued the possible origin of the CCN2 in platelets. First, we examined if the CCN2 in platelets was produced by megakaryocyte progenitors during differentiation. Unexpectedly, neither megakaryocytic CMK cells nor megakaryocytes that had differentiated from human haemopoietic stem cells in culture showed any detectable CCN2 gene expression or protein production. Together with the fact that no appreciable CCN2 was detected in megakaryocytes in vivo, these results suggest that megakaryocytes themselves do not produce CCN2. Next, we suspected that mesenchymal cells situated around megakaryocytes in the bone marrow were stimulated by the latter to produce CCN2, which was then taken up by platelets. To evaluate this hypothesis, we cultured human chondrocytic HCS-2/8 cells with medium conditioned by differentiating megakaryocyte cultures, and then monitored the production of CCN2 by the cells. As suspected, CCN2 production by HCS-2/8 was significantly enhanced by the conditioned medium. We further confirmed that human platelets were able to absorb/uptake exogenous CCN2 in vitro. These findings indicate that megakaryocytes secrete some unknown soluble factor(s) during differentiation, which factor stimulates the mesenchymal cells to produce CCN2 for uptake by the platelets. We also consider that, during bone growth, such thrombopoietic-mesenchymal interaction may contribute to the hypertrophic chondrocyte-specific accumulation of CCN2 that conducts endochondral ossification.     

Oxidative stress plays a role in high glucose-induced activation of pancreatic stellate cells

The activation of pancreatic stellate cells (PSCs) is thought to be a potential mechanism underlying islet fibrosis, which may contribute to progressive β-cell failure in type 2 diabetes. Recently, we demonstrated that antioxidants reduced islet fibrosis in an animal model of type 2 diabetes. However, there is no in vitro study demonstrating that high glucose itself can induce oxidative stress in PSCs. Thus, PSCs were isolated and cultured from Sprague Dawley rats, and treated with high glucose for 72 h. High glucose increased the production of reactive oxygen species. When treated with high glucose, freshly isolated PSCs exhibited myofibroblastic transformation. During early culture (passage 1), PSCs treated with high glucose contained an increased number of α-smooth muscle actin-positive cells. During late culture (passages 2–5), PSCs treated with high glucose exhibited increases in cell proliferation, the expression of fibronectin and connective tissue growth factor, release of interleukin-6, transforming growth factor-β and collagen, and cell migration. Finally, the treatment of PSCs with high glucose and antioxidants attenuated these changes. In conclusion, we demonstrated that high glucose increased oxidative stress in primary rat PSCs, thereby facilitating the activation of these cells, while antioxidant treatment attenuated high glucose-induced PSC activation.     

The expression of chicken NOV, a member of the CCN gene family, in early stage development

The nephroblastoma overexpressed gene, NOV, is a member of the CCN gene family. We investigated the NOV gene expression pattern in the chicken during early stage embryogenesis. Several embryonic structures showed a distinct expression pattern. The initial expression was detected in Hensen's node (Hamburger and Hamilton stage (HH) 5). The expression was noted in the presumptive notochord and floor plate forming cells. The expression on the left side was more elongated posteriorly, a type of left-right asymmetry. Chicken NOV gene expression in the forming notochord and floor plate was observed until HH 18. The expression was also detected in the ventral area of the mesencephalon and isthmus at HH 14-16.