Role of transforming growth factor-beta 1 and connective tissue growth factor levels in coronavirus disease-2019-related lung Injury: a prospective, observational, cohort study

ABSTRACT Background: Coronavirus disease-2019 (COVID-19) results in acute lung injury. This study examined the usefulness of serum transforming growth factor-beta 1 (TGF-β1) and connective tissue growth factor (CTGF) levels in predicting disease severity in COVID-19 patients with pulmonary involvement. Methods: Fifty patients with confirmed COVID-19 and pulmonary involvement between September 2020, and February 2021 (Group 1) and 45 healthy controls (Group 2) were classified into three subgroups based on clinical severity: moderate, severe, and critical pneumonia. Serum TGF-β1 and CTGF concentrations were measured on days 1 and 7 of admission in Group 1 using an enzyme-linked immunosorbent assay. These concentrations were also measured in control cases. The mean serum TGF-β1 and CTGF levels were then compared among COVID-19 patients, based on clinical severity. Results: Significantly higher mean serum TGF-β1 and CTGF levels were observed on both days in Group 1 than in the control group. The mean serum TGF-β1 and CTGF levels on day 7 were also significantly higher than those on day 1 in Group 1. The critical patient group had the highest serum TGF-β1 and CTGF levels on both days, and the difference between this group and the moderate and severe pneumonia groups was significant. Cutoff values of 5.36 ng/mL for TGF-β1 and 626.2 pg/mL for CTGF emerged as predictors of COVID-19 with pulmonary involvement in receiver-operating characteristic curve analysis. Conclusions: TGF-β1 and CTGF are potential markers that can distinguish COVID-19 patients with pulmonary involvement and indicate disease severity. These findings may be useful for initiating treatment for early-stage COVID-19.

A smart O2-generating nanocarrier optimizes drug transportation comprehensively for chemotherapy improving

@#Drug transportation is impeded by various barriers in the hypoxic solid tumor, resulting in compromised anticancer efficacy. Herein, a solid lipid monostearin (MS)-coated CaO2/MnO2 nanocarrier was designed to optimize doxorubicin (DOX) transportation comprehensively for chemotherapy enhancement. The MS shell of nanoparticles could be destroyed selectively by highly-expressed lipase within cancer cells, exposing water-sensitive cores to release DOX and produce O2. After the cancer cell death, the core-exposed nanoparticles could be further liberated and continue to react with water in the tumor extracellular matrix (ECM) and thoroughly release O2 and DOX, which exhibited cytotoxicity to neighboring cells. Small DOX molecules could readily diffuse through ECM, in which the collagen deposition was decreased by O2-mediated hypoxia-inducible factor-1 inhibition, leading to synergistically improved drug penetration. Concurrently, DOX-efflux-associated P-glycoprotein was also inhibited by O2, prolonging drug retention in cancer cells. Overall, the DOX transporting processes from nanoparticles to deep tumor cells including drug release, penetration, and retention were optimized comprehensively, which significantly boosted antitumor benefits.

Changes of fibrotic factors in the recovery process of skeletal muscle strain / 中国组织工程研究

BACKGROUND: Skeletal muscle injury is the most common sports injury, which can develop fibrosis and then impair muscle function. Varieties of fibrotic factors are involved in the repair of skeletal muscle and effectively regulate the process of tissue fibrosis. OBJECTIVE: To establish a rat model of acute gastrocnemius strain, observe the changes of fibrotic factors in the recovery of injury and explore the possible roles of fibrotic factors. METHODS: Totally 112 male Sprague-Dawley rats were randomly divided into control group (n=56) and injury group (n=56). The left gastrocnemius muscle was injured by a rat gastrocnemius pull-off device in the injury group. Each group was further subdivided into immediately group, 2 days group, 4 days group, 7 days group, 14 days group, 21 days group and 28 days group depending on the sampling time points. There were eight rats in each subgroup. The control group did not receive any intervention. The left gastrocnemius muscle of the injury group was strained by specialized equipment. Muscle fiber injury and inflammation were observed by hematoxylin-eosin staining. The expression of transforming growth factor β1 (TGF-β1), connective tissue growth factor (CTGF), matrix metalloproteinase-1 (MMP-1), and matrix metalloproteinase tissue inhibitor 1 (TIMP-1) were detected by western blot. The experiment was approved by the Sports Science Experimental Ethics Committee of Beijing Sport University. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the muscle fibers of the gastrocnemius muscle arranged disorderedly and inflammatory cells were aggregated in the injury group. Compared with the control group, protein expression levels of TGF-β1 and CTGF significantly increased in the injury group from the 2nd day throughout the recovery (P 0.05). TIMP-1 expression significantly increased from the 7th day until the 28th day (P < 0.05). These findings indicate that TGF-β1/CTGF signal pathway may be successively activated during the recovery of skeletal muscle strain, triggering the occurrence and development of skeletal muscle fibrosis. Meanwhile, MMP-1 and TIMP-1 protein may be activated to participate in the recovery of injury.

Early intervention with Di-Dang Decoction prevents macrovascular fibrosis in diabetic rats by regulating the TGF-β1/Smad signalling pathway / 中国天然药物

Macroangiopathy is a complication of Type II Diabetes Mellitus (T2DM), which is mainly caused by fibrosis of blood vessels. Using T2DM rat models, we investigated whether the traditional Chinese medicine, Di-Dang Decoction (DDD), exhibited anti-fibrotic actions on great vessels. T2DM rats were randomly divided into non-intervention group, early-, middle-, late-stage DDD intervention groups and control groups, including pioglitazone group and aminoguanidine group. After administration of DDD to T2DM rats at different times, we detected the amount of extracellular matrix (ECM) deposition in the thoracic aorta. The results showed that early-stage intervention with DDD could effectively protect great vessels from ECM deposition. Considering that TGF-β1 is the master regulator of fibrosis, we further validated at the molecular level that, compared to middle- and late-stage intervention with DDD, early-stage intervention with DDD could significantly decrease the expression levels of factors related to the activated TGF-β1/Smad signalling pathway, as well as the expression levels of downstream effectors including CTGF, MMP and TIMP family proteins, which were directly involved in ECM remodelling. Therefore, early-stage intervention with DDD can reduce macrovascular fibrosis and prevent diabetic macroangiopathy.

lncRNA H19 acts as a ceRNA to regulate the expression of CTGF by targeting miR-19b in polycystic ovary syndrome

The etiology of polycystic ovary syndrome (PCOS) is complex and the pathogenesis is not fully understood. Some studies have shown that dysregulation of ovarian granulosa cells may be related to abnormal follicles and excessive androgen in women with PCOS. Our team has also confirmed the high expression status of H19 in PCOS patients in the early stage. However, the relationship between H19 and miR-19b in the development of PCOS is still unknown. Therefore, we used bioinformatics to predict the binding sites of human H19 and miR-19b, and of miR-19b and CTGF genes. After the silencing and overexpression of H19, real-time polymerase chain reaction (PCR) was used to detect the expressions of H19, miR-19b, and CTGF. Western blotting was used to detect CTGF protein. Proliferation of KGN cells after H19 silencing was detected by CCK8. Flow cytometry was used to detect the apoptosis of KGN cells after H19 silencing. After the overexpression of H19, it was found that the expression of miR-19b gene decreased and the expression of CTGF increased, whereas silencing of H19 did the opposite. In addition, H19 could promote cell proliferation and decrease cell apoptosis. Finally, luciferase reporter assays showed that the 3′-end sequences of lncRNA H19 and CTGF contained the binding site of miR-19b. In conclusion, our study indicated that lncRNA H19 acted as a ceRNA to bind to miR-19b via a "sponge" to regulate the effect of CTGF on KGN cells, which may play a vital role in PCOS.

Regulatory Mechanism of MicroRNA-145 in the Pathogenesis of Acute Aortic Dissection

PURPOSE: Previous studies have confirmed that microRNAs play important roles in the pathogenesis of acute aortic dissection (AAD). Here, we aimed to explore the role of miR-145 and its regulatory mechanism in the pathogenesis of AAD. MATERIALS AND METHODS: AAD tissue samples were harvested from patients with aortic dissection and normal donors. Rat aortic vascular smooth muscle cells (VSMCs) were transfected with miR-145 mimic/inhibitor or negative control mimic/inhibitor. Gene and protein expression was measured in human aortic dissection tissue specimens and VSMCs by qRT-PCR and Western blot. Luciferase reporter assay was applied to verify whether connective tissue growth factor (CTGF) was a direct target of miR-145 in VSMCs. Methyl thiazolyl tetrazolium assay was used to detect VSMC viability. RESULTS: miR-145 expression was downregulated in aortic dissection tissues and was associated with the survival of patients with AAD. Overexpression of miR-145 promoted VSMC proliferation and inhibited cell apoptosis. Moreover, CTGF, which was increased in aortic dissection tissues, was decreased by miR-145 mimic and increased by miR-145 inhibitor. Furthermore, CTGF was confirmed as a target of miR-145 and could reverse the promotion effect of miR-145 on the progression of AAD. CONCLUSION: miR-145 suppressed the progression of AAD by targeting CTGF, suggesting that a miR-145/CTGF axis may provide a potential therapeutic target for AAD.

Effects of connective tissue growth factor gene-silencing on cell growth and cell cycle of hepatic stellate cells / 上海交通大学学报（医学版）

Objective • To investigate the effects of silencing connective tissue growth factor (Ctgf) gene on the growth, cell cycle and the expression of TGF-β1, Smad3 and Smad7 of rat hepatic stellate cell line HSCT6. Methods • The recombinant lentivirus vector pCDH/Ctgf-shRNA of Ctgf gene was constructed by RNA interference. The recombinant vector was packaged to obtain highly infectious pCDH/Ctgf-shRNA lentivirus particles for HSCT6 infection. The expression of green fluorescent protein (GFP) in the transfected HSCT6 cells was observed under fluorescence microscope. The effects of Ctgf-shRNA lentivirus on the growth of HSCT6 cells were tested by CCK-8. The effects of Ctgf-shRNA lentivirus on the cell cycle of HSCT6 cells were analyzed by flow cytometry (FCM). The effects of Ctgf-shRNA lentivirus on the expression of mRNA of Ctgf, Tgf-β1, Smad3 and Smad7, and their proteins in HSCT6 cells were detected by real-time PCR and Western blotting, respectively. Results • The lentiviral vector pCDH/Ctgf-shRNA has been constructed successfully. The HSCT6 cells transfected by Ctgf-shRNA lentivirus significantly expressed GFP under fluorescence microscope. The results of CCK-8 confirmed that the growth of HSCT6 cells transfected by Ctgf-shRNA lentivirus was slower than that of controls and the differences were statistically significant after being cultured for 72 h (P<0.05). The results of FCM revealed that the growth of HSCT6 cells transfected by Ctgf-shRNA lentivirus was blocked in the S phase of cell cycle. The results of real-time PCR and Western blotting showed that the Ctgf-shRNA lentivirus effectively silenced Ctgf gene, down-regulated the expression of genes and encoding proteins of TGF-β1, and Smad3 of HSCT6 and up-regulated the expression of genes and encoding proteins of Smad7 of HSCT6 cells. The differences between transfected cells and controls were statistically significant (P<0.05). Conclusion • Silencing Ctgf gene can effectively inhibit the growth of HSCT6 cells, down-regulate the expression of TGF-β1 and Smad3 and up-regulate the expression of Smad7. The inhibition of the growth of HSCT6 cells may be closely related to interference of the TGF-β1/Smads (Smad3 and Smad7) signaling pathway.

Control of TGF-βl/Smads signaling pathway in diabetic nephropathy rats by Danzhijiangtang capsule / 中国药理学通报

Aim To investigate the preventive and therapeutic effects of Danzhijiangtang capsule (DZJT) on diabetic nephropathy in rats and its mechanism. Methods Twelve rats were randomly selected as the normal group from 100 male SD rats, the remaining was induced by high fat diet and streptozotocin (34 mg • kg-1, ip), and 65 successful modeling rats were randomly divided into model group, DZJT high, medium, low dose group and pioglitazone group. Gavage was administered for 8 weeks. The blood glucose, renal function and the expression of TGF-β1 in serum were measured. The pathological changes of renal tissue were observed. The expressions of TGF-β1 Smad2, Smad3, Smad7 and CTGF in renal tissues were detected. Results Compared with normal group, the blood glucose of rats in model group significantly increased, renal and glomerular hypertrophy, basement membrane thickening, extracellular matrix increased, and inflammatory cell infiltration, fibrous tissue hyper plasia, kidney pathological score increased significantly; the expression of TGF-β1, SmacB and CTGF in renal tissues increased significantly, and the expression of Smad2 and Smad7 decreased markedly. Compared with model group, the above indicators in each treatment group decreased in different degrees, and the pathological morphology of kidney was improved obviously; TGF-β1, Smad3 and CTGF decreased, while the levels of Smad2 and Smad7 increased. Conclusions The capsule can effectively improve the symptoms of diabetes and alleviate the kidney damage in rats, and its mechanism may be related to the regulation of TGF-β1/Smads signaling pathway and its downstream CTGF expression.

CTGF and TGF-β1 detection levels in serum and bronchoalveolar lavage fluid of patients with chronic obstructive pulmonary disease and significance / 国际检验医学杂志

Objective To explore connective tissue growth factor(CTGF)and transforming growth factor β1(TGF-β1)detection levels in serum and bronchoalveolar lavage fluid of patients with chronic obstructive pulmonary disease(COPD)and significance.Methods Totally 65 patients with COPD treated in the hospital from March 2016 to March 2017 were selected as the subjects,the patients were divided into acute exacerba-tion group(32 cases)and stable phase group(33 cases)according to the severity of the disease.Another 35 cases of physical examination in the hospital were selected as the control group.Serum and bronchoalveolar lavage fluid CTGF and TGF-β1 levels were detected by using enzyme-linked immunosorbent assay(ELISA) method,and lung function index,such as forced vital capacity(FVC),forced expiratory volume(FEV1),and FEV1/FVC of patients with COPD were detected,and the relationship was analyzed of CTGF and TGF-β1 in serum and bronchoalveolar lavage fluid and lung function.Results CTGF and TGF-β1 levels in serum and bronchoalveolar lavage fluid in acute exacerbation group were significantly higher than those in stable group and control group(P<0.05).CTGF and TGF-β1 levels in serum and bronchoalveolar lavage fluid of the sta-ble group were significantly higher than those in the control group(P<0.05).Lung function results showed that lung function index FVC,FEV1 and FEV1/FVC value in acute exacerbation group decreased significantly when compared with those in stable group and the control group(P<0.05).FEV1 and FEV1/FVC in the sta-ble group was significantly lower than that in the control group(P<0.05).CTGF expression level was posi-tively correlated in serum and bronchoalveolar lavage fluid of acute exacerbation group,and TGF-β1 was also positively correlated.Serum CTGF and TGF-β1 levels were positively correlated,and CTGF and TGF-β1 in alveolar lavage fluid were also positively correlated.Serum CTGF and TGF-β1 levels were negatively correla-ted with FVC,FEV1,FEV1/FVC,and CTGF and TGF-β1 in alveolar lavage fluid were negatively correlated with FVC,FEV1 and FEV1/FVC respectively.Conclusion The high expression of CTGF and TGF-β1 is closely related to the occurrence and development of COPD,and it can be used as an index for monitoring the condition of COPD.

A Study on the Effects of Occlusal Function on CTGF Expression in Orthodontics / 昆明医科大学学报

Objective To investigate CTGF changes in the expression of the gum tissue before and after orthodontic treatment, and to preliminarily explore the modification mechanism of gingival tissue and the effectiveness of the intervention measures. Methods 72 male Sprague-Dawley rats of 12-week-old, weight about 250 g-300 g, randomly divided into 6 groups: blank control group (A), healthy teeth orthodontic group (B), low functional group (C), low occlusal function teeth orthodontic group (D), combined intervention group (E), bite orthodontic intervention group (F). The results of the study were compared at 1 w, 2 w, 4 w, 6 w.Results (1) HE staining results showed the atrophy of the gingival tissue, which suggested that occlusal hypofunction SD rats model were successfully established. (2) Fluorescence quantitative results of CTGF in gingival tissue: 6 w: group B was higher than group D and group F (P＜0.05). Conclusion (1) the expression of CTGF in the low occlusion group was lower than that of the normal control group with time, and the gum tissue was vulnerable to atrophy. (2) It remains to be further studied whether the bite force recovery is effective.

Research progress of CTGF/CCN2 in the development and targeted therapy of malignant tumor / 实用肿瘤学杂志

The current incidence of malignant tumors is increasing.Although we have been exploring the mechanism of tumor development and its treatment,its efficacy is little.Malignant tumor development mechanism is very complex,and a variety of proteins and genes involved.Connective tissue growth factor(CTGF,also known as CCN2)is a secreted protein,a member of the CCN family,which plays a very important role in the development and progression of tumors.This article summarizes the structure and function of CTGF /CCN2 protein,the mechanism of action in the development of malignant tumors and the latest research progress in targeted therapy.

Effects of recombinant human relaxin-2 on hepatic stellate cell proliferation, synthesis of Collagen Ⅰ,and mRNA expressions of TGF-β1 and CTGF in vitro / 西安交通大学学报(医学版)

ABSTRACT:Objective To validate that relaxin can resist hepatic fibrosis at the cellular level and explore its molecular mechanism in order to provide experimental basis for the treatment of liver cirrhosis.Methods Cultured HSC-T6s were treated with different concentrations (20,50 and 100 ng/mL)of recombinant human relaxin-2 (RLX-2).The proliferation of HSC-T6 was measured by MTT colorimetric assay.The content of type Ⅰcollagen in the cell culture supernatant of each group was detected by ELISA at 48 h of drug intervention;RT-PCR was used to detect the mRNA expressions of CTGF and TGF-β1 in HSC-T6 at 48 h of drug intervention.Results RLX-2 inhibited the proliferation of HSC and reduced type Ⅰ collagen content of HSC cells.It also inhibited the CTGF mRNA expression of HSC,but did not have a significant effect on the expression of TGF-β1 mRNA. Conclusion In the experiment of culturing HSC-T6 in vitro,RLX-2 may play a role in rat liver fibrosis by inhibiting cell proliferation and type Ⅰ collagen and CTGF mRNA expressions.

Effect of Palmitoyl-Pentapeptide (Pal-KTTKS) on Wound Contractile Process in Relation with Connective Tissue Growth Factor and α-Smooth Muscle Actin Expression

To evaluate whether Palmitoyl-pentapeptide (Pal-KTTKS), a lipidated subfragment of type 1 pro-collagen (residues 212–216), plays a role in fibroblast contractility, the effect of Pal-KTTKS on the expression of pro-fibrotic mediators in hypertropic scarring were investigated in relation with trans-differentiation of fibroblast to myofibroblast, an icon of scar formation. α-SMA was visualized by immunofluorescence confocal microscopy with a Cy-3-conjugated monoclonal antibody. The extent of α-SMA-positive fibroblasts was determined in collagen lattices and in cell culture study. Pal-KTTKS (0–0.5 µM) induced CTGF and α-SMA protein levels were determined by western blot analysis and fibroblast contractility was assessed in three-dimensional collagen lattice contraction assay. In confocal analysis, fibroblasts were observed as elongated and spindle shapes while myofibroblast observed as squamous, enlarged cells with pronounced stress fibers. Without Pal-KTTKS treatment, three quarters of the fibroblasts differentiates into the myofibroblast; α-SMA-positive stress fibers per field decreased twofold with 0.1 µM Pal-KTTKS treatment (75 ± 7.1 vs 38.6 ± 16.1%, n = 3, p<0.05). The inhibitory effect was not significant in 0.5 µM Pal-KTTKS treatment. Stress fiber level and collagen contractility correlates with α-SMA expression level. In conclusion, Pal-KTTKS (0.1 µM) reduces α-SMA expression and trans-differentiation of fibroblasts to myofibroblast. The degree of reduction is dose-dependent. An abundance of myofibroblast and fibrotic scarring is correlated with excessive levels of α-SMA and collagen contractility. Delicate balance between the wound healing properties and pro-fibrotic abilities of pentapeptide KTTKS should be considered for selecting therapeutic dose for scar prevention.

Effects of total flavonoids of litchi and rosiglitazone on expression of PPAR-γ and CTGF in HSC-T6 rat hepatic stellate cells / 实用医学杂志

Objective To investigate effects of total flavonoids of litchi (TFL) on the proliferation of rat hepatic stellate cells (HSC-T6) in comparison with western medicine rosiglitazone, and to explore the mechanism of anti hepatic fibrosis of TFL. Methods Effect of TFL on proliferation of HSC-T6 was examined by MTT. The expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) mRNA, connective tissue growth factor (CTGF) mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR. Effects on HSC-T6 CTGF protein from TFL and rosiglitazone were detected by Western bloting. Results The expression of PPAR-γ mRNA was upregulated and the expression of CTGF mRNA and protein was downregulated after exposure to TFL and rosiglitazone for 72 hours. And the effect of TFL increased with the increase of concentration. Conclusion TFL can inhibit the proliferation of HSC-T6 and antagonizing liver fibrosis. This mechanism may be associated with the upregulation of PPAR-γ expression and the downregulation of CTGF expression.

Research progress on the relationship between CTGF and the formation of congenital hydronephro-sis / 中国医师杂志

Congenital hydronephrosis is a common disease in children,causes and pathogenesis re-mains unclear.Hydronephrosis formation process is a slow and gradual development to the dynamic process of renal fibrosis.It involves a variety of cells,cytokines and ECM,more than aspects of interaction and mu-tual adjustment.The study found abroad that CTGF is closely related to the formation of congenital hydrone-phrosis.This article reviews the recent progress made CTGF relationship with congenital hydronephrosis formed on.

Yap1 plays a protective role in suppressing free fatty acid-induced apoptosis and promoting beta-cell survival

Mammalian pancreatic β-cells play a pivotal role in development and glucose homeostasis through the production and secretion of insulin. Functional failure or decrease in β-cell number leads to type 2 diabetes (T2D). Despite the physiological importance of β-cells, the viability of β-cells is often challenged mainly due to its poor ability to adapt to their changing microenvironment. One of the factors that negatively affect β-cell viability is high concentration of free fatty acids (FFAs) such as palmitate. In this work, we demonstrated that Yes-associated protein (Yap1) is activated when β-cells are treated with palmitate. Our loss- and gain-of-function analyses using rodent insulinoma cell lines revealed that Yap1 suppresses palmitate-induced apoptosis in β-cells without regulating their proliferation. We also found that upon palmitate treatment, re-arrangement of F-actin mediates Yap1 activation. Palmitate treatment increases expression of one of the Yap1 target genes, connective tissue growth factor (CTGF). Our gain-of-function analysis with CTGF suggests CTGF may be the downstream factor of Yap1 in the protective mechanism against FFA-induced apoptosis.

Effects of FAR on myocardial fibrosis of diabetic rats / 中国药理学通报

Aim To study the effects of Free Anthra-quinone from Rhubarb (FAR)on myocardial CTGF and collagen expression and interstitial fibrosis in dia-betic rats.Methods The male SD rats were randomly divided into normal group (CON),diabetic cardiomy-opathy group (DCM) and FAR treatment group (FAR).Streptozocin was intraperitoneally injected in-to the animals in the latter 2 groups to induce diabetic rat model.The model was expected to be stable for 2 weeks before the treatment.At the end of the 8th week in treatment,fasting plasma glucose and heart mass in-dex were measured.Masson staining was used to ob-serve the myocardial fibrosis.RT-PCR was used to de-tect the mRNA levels of CTGF,procollagen type Ⅰand collagen type Ⅲ.Immunohistochemical method was used to detect the content of CTGF.ELISA was used to detect the depositions of collagen type I and collagen type Ⅲ. Results Compared with CON group,fasting plasma glucose,heart mass index,the degree of myocardial fibrosis,and the expressions of CTGF,collagen type I and collagen type Ⅲ in left ven-tricular myocardial tissue of DCM group were signifi-cantly increased. However, compared with DCM group,fasting plasma glucose,heart mass index,the degree of myocardial fibrosis,and the expressions of CTGF,collagen type I and collagen type Ⅲ in left ven-tricular myocardial tissue of FAR-treated rats were sig-nificantly decreased.Conclusion FAR retards the process of myocardial fibrosis in diabetic rats by down-regulating the expression of CTGF,reducing the syn-thesis and depositions of collagen type I and collagen type Ⅲ.

Effects of shRNA-CTGF on the expression of collagen-Ⅰ in keloid / 中华医学美学美容杂志

Objective To explore the effects of recombinant plasmids of pGPU6/GFP/NeoshRNA-CTGF (shRNA-CTGF) on the type Ⅰ collagen (COL-Ⅰ) protein expression in keloid,through RNA interference on connective tissue growth factor (CTGF) in vivo and in vitro.Methods Recombinant plasmids were designed and constructed by specific shRNA-CTGF; After transfeced human keloid fibroblast with shRNA-CTGF in vitro,RT-PCR was used to detect the CTGF mRNA level,and Western blot to detect the secretion of COL-Ⅰ.After transfected the keloid of nude mice with shRNA-CTGF in vivo,RT-PCR was used to detect the CTGF and COL-Ⅰ mRNA level,and Western blot was used to detect the protein expression of COL-Ⅰ.Results Recombinant plasmids of CTGF were successfully constructed,and the CTGF gene expression was significantly decreased in vivo and in vitro by 86.8％ and 54.1 ％,respectively; Down-regulation of CTGF in vitro significantly inhibited the mRNA and protein level of COL-Ⅰ by 76.8％ and 65.6％,respectively; Down-regulation of CTGF in vivo significantly reduced the COL-Ⅰ mRNA and protein level by 52.7％ and 48.0％,respectively.Conclusions CTGF gene expression is successfully down-regulated by the recombinant plasmid of shRNA-CTGF in vivo and in vitro.shRNA-CTGF significantly reduces the COL-Ⅰ protein level in keloid.It implies that CTGF gene is a potential target in the therapy of pathological scar.

Effect of combination of ganoderma lucidum polysaccharide and metformin on AGEs and CTGF of cardiac muscle in type 2 diabetic rats and the mechanism / 中国药理学通报

Aim To study the effects of ganoderma lu-cidum polysaccharides and metformin on myocardial fi-brosis of type 2 diabetic rats and its mechanism. Methods SD rats were fed with high fat diet for 4 weeks, and then were injected with streptozotocin (30mg·kg-1 ) to replicate type 2 diabetic model. The diabetic rats were randomized into normal control group,diabetes group, ganoderma lucidum polysaccha-rides group ( 600 mg · kg-1 ) , metformin group ( 600 mg·kg-1 ) , and combination group( ganoderma lucid-um polysaccharides 300 mg·kg-1 +metformin 300 mg ·kg-1 ) . After 12 weeks’ treatment,the levels of fast-ing serum glucose were determined and the extent of myocardial fibrosis was observed by Picro-sirius red staining. The contents of AGEs in serum were deter-mined by fluorescence spectrophotometer. The activities of CAT and GSH-Px in myocardium were detected. Im-munohistochemical method and Western blot were used to detect myocardial tissue AGEs and CTGF protein ex-pression. Results Combination group could repress patho-proceeding of myocardial fibrosis efficiently, im-prove the activity of CAT and GSH-Px in myocardium and lower the concentration of AGEs in serum, as well as reduce the expression of AGEs and CTGF in myo-cardium. Conclusions Ganoderma lucidum polysac-charides and metformin could prevent myocardial fibro-sis. The possible mechanism may be related to repress-ing oxidative stress of myocardium, lowering serum AGEs and down regulating AGEs and CTGF of myocar-dium.

Effect of AstragalosideⅣ on Expression of High Glucose Dialysis Solution-Induced Fibrosis Cytokines in Human Peritoneal Mesothelial Cells / 中国中医药信息杂志

Objective To investigate the effects of AstragalosideⅣ (AST) on the expression of high glucose peritoneal dialysis solution (PDS)-induced transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF) in cultured human peritoneal mesothelial cells. To discuss the regulating effect of AST on induced fibrosis cytokines.Methods The HMrSV5 cell line was cultivated to the fifth generation and divided into normal group (10%FBS cultivation solution), model group (4.25% PDS and 10% FBS cultivation solution) and the low, medium and high doses AST groups (4.25% PDS with a respective 10, 20, 40 μg/mL AST). MTT colorimetric assay was employed to detect cell activity and ELISA was applied to detect expression of TGF-β1, CTGF and VEGF in cultured supernatants.Results Except for 5 μg/mL group, HPMCs activity of high glucose plus different concentration AST groups were enhanced in different degrees, especially with 40, 45, and 50 μg/mL (P<0.05). The expression of TGF-β1, CTGF, and VEGF in model group increased. Compared with the control group, expression of the three AST groups significantly decreased and showed dose-effect relationship (P<0.05). Conclusion AST could reduce the expression of TGF-β1, CTGF and VEGF in high glucose-induced HPMCs and was useful in slowing down the progress of peritoneal fibrosis.