Spatial signalling mediated by the transforming growth factor-β signalling pathway during tooth formation

Tooth development relies on sequential and reciprocal interactions between the epithelial and mesenchymal tissues, and it is continuously regulated by a variety of conserved and specific temporal-spatial signalling pathways. It is well known that suspensions of tooth germ cells can form tooth-like structures after losing the positional information provided by the epithelial and mesenchymal tissues. However, the particular stage in which the tooth germ cells start to form tooth-like structures after losing their positional information remains unclear. In this study, we investigated the reassociation of tooth germ cells suspension from different morphological stages during tooth development and the phosphorylation of Smad2/3 in this process. Four tooth morphological stages were designed in this study. The results showed that tooth germ cells formed odontogenic tissue at embryonic day （E） 14.5, which is referred to as the cap stage, and they formed tooth-like structures at E16.5, which is referred to as the early bell stage, and E18.5, which is referred to as the late bell stage. Moreover, the transforming growth factor-β signalling pathway might play a role in this process.

Total flavone of Abelmoschus manihot suppresses epithelial-mesenchymal transition via interfering transforming growth factor-β1 signaling in Crohn's disease intestinal fibrosis

AIM To explore the role and mechanism of total flavone of Abelmoschus manihot(TFA) on epithelial-mesenchymal transition(EMT) progress of Crohn's disease(CD) intestinal fibrosis.METHODS First,CCK-8 assay was performed to assess TFA on the viability of intestinal epithelial(IEC-6) cells and select the optimal concentrations of TFA for our further studies.Then cell morphology,wound healing and transwell assays were performed to examine the effect of TFA on morphology,migration and invasion of IEC-6 cells treated with TGF-β1.In addition,immunofluorescence,real-time PCR analysis(q RT-PCR) and western blotting assays were carried out to detect the impact of TFA on EMT progress.Moreover,western blotting assay was performed to evaluate the function of TFA on the Smad and MAPK signaling pathways.Further,the role of co-treatment of TFA and si-Smad or MAPK inhibitors has been examined by q RTPCR,western blotting,morphology,wound healing andtranswell assays.RESULTS In this study,TFA promoted transforming growth factor-β1(TGF-β1)-induced(IEC-6) morphological change,migration and invasion,and increased the expression of epithelial markers and reduced the levels of mesenchymal markers,along with the inactivation of Smad and MAPK signaling pathways.Moreover,we revealed that si-Smad and MAPK inhibitors effectively attenuated TGF-β1-induced EMT in IEC-6 cells.Importantly,co-treatment of TFA and si-Smad or MAPK inhibitors had better inhibitory effects on TGF-β1-induced EMT in IEC-6 cells than either one of them.CONCLUSION These findings could provide new insight into the molecular mechanisms of TFA on TGF-β1-induced EMT in IEC-6 cells and TFA is expected to advance as a new therapy to treat CD intestinal fibrosis.

Regulation of transforming growth factor-β signaling as a therapeutic approach to treating colorectal cancer

According to data from 2020,Slovakia has long been among the top five countries with the highest incidence rate of colorectal cancer(CRC)worldwide,and the rate is continuing to rise every year.In approximately 80%of CRC cases,allelic loss(loss of heterozygosity,LOH)occurs in the long arm of chromosome 18q.The most important genes that can be silenced by 18q LOH or mutations are small mothers against decapentaplegic homolog(SMAD)2 and SMAD4,which are intracellular mediators of transforming growth factor(TGF)-βsuperfamily signals.TGF-βplays an important role in the pro-oncogenic processes,including such properties as invasion,epithelial-mesenchymal transition(commonly known as EMT),promotion of angiogenesis,and immunomodulatory effects.Several recent studies have reported that activation of TGF-βsignaling is related to drug resistance in CRC.Because the mechanisms of drug resistance are different between patients in different stages of CRC,personalized treatment is more effective.Therefore,knowledge of the activation and inhibition of factors that affect the TGF-βsignaling pathway is very important.

Effects of the Nocth signaling pathway on expression of inflammatory factors and transforming growth factors in diabetic foot ulcers

Objective:persistent hyperinflammation is an important reason for the development of diabetic foot ulcer.Notch signaling is an important signaling pathway involved in the inflammatory response and cell proliferation in diabetic foot ulcer rats.This paper aims to explore the effect of Notch signaling on inflammatory factors,chemokines and growth factors through the intervention of Notch signaling in diabetic foot ulcer rats.Methods:the experimental model was made by using high-fat feed combined with streptozotocin(STZ)to cause diabetes,and the experimental model of diabetic foot ulcer was established by constant temperature and constant pressure scald apparatus.The normal ulcer model was used as a control.The intervention controls of the experimental model included normal saline,western medicine growth factor,Notch agonist Jagged1,Notch signaling inhibitor ly-411575,and the intervention of traditional Chinese medicine Zizhu ointment for 7 days.Serum il-1,il-6,TNF-radiation,and il-17 were detected by ELISA.Real-time PCR was used to detect the inflammatory factors,chemokines,and growth factors associated with Notch signaling in wound tissues:tnf-uum,il-1,il-6,il-17,interleukin-8,ip-10,McP-1,TGF-uum,TGF-livelihood.Results:serum levels of il-1,il-6,TNF-radiation and il-17 in diabetic foot ulcer rats were significantly higher than that in normal ulcer rats.The contents of il-1,il-6,TNF-radiation and il-17a in ly-411575 group and Zizhu ointment group were significantly reduced.Real-time PCR results of wound tissue showed that the levels of inflammatory cytokines il-1,il-6,TNF-radiation,il-17 and chemokines ip-10,il-8 and McP-1 in the wound tissue of diabetic foot ulcer rat model were significantly higher than that of normal ulcer model,and the levels of growth factor TGF-exposure were lower than that of normal ulcer model.LY-411575 significantly reduced il-1,il-6,TNF-maxima,il-17,and the chemokines ip-10,il-8,and McP-1 in diabetic foot ulcer rats,and reduced the expression of TGF-,TGF-earth.Jagged1 can increase the expression of TGF--,TGF---,suggesting that inhibition of the Notch signaling pathway can reduce the expression of the inflammatory factors il-1,il-6,TNF--,il-17a,il-8,and the growth factors TGF--,TGF---.Zizhu ointment can reduce the levels of il-1,il-6,TNF-benand,il-17,and the chemokines ip-10,il-8,and McP-1 on the wound surface of diabetic foot rats,and improve the expression of TGF-benand TGF-SUNS.Ly-411575 inhibited the expression of TGF-bento and TGF-promoting of Zizhu ointment.Conclusion:the expression of inflammatory cytokines and chemokines was higher and the expression of growth factors was lower in diabetic foot ulcer rats than in normal ulcer rats.Inhibition of Notch signaling pathway can reduce the expression of inflammatory factors,chemokines and growth factors in experimental model rats,and Notch signaling pathway can promote inflammation and cell proliferation.Zizhu ointment can reduce the levels of inflammatory cytokines and chemokines in diabetic foot ulcer rats,improve the expression of growth factors,and reduce wound inflammation,which may be related to the inhibition of Nocth signal expression.

Dihydroergotamine ameliorates liver fibrosis by targeting transforming growth factor β type Ⅱ receptor

BACKGROUND The transforming growth factor β(TGFβ) signaling pathway plays a crucial role in the development of liver fibrosis by activating TGFβ type Ⅱ receptor(TGFβR2), followed by the recruitment of TGFβR1 finally triggering downstream signaling pathway.AIM To find drugs targeting TGFβR2 that inhibit TGFβR1/TGFβR2 complex formation, theoretically inhibit TGFβ signaling pathway, and thereby ameliorate liver fibrosis.METHODS Food and Drug Administration-approved drugs were screened for binding affinity with TGFβR2 by virtual molecular docking. We identified 6 candidates and further explored their potential by Cell Counting Kit-8(CCK-8) cell cytotoxic experiment to validate toxicity and titrated the best cellular working concentrations. Next, we further demonstrated the detailed molecular working mechanisms using mutagenesis analysis. Finally, we used a mouse model to investigate its potential anti-liver fibrosis effect.RESULTS We identified 6 drug candidates. Among these 6 drugs, dihydroergotamine(DHE) shows great ability in reducing fibrotic gene expressions such as collagen, p-SMAD3, and α-SMA in TGFβ induced cellular model of liver fibrosis in LX-2 cells. Furthermore, we demonstrated that DHE binds to TGFβR2. Moreover, mutation of Leu27, Phe30, Thr51, Ser52, Ile53, and Glu55 of TGFβR2 disrupted the binding of TGFβR2 with DHE. In addition, DHE significantly improved liver fibrosis, as evidenced by Masson’s trichrome staining of liver sections. This is further supported by the width and the velocity of the portal vein, and serum markers of liver function. In line with those observations, DHE also decreased macrophages infiltration and extracellular matrix deposition in the liver.CONCLUSION DHE alleviates liver fibrosis by binding to TGFβR2 thereby suppressing TGFβ signaling pathway. We show here that as far as drug repurposing, DHE has great potential to treat liver fibrosis.

Cross-talk between microRNA-let7c and transforming growth factor-β2 during epithelial-to-mesenchymal transition of retinal pigment epithelial cells

AIM: To explore the roles of microRNA-let7 c(miR-let7 c) and transforming growth factor-β2(TGF-β2) and cellular signaling during epithelial-to-mesenchymal transition(EMT) of retinal pigment epithelial cells. METHODS: Retinal pigment epithelial(ARPE-19) cells were cultured with no serum for 12 h, and then with recombinant human TGF-β2 for different lengths of time. ARPE-19 cells were transfected with 1×106 TU/mL miR-let7 c mimcs(miR-let7 cM), miR-let7 c mimcs negative control(miR-let7cMNC) and miR-let7 c inhibitor(miR-let7 cI) using the transfection reagent. The expression of keratin-18, vimentin, N-cadherin, IKB alpha, p65 were detected by Western blot, quantitative polymerase chain reaction and immunofluorescence. RESULTS: The expression of miR-let7c was dramatically reduced and the nuclear factor-kappa B(NF-κB) signaling pathway was activated after induction by TGF-β2(P<0.05). In turn, overexpressed miR-let7 c significantly inhibited TGF-β2-induced EMT(P<0.05). However, miR-let7 c was unable to inhibit TGF-β2-induced EMT when the NF-κB signaling pathway was inhibited by BAY11-7082(P<0.01). CONCLUSION: The miR-let7 c regulates TGF-β2-induced EMT through the NF-κB signaling pathway in ARPE-19 cells.

Targeting key signalling pathways in oesophageal adenocarcinoma:A reality for personalised medicine?

Cancer treatments are rapidly changing.Curative treatment for oesophageal adenocarcinoma currently involves surgery and cytotoxic chemotherapy or chemoradiotherapy.Outcomes for both regimes are generally poor as a result of tumor recurrence.We have reviewed the key signalling pathways associated with oesophageal adenocarcinomas and discussed the recent trials of novel agents that attempt to target these pathways.There are many trials underway with the aim of improving survival in oesophageal cancer.Currently,phase 2 and 3 trials are focused on MAP kinase inhibition,either through inhibition of growth factor receptors or signal transducer proteins.In order to avoid tumor resistance,it appears to be clear that targeted therapy will be needed to combat the multiple signalling pathways that are in operation in oesophageal adenocarcinomas.This may be achievable in the future with the advent of gene signatures and a combinatorial approach.

Modified Xiaochaihu Decoction(小柴胡汤)Prevents the Progression of Chronic Pancreatitis in Rats Possibly by Inhibiting Transforming Growth Factor-β 1/Sma-and Mad-Related Proteins Signaling Pathway

Objective： To investigate the effect of modified Xiaochaihu Decoction （小柴胡汤, MXD） on transforming growth factor- [3 1/Sma- and Mad-related proteins （TGF- 13 1/Smads） signaling pathway in rats with chronic pancreatitis （CP） induced by dibutyltin dichloride. Methods： Thirty healthy male Wistar rats were randomly divided into the normal control group, CP group and CP＋MXD-treated group. CP was induced by injection of dibutyltin dichloride （DBTC, 7 mg/kg of body weight） into the right caudal vein, and the control rats were treated with vehicle. MXD was given daily by gavage at a dose of 10 g/kg of body weight, starting from the day after CP induction. After 28-day treatment, the n-benzoyl-tyrosyl para-aminobenzoic acid （NBT-PABA） test was carried out to evaluate exocrine pancreatic function. Then, rats were sacrificed, and pancreatic tissues were harvested for histological evaluation. In addition, the mRNA expression of TGF- β 1, TGF- β 1 type Ⅱ receptor （TGF β R 11 ）, Smad3 and Smad7 was determined in pancreatic tissues by using real-time polymerase chain reaction. Results： Treatment of CP with MXD improved the PABA recovery, decreased the histological lesion, and reduced the mRNA expression of TGF- β 1, TGF β R 11 and Smad3 （P〈0.05）. However, MXD had no effect on Smad7 mRNA level. Conclusions： MXD could protect the pancreas against chronic injury and improve pancreatic exocrine function in DBTC induced rat CP model. Its mechanism may involve inhibition of the TGF-β 1/Smads signaling pathway.

Interplay between micro RNA-17-5p, insulin-like growth factor-Ⅱ through binding protein-3 in hepatocellular carcinoma

AIM: To investigate the effect of microR NA on insulinlike growth factor binding protein-3(IGFBP-3) and hence on insulin-like growth factor-Ⅱ(IGF-Ⅱ) bioavailability in hepatocellular carcinoma(HCC).METHODS: Bioinformatic analysis was performed using microrna.org, DIANA lab and Segal lab softwares. Total RNA was extracted from 23 HCC and 10 healthy liver tissues using mir Vana mi RNA Isolation Kit. microR NA-17-5p(miR-17-5p) expression was mimicked and antagonized in Hu H-7 cell lines using Hi Per Fect Transfection Reagent, then total RNA was extracted using Biozol reagent then reverse transcribed into cD NA followed by quantification of mi R-17-5p and IGFBP-3 expression using Taq Man real-time quantitative PCR. Luciferase reporter assay was performed to validate the binding of miR-17-5p to the 3'UTR of IGFBP-3. Free IGF-Ⅱ protein was measured in transfected Hu H-7 cells using IGF-Ⅱ ELISA kit. RESULTS: Bioinformatic analysis revealed IGFBP-3 as a potential target for miR-17-5p. Screening of miR-17-5p and IGFBP-3 revealed a moderate negative correlation in HCC patients, where mi R-17-5p was extensively underexpressed in HCC tissues(P = 0.0012), while IGFBP-3 showed significant upregulation in the same set of patients(P = 0.0041) compared to healthy donors. Forcing mi R-17-5p expression in Hu H-7 cell lines showed a significant downregulation of IGFBP-3 mR NA expression(P = 0.0267) and a significant increase in free IGF-Ⅱ protein(P = 0.0339) compared to mock untransfected cells using unpaired t-test. Luciferase assay validated IGFBP-3 as a direct target of mi R-17-5p; luciferase activity was inhibited by 27.5% in cells co-transfected with miR-17-5p mimics and the construct harboring the wild-type binding region 2 of IGFBP-3 compared to cells transfected with this construct alone(P = 0.0474).CONCLUSION: These data suggest that regulating IGF-Ⅱ bioavailability and hence HCC progression can be achieved through targeting IGFBP-3 via manipulating the expression of miR NAs.

TGF-β1/SMAD SIGNALING PATHWAY MEDIATES p53-DEPENDENT APOPTOSIS IN HEPATOMA CELL LINES

Objective To determine whether transforming growth factor betal (TGF-β1)/Smad signaling pathway mediates p53-dependent apoptosis in hepatoma cell lines.Methods Three human hepatic carcinoma cell lines, HepG2, Huh-7, and Hep3B, were used in this study.TGF-β1-induced apoptosis in hepatic carcinoma cell lines was analyzed using TUNEL assay.For identifying the mechanism of apoptosis induced by TGF-β1, cell lines were transfected with a TGF-β1-inducible luciferase reportor plasmid containing Smad4 binding elements.After transfection, cells were treated with TGF-β1, then assayed for luciferase activity.Results The apoptosis rate of HepG2 cell lines (48.51%± 8.21%) was significantly higher than control ( 12.72%±2.18%, P<0.05).But TGF-β1 was not able to induce apoptosis of Huh-7 and Hep3B cell lines.The relative luciferase activity of TGF-β1-treated HepG2 cell lines (4.38) was significantly higher than control (1.00, P< 0.05).But the relative luciferase activity of TGF-β1-treated Huh-7 and Hep3B cell lines less increased compared with control.Conclusions HepG2 cells seem to be highly susceptible to TGF-β1-induced apoptosis compared with Hep3B and Huh-7 cell lines.Smad4 is a central mediator of TGF-β1 signaling transdution pathway.TGF-β1/Smad signaling pathway might mediate p53-dependent apoptosis in hepatoma cell lines.

Regulatory Effect of Shenge Yifei Capsule on TGF-β1/Smad Signaling Pathway in Rats with Chronic Obstructive Pulmonary Disease

[Objectives]This study aimed to study the effects of Shenge Yifei capsule on the TGF-β1/Smad signaling pathway in rats with chronic obstructive pulmonary disease(COPD).[Methods]Ten rats were randomly selected as the control group,and the other 40 rats were selected for modeling by fumigation combined with Klebsiella pneumoniae infection.A total of 38 rats were successfully modeled.They were randomly divided into model group(8 rats),low-dose Shenge Yifei capsule group(10 rats),high-dose Shenge Yifei capsule group(10 rats)and theophylline group(10 rats)in accordance with the principle of half male and half female.The rats in the model and control groups were given with distilled water by gavage,and the rats in the drug administration groups were given with corresponding drugs.The TGF-β1 level in the serum,and the expression levels of TGF-β1,Smad2,Smad3 and Smad7 and TGF-β1,Smad3 and Smad7 in airway tissues were detected.[Results]After 12 weeks,the serum TGF-β1 levels of the theophylline group and high-dose Shenge Yifei capsule group were lower than that of the low-dose Shenge Yifei capsule group(P<0.05).The expression levels of TGF-β1 and Smad3 in the theophylline group and high-dose Shenge Yifei capsule group were lower than that in the low-dose Shenge Yifei capsule group(P<0.05).The expression levels of TGF-β1 and Smad3 in the high-dose Shenge Yifei capsule group were lower than those in the low-dose Shenge Yifei capsule group and theophylline group(P<0.05).The expression levels of Smad7 and the proteins in the model group were lower than those in the other groups(P<0.05).The expression levels of Smad7 in the theophylline group and high-dose Shenge Yifei capsule group were higher than that in the low-dose Shenge Yifei capsule group(P<0.05).After 18 weeks,no significant difference was found in serum TGF-β1 level among the theophylline group and low and high-dose Shenge Yifei capsule groups(P>0.05).The expression levels of Smad7 and the proteins in the model group were lower than those in the other groups.The expression level of Smad7 in the high-dose Shenge Yifei capsule group was lower than that in the theophylline group(P<0.05).[Conclusions]Shenge Yifei capsule can regulate the TGF-β1/Smads signaling pathway.They can down-regulate the expression of TGF-β1,Smad2 and Smad3 and up-regulate the expression of Smad7,reducing the degree of airway modeling,delaying the development of COPD disease.Conventional high-dose Shenge Yifei capsule is more effective in inhibiting the expression of Smad2.

Regulation Mechanism of TFP on TGF-β1/STAT3 Signaling Pathway in Immune-mediated Liver Injury in Mice

[Objectives]To study the effect of total flavonoids extracted from Polygonum perfoliatum L.(TFP)on immune-mediated liver injury induced by bacillus Calmette-Guerin plus lipopolysaccharide(BCG+LPS)in mice,and to explore its action mechanism.[Methods]60 Kunming mice were divided into normal group,model group,control group(bifendate)and TFP low,medium and high dose groups according to random number table method,with 10 mice in each group.On the first day of modeling,mice were injected with 0.2 mL of BCG solution(12.5 mg/mL)through the tail vein,and on the eleventh day,0.2 mL of LPS(37.5μg/mL)were injected into the tail vein to prepare a mouse model of immune-mediated liver injury;from the first day of modeling,the normal group and the model group were administered intragastrically with the corresponding volume of distilled water,and the bifendate group and the TFP high,medium,and low dose groups were administered intragastrically with the corresponding doses once a day for 11 d.After the last time administration,fasting but giving water for 16 h,took blood from eyes,then collected the liver tissue.The levels of alanine transaminase(ALT)and aspartate transaminase(AST)in serum were detected by biochemical method;transforming growth factor-β1(TGF-β1),intercellular adhesion molecule-1(ICAM-1),interleukin-6(IL-6)and interleukin-1β(IL-1β)expression levels in liver tissue were detected by enzyme-linked immunosorbent assay(ELISA);phosphorylated protein tyrosine kinase JAK-2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)protein expression levels were detected by Western Blot method;the degree of liver tissue lesions was detected by HE staining.[Results]Compared with the model group,the levels of ALT and AST in the serum of mice in each dose group of TFP(high dose 600 mg/kg,medium dose 400 mg/kg,and low dose 200 mg/kg)were reduced,and the activities of T-SOD and GSH-Px were increased;the content or expression ofβ1,ICAM-1,IL-6,IL-1βdecreased,and the expression of p-JAK2 and p-STAT3 protein decreased;pathological sections showed that the degree of inflammatory necrosis and the degree of lesions in the liver tissues of each dose group of TFP were reduced by varying degrees.[Conclusions]TFP has a protective effect on BCG+LPS-induced immune-mediated liver injury in mice.The mechanism may be related to regulating the phosphorylation level of JAK2 and inhibiting the inflammatory reaction,thereby regulating the TGF-β1/STAT3 signaling pathway and improving the immune-mediated liver injury.

TGF-β1/Smad3信号通路调控MMP-13/TIMP-1蛋白表达在糖尿病膀胱纤维化中的作用研究

目的探讨转化生长因子β1(TGF-β1)/细胞信号转导分子3(Smad3)信号通路调控基质金属蛋白酶13(MMP-13)/基质金属蛋白酶抑制剂1(TIMP-1)蛋白表达在糖尿病膀胱纤维化中的作用。方法将80只雄性Wistar大鼠按随机数字表法分为糖尿病组和正常对照组,每组40只;另取20只糖尿病大鼠按随机数字表法分为TGF-β1敲除组和Smad3敲除组,每组10只。采用链脲佐素65 mg/kg一次性给药法建立糖尿病模型;采用Cas9 gene targeting技术分别敲除TGF-β1或Smad3基因。采用HE或VG染色法观察糖尿病组与正常对照组大鼠膀胱组织病理学形态,Western blot法检测并比较糖尿病组与正常对照组、糖尿病组与TGF-β1敲除组、糖尿病组与Smad3敲除组大鼠膀胱平滑肌组织中相关信号通路因子蛋白表达水平,包括TGF-β1、Smad3、MMP-13、TIMP-1、Ⅰ型胶原蛋白(colⅠ)、Ⅲ型胶原蛋白(colⅢ)等。结果HE及VG染色结果显示,糖尿病组大鼠膀胱组织胶原纤维明显增多。糖尿病组大鼠膀胱平滑肌组织中TGF-β1、Smad3、MMP-13、TIMP-1蛋白表达水平均明显高于正常对照组,MMP-13、TIMP-1、colⅠ、colⅢ蛋白表达水平均明显高于TGF-β1敲除组、Smad3敲除组,差异均有统计学意义(均P<0.05)。结论MMP-13、TIMP-1在糖尿病膀胱纤维化中呈高表达。TGF-β1/Smad3信号通路通过调控MMP-13/TIMP-1蛋白表达参与糖尿病膀胱纤维化的发生、发展。

Bioinformatic identification of key candidate genes and pathways in axon regeneration after spinal cord injury in zebrafish

Zebrafish and human genomes are highly homologous;however,despite this genomic similarity,adult zebrafish can achieve neuronal proliferation,regeneration and functional restoration within 6–8 weeks after spinal cord injury,whereas humans cannot.To analyze differentially expressed zebrafish genes between axon-regenerated neurons and axon-non-regenerated neurons after spinal cord injury,and to explore the key genes and pathways of axonal regeneration after spinal cord injury,microarray GSE56842 was analyzed using the online tool,GEO2R,in the Gene Expression Omnibus database.Gene ontology and protein-protein interaction networks were used to analyze the identified differentially expressed genes.Finally,we screened for genes and pathways that may play a role in spinal cord injury repair in zebrafish and mammals.A total of 636 differentially expressed genes were obtained,including 255 up-regulated and 381 down-regulated differentially expressed genes in axon-regenerated neurons.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment results were also obtained.A protein-protein interaction network contained 480 node genes and 1976 node connections.We also obtained the 10 hub genes with the highest correlation and the two modules with the highest score.The results showed that spectrin may promote axonal regeneration after spinal cord injury in zebrafish.Transforming growth factor beta signaling may inhibit repair after spinal cord injury in zebrafish.Focal adhesion or tight junctions may play an important role in the migration and proliferation of some cells,such as Schwann cells or neural progenitor cells,after spinal cord injury in zebrafish.Bioinformatic analysis identified key candidate genes and pathways in axonal regeneration after spinal cord injury in zebrafish,providing targets for treatment of spinal cord injury in mammals.

Toxicarioside A,isolated from tropical Antiaris toxicaria,blocks endoglin/TGF-βsignaling in a bone marrow stromal cell line

Objective:To investigate possible mechanism of toxicarioside A in HS-5 bone stromal cells.Methods:HS-5 bone stromal cells were cultured in media supplemented with various concentrations of toxicarioside A or control DMSO(not treatment).Endoglin and TGF-βwere detected by Northern and Western blot analysis and quantified in a standard method. Downstream molecules of endoglin and TGF-β(Smad1,Smad2 and their active phosphorylated counterparts,pSmad1 and pSmad2) were also detected and quantified by Western blot analysis. In addition,cell proliferation assay and small interfering RNA(siRNA) against endoglin were used to certificate the function of endolgin in the HS-5 cells.Results:Compared with the not treated(0μg/mL) or DMSO treated control HS-5 cells,HS-5 cells treated with toxicarioside A were found significant attenuation of endolgin and TGF-βexpression.Significant inhibition of cell proliferation was also found in the HS-5 cells treated with toxicarioside A.ALK1-related Smad1 and ALK5-related Smad2 were decreased in HS-5 cells treated with toxicarioside A.In addition,phosphorylated Smad1(pSmad1) and Smad2(pSmad2) were also found attenuation in toxicarioside A-treated HS-5 cells.RNA interference showed that blockage of endoglin by siRNA also decreased Smad1 and Smad2 expression in HS-5 cells.Conclusions:Our results indicate that toxicarioside A can influence bone marrow stromal HS-5’s function and inhibit HS-5 cell proliferation by alteration of endoglin-related ALK1(Smad1) and ALK5(Smad2) signaling.

氨基胍对肺纤维化大鼠MMP-13、TIMP-1及转化生长因子-β1/Smads信号通路的影响

目的:探讨氨基胍对肺纤维化(PF)大鼠基质金属蛋白酶-13(MMP-13)和组织基质金属蛋白酶抑制物-1(TIMP-1)表达的影响及其对转化生长因子-β1/信号转导分子(TGF-β1/Smads)信号通路的调控机制。方法:SD大鼠随机分为4组:正常组、模型组、实验组、地塞米松组(对照组),每组30只。模型组、实验组、对照组大鼠气管滴注博莱霉素构建PF模型,正常组大鼠滴注等剂量等浓度氯化钠溶液。滴注完毕后,实验组、对照组大鼠每日分别腹腔注射1 ml氨基胍溶液(20 mg/kg)和1 ml地塞米松溶液(20 mg/kg),正常组、模型组分别腹腔注射等剂量生理盐水。处死大鼠,qRT-PCR、免疫组化、Western blot检测各组大鼠肺组织病理学及纤维化改变、TGF-β1、Smad2和Smad7 mRNA表达、MMP-13和TIMP-1表达、TGF-β1、p-Smad2、pSmad7蛋白表达。结果:与正常组相比,模型组肺组织TGF-β1和Smad2 mRNA表达、TGF-β1、p-Smad2和TIMP-1表达明显升高,Smad7 mRNA和p-Smad7蛋白表达、MMP-13表达明显降低;与模型组相比,实验组和对照组大鼠肺组织TGF-β1和Smad2mRNA表达、TGF-β1、p-Smad2、TIMP-1表达明显降低,Smad7 mRNA和p-Smad7蛋白表达、MMP-13表达明显升高(P<0.05)。结论:氨基胍可保护博来霉素诱导的PF大鼠肺组织,其抗纤维化作用可能通过调控TGF-β1/Smads信号通路实现。

Mechanism of ELL-associated factor 2 and vasohibin 1 regulating invasion,migration,and angiogenesis in colorectal cancer

BACKGROUND As a novel endogenous anti-angiogenic molecule, vasohibin 1(VASH1) is not only expressed in tumor stroma, but also in tumor tissue. Moreover, studies have shown that VASH1 may be a prognostic marker in colorectal cancer(CRC). Knockdown of VASH1 enhanced transforming growth factor-β1(TGF-β1)/Smad3 pathway activity and type Ⅰ/Ⅲ collagen production. Our previous findings suggest that ELL-associated factor 2(EAF2) may play a tumor suppressor and protective role in the development and progression of CRC by regulating signal transducer and activator of transcription 3(STAT3)/TGF-β1 signaling pathway. However, the functional role and mechanism of VASH1-mediated TGF-β1 related pathway in CRC has not been elucidated.AIM To investigate the expression of VASH1 in CRC and its correlation with the expression of EAF2. Furthermore, we studied the functional role and mechanism of VASH1 involved in the regulation and protection of EAF2 in CRC cells in vitro.METHODS We collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein and VASH1 protein in patients with advanced CRC. Following, we investigated the effect and mechanism of EAF2 and VASH1 on the invasion, migration and angiogenesis of CRC cells in vitro using plasmid transfection.RESULTS Our findings indicated that EAF2 was down-regulated and VASH1 was upregulated in advanced CRC tissue compared to normal colorectal tissue. KaplanMeier survival analysis showed that the higher EAF2 Level group and the lower VASH1 Level group had a higher survival rate. Overexpression of EAF2 might inhibit the activity of STAT3/TGF-β1 pathway by up-regulating the expression of VASH1, and then weaken the invasion, migration and angiogenesis of CRC cells.CONCLUSION This study suggests that EAF2 and VASH1 may serve as new diagnostic and prognostic markers for CRC, and provide a clinical basis for exploring new biomarkers for CRC. This study complements the mechanism of EAF2 in CRC cells, enriches the role and mechanism of CRC cellderived VASH1, and provides a new possible subtype of CRC as a therapeutic target of STAT3/TGF-β1 pathway.

肝硬化的研究进展

肝硬化是多种原因引起的慢性肝损伤所致的病理变化,是肝细胞外基质(ECM)产生与降解不平衡导致的纤维组织的过度堆积。肝星状细胞(HSC)是ECM产生的主要效应细胞。HSC激活、增殖以及改变表型,进而分泌胶原在肝脏沉积;同时,多种生长因子、细胞因子、信号转导物质与该病理过程密切相关。明确肝硬化的发病机制对治疗有重要意义。

增生性玻璃体视网膜病变相关细胞因子研究进展

增生性玻璃体视网膜病变（PVR）是一种常见的致盲眼病,表现为受损的玻璃体及视网膜的异常修复.PVR的形成涉及多种细胞因子,包括血小板衍生生长因子（PDGF）、血管内皮生长因子、肝细胞生长因子、转化生长因子β、表皮生长因子、肿瘤坏死因子α、结缔组织生长因子等.视网膜色素上皮（RPE）细胞涉及Jagged-1/Notch信号通路和RhoA/ROCK信号通路发生细胞上皮-间充质形态改变.RPE细胞的增生、迁移与PKC/ERK信号通路和Akt/mTORCl信号通路相关.非PDGF间接激活PDGF受体α是PVR形成的主要通路.对PVR的治疗开展实验研究,联合抑制多种细胞因子及应用抗氧化剂N-乙酰半胱氨酸均取得一定疗效.本文对相关细胞因子在PVR形成中的作用机制、信号通路及治疗相关的实验研究进行综述.

桑叶生物碱对肝纤维化小鼠的改善作用及机制

目的:观察桑叶生物碱对转化生长因子-β1(transforming growth factor-β1,TGF-β1)/Smads信号通路及基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)、基质金属蛋白酶抑制剂-1(inhibitor of metalloproteinase-1,TIMP-1)mRNA表达的调节作用,探讨桑叶生物碱改善小鼠肝纤维化的作用机制。方法:采用腹腔注射体积分数10%CCl4橄榄油溶液(5 mL/kg mb)联合高脂饮食诱导小鼠肝纤维化模型。造模8周后,正常对照组和模型组给予蒸馏水,低、中、高剂量组分别灌胃50、100、200 mg/kg mb剂量的桑叶生物碱,阳性药物组给予100 mg/kg mb水飞蓟宾,持续45 d。采用酶联免疫试剂盒测定各组小鼠肝组织α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、I型胶原蛋白(collagen I,Col I)及III型胶原蛋白(collagen III,Col III)含量;采用实时定量聚合酶链式反应测定肝组织TGF-β1、Smad3、Smad4、Smad7、MMP-13、TIMP-1 mRNA表达水平。结果:与模型组比,灌胃高剂量桑叶生物碱(200 mg/kg mb)的小鼠肝组织中α-SMA、Col I、Col III含量分别降低19.55%、19.93%、13.84%(P<0.05);TGF-β1、Smad3、Smad4、TIMP-1 mRNA表达水平分别下降59.04%、56.73%、50.70%、49.09%(P<0.05),Smad7、MMP-13 mRNA表达水平分别上升48.29%、25.72%(P<0.05)。结论:桑叶生物碱能改善小鼠肝纤维化,其机制可能是对TGF-β1/Smads信号通路相关因子和TIMP-1、MMP-13的基因表达具有调控作用。