Effects of ulinastatin on intestinal mucosal barrier function in sepsis rats and its effect on wnt signal transduction pathway / 解剖学报

Objective To investigate the intestinal mucosal barrier function protective effect of ulinastatin in sepsis rats and its effect on Wnt/β-catenin signaling pathway. Methods One hundred SD rats were randomly divided into control group, sepsis group, ulinastatin group, XAV939+ulinastatin group and lithium chloride( LiCl) +ulinastatin group. The classical cecal ligation was used to duplicate sepsis model, and the jejunal mucosal injury was evaluated. The levels of inflammatory factors interleukin (IL)-6 and tumor necrosis factor(TNF)-α were detected by ELISA, and the expressions of β-catenin and cyclin D1 were detected by Real-time PCR and Western blotting. We also observed the effect of the Wnt signal pathway blockage by XAV939 or Wnt signal pathway activator by LiCl on ulinastatin protection of intestinal mucosa and proteins related to the Wnt signal pathway. Results The levels of IL-6, TNF-α and intestinal mucosal injury in the sepsis group were significantly higher than those in the ulinastatin group. The mRNA and protein expression levels of β- catenin and cyclin D1 in the sepsis group were significantly higher than those in the control group (P<0.05), After ulinastatin treatment, the expression levels of β-catenin and cyclin D1 mRNA and protein were significantly decreased, and the difference was significant (P<0.05). Compared with the ulinastatin group, combined treatment with XAV939 promoted the protective effect of ulinastatin on the intestinal mucosa of rats, and the protein expression of β-catenin and cyclin D1 was reduced (P<0.05). Combined treatment with LiCl weakened the protective effect of ulinastatin on the intestinal mucosa of rats, and the protein expression of β-catenin and cyclin D1 was increased (P<0.05). Conclusion Ulinastatin may inhibit the Wnt signaling pathway by down-regulating the expression of β-catenin, reduce the expression of inflammatory factors IL-6 and TNF-α, thereby promote repairing the intestinal mucosal barrier function damage.

Signal transduction mechanism and targeted therapy of melanoma stem cells / 肿瘤

Melanoma is the most serious type of skin cancer with high metastasis potential and very low survival rate. Although the molecular targeted therapy and immunotherapy have prolonged the survival time of some patients with melanoma in the past few years, a considerable number of patients still can not obtain a longer stable remission period. According to the recent study, melanoma stem cells (MSCs) are one of potential causes of tumor invasion, drug resistance, distant metastasis and recurrence. MSCs can express a variety of cellular markers, and participate into the regulation of tumor-related signal transduction. At the same time, these signaling pathways can retroregulate the expression of cellular markers, and promote the development of melanoma. Therefore, targeting MSCs markers and blocking the related signal transduction pathway can inhibit the growth of melanoma. In this review, the cellular markers, signal transduction mechanism and targeted therapy of MSCs have been discussed in order to provide new ideas for the treatment of melanoma.

Effects of FRZB mRNA in adjusting WNT signal pathway for prognosis of non-small cell lung cancer / 临床检验杂志

Objective@#This study aimed to reveal the potential clinical and biological functions of frizzled related protein (FRZB) mRNA expression in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). @*Methods@#We used the keyword “lung cancer” to search the data through Gene Expression Omnibus (GEO) database attached to NCBI(National Center of Biotechnology) and download the data of LUAD and LUSC from TCGA (The Cancer Genome Atlas) Database. A total of eight LUAD and six LUSC datasets were incorporated in this analysis. We defined cutoff value of FRZB using Cutoff Finder into the two groups to calculate hazard ratio (HR). @*Results@#We found that high expression level of FRZB mRNA in tumor tissues was a positive prognostic factor for overall survival in LUAD [pooled HR（95%CI）=0.54（0.46-0.64）,P＜0.05 in univariate analysis; pooled HR（95%CI）=0.66（0.54-0.79）,P＜0.05 in multivariate analysis]. Interestingly, there was no similar results in LUSC [pooled HR（95%CI）=1.11（0.67-1.84）,P＞0.05 in univariate analysis; pooled HR（95%CI）=1.13（0.71-1.78）,P＞0.05 in multivariate analysis]. We also found that FRZB may inhibit WNT signal pathway by t-SNE and correlation analysis. By enrichment analysis, FRZB and its most correlated genes were involved in multiple immune-related pathways, such as complement and coagulation cascades, humoral immune response, etc. @*Conclusion@#High expression of FRZB mRNA in LUAD was associated with better prognosis of lung adenocarcinoma. These results suggest that FRZB may be used as a potential marker for favorable prognosis of lung adenocarcinoma.

The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats / 中国应用生理学杂志

OBJECTIVES@#To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism.@*METHODS@#Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot.@*RESULTS@#Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05).@*CONCLUSIONS@#Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.

Disheveled 2 around the implant at an early stage of hyperlipidemic rats / 华西口腔医学杂志

OBJECTIVE@#This study aims to investigate the expression of disheveled 2 (Dvl2) around the implant of hyperlipidemic rats at an early stage after the implantation.@*METHODS@#A total of 24 Wistar rats were divided equally into the experimental group fed with high-fat diet group and control group fed with a normal diet. After 8 weeks, the serum lipid levels were detected, and rats received implants in the femur metaphysis. Rats were sacrificed at 1, 3, and 5 days after implantation, and the bones around implants were obtained. Methylene blue-acid fuchsin staining was performed to observe the implant-bone interface. Real-time polymerase chain reaction was performed on runt-related transcription factor 2 (Runx2), cathepsin K (CatK), and Dvl2. Dvl2 Western blot or immunoprecipitation, phosphorylation, and ubiquitination were also conducted.@*RESULTS@#In the experimental group, less osteoblasts, lower expression of Runx2 and Dvl2, and lower Dvl2 phosphorylation (P<0.05) than those of the control group were observed; furthermore, the CatK expression and Dvl2 ubiquitination were higher than those in the control group (P<0.05).@*CONCLUSIONS@#Hyperlipidemia may suppress bone remodeling around the implant at an early stage by Dvl2 down-regulation, phosphorylation, and up-regulated ubiquitination.

Preliminary study on the short－term and long－term effects of the inhibition of hypoxia inducible factor－1 alpha decomposition and activation of Wnt signaling pathway in brain injury in neonatal rats / 中国新生儿科杂志

Objective To investigate the protective effect of inhibition of hypoxia inducible factor－1 alpha (HIF－1α) and activation of Wnt signaling pathway on brain injury in neonatal rats.Method A total of 312 newborn SD rats were used to establish cerebral white matter damage model (WMD) on day 3.And they were randomly assigned into the model group (WMD group), post－modeling HIF－1αdecomposition inhibition group (PHI group), and post－modeling HIF－1αdecomposition inhibition with activation of Wnt signaling pathway group (PHI+activated Wnt group ) and post－modeling HIF－1αdecomposition inhibition with inhibition of the Wnt signaling pathway group (PHI+inhibition Wnt group).Brain tissues were taken on day 1, 3, 7 and 14 after modeling, respectively.The changes of oligodendrocyte precursor cell (OPC) and oligodendrocyte ( OL ) in brain tissues at different time points were observed by tissue immunofluorescence.The expression of HIF－1αprotein in the brain tissue of each time point was measured by western blot technique.The mRNA level of HIF－1αand Wnt7a in the brain tissue of each time point was detected by RT－qPCR technique.Behaviors of rats were tested by the suspension experiment , the open field experiment and the dark experiment , at 28 d after modeling.Result The numbers of OPC on 1 d and 3 d after modeling and the number of OL on 7 d and 14 d after modeling in PHI +activated Wnt group were significantly higher than the other three groups.The content of HIF－1αin WMD group was the least on 1, 3, 7 and 14 d, but the content of PHI+activated Wnt group was the highest , and the difference was statistically significant (P＜0.05).On 1, 3, 7 and 14 d after modeling, the expression level of HIF－1αand Wnt7a in PHI+activated Wnt group were higher than those in the other three groups , the difference was statistically significant (P＜0.05), and Wnt7a expression was positively correlated with the change of HIF－1α(P＜0.05).There was no significant difference of suspension experiment at 28 d after modeling between groups. Compared with other groups , WMD group had the lowest score on open field experiment ( P＜0.05).The PHI+activation Wnt group had better memory function , and then the PHI group , WMD group.The latent time of dark experiment were significantly shorter in PHI +acitivation Wnt group.There were more mistaken time of dark experiment in WMD group compared with PHI +activation Wnt group.Conclusion Inhibition of HIF－1αdecomposition and activation of Wnt signaling pathway have partially repair effect on brain injury in neonatal rats.

Progress in genetics of NPHP related signaling pathway / 国际儿科学杂志

Nephronophthisis(NPHP),an autosomal recessive cystic kidney disease,is the most frequent genetic cause for end stage renal failure in the first thirty years of life.NPHP can be caused by Mutations in 22 genes(NPHP1-20,NPHPL1,NPHPL2),with abnormal structure or function of primary cilia,involved in Hh,Wnt,Hippo,DDR signaling pathways.Elucidating the pathogenic genes and possible pathogenesis would make a difference in prevention,diagnosis,treatment,prognosis,and genetic counseling of NPHP.This article reviews the pathogenic genetics and related signaling pathways.

Molecular mechanism of total flavonoids in Isodon amethystoides on adjuvant arthritis in rats / 中国中药杂志

Our preliminary study showed that the total flavonoids in Isodon amethystoides(TFIA), a local medicinal herb in Suzhou, had a certain therapeutic effect on adjuvant arthritis, and this therapeutic effect may be achieved through the up-regulation of miR-152 expression. In this paper, the molecular mechanism of TFIA on the pathogenesis of adjuvant arthritis(AA) rats was further studied. AA rats were prepared with complete Freund's adjuvant, and then treated with TFIA by intragastric administration. Real-time qPCR was used to detect the effects of TFIA on the negative regulatory loop of miR-152, methylase DNMT1 and methyl-CpG binding protein MeCP2 in fibroblast like synoviocytes(FLS) of AA rats, as well as the effects of TFIA on the classic Wnt signaling pathway and the expression of fibronectin gene in AA rats. Intragastric administration of TFIA significantly inhibited the expression of DNMT1 and reversed the negative regulatory loop composed of miR-152, DNMT1 and MeCP2 in the pathology of AA rats. After transfection of miR-152 inhibitors into the FLS in treatment group, DNMT1 expression was significantly restored. TFIA significantly up-regulated the expression of SFRP4 and inhibited the expression of β-catenin, C-myc and ccnd1, the key genes of canonical Wnt signaling pathway. TFIA also significantly inhibited the expression of fibronectin, an AA gene. The effect of TFIA on the expression of SFRP4, β-catenin, C-myc, ccnd1 and fibronectin was reversed after transfection with miR-152 inhibitors in the treatment group FLS. TFIA may inhibit the DNMT1 expression, up-regulate the SFRP4 expression, inhibit the expression of classical Wnt signaling genes β-catenin, C-myc, and ccnd1 as well as the RA gene fibronectin expression through the up-regulation of miR-152 expression.

Function of Wnt/β-catenin signal pathway in renal tubular epithelial-mesenchymal transition regulation / 中华实用儿科临床杂志

Renal interstitial fibrosis is the common pathway for a variety of chronic renal disease to final stage renal failure,the main feature of it is the large accumulation of extra cellular matrix (ECM).Myofibroblasts is the major effector cells which lead to the synthesis of ECM,an important source of myofibroblasts is epithelial-mesenchymal transition(EMT).Wnt/β-catenin signaling pathway take part in the regulation of EMT,which make it become a potential site of anti-fibrosis therapy.

Effect of AsperosaponinⅥon adipocyte differentiation in ST-2 cells and its underlying mechanisms / 天津医药

Objective The effect of Asperosaponin Ⅵ(ASAⅥ)on adipocyte differentiation and the involvement of Wnt signal pathway was investigated. Methods The murine bone marrow stromal cell line ST-2 were divided into 6 groups:control group, adipocyte differentiation group, and 4 different doses of ASAⅥgroups. Control group was exposed to the vehicle, adipocyte differentiation group was exposed to adipogenic reagent, and those 4 ASAⅥgroups were treated with different concentration(10-7, 10-6, 10-5, 10-4 mol/L)of ASAⅥafter adipocyte differentiation induction. 5 days later, oil red O staining was performed to calculate adipocyte rate. Then mRNA transcription levels of PPARγ, FABP4 genes andβ-catenin that were Wnt/β-catenin signaling pathway proteins were examined by FQ-PCR. Then Wnt pathway inhibitor DKK1 was supplemented into ST-2 cells treated with 10-4 mol/L ASAⅥfor 5 days. After that FQ-PCR was used to detect whether tran?scription levels of PPARγ, FABP4 andβ-catenin in ST-2 cells were changed. Results Compared with adipocyte differenti?ation group 10-5 mol/L and 10-4 mol/L ASAⅥtreatments greatly down-regulated the number of lipid droplets and markedly inhibited transcription levels of adipocyte characterization transcription factors included PPARγ, FABP4 while up-regulat?ed transcription level ofβ-catenin in ST-2 cells. DKK1 can reverse the inhibitory effect of ASAⅥon adipocyte differentia?tion in ST-2 adipocyte. The transcription levels of PPARγand FABP4 were up-regulated significantly while transcription level ofβ-catenin was inhibited. Conclusion ASAⅥblocks adipocyte differentiation in ST-2 cells which might be medi?ated through activating Wnt/β-catenin signaling pathway.

Tumor associated gene PFTK1 / 国际肿瘤学杂志

PFTK1,an important member of cyclin dependent kinase Cdc2 family,is over expressed in many malignancies.PFTK1 involves in multiple processes,such as regulation of cell cycle,tumor invasion and metastasis,and the Wnt signal transduction pathway,which has a significant impact on prognosis of tumor.

Wnt3a-induced proliferation in human lens epithelial cell and its mechanism / 中华实验眼科杂志

Background Aberrant proliferation of residual lens epithelial cells (LECs) is one of main causes of posterior capsular opacification (PCO).Researches indicated that Wnt3a signaling pathway promote proliferation of epithelial cells,but its effect on LECs is still unclear. Objective The present study was to investigate the effects of Wnt3a on proliferation of human LECs and its mechanism and to provide a new gene target in the prevention and treatment of PCO. Methods Human LECs line (SRA01/04 cells ) was cultured and then incubated to 6-well plate at the density of 4×105/well.A human Wnt3a cDNA expressing vector targeted human LECs was constructed to increase the Wnt3a expression in SRA01/04 cells,and pcDNA3-HA expression vector was used as the control group.The expression of Wnt3a was identified by Western blot assay after transfected.The growth and proliferation of SRA01/04 cells were detected by MTT and flow cytometry (FCM).The expressions of β-catenin,cyclin D1 and c-myc in the cells were detected by Western blot assay.β-Catenin expression was localized using immunofluorescence assay,and the expression and localization of proliferating cell nuclear antigen ( PCNA ) were analyzed by immunocytochemistry for the exploration of the active mechanism of Wnt3a to proliferation of LECs.Results Human Wnt3a cDNA expression vector was designed successfully and transiently transfected to SRA01/04 cells,and Wnt3a/SRA01/04 cells and pcDNA3-HA/SRA01/04 cells were obtained.The expression of Wnt3a was verified in the Wnt3a transfected group compared with the control group.MTT indicated that the cell proliferating rate was significantly different between the Wnt3a transfected group and the control group ( Fgroup =15.235,P =0.005 ;Ftime =369.677,P =0.000),and that in various time points after transfected was significantly different (t =20.843,P=0.001 ;t =26.214,P＜0.001 ;t=25.177,P=0.001 ;t =35.516,P＜0.001 ;t =615.056,P＜0.001 ).The proportion of SRA01/04 cells in G1 phase was 51.74％ in the Wnt3a cDNA transfected group,with a significantly decrease in comparison with 79.44％ of the control group.However,the proportion of SRA01/04 cells in S phase in the Wnt3a cDNA transfected group was higher than that of the control group (36.23％ versus 12.34％ ).The positive expression rate of PCNA protein in SRA01/04 was (47.00％ ±7.58％ ) in the Wnt3a cDNA transfected group and ( 16.00％ ±3.61％ ) in the control group with a significant difference between them (t =8.256,P＜0.01 ).After 48 hours of transfection of the Wnt3a cDNA,the expression amount of β-catenin proteins was higher and the immunofluorescence was stronger in cell nucleus,and the expressions of cyclin D1 and c-myc proteins were elevated in Wnt3a/SRA01/04 cells. Conclusions The overexpression of Wnt3a activates the Wnt/β-catenin signaling pathway and downregulates the expression of a subset of target genes,including cyclin D1 and c-myc,which plays an important role in promoting the proliferation of human LECs.

The role of COX-2 in the regulation of Wnt signal pathway / 中华内科杂志

Objective Wnt signal is very important to control the gastrointestinal mucosal proliferation, but its role in the gastric mucosal proliferation after acidified ethanol injury is not clear yet,neither is the relationship with cyclooxygenase 2 (COX-2). Methods X-gal staining was used to measure the expression of Wnt signaling; Western blot and RT-PCR were used to measure the expression of TCF4 and P-Catenin in the gastric mucosa before and after acidified ethanol injury. We also used TOP/flash plasmid to further indentify the relationship between COX-2 and Wnt signal pathway. Results After acidified ethanol injury, the expression of LacZ signal, β-Catenin and TCF4 increased only in the wild type mouse. The expression of β-Catenin and TCF4 protein increased about (3.52 ±0.52) times and (3.02 ±0.62) times separately, and the expression of β-Catenin and TCF4 mRNA increased about (19. 85 ±3.63) times and (17.82 ±4.82) times separately. Without ethanol injury, the expression of TOP/flash plasmid was inhibited by COX-2 inhibitor (NS398) about 80% and promoted by prostaglandin E2(PGE2) about 50%.After 1% ethanol injury, the expression of plasmid was inhibited by NS398 about 25% ; on the contrary,PGE2 promoted the expression about 10%. Conclusion Wnt signal in gastric mucosa is activated after acidified ethanol injury. COX-2 may work through modulating Wnt signal to control the proliferation of gastric mucosa.

Expression and Clinical Significance of ?-catenin and COX-2 in Human Colorectal Carcinoma / 中国医师杂志

Objective To investigate the expression and clinical significance of ?-catenin and COX-2 in human colorectal carcinoma. Methods [WTBZ]The expression of ?-catenin and COX-2 in 153 colorectal tissue specimens was examined by immunohistochemical technique, and their clinical significance and correlation were statistically analyzed. Results In colorectal adenocarcinoma, the loss of ?-catenin in the cytomembrane was significantly correlated with the differentiated degree, lymph node metastasis and Dukes' stages, while COX-2 overexpression was associated with the Dukes' stages,lymph node metastasis and infiltrating depth. The ectopic expression of ?-catenin was not associated with the COX-2 overexpression. [WTHZ] Conclusion COX-2 and ?-catenin may play a role in the pathogenesis of colorectal adenocarcinoma. The ectopic expression of ?-catenin may be not the main reason of the COX-2 overexpression in colorectal adenocarcinoma. Detection of ?-catenin and COX-2 expression may be helpful for Dukes' staging and evaluating the risk of lymph node metastasis in colorectal adenocarcinoma.