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Objective:To examine the effects of liraglutide on the transforming growth factor-β1(TGF-β1)/Smads signaling pathway in renal tissues of elderly rats with type 2 diabetes mellitus(T2DM)and to explore the underlying mechanisms.Methods:A total of 75 healthy elderly male Sprague-Dawley rats aged 20 months and weighing(500±100)g were divided into the normal control group(Group N, n=25)and the model group( n=50)by using a random number table.Rats in the model group were given high-glucose and high-fat diets for 8 weeks and then were injected with a single dose(30 mg/kg)of 1% streptozotocin into the abdominal cavity.Forty-eight rats in the model group were successfully molded and were divided into the T2DM group(Group D, n=24)and the intervention group(Group LD, n=24). Rats in Group LD were abdominally injected with liraglutide in a dose of 200 μg·kg -1·d -1, and the other two groups were given an equal volume of saline.At the end of 4, 8 and 12 weeks, eight rats in each group were randomly selected and 24-hour urine collections were made to measure 24-hour urinary protein.Then the rats were anesthetized, blood samples were collected for biochemical tests, and renal tissues were removed for microscopic examination of pathological changes after HE staining.The expression of type Ⅳ collagen(Col-Ⅳ)was detected by using an immunohistochemical method, and the mRNA expression of TGF-β1, Smad3 and Smad7 was detected by using real-time fluorescence quantitative polymerase chain reaction.One-way analysis of variance was used for comparisons between the groups, and the LSD-t test was used for pairwise comparisons. Results:Compared with Group N, Group D showed thickening of the glomerular basement membrane, mesangial proliferation and interstitial fibrosis at each time-point, which grew worse with time, and the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ also increased significantly(12-week: 0.69±0.01 vs.0.15±0.01, 0.51±0.02 vs.0.02±0.01, 183.33±2.08 vs.221.67±2.08, t=89.22, 60.87 and 24.52, P<0.05), while Smad7 mRNA levels decreased( t=13.42, P<0.05). Compared with Group D, the degree of renal fibrosis was reduced, and the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ at 12-week significantly decreased( t=71.703, 37.58 and 20.04, P<0.05), while Smad7 mRNA increased( t=9.96, P<0.05)in Group LD.With prolonged intervention of liraglutide, the lesions were mitigated, the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ decreased, and Smad7 mRNA increased gradually( P<0.05)in Group LD. Conclusions:Liraglutide has anti-renal fibrosis effects via inhibiting the TGF-β1/Smads pathway, thereby reducing the production of Col-Ⅳ, and can delay the progression of renal lesions in elderly T2DM rats.
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Objective To investigate the correlation between peripheral serum TNF-α and TGF-β1 expression levels with the infiltration degree of lymphocytes in labial glands and interstitial lung disease(ILD) in the patients with primary Sjogren's syndrome (pSS).Methods Serum level of TNF-α and TGF-β1 in 116 patients with pSS and 20 persons undergoing physical examination were measured by ELISA.Meanwhile the labial gland samples in pSS patients were performed the pathological examination.The patients were grouped according to the lesion stage of ILD and pathology grade of labial glands.The serum TNF-α and TGF-β1 levels were statistically analyzed.Results Serum levels of TNF-α and TGF-β1 in the pSS-ILD group were significantly higher than those in the simple pSS group and control group(P＜0.01).The levels of serum TNF-α and TGF-β1 in the simple pSS group were significantly higher than those in the control group(P＜0.01).The levels of serum TNF-α and TGF-β1 in the labial glands lymphocytes infiltration group were significantly higher than those in the labial glands lymphocytes non-infiltration group and control group(P＜0.01),while the levels of serum TNF-α and TGF-β1 in the labial glands lymphocytes non-infiltration group were significantly higher than those in the control group(P＜0.01).The TNF-α and TGF-β1 expression was positively correlated with lymphocytes infiltration lesion grade(r=0.867,0.613,P=0.000);the expression of TNF-α and TGF-β1 was positively correlated with the lesion grade of ILD(r=0.814,0.864,P =0.000);the level of TNF-α was positively correlated with the TGF-β1 relative expression amount(r=0.857,P=0.000).Conclusion TNF-α and TGF-β1 are involved in the lymphocytes infiltration of labial glands and ILD occurrence and develvp ment,and their expression increase may be the cause of labial glands structure destruction and complicating ILD.
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Objective To study the effect of cardiac contractility modulation (CCM) on TGFβ1/Smad/CTGF signaling pathway.Methods A rabbit heart failure (HF) model was established by ligating the ascending aortic root.Thirty rabbits were divided into sham operation group (n=10),HF group (n=10) and CCM group (n=10).Myocardial tissue collagen Ⅰ and collagen Ⅲ were analyzed with Sirius red staining,myocardial tissue hydroxyproline level was measured by chromometry,and expressions of TGFβ1,Smad3,Smad7,CTGF were detected by Western blot in 3 groups.Results Collagen Ⅰ,collagen Ⅲ and hydroxyproline content were highcr in HF group than in sham operation group,while collagen Ⅰ,collagen Ⅲ and hydroxyproline conten were lower in CCM group than in sham operation group (0.69±0.05 μg/mg vs 0.98±0.04 μg/mg,P＜0.05).The expression levels of TGFβ1,Smad3,CTGF were higher while those of Smad7 were lower in HF group than in sham operation group (P＜0.05).The expression levels of TGFβ1,Smad3,CTGF were lower while those of Smad7 were higher in CCM group than in HF group (0.49±0.03 vs 0.67±0.04,0.43±0.06 vs 0.59±0.06,0.45±0.08 vs 0.75±0.09,P＜0.05;0.43±0.08vs 0.26±0.04,P＜0.05).Conclusion CCM can improve the myocardial fibrosis in HF rabbits by downregulating the expression of TGFβ1,Smad3,CTGF and upregulating the expression of Smad7.
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Objective To observe the effect of febuxostat on epithelial-to-mesenchymal transition (EMT) of kidney tubules and the levels of serum IL-6 nad transforming growth factor (TGF) β1 in hyperuricemic rats.Methods Forty male SD rats were divided into 4 groups:normal control group (NC group),oteracil potassium group (OP group),oteracil potassium with febuxostat group (OF group) and oteracil potassium with benzbromarone group (OB group).Each group had 10 rats and balanced in body weights.To induce hyperuricemia,rats were given oteracil potassium by gastric garage once a day for eight weeks.Rats in OF group and OB group were given either febuxostat or benbromarone starting with oteracil potassium,and rats in NC group was given saline only.Blood samples were taken before,and at the end of 4 and 8 weeks of the treatments and serum uric acid,creatinine,blood usea nitrogen (BUN),IL-6 and TGFβ1 contents were measured at each time point.Renal pathological changes were observed via HE and Masson staining,and the expression of α-SMA and E-cadherin were detected by immunohistochemistry.Results Compared with those in NC group,the levels of serum uric acid,creatinine,BUN,IL-6 and TGFβ1 in the another three groups were increased significantly (all P ＜ 0.01).However,the IL-6 and TGFβ1 contents in OF group were much lower than those in OP group (P ＜0.01).HE and Masson staining showed that OF group had less damage and tubulointerstitial fibrosis than OP group and OB group (P ＜0.01).Moreover,the expression of α-SMA was significantly down-regulated (P ＜ 0.01) and that of E-cadherin was significantly up-regulated in OF group compared with those in OP group.Conclusion Febuxostat treatment significantly inhibited EMT and reduced the levels of IL-6 and TGFβ1 in hyperuricemia rats.
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Objective To explore the effects of 1,25-dihydroxyvitamin D3 on renal expression of TGFβ1,Smad3 and Smad7.Methods 40 Sprague-Dawley mts were randomized into 4 groups:normal control rats (group A),diabetic nephropathy group (group B),small dose 1,25-dihydroxyvitamin D treatment group (0.5 μg · kg-1 · d-1,group C) and large dose 1,25-dihydroxyvitamin D treatment group (1 μg · kg-1 · d-1,group D),each group had 10 rats.After 12 weeks,the renal function,blood glucose,glycosylated hemoglobin and the urine trace albumin content of each rats were tested.Results The biochemical indicators in group B were higher than those in group A,(t =-16.566,P ＜0.05;t =-16.949,P ＜0.05;t =-11.844,P ＜0.05;t =-19.778,P ＜0.05;t =-14.013,P ＜ 0.05).Compared with group B,the biochemical indicators and the expression of TGFβ1,Smad3 mRNA reduced in group C and group D,and the expression of Smad7 mRNA increased (F =37.892,P ＜ 0.05;F =70.068,P ＜ 0.05;F =21.95,P ＜0.05;F =77.619,P ＜0.05;F =37.670,P ＜0.05;F =1062.562,P ＜0.05;F =2463.789,P ＜0.05;F =81.745,P ＜ 0.05).There were no significant differences between group C and group D (t =0.538,P＞0.05;t =1.737,P＞0.05;t =0.671,P＞0.05;t =1.763,P ＞0.05;t =0.997,P ＞0.05;t =1.653,P ＞0.05;t=1.543,P＞0.05;t =-1.313,P ＞0.05).Conclusion 1,25-dihydroxyvitamnin D3 has protective effect on diabetic nephropathy rats model,the mechanism may be associated with inhibiting the expression of TGF β1 and Smad3,increasing the expression of Smad7.
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Objective To investigate the possible regulating effect of integrin-linked kinase (ILK) towards matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 (MMP-9/TIMP-1) ratio in the process of transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) in human kidney proximal tubular epithelial (HK-2) cells.Methods HK-2 cells were cultured and stimulated with 10 ng/ml TGF-β1.Specific ILK-small interfering RNA (ILK-siRNA) was used to inhibit ILK expression.The characteristic epithelial marker (E-cadherin) and mesenchymal marker (α-SMA) of EMT were examined by Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and Western blot.The expressions of ILK,MMP-9,and TIMP-1 were also examined,to determine the regulating effect of ILK towards MMP-9/TIMP-1 ratio.Results In the HK-2 cells cultured with TGF-β1,the expression of E-cadherin decreased,and α-SMA expression increased;overexpression of ILK and an abnormal changing of MMP-9/TIMP-1 ratio were observed.ILK inhibition by ILK-siRNA could adjust MMP-9/TIMP-1 ratio to near normal.Meanwhile,the overexpressed ILK and α-SMA were decreased.Conclusions Our data indicates that ILK-siRNA successfully inhibits ILK expression,which regulates the MMP-9/TIMP-1 ratio in HK-2 cells.The inhibition of ILK expression suppresses TGF-β1-induced EMT partially.
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Background Scarring of surgical area,the most important factor,leads to the failure of glaucoma filtering surgeries.Therefore,more and more attentions are paid to the causes and process of scar formation.Objective This study was to compare the differences of proliferation and migrating abilities of fibroblasts between filtering bleb scar tissue and normal Tenon capsular tissue,and to investigate the inhibitory effects of miRNA-200a (miR-200a) on biological behavior of conjunctival fibroblasts.Methods Normal Tenon capsular tissue and filtering bleb scar tissue were collected during the strabismus surgery and glaucoma filtering surgery,respectively for the primarily culture of fibroblasts.The proliferation (absorbency,A) of the cells was assayed by cell counting kit-8 (CCK8) method;the relative migrating distance of the cells was measured by cell scratch test;and the relative expressions of transforming growth factor-β1 (TGF-β1)mRNA and miR-200a mRNA in the cells were detected by realtime fluorescence quantitative PCR.TGF-β1 mimic of 0,1,2 and 5 ng/ml was added in the medium of human normal Tenon capsular-derived fibroblasts (HTFs),and 0.00,0.25,0.50,1.00 μg/ml TGF-β1 inhibitor was added in the medium of human scarring-derived fibroblasts (HSFs) for 24 hours,respectively,and CCK8 was used to evaluate the proliferation of the cells.The relative migrating distance as well as the relative expressions of miR-200a mRNA were analyzed in the 2 ng/ml TGF-β1 mimic-or 1.00 μg/ml TGF-β1 inhibitor-treated cells.Results The primary conjunctival presented the spindle and star-like in shape with large body and oval nuclei.The cells showed the positive response for keratin and vimentin antibodies.The A values were 1.476±0.110 in the HSFs and 0.958±0.074 in the HTFs,with a significant difference between them (t =24.900,P=0.016).The relative expressions of TGF-β1 mRNA were significantly higher in the HSFs than those in the HTFs,and the relative expressions of miR-200a were evidently lower in the HSFs than those in the HTFs,showing significant differences between them (t =6.358,P =0.024;t=7.394,P =0.018).Compared with the 2 ng/ml TGF-β1 mimic-treated HTFs,the relative migrating distance increased,while the expression level of miR-200a mRNA was significantly reduced in the 2 ng/ml TGF-β1 mimictreated HSFs (all at P＜0.05);Compared with the 1.00 μg/ml TGF-β1 inhibitor-treated HTFs,the relative migrating distance decreased,but the expression level of miR-200a mRNA was significantly elevated in the 1.00 μg/ml TGF-β1 inhibitor-treated HSFs (all at P＜0.05).Conclusions The proliferation and migrating abilities are stronger in the HSFs than those in the HTFs,which probably is regulated by the expression of miR-200a in the cells.The miR-200a plays a negative feedback for the effect of TGF-β1 promoting proliferation and migration of fibroblasts.
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Objective To explore the correlation among serum immunoglobulin G4 (IgG4),transforming growth factor-β1 (TGF-β1),connective tissue growth factor (CTGF) and Hashimoto thyroiditis (HT) thyroid fibrosis.Methods Case-control study.A total of 159 patients with HT visiting the Wuhan Union Hospital were collected from May 2013 to March 2015.All patients were divided into IgG4 HT group (IgG4≥1.35 g/L,n =39) and non-IgG4 HT group (IgG4 ＜ 1.35 g/L,n =120).The serum IgG4,TGF-β1 and CTGF were determined by enzyme-linked immunosorbent assays.The levels of serum thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibodies (TgAb) were measured by electrochemiluminescence immunoassay.Meanwhile,ultrasound of the thyroid gland was performed.Statistical analysis was performed by use of SPSS 17.0 software.The Mann-Whitney U test was used to compare two independent samples of non-normal distribution data,Fisher's exact test was employed to analyze thyroid imaging differences,correlation test was performed to examine various correlations,multivariate Logistic regression analysis was used to evaluate thyroid fibrosis risk factors.Results Compared with that of non-IgG4 HT group,IgG4 HT group:TPOAb [(455.2 ± 169.7) vs.(186.5 ± 102.3),U =27.0,P=0.003],TgAb [(984.6±452.7) vs.(289.3 ±245.1),U=30.5,P=0.017],TGF-β1 [(1.45±0.97) vs.(0.30±0.22),U=119.0,P=0.035] andCTGF [(88.65±14.39) vs.(62.21± 8.76),U =69.0,P =0.039] were significantly higher,thyroid ultrasound showed obvious fibrosis (35/4 vs.32/88,x2 =48.03,P =0.000);significant positive linear correlation between IgG4 and TPOAb (r =0.719,P =0.000),CTGF and TGF-β1 (r =0.500,P ＜ 0.01) respectively.Logistic regression analysis indicated that all the serum IgG4,TPOAb,TGF-β1 and CTGF were independent risk factors of thyroid fibrosis [IgG4,odds ratio (OR) =1.968,P =0.014,95％ confidence interval (CI) =1.287-2.041;TPOAb,OR =2.537,P =0.012,CI =1.322-2.869;TGF-β1,OR =1.549,P =0.023,CI =1.105-1.498;CTGF,OR =1.185,P =0.046,CI =1.204-1.625].Conclusion The highlevel of circulating antibodies,IgG4,TGF-β1 and CTGF were significantly associated with thyroid fibrosis,and were independent risk factors of HT fibrosis.
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Objective To observe the protective effect of N-NAC on radiation-induced lung injury. Methods 86 cases of thoracic neoplasm patients were chosen and randomly divided into two groups,group RT +N(n =43)and group RT(n =43).Two groups were observed by CT after radiotherapy.Acute and chronical toxicities were graded by RTOG.TGF-β1,IL-1,IL-4,TNF were observed before and after the radiotherapy.Results After 3 monthsof radiotherapy,RTOG≥2 was 23.4%(RT +N),while RTOG≥2 was 53.1%(RT).there was significant differencebetween the two groups(P <0.01).At 6,9 and 12 months,fibrosis was present in 28.4%,25.4%,22.4% receivingRT vs 58.4%,54.4%,52.4% receiving RT +N,there was significant difference between the two groups(P <0.05).TGF-β1,IL-1,IL-4,TNF were observed which showed that The RT +N were lower than RT.Conclusion N-NAC can reduce incidence rate of lung injury in radiotherapy,and can reduce the content and the release of TGF-β1,IL-1,IL-4,TNF.
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Objective To observe the effect of specific immunotherapy on serum interleukin-4 (IL-4),interleukin-18 (IL-18) and transforming growth factor-beta 1 (TGF-β 1) for the treatment of bronchial asthma in children.Methods Seventy-five patients with bronchial asthma in children were divided into observation group(38 cases) and control group(37 cases) by random digits table method.The control group was received drug treatment,the observation group was given specific immunotherapy on the basis of the control group.After 12 months of treatment,the level of forced expiratory volume in 1 second (FEV1),FEV1/forced vital capacity (FVC),FVC,peak expiratory flow (PEF) such as lung function and serum IL-4,IL-18 and TGF-β1 between two groups was compared and the level of serum IL-4,IL-18 and TGF-β1 was compared among different efficacy.Results After 12 months of treatment,the level of FEV1,FEV/FVC,FVC,PEF was (2.82 ± 0.35) L,(81.65 ± 5.38)％,(3.46 ± 0.45) L,(5.61 ± 1.44) L/s in observation group,(2.17 ±0.29) L,(72.84 ±4.82)％,(3.24 ±0.41) L,(5.08 ± 1.35) L/s in control group,and there was significant difference (P ＜ 0.05).After 12 months of treatment,the level of serum IL-4,IL-18 and TGF-β1 was (5.94 ± 4.76) ng/L,(192.85 ± 54.06) ng/L,(6.17 ± 0.42) μ g/L in observation group,(7.26 ± 5.33) ng/L,(259.61 ± 67.83) ng/L,(6.83 ± 0.48) μ g/L in control group,and there was significant difference(P ＜ 0.05).There was significant difference in the level of serum IL-4,IL-18 and TGF-β1 among different efficacy(P ＜ 0.05).Conclusion IL-4,IL-18 and TGF-β 1 in children with bronchial asthma are involved in the inflammatory response,serum IL-4,IL-18 and TGF-β1 levels can provide the basis for the treatment and prognosis of bronchial asthma in children.
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Objective To explore the effect and mechanism of TGF beta1/smad3 signaling pathways on apoptosis in mouse pulmonary fibrosis.Methods Fifty-four healthy male C57BL/6 mice were randomly divided into three groups:normal control (n=18),pulmonary fibrosis model (n =18) and TGF-β1/smad3 inhibitor group (n=18).Six mice in each group were randomly killed on days 7,14 and 28.Hematoxyli~eosin and Masson staining were adopted to evaluate the severity of pulmonary inflammation and fibrosis.The content of hydroxyproline (Hyp) in the lung tissues was detected by alkaline hydrolysis technique.The apoptosis was observed by tunnel apoptosis assay kit.P-smad3 and caspase3 protein expressions were assessed via Western blot.Results Lung in model mice versus normal control showed alveolar inflammatory change in 7 days and significant pulmonary fibrosis in 28 days(P＜0.05).Meanwhile,apoptosis index,hydroxyproline content,caspase3,and phosphorylated Smad3 were obviously higher in model mice than in control group (P ＜ 0.05).Compared with model group,TGF-β1/smad3 inhibitor group showed that alveolitis and pulmonary fibrosis degree,hydroxyproline content,cell apoptosis index,the expressions of p-smad3 and caspase3 were decreased at same time point (P ＜ 0.05).Conclusions TGF beta1/smad3 signaling pathways may participate the abnormal apoptosis during the development of pulmonary fibrosis,and TGF-β1/smad3 inhibitor SB431542 could inhibit this process.
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Objective To evaluate the effects of moderate oxygen concentration (40％ O2) on A549 cells and expression of transforming growth factor-β1 (TGF-β1) mRNA and connective tissue growth factor (CTGF) mRNA in A549 cells.Methods Cultured A549 cells were seeded in 6-well plates (1 mYhole) or in 10 cm diameter dishes (5 ml/dish) with the density of 1 × 1O5 cells/ml and randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),40％ oxygen concentration group (group 40％ O2),and 95％ oxygen concentration group (group 95 ％ O2).A549 cells were exposed to air,40 ％ O2 and 95 ％ O2 for 48 h in C,40％ O2 and 95％ O2 groups,respectively.The necrosis and expression of TGF-β1 mRNA and CTGF mRNA were detected at the end of culture.Results Compared with group C,the necrosis rate was significantly increased,while the expression of TGF-β1 mRNA and CTGF mRNA was up-regulated in 40％ O2 and 95％ O2 groups (P ＜ 0.05).Compared with group 95 ％ O2,the necrosis rate was significantly decreased,while the expression of TGF-β1 mRNA was down-regulated (P ＜ 0.05),and there was no significant difference in CTGF mRNA expression in group 40 ％ O2 (P ＞ 0.05).Conclusion Exposure to moderate oxygen concentration for a certain time can induce damage to A549 cells,but the severity is lighter than that caused by high oxygen concentration,and up-regulation of TGF-β1 and CTGF gene expression may be involved in the mechanism.
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Objective To investigate whether transforming growth factor-β1 (TGF-β1) could induce the expression of twist protein in the human kidney tubular epithelial cells (HKCs).Methods Human proximal HKCs were cultured in vitro and divided into three groups as follows:control group,TGF-β1group (10 ng/ml),and TGF-β1 + wortmannin group (10 ng/ml,10nmol/l,respectively).After cultured for 48 hours,cell immunofluorescene was used to observe the expression of E-cadherin and alpha-smooth muscle actin (α-SMA).Meanwhile,Western blotting was used to detect the protein expression of E-cadherin,α-SMA,and twist.Results Compared to control group,the protein expressions of E-cadherin were significantly decreased in T group (3.54 ± 0.17 vs 16.06 ± 0.50,P =0.001),whereas the protein expressions of α-SMA and twist were significantly increased in T group (α-SMA:14.78 ± 0.48 vs 3.75 ± 0.50,P=0.001 ;twist:14.24 ±0.14 vs 3.06 ±0.15,P =0.001).Compared to T group,a significant increase of the E-cadherin protein expression was detected in TGF-β1 + wortmannin group (15.88 ± 0.36,3.46 ±0.19,P =0.001),whereas,the protein expressions of α-SMA and twist were significantly decreased in TGF-β1 + wortmannin group (α-SMA:3.50 ±0.39 vs 15.0 ±0.24,P =0.001 ;twist:3.09 ±0.1 vs 14.04± 0.16,P =0.001).Cell imunofluorescence showed that the expressions of E-cadherin and α-SMA were consistent with the results of Western blotting.Conclusions TGF-β1 can induce the expression of twist protein in HKCs through Akt pathways.
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Objective To investigate the effect of transforming growth factor β1 (TGF-β1) on the expression of matrix metalloproteinase 9 (MMP-9),tissue inhibitor of metalloproteinase 1 (TIMP-1),nuclear factor kappa B(NF-κB) and the possible signalling pathways in human amniotic cells WISH.Methods The WISH cell line was cultured.WISH cells were added with TGF-β1 of different concentrations (0,2,10 and 20 ng/ml,respectively) for 24 hours.Then,reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot.Results (1) The profile of TIMP-1 mRNA (0.413 ±0.036,0.623 ±0.058,1.392 ±0.124,1.387 ±0.102) in WISH cells elevated when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).In accordance with TIMP-1 mRNA,the expression of TIMP-1 also elevated with the increase of TGF-β1 (0.357 ± 0.031,0.596 ± 0.048,1.243 ± 0.097 and 1.359 ± 0.121,respectively).And when 2,10 or 20 ng/ml of TGF-β1 was added,the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-β1 was added(P ＜ 0.05).(2)In contrast with TIMP-1,MMP-9 mRNA (1.325 ±0.056,0.987 ±0.081,0.610 ±0.034,0.347 ±0.023) in WISH cells decreased when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).The MMP-9 protein (1.119 ±0.064,1.008 ±0.052,0.578 ±0.041,0.401 ±0.015) also decreased with the increase of TGF-β1.And when 2,10 or 20 ng/ml of TGF-β1 was added,the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-β1 was added (P ＜ 0.05).(3) The NF-κB protein (1.423 ±0.065,1.116 ± 0.045,0.796 ± 0.041,0.359 ± 0.021) was significandy reduced with the increase of TGF-β1 (0,2,10,20 ng/ml; P ＜ 0.05).Conclusions The mRNA and protein expression of TIMP-1 decreased when TGF-β1 was low in WISH cells,whereas those of MMP-9 elevated when TGF-β1 was low.The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane.And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.
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Objective To investigate the implications of ratio of the CD4+ and CD25+ positive regulatory T cells (CD4+CD25+Tregs) in peripheral blood mononuclear cells (PBMC) and its associated regulatory factors such as forkhead transcription factor 3 (Foxp3) mRNA transcriptional activity in PBMC,serum levels of transforming growth factor beta-1 (TGF-β1),and interleukin 10 (IL-10) in the immunopathology of patients with middle to late staged nasopharyngeal carcinoma (NPC) based on a clinical trial.Methods In this study,18 NPC cases at middle to late stage as observing group and 10 healthy persons as control group were included to detect their ratio of the CD4+CD25+Tregs in the PBMC with flow cytometry (FCM) technique,transcriptional activity of Foxp3 with RT-PCR procedure,and serum levels of TGF-β1 and IL-10 with enzyme-linked immunosorbent assay (ELISA) method.A comparative analysis was used to explore their implications in the immunopathological correlation of NPC cases with their lesion.Results The ratio of the CD4+CD25+Tregs to total CD4+T cells in PBMC was significantly increased [(4.23 ±0.53)％ vs (2.65 ±0.31)％,t =8.60,P ＜0.01],accompanied with significantly elevated levels of Foxp3 transcription in PBMC (3.699 ± 0.309 vs 1.109 ± 0.146,t' =31.08,P ＜ 0.05],and serum contents of TGF-β1 [(645.56 ± 39.61) pg/ml vs (488.82 ± 36.91) pg/ml,t =10.27,P ＜ 0.01] and IL-10 [(1.27 ± 0.21) pg/ml vs (0.68 ± 0.08) pg/ml,t' =10.61,P ＜ 0.05] in these patients,when compared with that of healthy controls.Conclusions It may be true that CD4 + CD25 + Tregs,transcriptional regulatory factor Foxp3,and cytokines TGF-β1 as well as IL-10 altogether were composed of a regulating system in a positive feedback way to promote the developing process of immunotolerance phenomena in the tumor microenvironment and the initiation of immunoescape among patients with middle to late staged nasopharyngeal carcinoma.
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ObjectiveTo investigate the clinical significance of transforming growth factor β1 (TGF-β1)and vascular endothelial growth factor (VEGF) expression in tissues of nasal inverted papilloma (NIP).MethodsThe clinical data of patients with NIP underwent surgical resection were retrospectively analyzed.The TGF-β1 and VEGF expression in NIP tissues and nasal polyps tissues were detected by immunohistochemistry method.100 patients with NIP were divided into benign lesions,atypical hyperplasia and malignant group according to result of pathological diagnosis,the nasal polyps was used as the control group.ResultsThe positive expression rate of TGF-β1 and VEGF in the NIP group were 46.0％ and 32.0％,compared with the control group the differences were significant(all P ＜ 0.05 ).In different pathological groups,the results of TGF-β1 and VEGF expression were malignant group ＞ atypical hyperplasia ＞ benign lesions.The positive expression rate of TGF-β1 and VEGF in the NIP group had significantly positive correlation(P ＜ 0.05).ConclusionThe TGF-β1 and VEGF expression were closely related to the the occurrence,development and malignant of NIP.TGF-β1 was highly expressed in the NIP tissues,and could increase the expression of VEGF and promote the formation of neovascularization of NIP.
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Objective To investigate the effect of propofol on transforming growth factor (TGF)-β1/Smad2 signaling pathway in lung tissues in rats with lipopolysaccharide (LPS)-induced acute lung injury (ALI).Methods Fifty-six male Wistar rats,aged 7-8 weeks,weighing 260-300 g,were randomly divided into 5 groups:control group (group A,n =8) ; LPS group (group B,n =12); 3 propofol groups (groups C,D,E,n =12).ALI was induced by intravenous LPS 8 mg/kg in groups B,C,D and E.In groups C,D,E,propofol 5 mg/kg was injectedintravenously before LPS administration and at 0 and 1 h after LPS administration,respectively,followed by infusion of propofol at 10 mg· kg-1 · h-1 until 5 h after LPS administration.Group A received the equal volume of normal saline.Arterial blood samples were collected immediately before LPS administration and 1,3 and 5 h after LPS administration for determination of pH value and PaO2.Then the animals were sacrificed and the lungs were immediately removed for calculation of the wet/dry lung weight ratio and for determination of the expression of TGF-β1-mRNA and Smad2 in lung tissues.Results Compared with group A,pH value and PaO2 were significantly decreased,wet/dry lung weight ratio was increased and the expression of TGF-β1 mRNA and Smad2 was up-regulated in groups B and E (P ＜ 0.05).Compared with group B,pH value and PaO2 were significantly increased,wet/dry lung weight ratio was decreased and the expression of TGF-β1 mRNA and Smad2 was down-regulated in groups Cand D,and PaO2 was significantly increased in group E (P ＜ 0.05).Conclusion The mechanism by which propofol alleviates ALI induced by LPS is related to inhibition of TGF-β1/Smad2 signaling pathway.
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Objective To investigate the changes of peripheral blood Foxp3+ regulatory T cell (Treg cell) in patients with ovarian cancer (OC).Methods In 46 patients with OC and 46 normal controls,the percentage of peripheral blood CD4+ Foxp3+ Treg cell was assessed by flow cytometry and Foxp3 mRNA level was detected by real-time quantitative reverse transcription polymerase chain reaction.The level of plasma.transforming growth factor β1 (TGF-ββ 1) was measured by enzyme linked immunosorbent assay.Results The percentages of CD4+ Foxp3+ Treg cell,Foxp3 mRNA and level of plasma TGF-β1 in patients with OC were statistically higher than those in normal controls [(11.42 ± 2.67)％ vs.(8.94 ± 1.98)％,0.59 ± 0.21 vs.0.37 ±0.14,(35 580 ±7274) ng/L vs.(28 610 ±5631) ng/L,P=0.0000].Conclusion The number and/or function of CD4+ Foxp3+ Treg cell in peripheral blood of patients with OC are abnormal,CD4+ Foxp3+Treg cell may participate in the occurrence of OC.
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Objective To investigate the efficacy of sophocarpine in rats with alcoholic liver disease and its effects on the expression of tumor necrosis factor (TNF)-α,interleukin (IL)-6 and transforming growth factor (TGF)-β1.Methods A total of 48 male Sprague-Dawley adult rats were evenly divided into healthy control group,model group,prevention group and treatment group.The rats in the healthy control group were gavaged with 0.9％NaCl every day for 12 weeks.The rats in the model group,prevention group and treatment group were gavaged with alcohol for 12 weeks to establish the model.The prevention group was injected with 20 mg · kg1 · d1 sophocarpine for 12 weeks.Since the fifth week,the treatment group was continuously injected with 20 mg · kg1 · d-1 sophocarpine for eight weeks.The histological changes were evaluated.The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase ( AST ), alkaline phosphatase (AKP),triglyceride (TG) and total cholesterol (TC) were examined.And the expression of TNF-α,IL-6 and TGF-β1 in liver tissue at mRNA and protein level were detected with immunohistochemistry and real-time polymerase chain reaction (PCR) assay.Comparison among groups was perform with single factor analysis of variance,pairwise comparisons with least significant difference method (LSD method),ranked data with Kruskal-Wallis H-test and multiple pairwise comparison with Nemenyi test.Results Compared with model group,hepatic steatosis and inflammatory cell infiltration were significantly improved in the treatment group and prevention group.The levels of ALT (41.40 U/L± 10.53 U/L and 40.75 U/L±6.94 U/L vs 58.37 U/I±5.35 U/L),AST(121.60 U/L±16.24 U/L and 109.50 U/L±9.23 U/L vs 156.63 U/L±32.47 U/L),AKP(114.88 U/L±40.37 U/L and 112.60 U/L±44.34 U/L vs 161.75 U/L±28.95 U/L),TG (4.19 mmol/L±0.99 mmol/L and 2.69 mmol/L± 1.35 mmol/L vs 4.50 mmol/L±0.99 mmol/L) and TC (1.48 mmol/L±0.28 mmol/L and 1.43 mmol/L±0.19 mmol/L vs 1.67 mmol/L±0.20 mmol/L) significantly decreased and the difference was statistically significant ( all P＜0.05).The expression of TNF-α,IL-6 and TGF-β1 at mRNA and protein level in liver tissue of model group were significantly higher than those of healthy control group,prevention group and treatment group.After treated with sophocarpine,the expression of TNF-α(mRNA:1.36 ± 0.08,1.16 ± 0.05 ; protein:3.38 ％ ± 0.82 ％,1.74 ％ ± 0.65 ％ ),IL-6 (mRNA:1.51 ± 0.05,1.39 ± 0.02; protein:5.89％ ± 0.96％,4.26％ ± 0.53％) and TGF-β1 (mRNA:1.39±0.04,1.37±0.02; protein:4.27％ ±0.97％,2.11％ ±0.83％) of treatment group and prevention group at mRNA and protein level significantly lower than those of model group (mRNA:1.81±0.16,1.95 ±0.13,1.84±0.22; protein:5.82％ ± 1.21％,7.63％ ±1.03％,5.33％± 1.12％) and the difference was statistically significant (all P＜0.01).Conclusion Sophocarpine significantly alleviates alcohol induced liver injury in rats,improves liver steatosis and inflammatory reaction degree,which may be related with the downregulation of TNF-α,TGF-β1 and IL-6 expression in liver tissue of ALD rats.
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Objective To investigate the expression and distribution of Cdc42-interacting protein 4 (CIP4) in renal fibrotic tissue,and the interaction between CIP4 and β-catenin in transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) model of HK-2 cell line.Methods In vivo,the model of renal fibrosis was induced by 5/6 subtotal nephrectomy in rat.Masson staining was used to evaluate the level of renal tissue fibrosis.The expression and distribution of CIP4 was detected by immunohistochemistry.In vitro,the EMT model of HK-2 cell line was induced by TGF-β1 (10 μg/L) for 72 h.Western blotting was used to observe the expression of E-cadherin,β-catenin,CIP4 and α-SMA.Colocalization and interaction of CIP4 and β-catenin were detected by immunofluorescence and immunoprecipitation respectively.Results Compared to sham group,CIP4 expression was increased in group of 5/6 subtotal nephrectomy,CIP4 was mainly distributed in basolateral side of renal tubular epithelia.In vitro,expressions of α-SMA and CIP4 were increased in HK-2 cells stimulated by TGF-β1 for 72 h (2.5and 1.8 folds,respectively) (all P＜0.05),expression of E-cadherin was decreased(P＜0.05).Partial colocalization between CIP4 and β-catenin was detected by immunofluorescence.In control group,CIP4 and β-catenin partially colocalized at the cell membrane.Mter the stimulation of TGF-β1,translocation to nucleus of CIP4 and β-catenin were increased,and partially colocalized in nucleus.The interaction between CIP4 and β-catenin was observed by immunoprecipitation in both control and TGF-β1 stimulated groups.Conclusions Expression of CIP4 in renal fibrotic tissue is increased,which is mainly distributed in basolateral side of renal tubular epithelia.CIP4 and βcatenin partially colocalize and interact with each other.CIP4 may play a role in EMT process through the interaction with β-catenin.