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Objective:To investigate the clinical significance of the transforming growth factor-β receptor I (TGF-βRⅠ) expression in na?ve CD4 + T cells from patients with systemic lupus erythematosus (SLE). Methods:Na?ve CD4 + T cells were purified using magnetic microbeads from peripheral blood mononuclear cells of SLE patients and healthy controls. Real-time quantitative PCR was used to detect TGF-βRⅠ mRNA level, and flow cytometry was used to detect the percentage of CD69 +CD4 + T cells. Data were analyzed by t test and Pearson correlation analysis. Results:The level of TGF-βR Ⅰ mRNA in na?ve CD4 + T cells from SLE patients was significantly lower than that in healthy controls [(0.674±0.873) vs (1.445±1.112), t=2.301, P<0.05]. The TGF-βR Ⅰ mRNA level was negatively correlated with systemic lupus erythematosus disease activity index (SLEDAI) ( r=-0.376, P<0.05), erythrocyte sedimentation rate (ESR) ( r=-0.376, P<0.05), serum creatinine ( r=-0.323, P<0.05) and 24 h urine protein ( r=-0.331, P<0.05), and positively correlated with serum com-plement C3 ( r=0.528, P<0.01). The level of TGF-βRⅠ mRNA level in na?ve CD4 + T cells in SLE patients with renal involvement was lower than that in SLE patients without renal involvement [(0.525±0.536) vs (1.071±1.007), t=2.198, P<0.05]. The TGF-βR Ⅰ mRNA level in the na?ve CD4 + T cells in anti-dsDNA antibody positive group was lower than that in the anti-dsDNA antibody negative group [(0.344±0.315) vs (0.958±1.076), t=2.277, P<0.05]. The expression of TGF-βRⅠ mRNA in na?ve CD4 + T cells from SLE patients was reduced after 24 h stimulation with anti-CD3/CD28 beads [(0.047±0.013) vs (1.008±0.129), t=14.38, P<0.01], which was partially reversed by dexamethasone treatment [(0.240±0.042) vs (0.047±0.013), t=7.845, P<0.01]. Meanwhile, dexamethasone significantly decreased the expression of CD69 in CD4 + T cells [(15.0±2.1)% vs(34.9±2.0)%, t=32.57, P<0.01]. Conclusion:The abnormally low expression of TGF-βRⅠ in na?ve CD4 + T cells may be involved in the pathogenesis of SLE. Glucocorticoid treatment can upregulate the expression of TGF-βRI and inhibit the activation of T cells, This suggests suggesting that TGF-βRⅠ may be a potential target for SLE treatment.
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Objective To investigate the effect of transforming growth factorβreceptor typeⅠ(TGFBRⅠ) on adipocyte differentiation by using a small interference RNA (siRNA). Methods The siRNA targeting TGFBRⅠwas synthesized as experimental group, and negative control siRNA was used as control group. The efficiency of TGFBRⅠdepletion and the expression levels of adipocyte-specific transcription factors CCAAT enhancer binding protein α (C/EBPα),peroxisome proliferator-activated receptor gamma (PPARγ) and adipocyte marker gene fatty acid binding protein 4 (FABP4) were detected by quantitative real-time PCR. After treating with adipocyte differentiation agents for 5 days, the cells were stained with oil red O, and the staining of adipocyte was observed and photographed by laser confocal microscope. In addition, with isopropanol extracted oil red O, optical density values of oil red O were measured at a wavelength of 520, and which were compared between groups. Results After transfection of TGFBRⅠ siRNA, gene expression levels of TGFBRⅠwere significantly reduced in ST2 cells, the number of differentiated adipocytes was significantly increased, and the mRNA levels of adipocyte specific transcription factor C/EBPαand PPARγand adipocyte marker gene FABP4 were enhanced compared with those of control group. After treating with adipocyte differentiation agents for 5 days,the number of lipid droplets of cells with transfection of TGFBRⅠsiRNA was increased than that of cells with transfection of control siRNA. The value of optical density was higher in cells with transfection of TGFBRⅠsiRNA than that of control siRNA group. Conclusion TGFBRⅠsiRNA can effectively facilitate adipocyte formation, which suggests that TGFBRⅠis an important regulator of adipogenic differentiation from progenitor cells.
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Objective Human bone marrow mesenchymal stem cells were transfected by Smad7 and labeled with green fluores-cent mark.BMMSCs were implanted into rabbit glaucoma operational model and observed surviving condition.Methods Through BP and LR reaction,Smad7 with green fluorescent mark was inserted into human bone marrow mesenchymal stem cells by bacterio-phage,filtered positive colony and picked out cell line.15 New Zealand white rabbits were enforced trabeculectomy with BMMSC, then following up cell survival condition.Results Smad7 expressed stable in human bone marrow mesenchymal stem cells with sat-isfactory green fluorescent mark.BMMSC survived in rabbit trabecula with stable green fluorescent and effective ocular press.Con-clusion Smad7 with green fluorescent mark could be inserted into human bone marrow mesenchymal stem cells stably,and has ef-fective results in rabbit model.
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Objective To evaluate clinical value of denaturing high performance liquid chromatography (DHPLC) used in detecting transforming growth factor beta receptor 3 (TGFBR-3) exons 11 and 12 polymorphism in women with idiopathic premature ovarian failure (POF).Methods From Feb.2009 to Dec.2011,110 patients with idiopathic POF undergoing treatment at Shenzhen Maternal & Child Health Institute affiliated to Southern Medical University were enrolled as POF group in this study.In the mean time,110 women under 40 years old with normal hormonal level and menstrual cycles as control group.The exons 11 and 12 of TGFBR-3 gene polymorphism were screened by using DHPLC,and results of DNA sequencing was as golden standard.Some related indexes were calculated,such as sensitivity,specificity,false negative value,false positive value,Youden index,positive predictive value,and negative predictive value.At the same time,20％ of the tested specimens were chosen randomly and detected by DHPLC again.The value of Kappa index were calculated by comparing the results between the first and second DHPLC analysis.Results The exon 11 of TGFBR-3 were not identified gene polymorphism and two nucleotide polymorphisms were identified in exon 12.For 2022 T/C polymorphism,the frequencies of CC with 0.9％ (1/110),TC with 22.7％ (25/110),TT with 76.4％ (84/110),Cwith12.3％ (27/220) and T with 87.7％ (193/220) in POF group were significantly different from CC with 0,TC with 9.1％ (10/110)and TT with 90.9％ (100/110),C with 4.5％ (10/220) and T with 95.5％ (210/220) in control group (all P ＜ 0.05).Allelic and genotypic frequencies of 2161-75 C/T were not differed significantly between the two groups (all P ＞ 0.05).As DNA sequencing as golden standard,DHPLC showed that the sensitivity was 100％,specificity was 97.9％,Youden index was 97.9％,positive predictive value was 96.3％,negative predictive value was 100％,and Kappa index was 0.888 (P ＜ 0.05).Conclusion DHPLC analysis is higher validity,reliability and practicability method in detecting TGFBR-3 polymorphism in idiopathic premature ovarian failure.
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The expression of transforming growth factor(TGF)-β supeffamily co-receptor (Tβ3R) Ⅲ is often lost in many kinds of cancers.Tβ3RⅢ plays a role of negative regulation in tumorigenesis.T3RⅢ could regulate the cellular invasion,migration,proliferation and apoptosis by multiple mechanisms,such as mediating the TGF-β signaling pathway,impacting the mitogen-activated protein kinase(MAPK),nuclear factor-kappa B (NF-κB) and Cdc42 and producing soluble TβR(sTβR) Ⅲ.Defining the mechanisms of absence and physiological functions of TβR Ⅲ in cancers has great significant for the diagnosis,treatment and prognosis of cancer.
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Objective T0 investigate the effects of transforming growth factorβ1 ( TGFβ1 ) and its receptorβ2 (TGFβR2) gene single nucleotide polymorphisms on the risk of intracranial hemorrhage in patients with brain arteriovenous malformation (BAVM).Methods Fifty-three BAVM patients of both sexes aged 18-64 yr who were genetically unrelated native HAN of Guangdong province were divided into 2 groups:patients with and without intracranial hemorrhage ( n =30:23).Venous blood samples were collected and anti-coagulated with ethylene diaminetetraacetic acid for genomic DNA extraction.TGFβ1-509C/T (rs1800469) and TGFβR2 875A/G (rs3087465) gene SNPs were genotyped by using PCR-RFLP.Results There were no significant differences in genotype and frequency between the 2 groups.The G carrier frequency of the TGFβR2 genotype was significantly higher in patients with intracranial hemorrhage than in patients without intracranial hemonrhage.The G carrier of the TGFβR2 genotype was associated with intrarcranial hemorrhage in patients with BAVM.Conclusion TGFβ1 gene polymorphism is not relevant to the intracranial hemorrhage in patients with BAVM,but polymorphisms of TGFβR2 could be a risk factor.
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ObjectiveTo investigate the effect of FUT8-siRNA on transforming growth factor β (TGF-β)-Smad2/3 signalling pathway in renal tubular epithelial cells.MethodsHK-2 cells were divided into six groups:normal group,negative control group,TGF-β1 group,TGF-β1 with FUT8 interference group,TGF-β1 with negative control group,FUT8 interference group.RNAi was performed to silence the expression of FUT8 gene,then immunofluorescent analysis was used to detect the expression of core fucose in the HK-2,immunoprecipitation and lectin blotting were performed to detect the core fucosylation of TGF-βR Ⅱ and ALK-5,and detect the change of Smad2/3 and p-Smad2/3 in HK-2 cells after FUT8 gene was silenced.ResultsCompared with the normal and negative control group,incubation with 5 μg/L TGF-β1 for 48 h could significantly up-regulate the core fucosylation of HK-2 cells,enhance the protein expression of TGF-βR Ⅱ and ALK-5 (P＜0.05),markedly increase the expression level of p-Smad 2/3 (P＜0.05) and cause it to nuclear translocation in HK-2 cells.While FUT8siRNA could inhibit the above up-regulation of TGF-βR Ⅱ and ALK-5(P＜0.05),suppress the increase of p-Smad 2/3(P＜0.05) and its nuclear translocation without disturbing the protein expression of TGF-βR Ⅱ and ALK-5.Conclusion FUT8-catalized core fucosylation of TGF-βR Ⅱ and ALK-5 is needed to fulfill their functions,and blocking core fucosylation of TGF-βR Ⅱ and ALK-5 leads to the inhibition of TGF-β-Smad2/3signalling pathway in HK-2 cells.
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ObjectiveTo investigate the influence of TGF-β receptor subtypes expression and their downstream signaling Smad proteins on rat renal interstitial fibrosis induced by unilateral ureteral obstruction(UUO).MethodsA total of 90 rats were randomly divided into three groups:normal control(CON),sham operation (SOR) and UUO group,and sacrificed 1,3,7,14 and 21 days after operation.Serum creatinine and urea nitrogen were detected to assess renal function.PAS and Masson staining were performed to observe histological damage in the kidneys.Quantitative RT-PCR was used to define expression of mRNA encoding TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney tubular cells.Real-time PCR,Western blotting and immunofluorescence were used to monitor the time-related expression of the TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney.ResultsCompared with the CON group,serum creatinine and urea nitrogen in UUO groups increased at day 3 after operation (P＜0.05) and reached their peak 21 days after operation (P＜0.01).Obvious inflammatory cell infiltration was observed in UUO group 3 days after operation,while renal tubular atrophy and renal interstitial fibrosis were observed in UUO group14 days after operation.The mRNA expressions of ALK-5,ALK-7 and TGF-βR Ⅱ increased significantly in UUO group 3 days after operation (all P＜0.05) and reached their peaks 14 days after operation (all P＜0.01).The mRNA expression of ALK-6 decreased significantly in UUO group 3 days after operation(P＜0.05) and reached its lowest level 14 days after operation (P＜0.01).The changes in the protein level of those receptors were consistent with their mRNA expressions.The protein expressions of Smad2/3 and p-Smad2/3 increased significantly in UUO group at day 3(all P＜0.05) and reached their peak at day 14 after operation(all P＜0.01).ConclusionExpressions of TGF-β receptor subtypes ALK-5,ALK-6,ALK-7,TGF-βR Ⅱ and their downstream signaling Smad2 and Smad3 proteins may influence the progress of renal interstitial fibrosis,tubular atrophy and inflammatory cell infiltration in UUO model rats.
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Objective To construct transforming growth factor β receptor Ⅱ (TGFβRⅡ) siRNA expression vector, and investigate the inhibitory effect on TGFβRⅡ in hepatic stellate cells (HSC-T6) ,thus offer a preliminary study for RNAi therapy in liver fibrosis. Methods The two sites of RNAi action were selected in TGFβRⅡ cDNA through online primer design software of Ambion company. The corresponding double-stranded DNA sequence was constructed into pSilencer-U6 plasmid, which could transcribe small interference RNA, the plasmid was transfected into HSC-T6 cells with Lipofectamine 2000. The expression of TGFβRⅡ mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. Results Expression siRNA plasmids of siRNA-a and siRNA-b that target TGFβRⅡ were successfully constructed. Compared with control group, the expression of TGFβRⅡ mRNA was reduced by 0. 89 ± 0. 06 and 0. 25 ± 0. 03 in two siRNA groups, and expression of TGFβRⅡ protein reduced to 0. 86± 0. 05 and 0. 23 ± 0. 02, respectively. TGFβRⅡ expression was inhibited at mRNA and protein levels after transfected with the siRNA-b plasmid( P <0. 01 ).. There was no difference in the TGFβRⅡ expression after transfected with the siRNA-a plasmid( P > 0. 05). Conclusion TGFβRⅡ-siRNA can effective inhibit expression of TGFβRⅡ in HSC-T6.
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Transforming growth factor-beta (TGF-beta) and its receptors have been suggested to play key roles in the pathogenesis of asthma. The aim of this study was to evaluate the effects of genetic variations in the TGF-beta receptor type III (TGFBR3) on asthma and on its related phenotypes in the general population. A cohort of 2,118 subjects aged from 10 to 18 years responded to a questionnaire concerning asthma symptoms and risk factors. Methacholine airway hyperresponsiveness (AHR), skin test responses to common aeroallergens, and serum total IgE levels were evaluated in the cohort. A total of 19 SNPs for TGFBR3 were found using direct re-sequencing in 24 healthy adults. Of these, informative SNPs [+44T>C (S15F) and +2753G>A at 3'UTR] were selected and scored using the high throughput single base extension method. Atopy was identified in subjects with 44T>C allele [P = 0.04, OR (95% CI) = 0.79 (0.62-0.99)] and in subjects with Ht1 (CG) more frequently than in subjects with other haplotypes [P = 0.04, OR (95% CI) = 1.27 (1.01-1.59)]. The A allele in 2753G>A was more common in subjects with non-atopic asthma [OR (95% CI) = 1.76 (1.01-3.05)]. A significant association was found between non-atopic asthma and 44T_2753A [OR (95% CI) = 2.16 (1.22-3.82)]. Genetic variations in TGFBR3 appear to be associated with a genetic predisposition to development of asthma and to phenotypes of asthma. Also, the minor allele 2753G and the haplotype TA in the TGFBR3 gene were associated with a pathogenesis of non-atopic asthma.
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Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Pueblo Asiatico/genética , Asma/etnología , Estudios de Casos y Controles , Estudios de Cohortes , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Variación Genética/fisiología , Genética de Población , Estudio de Asociación del Genoma Completo , Genotipo , Inmunoglobulina E/inmunología , Desequilibrio de Ligamiento , Proteoglicanos/genética , Receptores de Factores de Crecimiento Transformadores beta/genéticaRESUMEN
Objective: To investigate the relationship between the expressions of the transforming growth factor beta 1 (TGF-β1), Smad4, and TβR II in the different stages of gallbladder carcinoma and discuss their roles in the development of gallbladder carcinoma. Methods: Immunohistochemical SP method was used to examine the expression of TGF-β1, Smad4, and TβR II in 30 cases of gallbladder carcinoma, 11 cases of gallbladder adenoma, and 30 cases of cholecystitis. The relationship between the expressions of TGF-β1, Smad4 and TβR II and the clinicopathological features of gallbladder carcinoma were analyzed. Results: The positive rates of TGF-β1, Smad4 and TβR II was 73.3%, 20%, and 16.7% in 30 cases of gallbladder carcinoma; 90.9%, 63.7%, and 54.5% in 11 cases of gallbladder adenoma; 96.7%, 93.3%, and 90% in 30 cases of cholecystitis. The expression of TGF-β1 was significantly lower in gallbladder carcinoma than that in cholecystitis (P<0.05). The expressions of Smad4 and TPR H were significantly lower in gallbladder carcinoma than that in cholecystitis (P<0.05). Gallbladder carcinoma at stages I and II had lower TGF-β1 expression but higher Smad4 and TβR II expression than those at stage III and V (P<0.05). TGF-β1 expression was significantly higher (94.1%) in gallbladder carcinoma with metastasis than that without metastasis (P<0.05). Conclusion: The decreased expression of TGF-β1 in gallbladder carcinoma may be related with the cell malignant transformation and uncontrolled growth. Over-expression of TGF-β could not inhibit the proliferation of gallbladder carcinoma which may be due to the decreased expression of Smad4 and TβR II. The high expression of TGF-β1 is related with the progression, invasion, and metastasis of gallbladder carcinoma.
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Objective To compare the expression of renal transforming growth factor-?_1(TGF-?_1),its receptors type Ⅰ and Ⅱ(T?RⅠ and T?R Ⅱ) in two rat models of chronic nephropathy induced by CsA and FK506 respectively.Methods Rat models of chronic CsA-/FK506-induced nephropathy were established by administering sandimun Neoral and Prograf separately.Then their kidneys were dissected and the protein expression of TGF-?_1,T?RⅠand T?RⅡ,the mRNA levels of T?RⅠ and T?RⅡ was detected by using immunohistochemistry(IHC) and in situ hybridization(ISH) respectively.Results The IHC revealed that the integrated optical densities(IODs) of renal TGF-?_1,T?RⅠ and T?RⅡ were(605.24)?(140.24),(876.28)?(208.73) and(981.59)?(258.65) respectively in CsA-treated rats,(488.37)?(101.76),(586.63)?(131.21) and(543.34)?(105.26) respectively in FK506-treated ones.The ISH indicated that the IODs of renal T?RⅠ and T?RⅡ mRNA were(905.08)?(158.75) and(1090.92)?(242.73) respectively in CsA-treated rats,(661.37)?(205.65) and(716.27)?(195.55) respectively in FK506-treated ones.The difference of the above-mentioned five factors between the two groups was significant(P
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Objective: To study the effect of helicobacter pylori on gastric mucosal cell proliferation in gastritis. Methods: Fifty-six gastritis patients with or without Helicobacter pylori infection (Hp+ 27; Hp- 29) were selected. The expression of proliferation cell nuclear antigen (PCNA), epidermal growth factor receptor (EGFR), transforming growth factor beta receptor type Ⅰand typeⅡ(TGF?RⅠ, TGF?RⅡ) in gastric mucosa were examined by immunohistochemical method. Results: The PCNA and EGFR were significantly higher in Hp positive chronic gastritis patients than in Hp negative ones(P
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Objective To study the mRNA expression and function of TGF-?1, TGF-?RⅡ and CD105 in the lesional, non-lesional skin of psoriasis and of normal human skin. Methods The RNA of skin tissue was extracted using single-step method of RNA isolation by acid guanidinium thiocyanate. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure the expression of mRNA. Results The mRNA expression of TGF-?1 and TGF-?RⅡ was lower in the epidermis of psoriatic lesion than that of non-lesional psoriatic and normal human skin(P
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Objective To investigate the expression and significance of transforming growth factor beta receptors (TGF?RⅠ, TGF?RⅡ) in basal cell carcinoma and squamous cell carcinoma. Methods The expression of TGF?RⅠand TGF?RⅡwere assessed by quantitative real-time PCR and streptavidin -peroxidase (SP) immunohistochemical techniques in specimens of basal cell carcinoma, squamous cell carcinoma and normal control skin. Results Eighteen patients with basal cell carcinoma and twenty-four with squamous cell carcinoma were enrolled in the study. The expression levels of TGF?RⅠand RⅡmRNA were significantly down-regulated in basal cell carcinoma and squamous cell carcinoma in comparison with those in control skin specimens (P
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Objective To investigate the effects of the serum containing Fufangbiejiaruanganfang (FFBJRGF) on cell cycle and apoptosis of hepatic stellate cells(HSC). Methods The influence of TGF-? 1 on cell cycle and rate of apoptosis of HSC in vitro was examined by using flow cytometer, furthermore, the effects of FFBJRGF drug sera on HSC were observed compared with IFN-? serum and normal rat serum controls. Results The numbers in phase of G 0 and G 1 were increased when HSCs were incubated by adding TGF-? 1, IFN-? serum and different dosages of FFBJRGF drug serum respectively, especially being predominant in the FFBJRGP groups. Meanwhile, the intervenient roles in phase of G 2?M?S of HSC were found in FFBJRGF drug serum treatments. Compared with normal serum control group, there were not notable effects on the cellular apoptosis of HSC in the different dosages of FFBJRGF drug serum groups as well as IFN-? group, however, TGF-? 1 could significantly increased HSC apoptosis rate. Conclusion Our results indicates FFBJRGF has anti-hepatic fibrosis efficiency through inhabiting the proliferative cycle of HSC, not up-regulating HSC apoptosis in vitro.
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Objective To study the effects of TGF-? 1 on the proliferation of hepatic stellate cell(HSC) and the interfering effect of serum containing Fufangbiejiaruanganfang (FFBJRGF ) drug in vitro. Methods TGF-? 1 was added to the culture of HSC in vitro and the proliferation of HSC was detected by MTT method. The effects of the serum containing different dosage FFBJRGF on the HSC proliferation was observed by using modified method of serum pharmacology of the traditional Chinese medicine and MTT method. Results The proliferation of HSC was decreased under adding exogenous TGF-? 1, which effect was presented at 72 hours after TGF-? 1 complement. The HSC proliferative reactions were obviously inhibited by adding sera containing high, moderate, low dosage of FFBJRGF as well as IFN-?serum compared with the normal control HSC culture (P
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Objective To observe the chronological and spatial expression of fibronectin(Fn)and TGF-?RⅡ in developing mouse glomeruli.Methods The expressions of Fn and TGF-?RⅡ were examined by immunohistochemical and stereological method in embryos(including fetal age of 12,14,16 and 18d)and postnatal mouse kidneys(including the age of 1,3,7,14,21,28 and 42d).Results Immunohistochemical results showed that at fetal age of 14d,Fn was stained in S-shaped corpuscles,but not stained in the comma-shaped corpuscles.After fetal age 16d,Fn was expressed in all stages of glomeruli except the comma-shaped corpuscles.TGF-?RⅡ was stained in all stages of glomeruli from fetal age 14d.Fn and TGF-?RⅡ were localized respectively in basement membrane and cell plasma of glomeruli.Stereological analysis showed that the expression of Fn was increased gradually in all stages of glomeruli with renal development,except in the comma-shaped corpuscles;also,the expression of TGF-?RⅡ increased gradually in all stages of glomeruli with renal development.Conclusion The expressions of Fn and TGF-?RⅡ in embryonic and postnatal mouse kidney show chronological and spatial sequence,implying that they might play a role in the development and maturation of renal glomeruli of mouse.
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AIM: To determine the relationship between the mutation of the RII gene and RER status in the tumorigenesis of sporadic colorectal cancer. METHODS: We screened RER status and mutation of the RII gene from 50 sporadic colorectal cancers (19 in the proximal colon, 31 in the distal colorectum). RESULTS: RER was found in 13 cases (8 in the proximal colon, 5 in the distal colorectum), and 5 of them showed mutations of the RII gene. All 5 cancers carrying a TGF-? RII gene mutation showed RER+, but there wasn't any mutation of RII gene in RER(-) cases. Four of 5 RII mutation were located at the cecum. CONCLUSION: These data indicate that the TGF-? RII gene is a major target of microsatellite instability and mutation of the RII gene play an important role in carcinogenesis of sporadic colorectal cancer with microsatellite instability, especially at the cecum. [
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AIM:To investigate the role of danshensu on Smad signal transduction in rat hepatic stellate cells(HSCs)stimulated with transforming growth factor(TGF-?1).METHODS:The rat HSCs was isolated with collagenase by in situ-liver recirculation perfusion and cultured in vitro.MTT colorimetric assay was used to detect proliferation of HSCs treated with different concentration of danshensu.The expressions of ?-SMA and T?R I and II were observed by immunocytochemitry,indirect immunofluorescent staining and Western blotting when HSCs stimulated with TGF-?1 and with different concentrations of danshensu for 24 h.RESULTS:(1)Danshensu at the concentration from 0.0625 mmol/L to 1 mmol/L prevented the proliferation of HSCs in a dose-dependent manner(P