转染外源性转化生长因子β_1基因对兔椎间盘髓核细胞成纤维细胞生长因子表达的影响

①目的探究体内转染腺病毒载体转化生长因子β1(Ad/CMV-hTGF-β1)对椎间盘组织退变的保护作用机制。②方法纯种成年新西兰大白兔10只,随机分为实验组和对照组,每组5只。采取腹部手术入路,暴露腰椎间盘,取20μL本院骨科实验室构建的以腺病毒为载体的Ad/CMV-hTGF-β1,其滴度为6×106pfu,注射于腰3、4椎间盘髓核内;对照组仅进行相应的外围手术操作,椎间盘未做任何处理。手术后3周取出椎间盘,通过苏木精-伊红(HE)染色观察髓核组织细胞形态;用免疫组织化学方法观察髓核细胞成纤维细胞生长因子(FGF)的表达;并用VIDAS图像分析系统进行半定量分析。③结果普通HE染色结果显示,2组椎间盘髓核组织形态基本相同。免疫组织化学染色显示,实验组椎间盘髓核细胞FGF的表达比对照组明显增高,差异有统计学意义(t=3.48,P<0.05)。④结论体内转染腺病毒载体Ad/CMV-hTGF-β1具有提高椎间盘髓核细胞FGF表达的作用。

转染外源性转化生长因子β_1基因对兔髓核细胞凋亡的影响

①目的 探讨转染转化生长因子 β1 (hTGF β1 )基因对兔髓核细胞凋亡的影响 ,为活化椎间盘细胞功能、预防椎间盘组织退变提供依据。②方法 取成年新西兰大白兔 5只 ,将外源性hTGF β1 基因直接注射于L3～ 4椎间隙 ,以注射同样剂量磷酸缓冲液的L4～ 5作为实验对照。术后 3周取出相应椎间盘组织 ,利用TUNEL免疫荧光组织化学方法观察髓核细胞凋亡情况。应用Vidas图像分析系统进行免疫阳性细胞半定量分析。③结果 注射hTGF β1 基因的椎间盘凋亡细胞数明显少于对照组 ,测得OPTDM值分别为 0 .0 0 2± 0 .0 0 1、0 .1 5 9± 0 .0 4 1 ,二者差异有显著性 (t=2 7.83,P <0 .0 1 )。④结论 转染hTGF β1

转化生长因子β_1在肝细胞癌中的表达及意义

目的探讨转化生长因子(TGFβ1)在肝细胞癌中可能的作用机制。方法用免疫组化SABC方法检测45份石蜡包埋人肝细胞癌标本及36例癌旁组织中TGFβ1蛋白表达的情况。结果45例肝癌组织中TGFβ1阳性表达率和为62.2%.肝细胞癌组织中TGFβ1表达在肝细胞癌中明显升高,与癌旁组织相比较差异有显著性(P<0.05),与肝细胞癌的组织分化程度(按Edmondson-Steiner分级标准),包膜是否完整及有无癌栓有相关性(P<0.05)。结论肝细胞癌组织中TGFβ1呈过表达。TGFβ1的过表达对诱发肝细胞癌及对肝细胞癌的转移都起到一定的作用,可作为肝细胞癌的预后指标。

Effect of porous titanium coated with IGF-1 and TGF-β_1 loaded gelatin microsphere on function of MG63 cells

Porous titanium with porosity of 60% was prepared by metal injection molding（MIM）,and coated with gelatin sustained-release microspheres which were made by improved emulsified cold condensation method.The effects of porous titanium coated with insulin-like growth factor-1（IGF-1） and transforming growth factor-β1（TGF-β1） gelatin microspheres on the function of MG63 cells were evaluated in vitro.The results show that porous titanium coated with gelatin sustained-release microspheres has no cytotoxicity.The IGF-1 and TGF-β1 loading concentrations are positively correlative with the proliferation and differentiation of MG63 after co-culturing with the concentrations of IGF-1 and TGF-β1 gelatin microspheres in the range of 0.1-10 ng/mg and 0.25-2.5 ng/mg,respectively.The MG63 cells exhibit the best proliferation and differentiation with the IGF-1 and TGF-β1 loading concentrations of 10 ng/mg and 2.5 ng/mg,respectively.The joint application of IGF-1 and TGF-β1 group,which promote adhesion,proliferation and differentiation of MG63 cells,is superior to a single application group.

CONSTRUCTING ADENO-ASSOCIATED VIRUS-TGFβ_3 AND COMPARING ITS BIOLOGICAL EFFECT ON PROTEOGLYCAN SYNTHESIS IN DEDIFFERENTIATED NUCLEUS PULPOUS CELLS WITH ADENOVIRUS-TGFβ_1

Objective To construct adeno-associated virus (AAV) expression system for transforming growth factor β3 (TGFβ3) and detect its biological effect on proteoglycan synthesis of the earlier and later dedifferentiated rabbit lumbar disc nucleus pulpous (NP) cells, which was compared with that of adenovirus (AV) expression system for TGFβ1. Methods TGFβ3 gene was obtained using PCR. Its upstream contained restriction enzyme site Kpn Ⅰ, and its downstream contained restriction enzyme site SalⅠ. Using the restriction enzyme sites of PCR product of TGFβ3 and the corresponding multiple cloning site (MCS) in plasmid AAV, TGFβ3 was subcloned into AAV. The recombinant plasmid AAV-TGFβ3 was transfected into H293 cells with LipofectamineTM 2000, and the expression of TGFβ3 gene was detected using immunofluorescent analysis. After AAV-TGFβ3 virus particle with infectious activity was packaged, TGFβ3 expression in NP cells was detected by immunoblotting, and its biological effect on proteoglycan synthesis was detected by antonopulos method and compared with that of AV-TGFβ1 in the earlier and later dedifferentiated NP cells. Results For the earlier dedifferentiated NP cells, AAV-TGFβ3 slowly and stably enhanced proteoglycan synthesis, but AV-TGFβ1 rapidly and transiently enhanced its synthesis. For the later dedifferentiated NP cells, AAV-TGFβ3 stably enhanced proteoglycan synthesis, but AV-TGFβ1 inhibited its synthesis. Conclusion AAV expression system can mediate TGFβ3 gene to be expressed stably, and AAV-TGFβ3 can enhance proteoglycan synthesis of the earlier and later dedifferentiated NP cells.

TGF-β_1 inhibits connexin-43 expression in cultured smooth muscle cells of human bladder

Objective: In this research, we studied the TGF-β1 effects on connexin-43 expression in cultured human bladder smooth muscle cells. Methods: Human bladder smooth muscle cells primary cultures, with bladder tissue obtained from patients undergoing cystectomy, were intervened by recombinant human TGF-β1. Connexin-43 expression in human bladder smooth muscle cells was then examined by Western blotting and immunocytochemistry. Results: Stimulation with TGF-β1 led to significant reduction of connexin-43 immunoreactivity and coupling (P<0.0001). Connexin-43 protein expression was significantly downregulated (P<0.05). Simultaneously, low phosphorylation species of connexin-43 were particularly affected. Conclusion: Our experiments demonstrated a significant downregulation of connexin-43 by TGF-β1 in cultured human bladder smooth muscle cells. These findings support the view that TGF-β1 is involved in the pathophysiology of urinary bladder dysfunction.

PADRE-TGFβ1融合蛋白的原核表达及鉴定

构建PADRE-TGFβ1原核表达质粒,并对其进行诱导表达和反应原性分析。对PADRE、联接序列(Linker)及TGFβ1的基因序列进行密码子优化,采用全基因合成的方法人工合成全长PADRE-Linker-TGFβ1序列,合成产物克隆至pET-28a(+)质粒。将构建好的质粒转化至E.coliBL21(DE3)感受态细胞,经1 mmol/L IPTG诱导表达,表达产物经SDS-PAGE分析,在约19.87 ku处出现与预期目的蛋白一致的新蛋白条带,Western blot鉴定结果显示,该蛋白能够与TGFβ1抗体发生特异性结合。表明成功构建了pET28-PADRE-TGFβ1原核表达质粒,表达的融合蛋白质具有TGFβ1的反应原性,为TGFβ1自体疫苗的制备及应用奠定了基础。

Neuropeptide Y promotes TGF-β1 production in RAW264.7 cells by activating PI3K pathway via Y1 receptor

Objective To examine the effect of neuropeptide Y （NPY） on TGF-β1 production in RAW264.7 macrophages. Methods Enzyme linked immunosorbent assay （ELISA） was used to detect TGF-β1 production. Cell counting kit 8 （CCK-8） was used to assay the viability of RAW264.7 cells. Western blot was used to detect the phosphorylation of PI3K p85. Results NPY treatment could promote TGF-β1 production and rapid phosphorylation of PI3K p85 in RAW264.7 cells via Y1 receptor. The elevated TGF-β 1 production induced by NPY could be abolished by wortrnannin pretreatment. Conclusion NPY may elicit TGF-β production in RAW264.7 cells via Y1 receptor, and the activated PI3K pathway may account for this effect.

TRANSFORMING GROWTH FACTOR-β1 AND SMAD4 SIGNALING PATHWAY DOWN-REGULATES RENAL EXTRACELLULAR MATRIX DEGRADATION IN DIABETIC RATS

Objective To investigate the role of transforming growth factor-131 （TGF-β1）/Smad4 pathway in development of renal fibrosis in streptozotocin （STZ）-induced diabetic nephropathy （DN） rats and explore its possible mechanism. Methods Male Wistar rats weighing 180-220 g were divided into 5 groups： group A （ normal control）, group B [ diabetes mellitus （DM） 2 weeks ], group C （ DM 4 weeks）, group D （ DM 8 weeks）, and group E （ DM 16 weeks）. Except for the normal control group, other groups were induced DM by single injection of STZ （55 mg/kg） respectively. Blood glucose level, serum creatinine, and 24-hour urine protein were examined. Expressions of TGF-β1 and Smad4 protein and mRNA in kidney were detected using immunohistochemical technique, Western blot, and real-time PCR. mRNA expressions of stromelysin-1 （ MMP-3 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and collagen Ⅲ in kidney were also detected by real-time PCR. Results The levels of blood glucose, serum creatinine, and 24-hour urine protein in rats of group B, C, D, and E were higher than those of the control group. With the progression of renal fibrosis, the expressions of TGF-β1 and Smad4 protein and mRNA in kidney of diabetic rats elevated. In addition, the renal MMP-3 mRNA expression diminished in diabetic rats, while TIMP-1 and collagen Ⅲ mRNA increased. Conclusions In STZ-induced diabetic rats, the TGF-β1/Smad4 appears to play an important role in renal fibrosis of DN. The increased expression of TGF-β1 and Smad4 might result in the transcriptional regulation of downstream target genes of TGF-β1/Smad4 pathway, which contributes to the progression of renal fibrosis in diabetic rats.

Transforming growth factor-β1 gene polymorphisms are associated with progression of liver fibrosis in Caucasians with chronic hepatitis C infection

AIM: Considerable attention is focused on polymorphisms in the gene encoding transforming growth factor-pi (TGF-β1), a multifunctional cytokine that is in turn a potent growth inhibitor involved in wound healing and differentiation. In humans, it promotes the pathogenesis of organ fibrosis, atherosclerosis, cancer, autoimmune and inflammatory diseases, keloid disease, and hypertrophic scarring. For this reason, much emphasis has been placed on studies elucidating the impact of TGF-β1 and its gene variations for the susceptibility and pathogenesis of these diseases. Unfortunately, some studies have serious limitations. METHODS: We have recently described a high-throughput method for investigation the Arg25Pro polymorphism of human TGF-β1 gene and showed that the frequency of the Pro25 allele is significantly associated with hepatic fibrogenesis. In this report, we describe two novel LightCyder (LC) techniques that facilitate the examination of the two other known alterations in the coding region of TGF-β1. We investigated whether these polymorphisms contribute to hepatitis-induced progression of fibrogenesis in Chinese and Caucasians. RESULTS: In the Chinese ancestry, the gene polymorphisms at codons 25 and 263 were not found and the genetic variant at codon 10 is unlikely to confer susceptibility to hepatic fibrosis. Contrarily, in Caucasians TGF-β1 allelic variations are more frequent and the presence of prolines either in codon 25 or 10 is associated with the interindividual variability in developing more severe fibrosis during chronic hepatitis C infection. CONCLUSION: In summary, these results confirm the hypothesis that TGF-β1 polymorphisms are associated with fibrosis progression in Caucasians chronically infected with hepatitis C.

Relationship between transforming growth factorβ1 and antifibrotic effect of interleukin-10

AIM： To study the effect of interleukin-10 （IL-10） on the expression of transforming growth factor β1 （TGF-β1） in hepatic fibrosis rats and the anti-fibrotic role of exogenous IL-10. METHODS： Hepatic fibrosis was induced by carbon tetrachloride administered （CCh） intraperitoneally. The experiment was performed in two stages. In the first stage, 60 SD rats were divided randomly into normal control group I（GNI, n = 8）, hepatic fibrosis group（GC, n = 28）and IL-10 intervened group（GI, n = 24）. At the beginning of the 7^th and 11^th wk, hepatic stellate cells （HSCs） were isolated, reverse transcription-polymerase chain reation （RT-PCR） and immunocytochemistry were performed to detect the expression of TGF-β1 in HSCs. Histological examination was used to determine the degree of hepatic fibrosis. In the second stage, 47 SD rats were divided randomly into normal control group 2 （GN2, n = 6）and CCh group（GZ, n = 41）. At the end of the 9th week, rats in GZ group were allocated randomly into model group（GM, n = 9）, IL-10 treatment group（GT, n = 9） and recovered group （GR, n = 9）. At the end of the 12^th week, all rats were sacrificed. RT-PCR and immuno- histochemistry were performed to detect the expression of TGF-β1 in liver tissue. ELISA was used to assay serum TGF-β1 levels. RESULTS： Hepatic fibrosis developed in rats with the increase of the injection frequency of CCI4. In the first stage, hepatic fibrosis developed and HSCs were isolated successfully. At the 7^th and 11^th week, TGF-β1 mRNA in GC group increased significantly compared with that in GN1（P = 0.001/0.042） and GI groups（P = 0.001/0.007）, whereas there was no significant difference between the two groups. The levels of TGF-β1 at the beginning of the 7^th wk was higher than that of the 11^th wk （P = 0.049）.Immunocytochemistry results of TGF-β1 were consistent with the above findings. In the second stage, TGF-β1 increased significantly in GM group compared to GN2. Alter treatment with IL-10, TGF-β1 declined obviously. The expression of TGF-β1 decreased in GR group but was still higher than that in GT group. CONCLUSION： The levels of TGF-β1 are increased in hepatic fibrosis rats and decreased alter treatment with exogenous IL-10. IL-10 may play an anti-fibrotic role by suppressing TGF-β1 expression.

Effect of pegylated interferon alpha 2b plus ribavirin treatment on plasma transforming growth factor-β1,metalloproteinase-1,and tissue metalloproteinase inhibitor-1 in patients with chronic hepatitis C

AIM： To evaluate the effect of antiviral treatment on plasma levels of transforming growth factor-β1 （TGF-β1）, metalloproteinase 1 （MMP-1）, and tissue inhibitor of metaUoproteinase-1 （TIMP-1） in patients with chronic hepatitis C. METHODS： TGF-β1, MMP-1, and TIMP-1 plasma concentrations were measured by an enzyme immunoassay in 28 patients, during 48 wk of treatment with pegylated interferon-alpha 2b （PEG-IFN-α2b） plus ribavirin （RBV） and after 24 wk of follow-up. Patients were divided into two groups： responders （R） and non-responders （NR） related to achieved sustained virologic response. Normal values were evaluated in plasma samples of 13 healthy volunteers. RESULTS： Baseline plasma concentrations of TGF-β1 and TIMP-1 （30.9±3.7 and 1 506±61 ng/mL respectively） measured in all subjects significantly exceeded the normal values （TGF-β1： 18.3±1.6 ng/mL and TIMP-1： 1 102±67 ng/mL）. In contrast, pretreatment MMP-1 mean level （6.5±0.9 ng/mL） was significantly lower than normal values （11.9±0.9 ng/mL）. Response to the treatment was observed in 12 patients （43%）. TGF-β1 mean concentration measured during the treatment phase decreased to the control level in both groups. However at wk 72, values of NR patients increased and became significantly higher than in R group. TIMP-1 concentrations in R group decreased during the treatment to the level similar to normal. In NR group, TIMP-1 remained significantly elevated during treatment and follow-up phase and significant difference between both groups was demonstrated at wk 48 and 72. MMP-1 levels were significantly decreased in both groups at baseline. Treatment caused rise of its concentration only in the R group, whereas values in NR group remained on the level similar to baseline. Statistically significant difference between groups was noted at wk 48 and 72. CONCLUSION： These findings support the usefulness of TGF-β1, TIMP-1, and MMP-1 in the management of chronic hepatitis C. Elevated TIMP-1 and low MMP-1 plasma concentrations during antiviral therapy may indicate medication failure.

Effects of Chinese traditional compound, JinSanE, on expression of TGF-β1 and TGF-β1 type II receptor mRNA, Smad3 and Smad7 on experimental hepatic fibrosis in vivo

AIM:The transforming growth factor-beta (TGF-β)/Smad signaling pathway system plays a prominent role in the control of cell growth and extracellular matrix formation in the progression of liver fibrogenesis. Smad proteins can either positively or negatively regulate TGF-β responses. In this study, the therapeutic effects of Chinese traditional compound decoction, JinSanE, and the changes of TGF-β/Smad signaling pathway system in carbon tetrachloride (CCl4)-induced rat experimental liver fibrosis were examined. METHODS: Seventy-two healthy Wistar rats were assigned to groups including normal control group, CCl4 model group, JinSanE treatment group I and JinSanE treatment group II. Each group contained 18 rats. All groups, except the normal control group, received CCl4 subcutaneous injection for 8 wk. Rats in JinSanE groups I and II were orally treated with JinSanE daily at the 1st and 5th wk, respectively, after exposure to CCl4. The expression of TGF-β1 and TGF-β1 type II receptor (TRII) mRNA in the liver was determined by reverse transcription polymerase chain reaction, and the expression of TGF-β1, Smad3 and Smad7 by immunohistochemistry. The liver histopathology was also examined by HE staining and observed under electron microscope. The activities of several serum fibrosis-associated enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), the levels of serum hyaluronic acid (HA) were assayed. RESULTS: Hepatic fibrosis caused by CCl4 was significantly inhibited in the JinSanE-treated groups. The degrees of necrosis/degeneration and fibrosis scores were significantly lower in the JinSanE-treated groups than in the model control group. The expression of TGF-β1, TRII and Smad3 was significantly higher in the model group than that in the JinSanE-treated groups, and the active/total TGF-β1 ratio in the JinSanE groups was suppressed. Expression of TRII mRNA and Smad3 proteins snowed a distribution pattern similar to that of TGF-β1 with a direct correlation in terms of the degree of hepatic fibrosis. The amount of positive staining Smad7 cells was significantly less in the model group than in the JinSanE-treated groups and the normal group. The contents of ALT, AST and HA were significantly lower in the JinSanE-treated groups than those in the model group. CONCLUSION: Traditional Chinese medicine, JinSanE, prevents the progression of hepatic damage and fibrosis through the inhibition of TGF-β1, TRII and Smad3 signal proteins, and increases expression of Smad7 signal protein in vivo.

Promoter polymorphism of transforming growth factor-β1 gene and ulcerative colitis

AIM: To elucidate the possible difference in two promoter polymorphisms of the transforming growth factor-β1 (TGF-β1) gene (-800G > A, -509C > T) between ulcerative colitis (UC) patients and normal subjects.METHODS: A total of 155 patients with established ulcerative colitis and 139 normal subjects were selected as controls. Two single nucleotide polymorphisms within the promoter region of TGF-β1 gene (-509C > T and -800G > A) were genotyped using PCR-RFLP. RESULTS: There was a statistically significant difference in genotype and allele frequency distributions between UC patients and controls for the -800G > A polymorphism of the TGF-β1 gene (P < 0.05). The frequency of the TGF-β1 gene polymorphism at position -800 showed that the AA genotype and the allele A frequencies significantly differed between the patients and healthy controls (P < 0.05). At position -509, there was no statically significant difference in genotype and allele frequency between the patients and control subjects.CONCLUSION: The results of our study indicate that there is a significant difference in both allele and genotype frequency at position -800G > A of TGF-β1 gene promoter between Iranian patients with UC and normal subjects.

Effect of transforming growth factor-β1 on human intrahepatic cholangiocarcinoma cell growth

AIM： To elucidate the biological effects of transforming growth factor-β1 （TGF-β1） on intrahepatic cholangiocarcinoma （ICC）.METHODS： We investigated the effects of TGF-β1 on human ICC cell lines （HuCCT1, MEC, and HUH-28） by monitoring the influence of TGF-β1 on tumor growth and interleukin-6 （IL-6） expression in ICC cells.RESULTS： All three human ICC cell lines produced TGF-β1 and demonstrated accelerated growth in the presence of TGF-β1 with no apoptotic effect. Studies on HuCCT1 revealed a TGF-β1-induced stimulation of the expression of TGF-β1, as well as a decrease in TGF-β1 mRNA expression induced by neutralizing anti-TGF-β1 antibody. These results indicate that TGF-β1 stimulates the production and function of TGF-β1 in an autocrine fashion. Further, IL-6 secretion was observed in all three cell lines and exhibited an inhibitory response to neutralizing anti-TGF-β1 antibody. Experiments using HuCCT1 revealed a TGF-β1-induced acceleration of IL-6 protein expression and mRNA levels. These findings demonstrate a functional interaction between TGF-β1 and IL-6. All three cell lines proliferated in the presence of IL-6. In contrast, TGF-β1 induced no growth effect in HuCCT1 in the presence of small interfering RNA against a specific cell surface receptor of IL-6 and signal transducer and activator of transcription-3.CONCLUSION： ICC cells produce TGF-β1 and confer a TGF-β1-induced growth effect in an autocrine fashion.TGF-β1 activates ILo6 production, and the functional interaction between TGF-β1 and IL-6 contributes to ICC cell growth by TGF-β1.

The expression and antigenicity identification of recombinant rat TGF-β1 in bacteria

In order to study structure-function details of TGF-beta1, the recombinant mature form of rat TGF-beta1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-beta1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. This system allowed an active and selective synthesis of recombinant TGF-beta1. The molecular weight of expressed TGF-alpha1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF-beta1 antibody. The mature recombinant rat TGF-beta1 expressed in this study provides a useful tool for future detailed structural and functional studies.

Down-regulation of transforming growth factor β1/activin receptor-like kinase 1 pathway gene expression by herbal compound 861 is related to deactivation of LX-2 cells

AIM: To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-β1 (TGFβ1)/ activin receptor-like kinase 1 (ALK1, type Ⅰ receptor) signaling-pathway-related gene expression in the LX-2 cell line, and the inhibitory mechanism of Cpd861 on the activation of LX-2 cells. METHODS: LX-2 cells were treated with TGFβ1 (5 ng/mL) Cpd861 (0.1 mg/mL), TGFβ1 (5 ng/mL) plus Cpd861 (5 ng/mL) for 24 h to investigate the effect of Cpd861 on the TGFβ1/ALK1 pathway. Real-time PCR was performed to examine the expression of α-SMA (α-smooth muscle actin), ALK1, Id1 (inhibitor of differentiation 1). Western blotting was carried out to measure the levels of α-SMA and phosphorylated Smad1, and immunocytochemical analysis for the expression of α-SMA. RESULTS: In LX-2 cells, TGFβ1/ALK1-pathway-related gene expression could be stimulated by TGFβ1, which led to excessive activation of the cells. Cpd861 decreased the activation of LX-2 cells by reducing the expression of α-SMA mRNA and protein expression. This effect was related to inhibition of the above TGFβ1/ALK1-pathway- related expression of genes such as Id1 and ALK1, and phosphorylation of Smad1 in LX-2 cells, even with TGFβ1 co-treatment for 24 h. CONCLUSION: Cpd861 can restrain the activation of LX-2 cells by inhibiting the TGFβ1/ALK1/Smad1 pathway.

Effects of saponin from the seed of Litchi chinensis Sonn on TGF-β1, FN and SOCS-1 in renal tubular epithelial cells under high glucose

Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12 medium supplemented with 30 mmol/L glucose and treated with or without SLS.In the normal group,isometric DMEM/F12 medium with 5.5mmol/L glucose was added.The secretion of TGF-β1 and fibronectin(FN)were detected by ELISA.Cell apoptosis was detected by the method of Annexin V-FITC/PI double staining.Western blot was used to detect the level of suppressor of cytokine signaling-1(SOCS-1).Results:The result of ELISA showed that the secretion of TGF-β1 and FN was decreased in SLS groups compared with those in 30 mmol/L glucose treated group(P<0.05).There were more cells apoptosis in 30 mmol/L glucose treated group than that in the normal group(P<0.01).Compared with the 30 mmol/L glucose treated group,the apoptosis of HKC were significantly decreased in SLS groups(P<0.01).Western blot showed that the level of SOCS-1 in high glucose+SLS group was decreased(P<0.01),compared with the high glucose group.Conclusion:SLS can reduce the secretion of TGF-β1 and FN in HKC by reducing the deposition of extracellular matrix.SLS also significantly reduced the apoptosis of HKC by inhibiting the level of SOCS-1.These results suggest the roles of SLS in preventing the progress of glomerular sclerosis.

Influence of HBcAg in liver cell plasma on expression of transforming growth factor-beta 1 in liver tissue of low-grade chronic hepatitis B patients

AIM： To study the influence of HBcAg on the expression of transforming growth factor-beta 1 （TGF-131） in liver tissue of low-grade chronic hepatitis B （CHB） patients. METHODS： The expression of T（3F-β1 and HBcAg in liver samples from 93 low-grade CHB patients was detected by immunohistochemistry and valuated by semi-quantitative scoring. RESULTS： In the 93 low-grade CHB patients, HBcAg was expressed in cell plasma but not in the liver tissue. There was no significant difference between the two groups. CONCLUSION： The expression of TGF-β1 is not related with HBcAg expressed as plasma type in the tissues of low-grade CHB patients.

A study on the expressions and the correlation of TGF-β1 and α-SMA in healing process of bile duct trauma

Objective: To explore the formation mechanism of benign biliary stricture. Methods: A model of trauma of common bile duct was established in 28 dogs and then repaired. The anasomosis tissues were taken on the 1st week, 3rd week and the 3rd month, 6th month respectively after operation and examined by using light microscopy and elec-tromicroscopy. Macrophage, TGF-p, and a-SMA were studied immunohistochemically. Results: The mucosal epithelium of common bile duct restored poorly, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix overdeposited; and myofibroblasts functioned actively and existed during the whole healing process. Immunohistochemical test showed a high expression of macrophage, TGF-β1 and a-SMA during healing process lasting a long duration. Macrophages were found in the lamina propria under mucosa, TGF-β1 in the granulation tissue, fibroblasts and endothelial cells of blood vesssels, while a-SMA in the myofiroblasts and smooth muscle tissue. Conclusion: The healing of bile duct is in the mode of overhealing. Myofibroblast is the main cause for contracture of scar and stricture of bile duct. The high expression of macrophage, TGF-β1 and a-SMA is closely related to active proliferation of fibroblasts, extracelluar matrix overdeposition and scar contracture of bile duct.