Ginsenoside Rb1, a panoxadiol saponin against oxidative damage and renal interstitial fibrosis in rats with unilateral ureteral obstruction

Objective:To investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction(UUO). Methods:In total,80 male rats were randomly divided into 4 groups,20 in each group:the sham operated group (SOR),UUO group,UUO with ginsenoside Rb1 treatment group(treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group(as the positive control,treated with 20 mg/kg by ga...     

Role of transforming growth factor-β1 in the process of fibrosis of denervated skeletal muscle

In order to investigate the biological function of transforming growth factor-β1(TGF-β1) during fibrosis in denervated skeletal muscle,we recruited sciatic nerve injury model of SD rats in which denervated gastrocnemius was isolated for analysis.At different time points after operation,denervated muscle was examined by several methods.Masson trichrome staining showed morphological changes of denervated skeletal muscle.Quantitative RT-PCR detected the rapid increase of TGF-β1 expression at mRNA level after nerve injury.It was found that a peak of TGF-β1 mRNA expression appeared one week post-operation.The expression of collagen Ⅰ(COL Ⅰ) mRNA was up-regulated in the nerve injury model as well,and reached highest level two weeks post-injury.Immunoblot revealed similar expression pattern of TGF-β1 and COL Ⅰ in denervated muscles at protein level.In addition,we found that the area of the gastrocnemius muscle fiber was decreased gradually along with increased interstitital fibrosis.Interestingly,this pathological change could be prevented,at least partly,by local injection of TGF-β1 antibodies,which could be contributed to the reduced production of COL Ⅰ by inhibiting function of TGF-β1.Taken together,in this study,we demonstrated that the expression of TGF-β1 was increased significantly in denervated skeletal muscle,which might play a crucial role during muscle fibrosis after nerve transection.     

All-trans retinoic acid diminishes collagen production in a hepatic stellate cell line via suppression of active protein-1 and c-Jun N-terminal kinase signal

Following acute and chronic liver injury,hepatic stellate cells (HSCs) become activated to undergo a phenotypic transformation into myofibroblast-like cells and lose their retinol content,but the mechanisms of retinoid loss and its potential roles in HSCs activation and liver fibrosis are not understood.The influence of retinoids on HSCs and hepatic fibrosis remains controversial.The purpose of this study was to evaluate the effects of all-trans retinoid acid (ATRA) on cell proliferation,mRNA expression of collagen genes [procollagen α1 (Ⅰ),procollagen α1 (Ⅲ)],profibrogenic genes (TGF-β 1,CTGF,MMP-2,TIMP-1,TIMP-2,PAI-1),fibrolytic genes (MMP-3,MMP-13) and the upstream element (JNK and AP-1) in the rat hepatic stellate cell line (CFSC-2G).Cell proliferation was evaluated by measuring BrdU incorporation.The mRNA expression levels of collagen genes [procollagen α1 (Ⅰ),procollagen α1 (Ⅲ)],profibrogenic genes (TGF-β 1,CTGF,MMP-2,TIMP-1,TIMP-2,PAI-1),and fibrolytic genes (MMP-3,MMP-13) were quantitatively detected by using real-time PCR.The mRNA expression of JNK and AP-1 was quantified by RT-PCR.The results showed that ATRA inhibited HSCs proliferation and diminished the mRNA expression of collagen genes [procollagen α1 (Ⅰ),procollagen α1 (Ⅲ)] and profibrogenic genes (TGF-β 1,CTGF,MMP-2,TIMP-1,TIMP-2,PAI-1),and significantly stimulated the mRNA expression of MMP-3 and MMP-13 in HSCs by suppressing the mRNA expression of JNK and AP-1.These findings suggested that ATRA could inhibit proliferation and collagen production of HSCs via the suppression of active protein-1 and c-Jun N-terminal kinase signal,then decrease the mRNAs expression of profibrogenic genes (TGF-β 1,CTGF,MMP-2,TIMP-1,TIMP-2,PAI-1),and significantly induce the mRNA expression of MMP-3 and MMP-13.     

The role of TGF-β1 in mice hepatic fibrosis bySchistosomiasis Japonica

Summary To investigate the role of transforming growth factor-β₁ (TGF-β₁) in mice with hepatic fibrosis caused bySchistosomiasis Japonica, ELISA, VG staining and multimedia color hieroglyph quantitative analysis were used to study the change of the serum TGF-β₁, liver collagen fiber and reticular fiber in mice. The level of serum TGF-β₁ in experimental group was significantly higher than that in control group (P < 0. 01 orP < 0. 05) 8, 10, 12 weeks after infected by schistosomiasis. After infection, the level of liver collagen fiber and reticular fiber, and that of TGF -β₁ increased over time (P < 0. 01 orP < 0. 05). In mice infected bySchistosomiasis Japonica, the level of TGF-β₁ increased with prolongation of infection time, and with the increase of liver collagen fiber and reticular fiber. TGFβ₁ plays an important role of immunomodulation in hepatic fibrosis formation caused bySchistosomiasis Japonica.     

Effect of huan-shuai recipe on expression of extracellular matrix in 5/6 nephrectomized rats

Objective: To investigate the effect of Huan-shuai Recipe on the expression level of fibronectin (FN) and transforming growth factor-β₂ (TGF-β₂) in the remnant kidney of rats.Methods : Male Wistar rats, after 4 weeks of 5/6 nephrectomy, were divided into two groups at random, and treated with water and Huanshuai Recipe (HSR) respectively. Serum creatinine (SCr) and urinary protein of 24-hours were observed at beginning, 6th week and 12th week of the experiment, and FN and TGF-β2 were assayed by immunohistochemical technique and computer assisted image analysis as well.Results : Serum SCr and urinary protein of 24-hours were increased significantly in the animals after 5/6 nephrectomy. The increased parameters decreased in the HSR group after treatment, as compared with those in the control group, and the difference was significant (P < 0.05); Immunohistochemical measurement and computer assisted image analysis showed a marked suppression of FN and TGF-β₂ in animals of the HSR group with significant difference from those in the control group (P<0.05).Conclusion : HSR not only plays the role of renal function protection, but could also alleviate the glomerular pathological injury. The effect on TGF-β2 and FN may be the mechanism of HSR in slowing down the progression of chronic renal failure.     

Effect of Anluohuaxian Tablet Combined with γ-IFN on Schistosomal Liver Fibrosis

The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis, Fifty Kunming mice were randomly divided into 5 groups： normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment （anluohuaian tablet＋γ-IFN） group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type Ⅰ and Ⅲ collagen, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation （P〈0.05）. Furthermore, the expression of collagen Ⅰ and Ⅲ, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups （P〈0.05）. In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group （P〈0.05）. It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.     

Effect of Coiquhoumia Root on the Expression of Transforming Growth Factor-β in Mesangiai Proliferation Giomerulonephritis Model

Summary： To study the efficacy and the mechanism of Colquhoumia root （ Tripterygium hypoglaucure （Le,vL） Hutch） in the treatment of mesangial proliferation glomerulonephritis （MsPGN）, SD rats were injected with anti-thymoeyte serum （ATS） to make MsPGN model （anti-Thyl model）. The rats were then divided into 3 groups： normal control group, anti-Thyl model group and treatment group. Histopathologieal （HE, PAS）, immunohistoehemieal, RT-PCR technique and computer imaging analysis system were used to evaluate mesangial matrix production, the expression of TGF-β protein and mRNA in the tissues of kidney. Our result showed that proteinuria and the ratio of extraeellular matrix/glomerular capillaries area （ECM/CA） were increased significantly in model group. The expression of both TGF-β protein and mRNA in glomeruli was much higher in model group than in control group （P〈0.01）. After the treatment with Colquhoumia root, proteinuria, ECM/CA and the expression of both TGF-β1 protein and mRNA in glomeruli were significantly decreased in treatment group as compared with those in model group. It is concluded that Colquhoumia root is effective in reducing proteinuria and mesangial matrix proliferation in MsPGN and it may achieve these effects by inhibiting the expressions of TGF-β1 protein and mRNA of mesangial cells.     

Role of Connective Tissue Growth Factor in Extracellular Matrix Degradation in Renal Tubular Epithelial Cells

In order to investigate the effects of connective tissue growth factor (CTGF) antisense oligodeoxynucleotide (ODN) on plasminogen activator inhibitor-1 (PAI-1) expression in renal tubular cells induced by transforming growth factor β1 (TGF-β1) and to explore the role of CTGF in the degradation of renal extracellular matrix (ECM), a human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODN was transfected into HKC. After HKC were stimulated with TGF-β1 (5 μg/L), the mRNA level of PAI-1 was detected by RT-PCR. In-tracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 in the media was determined by Western blot. The results showed that TGF-β1 could induce tubular CTGF and PAI-1 mRNA expression. The PAI-1 mRNA expression induced by TGF-β1 was significantly inhib-ited by CTGF antisense ODN. CTGF antisense ODN also inhibited intracellular PAI-1 protein syn-thesis and lowered the levels of PAI-1 protein secreted into the media. It was concluded that CTGF might play a crucial role in the degradation of excessive ECM during tubulointerstitial fibrosis, and blocking the biological effect of CTGF may be a novel way in preventing renal fibrosis.     

氨氯地平对肺纤维化大鼠的影响研究

目的  研究氨氯地平对大鼠肺纤维化的保护作用和可能的分子机制,为肺纤维化的治疗寻找新的途径。方法  选取240只实验大鼠进行研究,随机分为空白组、模型组、吡非尼酮组和氨氯地平组,每组各60只。模型组、吡非尼酮组和氨氯地平组采用气管内灌注博来霉素建立肺纤维化大鼠模型,空白组则气管内灌注等量生理盐水;然后空白组和模型组大鼠给予相同量生理盐水处理,氨氯地平组大鼠则给予1/2量的氨氯地平和1/2量的生理盐水处理;吡非尼酮组给予1/2量的吡非尼酮和1/2量的生理盐水处理。采用单因素方差分析和u检验分析1、2及4周后4组大鼠肺泡炎、肺纤维化的评分,并分析大鼠的血小板反应蛋白1（TSP-1）、转化生长因子β1（TGF-β1）及I型胶原和III型胶原mRNA的含量变化情况。结果  氨氯地平组和吡非尼酮组大鼠各时间段的TSP-1、TGF-β1及I型胶原和III型胶原mRNA的含量均低于模型组（P <0.05）,且氨氯地平组较吡非尼酮组也降低（P <0.05）,大鼠的肺泡炎和肺纤维化评分结果显示氨氯地平组和吡非尼酮组低于模型组（P <0.05）,且氨氯地平组显著低于吡非尼酮组（P <0.05）。结论  氨氯地平可能通过抑制TGF-β1、TSP-1的生成水平和减少I型胶原和III型胶原mRNA的表达达到减轻大鼠肺组织肺泡炎及肺纤维化程度的临床效果。

     

Modulation of Matrix Metalloproteinase and TIMP-1 Expression by TGF-β1 in Cultured Human RPE Cells

Matrix metalloproteinases(MMPs)are a familyof zinc binding,calcium-dependent endopeptidases thatfunction by degrading extracellular matrix(ECM)components.The functional effects of these enzymesare in part controlled byinteractions withtissueinhibi-tors of metalloproteinases(TI MPs)acting as naturalMMPinhibitors.Aprecise balance between MMPandTI MPactivities may be i mportant for the integrity ofECMcomponents[1].It shows that the expression andactivity of MMPs could be regulated by va…     

Influence of High Level TGF-β1 on Scleral Thickness

In order to explore the role of TGF-β1 in scleral remodeling and the possible mechanism,the influence of high level TGF-β1 on scleral thickness and the expression of MMP-2 and TIMP-2 was investigated in a TGF-β1 transgenic mouse model.Alb/TGF-β1(Cys223,225Ser) TGF-β1 transgenic mice were used as experimental subjects and non-transgenic littermates as controls.Plasma levels of TGF-β1 were determined by ELISA.TGF-β1,MMP-2 and TIMP-2 levels in sclera were detected by using Western blot.The thickness of posterior sclera was measured by computerized image analysis of a midsagittal section.Mean difference was analyzed with independent t-test.The results showed plasma levels of TGF-β1 in transgenic mice were 1.68 times as much as that in the controls(P<0.01).TGF-β1 levels in the sclera of transgenic mice were 2.68 times of the controls(P<0.01).Posterior scleral thickness in transgenic mice were significantly thicker than in the controls.There was no significant difference in the MMP-2 levels between transgenic mice and controls,but the TIMP-2 levels were increased significantly in transsgenic mice as compared with those in the controls.It was suggested that high levels of TGF-β1 in transgenic mice could result in the increased scleral thickness by inducing the expression of TIMP-2 to suppress the activity of MMP-2,finally inhibiting the degradation of collagen.     

Effect of TGF-β1 on the expression of IL-12, IL-15, IL-18, IL-4 and IL-10 in heart transplantation rejection in rats

Summary To investigate the effect of TGF-β1 on the expressions of IL-12, IL-15, IL-18, IL-4 and IL-10 in heart transplantation rejection in rats, a model of rat cervical heterotopic heart transplantation was set up and the model rats were randomly divided into three groups: control group, transplant group and TGF-β1 group. The mRNA expression levels of IL-12, IL-15, IL-18, IL-4 and IL-10 were determined by RT-PCR at the 5th day after the transplantation. The mRNA expression levels of IL-12, IL-15, IL-18 were increased obviously and those of IL-4, IL-10 were significantly decreased in the transplant group as compared with the control group (P<0.01). In the TGF-β1 group, the mRNA expression levels of IL-12, IL-15, IL-18 were significantly decreased and those of IL-4, IL-10 were significantly increased as compared with the transplant group (P<0.01). The immunosuppressive effect of TGF-β1 on heart transplantation rejection was related to its inhibition of the expressions of Th1-type cytokines (IL-12, IL-15, IL-18 etc) and its promotion of the expressions of Th2-tpye cytokines (IL-4, IL-10).     

The Dynamic Change of TGF-β1 in the Myocardial Remodeling of Rat after Myocardial Infarction

Chronic heart failure is the leading cause of mortality and morbidity in most countries. Ventricular remodeling was the important pathophysiological process of heart failure. Mechanical overload, neurohormones and system nerve adrenal gland system can evoke remodeling. There are plentiful evidence indicating that inflammation plays an important role in the ischemic cardiac disease[1—3]. In this study we used the MI rat to observe the morpho- logical change of the ventricle, expression of the…     

Expression of TGF-β2 mRNA and PCNA, FN Protein in Lens Epithelial Cells in Age-related Nuclear and Cortex Cataract

Summary： By using RT PCR and immunohistochemistry, the expressions of transforming growth factor 132 （TGF-β2） mRNA, proliferating cell nuclear antigen （PCNA） and fibronection （FN） protein in lens epithelial cells （LECs） of age-related nuclear and cortex cataract were detected and compared. The results of RT-PCR revealed that the expression of TGF-β2 mRNA was higher in cortex cataract than in nuclear cataract. Immunohistochemistry demonstrated that the expression of PCNA protein was lower and the expression of FN protein was higher in cortex cataract than in nuclear cataract. It was suggested that TGF-β2 , PCNA and FN might take important parts in the process of age-related cataract. Cortex cataract was related to the transdifferentiation of LECs, and nuclear cataract to the proliferation of LECs.     

Effects of <Emphasis Type="Italic">Panax notoginoside</Emphasis> on the expression of TGF-β1 and Smad-7 in renal tissues of diabetic rats

In order to explore the effects of Panax notoginoside (PNS) on the expression of transforming growth factor β1 (TGF-β1) and Smad-7 in renal tissues of diabetes, a rat model of diabetic nephropathy was set up by intravenous injection of streptozotocin (STZ). Wistar rats were randomly divided into normal group, diabetic control group, group treated by PNS at low-dosage (PL), group treated by PNS at high-dosage (PH) and group treated by catopril (C), respectively. Fasting blood glucose (FBG), renal index, endogenous creatinine clearance rate (CCr) and urinary albumin (UAlb) in 24 h were examined after 6 weeks. Meanwhile, the expressions of TGF-β1 and Smad7 in renal tissues were immunohistochemically dectected. At the end of the sixth week, FBG, renal index, Ccr, UAlb were all elevated significantly in control group (P<0.01). The expression of TGF-β1 protein was increased while Smad7 protein decreased in renal tissue (P<0.01). However, the treatment with PNS reversed the aforementioned changes in renal tissues of diabetic rats. These results indicate that PNS possess a protective effect on the kidney of diabetic rats and it might protect kidney by inhibiting the expression of TGF-β1 protein and enhancing the expression of Smad7 protein.     

TGF-beta1 Transgenic Mouse Model of Thoracic Irradiation: Modulation of MMP-2 and MMP-9 in the Lung Tissue

TGF-β1has been shown to play a key role inthe development and progression of radiation-in-duced lung injury and subsequent fibrosis[1].Re-modeling of the lung is a hall mark of radiation-in-ducedlunginjury andinvolves extensive alterationsof lung extracellular matrix(ECM).MMPs are afamily of matrix degrading proteinases with struc-tural si milarities,and their activity is specificallyinhibited by the tissueinhibitors of metalloprotein-ases(TI MPs).In the present study,we intend toinvestig…     

Expression of TGF-β in region of bone defect repaired by collagen nano-beta-tricalcium phosphate composite artificial bone

Summary The distribution and function of transforming growth factor-beta (TGF-β) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated. Twenty-four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8.0 mm in diameter through the whole bone. On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group. At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out. Immunohistochemical staining was performed for TGF-β assay by avidin-biotin complex method (SABC). Simultaneously, TGF-β was quantitatively analyzed by HPIAS-1000 imaging analysis system. The immunohistochemical staining for TGF-β revealed that osteoblasts and immature osteocytes highly expressed TGF-β. Diffused TGF-β positive staining particles appeared in the mesenchymal and fibrous-tissue. There was no significant difference in the TGF-β positive staining between two groups in the medial region to original osseous beds at different time points (P>0.05). However, in distal original osseous bed of the defected region, the positive expression of TGF-β in the Co/N-TCP group was significantly stronger than in the control group (P<0.05 or 0.01). The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-β to aggregate and precipitate the healing of bone defect. LING Xiang, male, born in 1967, Doctor in Charge