Th17 cells influence intestinal muscle contraction during <Emphasis Type="Italic">Trichinella spiralis</Emphasis> infection

Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. The aim of the study was to use this experimental model to elucidate if Thl 7 cells are involved in the development of gastrointestinal hypermotility. Colonic smooth muscle contractility was investigated in response to acetylcholine. The levels of IL-17, IL-23 and TGF-β1 in colon were measured by Western blotting. Flow cytometric detection of intracellular IFN-7/IL-4/ IL- 17 cytokine production was used to analyze the proportions of CD4＋ T cells subsets in colon. Our results showed that colonic muscle contractility was increased 2 weeks post infection （PI） and stayed high 12 weeks PI when no discernible inflammation was present in the gut. The proportion of Th17 cells and the expression of IL-17 were up-regulated in colon 2 weeks PI and returned to normal 8 weeks PI. The content of IL-17 was correlated with the colonic smooth muscle hypercontracility 2 weeks PI. Meanwhile, TGF-β1 was increased 2 weeks PI, while IL-23 was normal. Our results suggest that Th17 cells affect the colonic muscle contractility in mice infected with Trichinella spiralis at intestine stage but not at muscle stage and the effect of Th17 cells on muscle contractility might be induced by TGF-β1. Other cytokines might be involved in the hypercontracility of colonic smooth muscle at muscle stage.     

Effects of different dose endothelin-1 on expession of peroxisome proliferator-γ in cultured vascular smooth muscle cells of adult rats

Objective： To investigate the effects of endothlin-1 （ET-1） on vascular smooth muscle cells（VSMCs） proliferation and the expression of Peroxisome proliferator-activated receptorγ （PPAR-γ） in VSMCs. Methods： VSMCs of 16-week-old wistar rats thoracic aorta were cultured. VSMCs were treated by ET-1 for 48 h and observed of the proliferation by MTr. The expression of PPAR-γ mRNA and protein in cultured VSMCs treated by different concentration of ET-1 for 48 h was detected by RT-PCR and Western blot. Results： Compared with control group, VSMCs treated by ET-1 proliferated with the increase concentration of ET-1. There was significant differences among different groups （P 〈 0.01）. Meanwhile,the expression of PPAR-γ both in mRNA and in protein levels deceased. The expression of PPAR-γ in VSMCs was gradually decreased along with the increase concentration of ET-1. There was significant differences among different groups （ P 〈0.01）. Compared with control group, the expression of both PPAR-γ mRNA and PPAR-γ in ET-1 treated groups were lower（ P 〈 0.01 ）. Conclusion： ET-1 could induce VSMCs proliferation and the expression of PPAR-γ in VSMCs, which demonstrates that high dose ET-1 obviously weakens the function of PPAR-γ to increase VSMCs proliferation.     

Clinical significance of interleukin-17 producing cell infiltration with TGF-β1 expression in glioma

Objective To investigate correlation between the amount of interleukin-17 (IL-17) producing cells and the expression of transforming growth factor (31 (TGF-β1) in glioma,and evaluate the clinical values of IL-17 and TGF-pl in predicting the prognosis of glioma. Methods The presence of IL-17 and TGF-pl was measured by immunohistochemistry in 135 human glioma (WHO Ⅰ 18,WHO Ⅱ 45,WHO Ⅲ 53,WHO Ⅳ 19) tissues and 15 normal brain tissues. Results There was no IL-17 positive staining in normal brain tissues. Of 135 glioma specimens showed low TGF-pl expression and 77 (57. 03% ) showed high TGF-pl expression. No TGF-β1 expression was detected in normal brain tissue. Furthermore,TGF-β1 expression was positively correlated with the amount of IL-17 producing cells in glioma tissues ( r=0.285, P<0.01). Compared with the low grade,the levels of IL-17 and TGF-pl positive cells were obviously increased in high grade. The Kaplan-Meier analysis showed that there were significant differences in overall survival (OS) between the IL-17 and TGF-pl high-expression and lowexpression group (P＜0.01, P＜0.05). The 3-year OS rates of IL-17 of high expression and low expression were 33.75% and 76. 36%. Multivariate Cox proportional hazards regression analysis indicated that age,KPS score, IL-17 were independent prognostic factor for OS (P＜0.01). Conclusion Intratumoral IL-17-producing cell density and the expression of TGF-β1 was associated with the malignancy of human glioma.     

Effects of different dose endothelin-1 on expession of peroxisome proliferator-γ in cultured vascular smooth muscle cells of adult rats

Objective: To investigate the effects of endothlin- 1 (ET- 1 ) on vascular smooth muscle cells(VSMCs ) proliferation and the expression of Peroxisome proliferator-activated receptorγ (PPAR-γ) in VSMCs. Methods: VSMCs of 16-week-old wistar rats thoracic aorta were cultured. VSMCs were treated by ET-1 for 48 h and observed of the proliferation by MTT. The expression of PPAR-γmRNA and protein in cultured VSMCs treated by different concentration of ET-1 for 48 h was detected by RT-PCR and Western blot.Results: Compared with control group, VSMCs treated by ET-1 proliferated with the increase concentration of ET-1. There was significant differences among different groups ( P ＜ 0.01). Meanwhile, the expression of PPAR-γ both in mRNA and in protein levels deceased. The expression of PPAR-γ in VSMCs was gradually decreased along with the increase concentration of ET-1. There was significant differences among different groups ( P ＜0.01). Compared with control group, the expression of both PPAR-γ mRNA and PPAR-γ in ET-1 treated groups were lower( P ＜ 0.01). Conclusion: ET-1 could induce VSMCs proliferation and the expression of PPAR-γ in VSMCs, which demonstrates that high dose ET-1 obviously weakens the function of PPAR-γ to increase VSMCs proliferation.     

Inhibition effect of small interfering RNA of connective tissue growth factor on the expression of vascular endothelial growth factor and connective tissue growth factor in cultured human peritoneal mesothelial cells

Background The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor β1 （TGF-β1 ） plays a key role in regulating tissue repair and remodelling after injury. Connective tissue growth factor （CTGF）, a downstream mediator of TGF-β1 inducing fibrosis, has been implicated in peritoneal fibrosis. Vascular endothelial growth factor （VEGF） plays a key role in angiogenesis that can hasten peritoneal fibrosis. In this study, we investigated the effect of small interfering RNA （siRNA） of CTGF by pRETRO-SUPER （PRS） retrovirus vector on the expression of CTGF and VEGF in human peritoneal mesothelial cells. Methods Retrovirus producing CTGF siRNA were constructed from the inverted oligonucleotides and transferred into packaging cell line PT67 with lipofectamine, and the virus supernatant was used to infect human peritoneal mesothelial cell （HPMC）. The cells were divided into seven groups： low glucose DMEM, low glucose DMEM ＋ TGF-β1 5 ng/ml, low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS-CTGF-siRNA1-4 and low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS. The expression of CTGF and VEGF were measured by semiquantitative RT-PCR and Western blot. Results Low levels of CTGF and VEGF were detected in confluent HPMCs. Following stimulation with TGF-β1 , the levels of CTGF and VEGF were significantly upregulated （P〈0.01）. Introduction of PRS-CTGF-siRNA1-4 resulted in the significant reduction of CTGF mRNA and protein, and VEGF mRNA （P〈0.01）, especially in groups PRS-CTGF-siRNA, and PRS-CTGF-siRNA4. The introduction of PRS void vector did not have these effects （P〉0.05）. Conclusions The expression of CTGF siRNA mediated by PRS retrovirus vector can effectively reduce the level of CTGF and VEGF induced by TGF-β1 in cultured HPMCs. This study may provide potential therapeutic strategies to prevent the peritoneal fibrosis.     

TGF-β1-induced LPP expression dependant on Rho kinase during differentiation and migration of bone marrow-derived smooth muscle progenitor cells

Lipoma preferred partner(LPP) has been identified as a protein which is highly selective for smooth muscle progenitor cells(SMPCs) and regulates differentiation and migration of SMPCs,but mechanisms of LPP expression are not elucidated clearly.The aim of the present study was to discuss the mechanisms by which LPP expression is regulated in the differentiation and migration of SMPCs induced by TGF-β1.It was found that TGF-β1 could significantly increase the expression of LPP,smooth muscle α-actin,smooth muscle myosin heavy chain(SM-MHC),and smoothelin in SMPCs.Moreover,inactivation of Rho kinase(ROK) with ROK inhibitors significantly inhibited LPP mRNA expression in TGF-β1-treated SMPCs and mouse aortic smooth muscle cells(MAoSMCs).At the same time,LPP silencing with short interfering RNA significantly decreased SMPCs migration.In conclusion,LPP appears to be a ROK-dependant SMPCs differentiation marker that plays a role in regulating SMPCs migration.     

Role of connective growth factor in plasminogen activator inhibitor-1 and fibronectin expression induced by transforming growth factor β1 in renal tubular cells

Background Connective tissue growth factor (CTGF) contributes greatly to renal tubulointerstitial fibrosis, which is the final event leading to end-stage renal failure. This study was designed to investigate the effects of CTGF antisense oligodeoxynucleotides (ODNs) on the expressions of plasminogen activator inhibitor-1 (PAI-1) and fibronectin in renal tubular cells induced by transforming growth factor β1 (TGF-β) in addition to the role of CTGF in the accumulation and degradation of renal extracellular matrix (ECM). Methods A human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipidmediated CTGF antisense ODNs were transfected into HKC cells. After HKC cells were stimulated with TGF-β1 (5 μg/L), the mRNA levels of PAI-1 and fibronectin were measured by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 and fibronectin in the medium were determined by Western blot and ELISA, respectively. Results TGF-β was found to induce tubular CTGF, PAI-1, and fibronectin mRNA expression. PAI-1 and fibronectin mRNA expression induced by TGF-β was significantly inhibited by CTGF antisenes. ODNs CTGF antisense ODNs also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 and fibronectin protein secreted into the medium. Conclusions CTGF may play a crucial role in the accumulation and degradation of excessive ECM during tubulointerstitial fibrosis, and transfecting CTGF antisense ODNs may be an effective way to prevent renal fibrosis.     

Effects of Biejia Ruangan Tablet(复方鳖甲软肝片)-Containing Serum on Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 Expression in Cultured Renal Interstitial Fibroblasts

Objective:To investigate the effects of Biejia Ruangan Tablet(复方鳖甲软肝片方,BRT)- containing serum on the expression of matrix metalloproteinase(MMP-9) and tissue inhibitor of metalloproteinase(TIMP-1) in cultured renal interstitial fibroblasts.Methods:Different BRT-containing sera were prepared by gastric gavages to rats with the high-dose(7 g/kg),mid-dose(3.5 g/kg),and low-dose(1.75 g/kg)BRT respectively.The expression of extracellular matrix in NRK-49 F cells was induced by treatment with human transforming growth factor-β1(recombined human TGF-β1),and BRT-containing serum.Western blotting and Northern blotting were used to measure type Ⅰ and Ⅲ procollagen,MMP-9,and TIMP-1.Results:The high dose BRT-containing serum could decrease the type Ⅰ and Ⅲ procollagen gene expression which boosted by TGF- β1,at the same time cut down TIMP-1 protein and gene expression which increased by TGF- β1(P0.05).Treatment of cells with recombined human TGF- β 1 had no significant effect on MMP-9 expression and BRTcontaining serum also had no effect on MMP-9 expression.Conclusions:High dose BRT has anti-fibrosis effects in NRK-49 F cells,as indicated by its inhibition of type Ⅰ and Ⅲ procollagen and TIMP-1 expression.     

Adenovirus-mediated TGF- β1 gene transfer to human degenerative lumbar int ervertebral disc cells

Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β(1) (TGF- β 1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerat ive lumbar disc disease. The cells were cultured and directly infected by two a denoviral constructs, Ad/CMV- EGFP containing the enhanced green fluorecence pro tein (EGFP) gene (marker gene) and Ad/CMV- TGF- β(1) containing the potentially therapeutic TGF- β(1) gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV- TGF- β(1)). Results Culture cells transducted by Ad/CMV- EGFP showed specific green fluorescence und er the fluoroscope and expression sustained for at least 4 weeks. When infe cted by Ad/CMV- TGF- β(1), approximally 30% of cultured cells were staind brown (+) with TGF- β(1) staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene , TGF- β(1), by using an adenoviral vector to human degenerative lumbar interve rtebral disc cells.     

Experimental Study of Plasmid TGF-β1 DNA Gene Transfer with Lipofectamine into Rabbit Corneal Epithelial Cells In Vitro

To investigate whether the TGF-β1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-β1in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMTGF-β1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-β1 protein expression specific for pMAMTGF-β1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23. 37 %. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-β1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.     

Effects of transforming growth factor-β1 and signal protein Smad3 on rat cardiomyocyte hypertrophy

Background SMAD proteins have recently been identified as the first family of putative transforming growth factor-β1 (TGF-β1) signal transducers. This study was to investigate the effects of TGF-β1 and signal protein Smad3 on rat cardiac hypertrophy.Methods The incorporation of [3H]-leucine was measured to determine the hypertrophy of cardiomyocyte incubated with different doses of TGF-β1 in cultured neonatal cardiomyocytes. The model of rat cardiac hypertrophy was produced with constriction of the abdominal aorta. At different times after the operation, rats were killed, and their left ventricular mass index (LVMI) determined.The mRNA expression of TGF-β1 and Smad3 of cultured cells and hypertrophic left ventricles were assessed by RT-PCR. The protein expression of Smad3 was assessed by Western blot.Results In cultured neonatal cardiomyocytes, TGF-β1 significantly promoted incorporation of [3H]-leucine. With the concentration of 3 pg/L, it increased the expression of Smad3 in mRNA and protein levels after 15 minutes, and continued for up to 8 hours of cultured cardiomyocytes. The LVMI and theexpression of TGF-β1 (mRNA) and Smad3 (mRNA and protein) of hypertrophic left ventricle were increased by day 3 after the operation and continued to the 4th week. The peak expression of these was in the second week after operation.Conclusion TGF-β1 has positive effects on could be related to the pathologic progressionrat cardiomyocyte hypertrophy. Signal protein Smad3 of rat cardiac hypertrophy.     

Differential expression of βig-h3 in human hepatoma cell lines

Objective：To observe the differential expression of TGF-β-induced gene human clone 3 （βig- h3） in human hepatoma cell lines. Methods：Human hepatoma cells HHCC, 7721, T7721 constructed by stably transfectiug HAb18G/CD147 cDNA into 7721 and normal human liver cell QZG were cultured as previously. RT-PCR and western blot were used to investigate the differential expression of βig-h3 in human hepatoma cell lines and normal liver cell. Results ：The results of RT-PCR suggested that the expression of βig-h3 mRNA in human hepatoma cells was higher than that in normal human liver cell QZG（P〈0.01）, and its expression level in human hepatoma cells in turn was HHCC〉T7721〉7721. Moreover, the similar results of βig-h3 protein expression were testified by western blot. Conclusion：This study demonstrates that the expression of βig-h3 in hepatoma cell lines is higher than that in normal liver cell QZG, which provides a sound basis for exploring the function of βig-h3 in processes of adhesion and metastasis of human hepatoma cells.     

RhoA/ROCK signaling in chondrogenic differentiation of MSCs induced by TGF-β1

Objective： To investigate the role of RhoA/ROCK in the process of chondrogenic differentiation of MSCs in vitro induced by TGF-β1. Methods ： MSCs were isolated from rat bone marrow, cultured and passaged. The cultured MSCs at the 3rd passage were induced to differentiate into chondrocytes in induction medium containing TGF-β1, the expressions of RhoA and ROCKI/2 in the induced MSCs cells were detected by Western-blot and RT-PCR. Results： In parallel to chondrogenic marker gene expressions of collagen Ⅱand aggrecan, ROCK inhibition by Y27632 in these cells caused a significant decrease in mRNA level of collagen Ⅱ. Conclusion： The results suggest that RhoA/ROCK pathway may play an important and complex role in regulation of chondrogenic differentiation and gene expression.     

MicroRNA-34a inhibits human brain glioma cell growth by down-regulation of notch1

The effects of microRNA-34a (miR-34a)-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups (P<0.05). Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups (P<0.05), but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.     

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.     

Changes of peroxide dismutase activity and malondialdehyde level in the smooth muscle cells of human fetal aorta cultured in vitro by low density lipoprotein conditional medium

Here are reported the changes of superoxide dismutase(SOD)activity andmalondialdehyde(MDA)in the smooth muscle cells of human fetal aorta cultured in vitro with lowdensity lipoprotein(LDL)conditional medium.The results showed that a single concentration of hu-man LDL(50μg/ml)stimulated proliferation of smooth muscle cells,and the SOD activityof the cells in the experimental group was higher,from the first to the fifth cultured day whenthe cells had a active proliferation,than that of the control cells.This suggests that LDL might in-duce the increase of SOD activity.At the seventh day,as the cells were in inactive proliferation,SOD activity was low and the difference was significant as compared with that at the fifth day ofthe same group.This also indicates that the SOD activity may be related to the cell proliferation.MDA level within the cells of the esperimental group was lowered with the cell active proliferationand the increase of SOD activity,but when the cells were in inactive proliferation and the SOD ac-tivity decreased,it will remained low.     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Expression of Proliferating Cell Nuclear Antigen and Vascular Endothelial Growth Factor in Primary Culture Cells Derived from Human Ovarian Mucinous Cystadenocarcinoma

The effects of antisense oligodeoxynucleotide （antisense ODN） to follicle-stimulating hormone receptor （FSHR） and follicle-stimulating hormone （FSH） on the expression of proliferating cell nuclear antigen （PCNA） and vascular endothelial growth factor （VEGF） were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma （OMC）. The primary （）MC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co-cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups （P〈0.05 or P〈 0.01）, while decreased significantly in antisense ODN groups （P〈0.05 or P〈 0.01） and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly （P〈0.01）. It was suggested that FSH might promote the development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH.     

Role of nitric oxide and inducible nitric oxide synthase in human abdominal aortic aneurysms: a preliminary study

Background Nitric oxide （NO） is an important mediator in the pathophysiology of many vascular diseases. However, the definite role of NO in human abdominal aortic aneurysm （AAA） formation is unclear. The aim of this study was to investigate production of NO and expression of inducible nitric oxide synthase （iNOS）, and their possible role in AAA. Methods A total of 28 patients with AAA, 10 healthy controls, and 8 patients with arterial occlusive disease were enrolled into this study. Standard colorimetric assay was used to examine NO concentration in plasma from patients with AAA and normal controls, and in cultured smooth muscle cells （SMCs）. Expression of iNOS in aortas and cultured SMCs were detected by immunochemistry. The correlation of iNOS expression with age of the patient, size of aneurysm, and degree of inflammation was also investigated by Cochran-Mantel-HaenszelX^2 test and Kendall＇ Tau correlation. Results Expression of iNOS increased significantly in the wall of aneurism in the patients with AAA compared to the healthy controls （P〈0.05） and the patients with occlusive arteries （P〈0.05）. iNOS protein and media NOx （nitrite＋nitrate） also increased in cultured SMCs from human AAA （n=4, P〈0.05）, while plasma NOx decreased in patients with AAA （n=25） compared to the healthy controls （n=20）. There was a positive correlation between iNOS protein and degree of inflammation in aneurismal wall （Kendall coefficient=0.5032, P=0.0029） Conclusions SMCs and inflammatory cells were main cellular sources of increased iNOS in AAA, and NO may play a part in pathogenesis in AAA through inflammation.     

Upregulated DJ-1 promotes renal tubular EMT by suppressing cytoplasmic PTEN expression and Akt activation

Recently,phosphatase and tensin homolog deleted on chromosome 10(PTEN) is suggested as a new agent in the fighting against fibrogenesis.In tumor,DJ-1 is identified as a negative regulator of PTEN.But the expression of DJ-1 and the regulation of PTEN in fibrosis are unclear.Renal fibrosis was induced in 5/6 subtotal nephrectomy rat model.Human proximal tubular epithelial cells(HKC) were treated with transforming growth factor-beta 1(TGF-β1),or transfected with DJ-1 or PTEN.Confocal microscope was used to investigate the localization of DJ-1 and PTEN.The selective phosphoinositide-3 kinase(PI3K) inhibitor,LY294002,was administered to inhibit PI3K pathway.The DJ-1 and PTEN expression,markers of epithelial-mesenchymal transition(EMT) and Akt phosphorylation were measured by RT-PCR,Western blotting or immunocytochemistry.In vitro,after HKC cells were stimulated with 10 ng/mL TGF-β1 for 72 h,the expression of DJ-1 was increased,and that of PTEN was decreased.In vivo,the same results were identified in 5/6-nephrectomized rats.In normal HKC cells,most of DJ-1 protein localized in cytoplasm,and little in nucleus.TGF-β1 upregulated DJ-1 expression in both cytoplasma and nuclei.In contrary,TGF-β1 emptied cytoplasmic PTEN protein into nucleus.Overexpression of DJ-1 decreased the expression of PTEN,promoted the activation of Akt and the expression of vimentin,and also led to the loss of cytoplasmic PTEN.Contrarily,overexpression of PTEN protected HKC cells from TGF-β1-induced EMT.In conclusion,DJ-1 is upregulated in renal fibrosis and DJ-1 mediates EMT by suppressing cytoplasmic PTEN expression and Akt activation.