Interleukin-1 beta up-regulates tissue inhibitor of matrix metalloproteinase-1 mRNA and phosphorylation of c-jun N-terminal kinase and p38 in hepatic stellate cells

AIM： To study the relationship between interleukin-lbeta （IL-1β） up-regulating tissue inhibitor of matrix metalloproteinase-1 （TIMMP-1） mRNA expression and phosphorylation of both c-jun N-terminal kinase （INK） and p38 in rat heffatic stellate cells （HSC）. METHODS： RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC. RESULTS： TIMMP-1 mRNA expression （1.191± 0.079） was much higher after treatment with IL-1β （10 ng/mL） for 24 h than in control group （0.545±0.091） （P〈0.01）. IL-1β activated INK and p38 in a time-dependent manner. After stimulation with IL-1β for 0, 5, 15, 30, 60 and 120 min, the INK activity was 0.982±0.299, 1.501±0.720, 2.133±0.882, 3.360±0.452, 2.181±0.789, and 1.385 ± 0.368, respectively. There was a significant difference in JNK activity at 15 min （P〈 0.01）, 30 min （P〈 0.01） and 60 min （P〈0.01） in comparison to that at 0 min. The p38 activity was 1.061±0.310, 2.050±0.863, 2.380±0.573, 2.973±0.953, 2.421±0.793, and 1.755 ± 0.433 at the 6 time points （0, 5, 15, 30, 60 and 120 min） respectively. There was a significant difference in p38 activity at 5 min （P〈0.05）, 15 min （P〈0.01）, 30 min （P〈0.01） and 60 min （P〈0.01） compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 （10 μmol/L, 1.022±0.113; 20 μmol/L, 0.869±0.070; 40 μmol/L, 0.666±0.123）. Their decreases were all significant （P〈0.05, P〈0.01, P〈0.01） in comparison to control group （without SP600125 treatment, 1.163±0.107）. In the other 3 groups pretreated with different concentrations of SB203580 （10 μmol/L, 1.507±0.099; 20 μmol/L, 1.698±0.107; 40 μmol/L, 1.857±0.054）, the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group （without SB203580 treatment, 1.027 ± 0.061） with a significant statistical significance （P〈 0.01）. CONCLUSION： IL-1β has a direct action on hepatic fibrosis by up-regulating TIMMP-1 mRNA expression in ratessionin in rate HSC.JNK and p38 mitogen-activated protein kinases （MAPKs） are involved in IL-1β-induced TIMMP-1 gene expression, and play a distinct role in this process, indicating that p38 and .INK pathways cooperatively mediate TIMP-1 mRNA expression in rat HSC.

Bromocriptine protects perilesional spinal cord neurons from lipotoxicity after spinal cord injury

Recent studies have revealed that lipid droplets accumulate in neurons after brain injury and evoke lipotoxicity,damaging the neurons.However,how lipids are metabolized by spinal cord neurons after spinal cord injury remains unclear.Herein,we investigated lipid metabolism by spinal cord neurons after spinal cord injury and identified lipid-lowering compounds to treat spinal cord injury.We found that lipid droplets accumulated in perilesional spinal cord neurons after spinal cord injury in mice.Lipid droplet accumulation could be induced by myelin debris in HT22 cells.Myelin debris degradation by phospholipase led to massive free fatty acid production,which increased lipid droplet synthesis,β-oxidation,and oxidative phosphorylation.Excessive oxidative phosphorylation increased reactive oxygen species generation,which led to increased lipid peroxidation and HT22 cell apoptosis.Bromocriptine was identified as a lipid-lowering compound that inhibited phosphorylation of cytosolic phospholipase A2 by reducing the phosphorylation of extracellular signal-regulated kinases 1/2 in the mitogen-activated protein kinase pathway,thereby inhibiting myelin debris degradation by cytosolic phospholipase A2 and alleviating lipid droplet accumulation in myelin debris-treated HT22 cells.Motor function,lipid droplet accumulation in spinal cord neurons and neuronal survival were all improved in bromocriptine-treated mice after spinal cord injury.The results suggest that bromocriptine can protect neurons from lipotoxic damage after spinal cord injury via the extracellular signal-regulated kinases 1/2-cytosolic phospholipase A2 pathway.

NOX4 promotes tumor progression through the MAPK-MEK1/2-ERK1/2 axis in colorectal cancer

BACKGROUND Metabolic reprogramming plays a key role in cancer progression and clinical outcomes;however,the patterns and primary regulators of metabolic reprogramming in colorectal cancer(CRC)are not well understood.AIM To explore the role of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)in promoting progression of CRC.METHODS We evaluated the expression and function of dysregulated and survival-related metabolic genes using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes.Consensus clustering was used to cluster CRC based on dysregulated metabolic genes.A prediction model was constructed based on survival-related metabolic genes.Sphere formation,migration,invasion,proliferation,apoptosis and clone formation was used to evaluate the biological function of NOX4 in CRC.mRNA sequencing was utilized to explore the alterations of gene expression NOX4 over-expression tumor cells.In vivo subcutaneous and lung metastasis mouse tumor model was used to explore the effect of NOX4 on tumor growth.RESULTS We comprehensively analyzed 3341 metabolic genes in CRC and identified three clusters based on dysregulated metabolic genes.Among these genes,NOX4 was highly expressed in tumor tissues and correlated with worse survival.In vitro,NOX4 overexpression induced clone formation,migration,invasion,and stemness in CRC cells.Furthermore,RNA-sequencing analysis revealed that NOX4 overexpression activated the mitogen-activated protein kinase-MEK1/2-ERK1/2 signaling pathway.Trametinib,a MEK1/2 inhibitor,abolished the NOX4-mediated tumor progression.In vivo,NOX4 overexpression promoted subcutaneous tumor growth and lung metastasis,whereas trametinib treatment can reversed the metastasis.CONCLUSION Our study comprehensively analyzed metabolic gene expression and highlighted the importance of NOX4 in promoting CRC metastasis,suggesting that trametinib could be a potential therapeutic drugs of CRC clinical therapy targeting NOX4.

Cross-talk between Smad4 and P38 Proteins in Non-small Cell Lung Cancer

Objective： Impaired signal transduction is associated with tumorigenesis and progression of various kinds of human cancers. Transforming growth factor （TGF）-beta/Smad and ras-mitogen activated protein kinase （MAPK） are two major signal transduction pathways for adjusting cell proliferation and differentiation. Little is known about TGF-beta/Smad4 in non-small cell lung cancer （NSCLC）. Hereby, we investigated the expression of Smad4 in NSCLC, its correlation with MAPK proteins （including p38, ERK1 and JNK1 proteins） and their clinical significance in NSCLC. Methods： The expressions of Smad4, p38, ERK1 and JNK1 were detected at protein level with Western blotting and immunohistochemistry, at transcription level with RT-PCR. Statistical analysis was performed for the comparisons of expressions of Smad4, p38, ERK1 and JNK1, and their correlation with various clinicopathological parameters and the prognosis of NSCLC. Results： The levels of protein and mRNA expression of Smad4 in lung cancer tissues were significantly lower than in normal tissues （P〈0.05）. All these four proteins were associated with TNM staging. There was a strongly negative correlation between p38 and Smad4. Expressions of Smad4, p38 and JNK1, as well as tumor differentiation and staging were significantly correlated with the prognosis of NSCLC by univariate analysis. By multivariate analysis, only Smad4, p38, tumor differentiation and staging were correlated with the prognosis. Taken together, the negative expression of p38 and positive expression of Smad4 were associated with a better prognosis of NSCLC. Conclusion： Smad4 could be of vital importance for the initiation and development of NSCLC. The expression of Smad4 might be inhibited by p38, supporting a cross-talk between main proteins of TGF-beta/Smad and ras-MAPK signal transduction pathways. Smad4 and p38 could be possible prognostic factors for NSCLC.

Puerarin reduces apoptosis in rat hippocampal neurons cultured in high glucose medium by modulating the p38 mitogen activated protein kinase and c-Jun N-terminal kinase signaling pathways

OBJECTIVE: To investigate the neuroprotective effect of puerarin on rat hippocampal neurons cultured in high glucose medium, and to examine the role of the p38 mitogen activated protein kinase(p38 MAPK) and c-Jun N-terminal kinase(JNK) signaling pathways in this effect.METHODS: Primary cultures of hippocampal neurons were prepared from newborn Sprague Dawley rats. Neuron-specific enolase immunocytochemistry was used to identify neurons. The neurons were cultured with normal medium(control group) or with high-glucose medium(high-glucose group),and puerarin(puerarin group), a p38 MAPK inhibitor(SB239063; p38 MAPK inhibitor group) or a JNK inhibitor(SP600125; JNK inhibitor group) were added. After 72 h of treatment, terminal deoxynucleotidyl transferase-mediated d UTP nick end labeling assay was performed to detect apoptosis, and western blotting was used to assess protein levels of p-p38, p38, p-JNK and JNK.RESULTS: In the high-glucose group, the neuronal apoptosis rate and the p-p38/p38 and p-JNK/JNK ratios were higher than in the control group. The p38 MAPK and JNK inhibitors prevented this increase in the apoptosis rate. The apoptosis rates in the puerarin group, the p38 MAPK inhibitor group and the JNK inhibitor group were significantly decreased compared with the high-glucose group.Moreover, protein levels of p-p38 and p-JNK were significantly reduced, and the p-p38/p38 and p-JNK/JNK ratios were decreased in the puerarin group compared with the high-glucose group. In addition, compared with the high-glucose group,p-p38 levels and the p-p38/p38 ratio were reduced in the p38 MAPK inhibitor group, and p-JNK levels and the p-JNK/JNK ratio were decreased in the JNK inhibitor group.CONCLUSION: Puerarin attenuates neuronal apoptosis induced by high glucose by reducing the phosphorylation of p38 and JNK.

Protective effect of Chushizi(Fructus Broussonetiae)on acetaminophen-induced rat hepatitis by inhibiting the Toll-like receptor 3/c-Jun N-terminal kinase/c-jun/c-fos/janus protein tyrosine kinase/activators of transcription 3 pathway

OBJECTIVE:To observe the intervention of Chushizi(Fructus Broussonetiae)(CSZ)on drug-induced liver injury(DILI)in rats,as well as indicators of liver function,serum levels of inflammatory cytokines,and expression of proteins and m RNA associated with toll-like receptor 3(TLR3)and the signal transducer and activator of transcription 3(STAT3)pathway in the liver[TLR3,janus protein tyrosine kinase 2(JAK2),c-jun,c-fos,c-Jun N-terminal kinase 2(JNK2),and STAT3].METHODS:Forty specified pathogen free grade Sprague-Dawley rats were randomly divided into the control group,the model group,the silybin group and the CSZ group.Rats were given acetaminophen(APAP)to trigger DILI.Histopathology of the liver was observed by hematoxylin-eosin staining.The levels of alanine aminotransferase(ALT),aspartate transaminase(AST),direct bilirubin(DBIL),and total bilirubin(TBIL)in serum were detected by a semi-automatic biochemical instrument.Content of tumor necrosis factor alpha(TNF-α),interleukin(IL)-6,IL-13,and IL-22 in serum were detected by the enzyme-linked immunosorbent assay,the expression of TLR3,phosphorylation of JAK2(p-JAK2),while c-jun and c-fos proteins in the liver were determined by immunohistochemistry;expression of JNK2,and STAT3 in the liver were assayed by Western blot and real-time quantitative polymerase chain reaction.P-JNK2 and p-STAT3 in the liver were assayed by Western blot.RESULTS:After treatment,the activity of ALT,AST,and concentrations of TBIL,DBIL,TNF-α,IL-6,as well as IL-13 in serum,were lower than those in the model group,and expression of p-JAK2,TLR3,c-jun,c-fos,p-STAT3,and p-JNK2 could be downregulated.CONCLUSION:Our findings suggest that CSZ is a valid medicine to alleviate APAP-induced DILI,while its partial mechanism may regulate the TLR3/JNK/c-jun/c-fos/JAK/STAT3 pathway.

Down-regulation of HIV-1 Infection by Inhibition of the MAPK Signaling Pathway

The human immunodeficiency virus type 1 （HIV-1） can interact with and exploit the host cellular machinery to replicate and propagate itself. Numerous studies have shown that the Mitogen-activated protein kinase （MAPK） signal pathway can positively regulate the replication of HIV-1, but exactly how each MAPK pathway affects HIV-1 infection and replication is not understood. In this study, we used the Extracellular signal-regulated kinase （ERK） pathway inhibitor, PD98059, the Jun N-terminal kinase （JNK） pathway inhibitor, SP600125, and the p38 pathway inhibitor, SB203580, to investigate the roles of these pathways in HIV-1 replication. We found that application of PD98059 results in a strong VSV-G pseudotyped HIV-1NL4-3 luciferase reporter virus and HIV-1NL4-3 virus inhibition activity. In addition, SB203580 and SP600125 also elicited marked VSV-G pseudotyped HIV-INL4-3 luciferase reporter virus inhibition activity but no HIV-1NL4-3 virus inhibition activity. We also found that SB203580 and SP600125 can enhance the HIV-1 inhibition activity of PD98059 when cells were treated with all three MAPK pathway inhibitors in combination. Finally, we show that HIV-1 virus inhibition activity of the MAPK pathway inhibitors was the result of the negative regulation of HIV-1 LTR promoter activity.

Oxidative stress and mitochondrial dysfunction of retinal ganglion cells injury exposures in long-term blue light

AIM:To investigate the phototoxic effect of long-term excessive narrow-band blue light in staurosporine-induced differentiated retinal ganglion cells-5(SSRGC-5).METHODS:SSRGC-5 cells were divided into two groups,blue light group(BL group)and control group.Cell viability was assessed by using CCK-8 assay.Metabolic profile analysis was performed by using Seahorse extracellular flux analyzer.Mitochondria ultrastructure were studied via transmission electron microscope(TEM).Mitochondria contents and oxidative stress was evaluated by flow cytometry.Western blotting was performed to monitor the changes in mitogen-activated protein kinases(MAPK)pathway and PI3 K/AKT pathway.RESULTS:Blue light caused morphological changes of SSRGC-5 cells.The cell viability was significantly decreased from 3 h in BL group.Intercellular ROS and mitochondrial superoxide levels were increased following blue light exposure.Metabolic profiling identified blue light induced SSRGC-5 cells to have severely compromised mitochondrial function.This was accompanied by impaired mitochondrial ultrastructure and remodeling,increased expression of the mitochondrial related proteins,and increased glycolysis as compensation.Moreover,the results showed that blue light induced higher expression of p-p38,p38,p-JNK,p-ERK,p-cJun,c-Jun,and p-AKT.CONCLUSION:These findings indicate that excessive narrow-band blue light induces oxidative stress and mitochondrial metabolic remodeling dysregulate in SSRGC-5 cells.Activated MAPK and AKT signaling pathways are involved in this process.

Protein kinase-based neural signaling pathways for ginsenosides:a retrospective review

Ginsenosides are the main active components of ginseng,which have been reported to target brain tissues and produce multiple neuroprotective effects.Ginsenosides have been shown to improve learning ability and memory in normal aged animals,and in an animal model of memory impairment.However,its underlying pharmacological mechanisms are very complicated,especially with regard to its effects on the activation of protein kinases in neurons.Previous reports have shown that some protein kinases may be affected by ginsenosides,including protein kinase C,calcium/calmodulin-dependent protein kinase Ⅱ,c-Jun-N terminal kinase,and protein tyrosine kinase.In this paper,protein kinases that may underlie the mechanisms of ginsenosides will be discussed.

JNK、p-JNK在散发性大肠管状腺瘤癌变过程中的表达及意义

目的探讨JNK、p-JNK在大肠管状腺瘤癌变过程中的作用。方法采用免疫组化检测87例腺瘤伴不同程度异型增生及癌变组织中JNK和p-JNK的表达。结果 JNK、p-JNK在正常黏膜、腺瘤伴异型增生及癌变中的阳性表达率均呈增高趋势,JNK和p-JNK在腺瘤伴异型增生和癌变组中的阳性表达率均高于正常黏膜组;JNK和p-JNK的表达存在一致性。结论随着腺瘤异型增生程度的增高及癌变的出现,JNK呈逐渐增高趋势,JNK在大肠腺瘤癌变过程中可能起一定作用。

Efficacy of Cigu Xiaozhi pill(慈菇消脂丸) on non-alcoholic steatohepatitis-associated lipoapoptosis through stress-activated c-Jun N-terminal kinase signalling pathway

OBJECTIVE: To investigate the efficacy of Cigu Xiaozhi pill(慈菇消脂丸, CGXZ) on non-alcoholic steatohepatitis(NASH)-associated lipoapoptosis through the stress-activated c-Jun N-terminal kinase(JNK)/stress-activated protein kinase signalling pathway.METHODS: Sixty male Sprague-Dawley rats were randomly divided into the following groups(10 rats each): blank control, model, low-dose CGXZ,medium-dose CGXZ, high-dose CGXZ, and positive control(treated with SP600125, a JNK inhibitor).The NASH model was established and the histomor-phological characteristics of haematoxylin and eosin-stained liver tissues were examined under a light microscope. Cell apoptosis in liver tissues was assessed via terminal deoxynucleotidyl transferase d UTP nick-end labelling assay. In addition, the m RNA and protein expression levels of p-JNK,p-c-Jun, caspase-8, Fas, and Fas-L were determined via fluorescence-based quantitative real-time PCR,immunohistochemical and Western blot assays.RESULTS: Histopathological examination of the liver showed that the model rats had moderate-to-severe steatosis with infiltration of inflammatory cells as well as significantly higher expression levels of the p-JNK, p-c-Jun, caspase-8, Fas, and Fas-L proteins, compared with those in the blank control group(P < 0.01). Hepatic lobules of the rats in the treatment groups showed significantly reduced vacuolar degeneration and steatosis as well as alleviated inflammatory cell infiltration. The high and medium-dose CGXZ groups exhibited significantly lower m RNA and protein expression levels of p-JNK, p-c-Jun, caspase-8, Fas, and Fas-L, compared with those in the model group(P < 0.05 or P <0.01).CONCLUSION: CGXZ pill inhibited the onset of hepatocyte apoptosis by regulating the expression of p-JNK, p-c-Jun, caspase-8, Fas, and Fas-L, thereby exerting therapeutic effects against NASH.

c-Jun, at the crossroad of the signaling network

c-Jun,the most extensively studied protein of the activator protein-1(AP-1)complex,is involved in numerous cell activities,such as proliferation,apoptosis,survival,tumorigenesis and tissue morphogenesis.Earlier studies focused on the structure and function have led to the identification of c-Jun as a basic leucine zipper(bZIP)transcription factor that acts as homo-or heterodimer,binding to DNA and regulating gene transcription.Later on,it was shown that extracellular signals can induce post-translational modifications of c-Jun,resulting in altered transcriptional activity and target gene expression.More recent work has uncovered multiple layers of a complex regulatory scheme in which c-Jun is able to crosstalk,amplify and integrate different signals for tissue development and disease.One example of such scheme is the autocrine amplification loop,in which signal-induced AP-1 activates the c-Jun gene promoter,while increased c-Jun expression feedbacks to potentiate AP-1 activity.Another example of such scheme,based on recent characterization of gene knockout mice,is that c-Jun integrates signals of several developmental pathways,including EGFR-ERK,EGFR-RhoA-ROCK,and activin B-MAP3K1-JNK for embryonic eyelid closure.After more than two decades of extensive research,c-Jun remains at the center stage of a molecular network with mysterious functional properties,some of which are yet to be discovered.In this article,we will provide a brief historical overview of studies on c-Jun regulation and function,and use eyelid development as an example to illustrate the complexity of c-Jun crosstalking with signaling pathways.

Hydroxychavicol,a polyphenol from Piper betle leaf extract,induces cell cycle arrest and apoptosis in TP53-resistant HT-29 colon cancer cells

This study aims to elucidate the antiproliferative mechanism of hydroxychavicol(HC).Its effects on cell cycle,apoptosis,and the expression of c-Jun N-terminal kinase(JNK)and P38 mitogen-activated protein kinase(MAPK)in HT-29 colon cancer cells were investigated.HC was isolated from Piper betle leaf(PBL)and verified by high-performance liquid chromatography(HPLC),nuclear magnetic resonance(NMR),and gas chromatography-mass spectrometry(GC-MS).The cytotoxic effects of the standard drug 5-fluorouracil(5-FU),PBL water extract,and HC on HT-29 cells were measured after 24,48,and 72 h of treatment.Cell cycle and apoptosis modulation by 5-FU and HC treatments were investigated up to 30 h.Changes in phosphorylated JNK(pJNK)and P38(pP38)MAPK expression were observed up to 18 h.The half maximal inhibitory concentration(IC_(50))values of HC(30μg/mL)and PBL water extract(380μg/mL)were achieved at 24 h,whereas the IC_(50)of 5-FU(50μmol/L)was obtained at 72 h.Cell cycle arrest at the G0/G1 phase in HC-treated cells was observed from12 h onwards.Higher apoptotic cell death in HC-treated cells compared to 5-FU-treated cells(P<0.05)was observed.High expression of pJNK and pP38 MAPK was observed at 12 h in HC-treated cells,but not in 5-FU-treated HT-29 cells(P<0.05).It is concluded that HC induces cell cycle arrest and apoptosis of HT-29 cells,with these actions possibly mediated by JNK and P38 MAPK.

Efficacy of decoction from Jieduan Niwan formula on rat model of acute-on-chronic liver failure induced by porcine serum

OBJECTIVE:To dynamically observe the efficacy of Jieduan Niwan formula(JDNW)on a rat model of acute-on-chronic liver failure(ACLF).METHODS:Seventy Wistar rats were divided into control group(6 rats),model group(22 rats),JDNW group(21 rats),and SP600125 group(21 rats).13 weeks'porcine serum injection followed with D-galactosamine and lipopolysaccharide joint acute attack was used to establish ACLF model.Rats in JDNW group were orally given JDNW formula for 3 days before acute attack;rats in SP600125 group were injected with SP60012530 min ahead of acute attack.Rats were sacrificed respectively at 4,8 and 12 h after model established.Alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),Creatinine(CR),blood urea nitrogen(BUN),prothrombin activity(PTA)were examined by biochemical process,Tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),transformed growth factor-beta 1(TGF-β1),High mobility group box-1(HMGB-1),CD3,CD4,CD8 were analyzed by enzyme-linked immunosorbent assay,apoptotic index(AI)was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining,expression of Bad,phosphorylated Jun N-terminal kinases(p-JNK)and Cytochrome C(Cyt C)were detected by immunohistochemical analysis,Bax and Bid were detected by Western blot analysis.RESULTS:In model group,the levels of ALT,AST,TBIL,CR,BUN,IL-1β,IL-6,IL-10,TGF-β1 and HMGB-1 remarkably increased and PTA decreased compared with control group(P<0.05),as time goes on,ALT,AST,TBIL,CR,BUN,continued to grow,while IL-1β,IL-6,IL-10,HMGB-1,TGF-β1 and PTA gradually decreased;massive necrosis could be seen;the levels of TNF-α,CD3,CD4,CD8,AI,p-JNK,Bax,Bad,Bid and Cyt C increased at 4 h and peaked at 8 h,but decreased at 12 h(P<0.05).JDNW group,by contrast,showed less pathological injury,increased PTA level,and reduced ALT,AST,TBIL,TNF-α,IL-1β,IL-6,IL-10,TGF-β1,HMGB-1,CD3,CD4 and CD8 levels(P<0.05),moreover,the AI and expression of p-JNK,Bax,Bad,Bid and Cyt C were lower than model group at 4 and 8 h but were higher at 12 h(P<0.05).Similar results were observed in SP600125 group.CONCLUSION:An ACLF rat model with low mortality can be established by porcine serum joint with D-galactosamine+lipopolysaccharide induction;JDNW decoction can effectively suppress the inflammatory reaction,improve the immune system,and protect the liver of ACLF rats,the mechanism might involve the inhibition of the JNK-induced mitochondrial apoptotic pathway.

Shenweifang-containing serum inhibits transforming growth factor-β1–induced myofibroblast differentiation in normal rat kidney interstitial fibroblast cells

OBJECTIVE:To investigate the efficacy of Shenweifang(SWF)-containing serum on transforming growth factor(TGF)-β1–induced fibroblast-myofibroblast transition in normal rat kidney interstitial fibroblast cells(NRK-49 F).METHODS:Sprague-Dawley rats were gavaged with one of five solutions:(a)saline;(b)saline plus low-dose SWF;(c)saline plus medium-dose SWF;(d)saline plus highdose SWF;and(e)saline plus valsartan.NRK-49 F cells were treated with TGF-β1 and cultured using serum from the gavaged rats.RESULTS:TGF-β1 treatment increased the expression ofα-smooth muscle actin,proliferating cell nuclear antigen,collagenⅠ,Smad3,mitogen-activated protein kinase(MAPK)10,and c-Jun N-terminal kinase(JNK)3 and induced abnormalities in cell morphology,cell cycle progression,and cell proliferation.CONCLUSIONS:SWF-or valsartan-containing serum corrected(or partially corrected)TGF-β1–induced abnormal changes in this in vitro system.SWF-containing serum reversed abnormalities in morphology,cell cycle progression,and proliferation in TGF-β1–treated NRK-49F cells,probably by blocking the TGF-β1/Smads and TGF-β1/MAPK/JNK pathways.