Visual Analysis of Knowledge Map of Traditional Chinese Medicine in Prevention and Treatment of Pulmonary Interstitial Fibrosis Based on CiteSpace

Objective: To analyze the relevant research literature on the prevention and treatment of pulmonary interstitial fibrosis with traditional Chinese medicine (TCM), understand the current research status, hot spots and future development trend in this field, and provide basis and feasible suggestions for further research in this field. Methods: The journal literatures related to the prevention and treatment of pulmonary interstitial fibrosis with TCM in recent 20 years in CNKI database were searched and passed through CiteSpace 5.8.R3 generates the knowledge map of relevant literature authors, document issuing institutions and keywords, and makes visual analysis. Results: A total of 1,576 documents were included, and the annual number of documents showed a fluctuating upward trend, forming a relatively stable research team represented by authors such as LYU Xiaodong, PANG Lijian and LIU Chuang;According to the atlas of document issuing institutions, Shandong University of Traditional Chinese Medicine and its affiliated hospitals ranked first in the number of documents issued, and the cooperation between institutions is dominated by the University of traditional Chinese medicine and its affiliated hospitals;Keyword cluster analysis shows that a large number of studies have been carried out in the field of etiology and pathogenesis, TCM compound, clinic and experiment. Conclusion: The research on the prevention and treatment of pulmonary interstitial fibrosis with TCM has a high degree of attention, but the cooperation network between the research authors and institutions needs to be strengthened. The research on the pathogenesis and improving the quality of life of patients is the trend of development in the future.

Mechanisms of renal interstitial fibrosis: cross-talk between mitophagy and NLRP3 inflammasome

Renal interstitial fibrosis(RIF)is the main pathological basis leading to end-stage renal disease,and is closely related to the prognosis of patients with kidney disease.Increasing evidence as shown that mitophagy and NLRP3 inflammasome play important roles in the pathogenesis of RIF.Studies suggest that inhibiting NLRP3 inflammasome by activating mitophagy can prevent and alleviate RIF.This review summarizes role played by cross-talk between mitophagy and NLRP3 inflammasome in promoting RIF,so as to offer new perspectives on more effective slow the progression of renal diseases and fibrosis prevention.

Research progress of renal interstitial fibrosis

Renal interstitial fibrosis （RIF） is a common pathological process of chronic kidney disease that progresses toend-stage renal failure. The degree of RIF is closely related to renal function. The study of the pathogenesis of renalinterstitial fibrosis, exploration of effective prevention measures to delay the progress of end stage renal disease andprolong the life of patients is significant. The pathology of RIF has complicated extracellular and intercellularmechanisms, involving many cells and cytokines, resulting in an incomplete mechanistic understanding of thedisease. Finding effective herbs or herbal extracts for prevention and treatment of RIF is crucial because currentmedical approaches do not reliably slow or reverse RIF. The research progress of RIF in recent years issummarized as follows.

Exploring the mechanism of Qinghaobiejiatang in treating renal interstitial fibrosis based on network pharmacology

To explore the potential mechanism of the classic ancient prescription Qinghaobiejiatang(QHBJT)in treating renal interstitial fibrosis.Methods:Obtain the active ingredients of Qinghao(Artemisiae annuae herba),Biejia(Trionycis carapax),Dihuang(Rehmanniae radix),Zhimu(Anemarrhenae rhizoma),and Mudanpi(Moutan cortex)through TCMSP and TCMID databases.Collect disease targets through the GENECARDS database.Use Venny2.1.0 platform to draw Venn diagrams to map drugs and disease targets.Import the key targets into Cytoscape 3.7.2 software to draw a network diagram of“drugs-active ingredients-diseases-key targets”.The STRING11.0 database was used to construct the key target protein interaction network diagram.Use R language for Gene ontology function and kyoto encyclopedia of genes and genomes pathway enrichment analysis.Results:A total of 317 active ingredients were obtained through screening,involving 166 targets,and 102 active ingredients related to disease targets,mainly involved in the regulation of key targets such as FOS,IL6,MTOR,MAPK8,RELA,CCND1,TP53,EGFR,and CASP3 signal pathways related to viral infection,tumor-related,apoptosis,signal transduction,fluid shear stress and atherosclerosis play a synergistic role in the treatment of renal fibrosis.Conclusion:The effect mechanism of QHBJT in treating renal interstitial fibrosis is related to inflammation,oxidative stress,hypoxia,apoptosis,pathological activation and damage of renal tubular epithelial cells mediated by the above pathways.

Shen Qi Wan attenuates renal interstitial fibrosis through upregulating AQP1

Renal interstitial fibrosis(RIF)is the crucial pathway in chronic kidney disease(CKD)leading to the end-stage renal failure.However,the underlying mechanism of Shen Qi Wan(SQW)on RIF is not fully understood.In the current study,we investigated the role of Aquaporin 1(AQP1)in SQW on tubular epithelial-to-mesenchymal transition(EMT).A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo.Subsequently,the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown.The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine,increased the protein expression of E-cadherin and AQP1 expression,and decreased the expression of vimentin andα-smooth muscle actin(α-SMA).Similarly,treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells.The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1.AQP1 knockdown also increased the mRNA expression of vimentin andα-SMA,and decreased the expression of E-cadherin.The protein expression of vimentin increased,while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells.These results revealed that AQP1 knockdown promoted EMT.Furthermore,AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells.In sum,SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.

Correlation between the Expression of TGF-β1,Rhoa,SOX9 and Renal Interstitial Fibrosis in Rats with Chronic Kidney Disease

Objective:To study the correlation between the expression of transforming growth factor-β1(TGF-β1),Rho A,SOX9 and renal interstitial fibrosis in rats with chronic kidney disease.Methods:Forty specific pathogen-free(SPF)male SD rats were randomly divided into study group and control group,with 20 cases in each group.The study group was given adenine suspension by gavage,while the control group was given the same amount of saline by gavage.Blood,urine and renal tissue specimens were collected from all rats at 3rd and 6th weeks after modeling.The kidney weight,kidney weight/body weight,renal function indexes,the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues,Masson staining and renal interstitial fibrosis score were compared between the two groups.Pearson correlation was used to analyze the relationship between the renal interstitial fibrosis score and the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues of rats with chronic kidney disease.Results:The kidney weight and kidney weight/body weight of rats in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The quantitative levels of creatinine,urea nitrogen and 24-hour urinary protein in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The expression levels of TGF-β1,Rho A and SOX9 m RNA in renal tissues of rats in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The renal interstitial fibrosis score in the study group was higher than that in the control group at 3rd and 6th weeks after modeling(P<0.05).Pearson correlation analysis confirmed that the renal interstitial fibrosis score in rats with chronic kidney disease was positively correlated with the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues(P<0.05).Conclusion:The expression of TGF-β1,Rho A and SOX9 was abnormally high in rats with chronic kidney disease and was closely related to renal interstitial fibrosis,which may play a promoting role in the process of renal interstitial fibrosis.

Interventional mechanisms of herbs or herbal extracts on renal interstitial fibrosis

Renal interstitial fibrosis（RIF） is a common development in chronic renal diseases that can lead to uremia and be life-threatening. The RIF pathology has complicated extracellular and intercellular mechanisms, involving many cells and cytokines, resulting in an incomplete mechanistic understanding of the disease. Finding effective herbs or herbal extracts for prevention and treatment of RIF is crucial because current medical approaches do not reliably slow or reverse RIF. In recent years, many experts have worked to identify herbs or herbal extracts to combat RIF both in vivo and in vitro, with some success. This review attempts to summarize the possible interventional mechanisms of herbs or herbal extracts involved in protecting and reversing RIF. The authors found some herbs and their extracts that may ameliorate renal impairments through anti-inflammation, anti-fibrogenesis and stabilization of extra cellular matrix. Among them, tetramethylpyrazine/ligustrazine, curcumin and polyglucoside of Tripterygium have experimentally shown good potential for improving RIF. However, conclusive evidence is still needed, especially in randomized controlled clinical trials. We expect that herbs or herbal extracts will play an important role in RIF treatment and reversal in the future.

Fuzheng Huayu Formula(扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

Objective：To investigate the mechanism of action of Fuzheng Huayu Formula（扶正化瘀方,FZHY）against renal interstitial fibrosis（RIF）relating to oxidative injury and nuclear factor-kappa B（NF-κB）activity.Methods：Thirty-two Sprague-Dawley rats were randomly divided into 3 groups：normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline（Hyp）and collagen deposition in kidney were observed.The activities of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）,the content of glutathione（GSH）and malondialdehyde（MDA）of kidney were tested.The expressions of inhibitor-κappa B（IκB）,phospho-IκB（p-IκB）,tumor necrosis factor-α（TNF-α）,matrix metalloproteinase-2（MMP-2）andα-smooth muscle actin（α-SMA）were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results：Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney（P〈0.01）.FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats（P〈0.01）.GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats（P〈0.01）.FZHY significantly increased GSH content and decreased MDA content in the model rats（P〈0.01）.The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression（P〈0.01）.The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group（P〈0.01）,FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions（P〈0.01）.Conclusions：FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.

Fuzheng Huayu recipe, a traditional Chinese compound herbal medicine, attenuates renal interstitial fibrosis via targeting the miR-21/PTEN/AKT axis

Objective: MicroRNAs(miRNAs) may be viable targets for treating renal interstitial fibrosis(RIF). Fuzheng Huayu recipe(FZHY), a traditional Chinese compound herbal medicine, is often used in China to treat fibrosis. This study sought to assess the mechanisms through which FZHY influences miRNAs to treat RIF.Methods: RIF was induced in rats by mercury chloride and treated with FZHY. Hydroxyproline content,Masson’s staining and type I collagen expression were used to evaluate renal collagen deposition.Renal miRNA profiles were evaluated using a miRNA microarray. Those miRNAs that were differentially expressed following FZHY treatment were identified and subjected to bioinformatic analyses. The miR-21 target gene phosphatase and tensin homolog(PTEN) expression and AKT phosphorylation in kidney tissues were assessed via Western blotting. In addition, HK-2 human proximal tubule epithelial cells were treated using angiotensin II(Ang-II) to induce epithelial-to-mesenchymal transition(EMT), followed by FZHY exposure. miR-21 and PTEN expressions were evaluated via quantitative reverse transcriptionpolymerase chain reaction(qRT-PCR), while E-cadherin and a-smooth muscle actin(a-SMA) expressions were assessed by immunofluorescent staining and qRT-PCR. Western blotting was used to assess PTEN and AKT phosphorylation.Results: FZHY significantly decreased kidney collagen deposition, hydroxyproline content and type I collagen level. The miRNA microarray identified 20 miRNAs that were differentially expressed in response to FZHY treatment. Subsequent bioinformatic analyses found that miR-21 was the key fibrosis-related miRNA regulated by FZHY. FZHY also decreased PTEN expression and AKT phosphorylation in fibrotic kidneys. Results from in vitro tests also suggested that FZHY promoted E-cadherin upregulation and inhibited a-SMA expression in Ang-II-treated HK-2 cells, effectively reversing Ang-II-mediated EMT. We also determined that FZHY reduced miR-21 expression, increased PTEN expression and decreased AKT phosphorylation in these cells.Conclusion: miR-21 is the key fibrosis-related miRNA regulated by FZHY. The ability of FZHY to modulate miR-21/PTEN/AKT signaling may be a viable approach for treating RIF.

Chinese herbal decoction astragalus and angelica exerts its therapeutic effect on renal interstitial fibrosis through the inhibition of MAPK,PI3K-Akt and TNF signaling pathways

Astragalus and Angelica decoction(A&A)has been clinically used as a classical traditional Chinese medicine(TCM)formula in China for many years for the treatment of kidney diseases,especially renal interstitial fibrosis(RIF).However,the mechanisms underlying the therapeutic effects of A&A on RIF remains poorly understood.In the present study,systematic network pharmacology and effective experimental verification were utilized for the first time to elucidate the pharmacological efficacy and potential mechanism.The outcomes indicated that 22 active components and 87 target genes of A&A were identified and cross-referenced with RIF-associated genes,contributing to confirmation of 74 target genes of A&A for RIF.Pathway and functional enrichment analyses revealed that A&A had substantial effects on MAPK,PI3K-Akt and TNF signaling pathways.In addition,seven core targets with relatively higher betweenness and degree were identified in the constructed Chinese medicine material-chemical component-target-signal pathway network.Moreover,we verified the potential therapeutic effect of A&A in vivo(using a mouse model of RIF),confirming that A&A could effectively protect the kidney by regulating these target genes.The therapeutic effect of A&A on RIF could be attributed to its role in regulating the cell cycle,limiting the apoptosis,and inhibiting the inflammation.

Huangqi decoction(黄芪汤) attenuates renal interstitial fibrosis via transforming growth factor-β1/mitogen-activated protein kinase signaling pathways in 5/6 nephrectomy mice

OBJECTIVE: To investigate the effect of Huangqi decoction( 黄芪汤) on renal interstitial fibrosis and its association with the transforming growth factor-β1(TGF-β1)/mitogen-activated protein kinase(MAPK) signaling pathway. METHODS: 120 C57/BL mice were randomly divided into six groups: sham group, Enalapril(20 mg/kg) group, 5/6 nephrectomy model group, and 5/6 nephrectomy model plus Huangqicoction(0.12, 0.36 and 1.08 g/kg respectively) groups. Detecting 24hours urinary protein, blood pressure, serum creatinine, urea nitrogen content changes. Periodic Acid-Schiff stain(PAS) and Masson’s trichrome staining was used to observe the renal tissue pathological changes. Protein expression of TGF-β1, Phosphorylated P38 mitogen activated protein kinases(P-P38), Phosphorylated c-jun N-terminal kinase(P-JNK), Phosphorylated extracellular regulated proteinhnase(PERK), Fibroblast-specific protein-1(FSP-1), Alpha smooth muscle actin(α-SMA), Type Ⅲ collagen(Collagen Ⅲ), Connective tissue growth factor(CTGF),Bcl-2 Assaciated X protein(Bax) and B cell lymphoma 2(Bcl-2) were measured with western blot and immunohistochemical. RESULTS: Both Huangqi decoction and Enalapril improved the kidney function, 24 h urinary protein and the fibrosis in 5/6 nephrectomy mice, Huangqi decoction downregulated the expressions of TGF-β1, FSP-1, α-SMA, Collagen Ⅲ and CTGF in a dose-dependent manner, and it has a significant difference(P < 0.01) compared with model group.Huangqi decoction downregulated the expressions of P-P38, P-JNK, P-ERK and Bcl-2 in a dose-dependent manner, while upregulated the expression of Bax. CONCLUSIONS: The protective effect of Huangqi decoction for renal interstitial fibrosis in 5/6 nephrectomized mice via the inhibition of EpithelialMesenchymal Transitions and downregulating the TGF-β1/MAPK signaling pathway.

Predicting outcomes after kidney transplantation: Can Pareto’s rules help us to do so?

Kidney transplantation is the best option for kidney replacement therapy,even considering that most of the times the grafts do not survive as long as their recipients.In the Khalil et al's experience,published in this issue of the Journal,they analyze their second kidney graft survival and describe those significant predictors of early loss.This editorial comments on the results and put in perspective that most of the times,long-term graft survival could be inadvertently jeopardized if the immunosuppressive therapy is reduced or withdrawn for any reason,and that it could happen frequently if the transplant physician intends to innovate with the clinical care without proper evidence-based data.

Z-Guggulsterone alleviated renal fibrosis and G_(2)/M cycle arrest through Klotho/P53 signaling

OBJECTIVE Chronic kidney disease(CKD)has become a global public health problem with 10%-15%incidence rate,and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD.Z-Guggulsterone(Z-GS),an active compound from derived from Commiphora mukul,has been proved to be effective in various diseases.The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis.METHODS Unilateral ureteral obstruction(UUO)mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo,respectively.The mice and cells were treated with different doses of Z-GS to observe the pharmacological action.Renal function,including Scr,BUN,and UA,were detected by commercial kits.H&E and Masson staining were performed to observe histopathological changes of kidney.Cell viability and LDH release of HK-2 cells were detected by commercial kits.Cell cycle distribution and apoptosis rate were analyzed by flow cytometry.Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis.Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting.RESULTS The results showed that Z-GS decreased the rise of Scr,BUN,and UA and lightened renal histopathological injury,which were induced by UUO.Besides,Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions ofα-SMA,TGF-βand collagenⅣ,and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate.Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells.In addition,hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS.The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level.Nevertheless,the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho.Moreover,siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis.CONCLUSION This study clarified that Z-GS alleviated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway.People who have suffered CKD may potentially benefit from treatment with Z-GS.

Effects of Yishenbupi (tonifying-kidney and invigorating-spleen) prescription on the expression of renal fibrosis-associated proteins in unilateral ureteral occlusion rats

Objective:To evaluate the effects and mechanism of the Yishenbupi(tonifying-kidney and invigorating-spleen)prescription on the expression of renal fibrosis-associated vimentin,α-SMA,and fibronectin in unilateral ureteral occlusion rats.Methods:A total of 48 SD(Sprague-Dawley)rats were randomly divided into the model,sham-operated(sham),irbesartan,and Yishenbupi groups,with 12 rats in each group.After the unilateral ureteral occlusion model was established,rats in the model and sham groups were administered normal saline,whereas rats in the Yishenbupi group were administered Yishenbupi prescription(18 g/kg/d)intragastrically and those in the irbesartan group were administered irbesartan(10 mg/kg/d)intragastrically.All rats were sacrificed 21 days later.Pathological changes in rat renal tissue were evaluated by H&E staining.The expression of vimentin,α-SMA,and fibronectin in renal tissues was detected by western blotting.Results:Compared with the sham group the model group had renal tubular epithelial cell atrophy,inflammatory cell infiltration accompanied with the proliferation of interstitial collagen fibers,fewer glomeruli,or glomerulosclerosis.Compared with the model group,significantly less renal tubular and glomerular damages,inflammatory cell infiltration,and collagen fibers were observed in different intervention groups,especially in the Yishenbupi group.Compared with the sham group,significantly higher expressions of fibrosis markers,including vimentin,α-SMA,and fibronectin,were observed in the model group.Compared with the model group,the expression of anti-fibrosis markers,including vimentin,α-SMA and fibronectin,was significantly decreased in both the irbesartan and Yishenbupi groups(P<0.01);however,the Yishenbupi group showed higher efficacy than the irbesartan group(P<0.05).Conclusion:The Yishenbupi prescription may improve renal fibrosis by reducing the expression of fibrosis-associated vimentin,α-SMA,and fibronectin.

Research progress of Chinese medicine in treatment of IPF(idiopathic pulmonary fibrosis)

Idiopathic pulmonary interstitial fibrosis is one of the respiratory refractory diseases,and the incidence rate is on the rise.At present,the effect of western medicine is not ideal and the side effects are obvious,while the traditional Chinese medicine shows good curative effect on the disease.This paper makes a summary on the traditional Chinese medicine theory in treating idiopathic pulmonary interstitial fibrosis in recent years.

The Expression of PDGF-B Chain mRNA in Lung Tissue from Rats Repeatedly Infected with Mycoplasma Pneumoniae

In order to investigate the role played by platelet derived growth factor-BB (PDGF-BB) in the pathogenesis of pulmonary interstitial fibrosis in rats repeatedly infected with mycoplasma pneumoniae (MP), a rat MP infection model was developed by infecting rats with MP for 9 times during a period of 24 weeks with a technique of ultrasonic nebulizing inhalation. Then in situ hybridization was performed with PDGF-B chain cDNA probe and the results were quantitatively analyzed to measure the changes in PDGF-B chain mRNA expression in the lung tissue. The results showed that: (1) MP polymerase chain reaction (PCR) tests showed positive results in the bronchoalveolar lavage fluid (BALF ) from all of the MP-infected rats (n = 4) while they were all negative in BALF from the control animals (n = 4, P<0. 05) and in BALF from those rats both infected with MP and, at the same time, treated with erythromycin (n = 4, P<0. 05). Bacterial cultures of the bronchial and lung tissue were negative in all three groups. The observation under a transmission electron microscope indicated that the interalveo-lar septa were widened with increased amount of collagen in the MP-infected rats while there were no obvious abnormalities in the other two groups. (2) Strong positive expression of PDGF-B chain mRNA was found in the plasma of mono-cytes and macrophages located in the locally widened interalveolar septa and alveolar spaces in the lung tissue from the MP-infected animals with the integral optical densities being 37. 42 ±9. 05 (n = 4) which was significantly higher than the values of control group (0. 42 + 0. 08, n = 4, P<0. 01) and of the group with MP-infec-tion plus erythromycin treatment (1. 62 ± 0. 40, n = 4, P<0. 01). These results suggest that PDGF-BB may be involved in the process of the development of pulmonary interstitial fibrosis caused by the repeated MP-infection. It may be an important growth factor for mediating the roles of monocytes and macrophages to promote the aggregation and proliferation of fibroblasts which can then secrete collagen in large quantity in the pulmonary interstitium.

Inhibition of Ubiquitin-specific Protease 4 Attenuates Epithelial-Mesenchymal Transition of Renal Tubular Epithelial Cells via Transforming Growth Factor Beta Receptor Type Ⅰ

Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.

mRNA Expression of Basic Fibroblast Growth Factor from A Single Intratracheal Instillation of Papain induced Emphysema in Rats

The relations between mRNA expression of basic fibroblast growth factor (bFGF) and the changes in collagen Ⅰand collagen Ⅲ in pulmonary tissues from a single intratracheal instillation of papain induced emphysema in rats were investigated. Wistar rats ( n =42) were randomly divided into normal group and emphysema model 1, 3, 5, 7, 15, 30 day groups ( n =6 in each group). The rat model of emphysema was induced by a single intratracheal instillation of papain. The results of immunohistochemistry SABC and in situ hybridization with bFGF probe were quantitatively analyzed to examine the changes of collagen Ⅰand collagen Ⅲ and bFGF mRNA expression in lung tissues and the percent of positive expression of bFGFmRNA in alveolar macrophages. The results were as follows: (1) In the emphysema model groups the optical densities of collagen Ⅰand collagen Ⅲ began to increase after 3 days, reached the highest at the 7 th day, and began to reduce at the 15 th day; (2) No expression of bFGFmRNA in pulmonary tissues was detectable in the normal group. The positive expression of bFGFmRNA was detectable in lung tissues one day after the intratracheal instillation of papain. The average optical densities reached the peak (41.895±7.017) at the 7th day, significantly higher than in the normal group (0.581±0.139, P <0.01). The positive expression of bFGFmRNA in lung tissues began to reduce at the 15th day; (3) Positive expression of bFGFmRNA in alveolar macrophages of instillation papain rats was detectable 3 days after the intratracheal instillation of papain, and reached the highest at the 7th day with the percent of positive expression of bFGF mRNA in alveolar macrophages being 70.13±11.21, higher than in the normal group (5.12±0.18, P <0.01); (4) The expression of bFGF mRNA in the lung tissues and macrophages was postively related with the changes in collagen Ⅰ and collagen Ⅲ ( P <0.01 or P <0.05) respectively. It was suggested that the up regulation of bFGF mRNA expression during the development of emphysema can lead pulmonary interstitial fibrosis, which may take part in the injury and repair and the lung tissue reconstruction.

Astragalus mongholicus ameliorates renal fibrosis by modulating HGF and TGF-β in rats with unilateral ureteral obstruction

Astragalus mongholicus （AM） derived from the dry root ofAstragalus membranaceus Bge. var. mongolicus （Bge.） Hsiao is a widely used traditional Chinese medicine. The present study investigated the potential role of AM on renal fibrosis on a rat model of unilateral ureteral obstruction （UUO）. We divided 48 Sprague-Dawley rats randomly into 4 groups： sham-operated group （Sham）, untreated UUO group, AM-treated （10 g/（kg.d）） UUO group, and losartan-treated （20 mg/（kg.d）） UUO group as positive control. Haematoxylin ＆ eosin （HE） and Masson staining were used to study the dynamic histological changes of the kidneys 7 and 14 d after operation. The expressions of fibronectin （FN）, type I collagen （coil）, hepatocyte growth factor （HGF）, transforming growth factor-β1 （TGF-β1）, and eL-smooth muscle actin （α-SMA） were analyzed by real-time polymerase chain reaction （PCR）, immunohistochemistry staining, and Western blot. Results show that, similar to losartan, AM alleviated the renal damage and decreased the deposition of FN and coil from UUO by reducing the expressions of TGF-β1 and α-SMA （P〈0.05）, whereas HGF increased greatly with AM treatment （P〈0.05）. Our findings reveal that AM could retard the progression of renal fibrosis. The renoprotective effect of AM might be related to inhibition ofmyofibroblast activation, inducing of HGF and reducing of TGF-β1 expression.

Involvement of hydrogen sulfide in the progression of renal fibrosis

Objective:Renal fibrosis is the most common manifestation of chronic kidney disease(CKD).Noting that existing treatments of renal fibrosis only slow disease progression but do not cure it,there is an urgent need to identify novel therapies.Hydrogen sulfide(H2S)is a newly discovered endogenous small gas signaling molecule exerting a wide range of biologic actions in our body.This review illustrates recent experimental findings on the mechanisms underlying the therapeutic effects of H2S against renal fibrosis and highlights its potential in future clinical application.Data sources:Literature was collected from PubMed until February 2019,using the search terms including"Hydrogen sulfide,""Chronic kidney disease"MRenal interstitial fibrosis,"Kidney disease,""Inflammation factor,""Oxidative stress,""Epithelialto-mesenchymal transition,""H2S donor,""Hypertensive kidney dysfunction,""Myofibroblasts,""Vascular remodeling,""transforming growth factor(TGF)-beta/Smads signaling,"and"Sulfate potassium channels."Study selection:Literature was mainly derived from English articles or articles that could be obtained with English abstracts.Article type was not limited.References were also identified from the bibliographies of identified articles and the authors5 files.Results:The experimental data confirmed that H2S is widely involved in various renal pathologies by suppressing inflammation and oxidative stress,inhibiting the activation of fibrosis-related cells and their cytokine expression,ameliorating vascular remodeling and high blood pressure,stimulating tubular cell regeneration,as well as reducing apoptosis,autophagy,and hypertrophy.Therefore,H2S represents an alternative or additional therapeutic approach for renal fibrosis.Conclusions:We postulate that H2S may delay the occurrence and progress of renal fibrosis,thus protecting renal function.Further experiments are required to explore the precise role of H2S in renal fibrosis and its application in clinical treatment.