Bone marrow mesenchymal stem cell-derived extracellular vesicles containing miR-181d protect rats against renal fibrosis by inhibiting KLF6 and the NF-κB signaling pathway.

Recent studies have investigated the ability of extracellular vesicles (EVs) in regulating neighboring cells by transferring signaling molecules, such as microRNAs (miRs) in renal fibrosis. EVs released by bone marrow mesenchymal stem cells (BMSCs) contain miR-181d, which may represent a potential therapy for renal fibrosis. miR-181d has been speculated to regulate Krüppel-like factor 6 (KLF6), which activates the nuclear factor-kappa B (NF-κB) signaling pathway. Luciferase assays were performed to confirm the relationship between miR-181d and KLF6. Gain- and loss-of-function studies in vivo and in vitro were performed to assess the effect of BMSC-derived EVs (BMSC-EVs), which contained miR-181d, on KLF6, NF-κB, and renal fibrosis. Transforming growth factor-ß (TGF-ß)-induced renal tubular epithelial HK-2 cells were treated with EVs derived from BMSCs followed by evaluation of collagen type IV α1 (Col4α1), Collagen I and α-smooth muscle actin (α-SMA) as indicators of the extent of renal fibrosis. Renal fibrosis was induced in rats by unilateral ureteral obstruction (UUO) followed by the subsequent analysis of fibrotic markers. BMSC-EVs had higher miR-181d expression. Overexpression of miR-181d correlated with a decrease in KLF6 expression as well as the levels of IκBα phosphorylation, α-SMA, Col4α1, TGF-ßR1 and collagen I in HK-2 cells. In vivo, treatment with miR-181d-containing BMSC-derived EVs was able to restrict the progression of fibrosis in UUO-induced rats. Together, BMSC-EVs suppress fibrosis in vitro and in vivo by delivering miR-181d to neighboring cells, where it targets KLF6 and inhibits the NF-κB signaling pathway.

Revealing Potential Diagnostic Gene Biomarkers Associated with Immune Infiltration in Patients with Renal Fibrosis Based on Machine Learning Analysis.

Chronic kidney disease is characterized by the development of renal fibrosis. The basic mechanisms of renal fibrosis have not yet been fully investigated despite significant progress in understanding the etiology of the disease. In this work, the researchers sought to identify potential diagnostic indicators for renal fibrosis. From the GEO database, we were able to acquire two gene expression profiles with publically available data (GSE22459 and GSE76882, respectively) from human renal fibrosis and control samples. 215 renal fibrosis specimens and 124 normal specimens were examined for differentially expressed genes (DEGs). The SVM-RFE and LASSO regression models were used to discover potential markers. CIBERSORT was applied to estimate the combined cohorts' immune cell fraction compositional trends in renal fibrosis. RT-PCR was used to examine the expression of ISG20 in renal fibrosis and healthy samples. In vitro experiments were applied to examine the function of ISG20 knockdown on the progression of renal fibrosis. In this study, we identified 24 DEGs. The result of LASSO and SVM-RFE identified nine critical genes. ROC assays confirmed the diagnostic value of the above nine genes for renal fibrosis. Immune cell infiltration analysis revealed that ISG20 and SERPINA3 were both found to be correlated with T cell follicular helper, neutrophils, T cell CD4 memory activated, eosinophils, T cell CD8, dendritic cell activated, B cell memory, monocytes, macrophage M2, plasma cells, T cell CD4 naïve, mast cell resting, B cell naïve, T cell regulatory, and NK cell activated. Finally, we observed that the expression of ISG20 and SERPINA3 was distinctly increased in renal fibrosis samples compared with normal samples. ISG20 siRNA significantly suppressed the progression of renal fibrosis in vitro. Overall, this study identified nine diagnostic biomarkers for renal fibrosis. ISG20 may be a novel therapeutic target of renal fibrosis.

Renoprotective effects of pirfenidone on chronic renal allograft dysfunction by reducing renal interstitial fibrosis in a rat model.

AIM: Pirfenidone (PFD) has been used as medication for idiopathic pulmonary fibrosis due to its ability in reducing lung fibrosis. However, the underlying mode of action in renal fibrosis during chronic renal allograft dysfunction (CRAD) requires further investigation. Therefore, the present study was conducted to explore the effects of PFD on renal injury induced by CRAD. MAIN METHODS: Initially, the CRAD rat model was established, followed by the intragastric administration of PFD to the rats. Urine and blood samples were collected and tested against indicators of renal functions. The renal tissues were microscopically observed to determine the changes in pathological morphology. The anti-inflammatory, anti-fibrotic and anti-oxidant properties of PFD were explored in the setting of CRAD. KEY FINDINGS: The success rate of model establishment was 92.31%, which was reflected by weight loss, appetite loss, faded fur, and retarded reaction, with the symptoms found to exacerbate with time. PFD treatment could improve renal function, ameliorate inflammation and renal fibrosis as well as promote the anti-oxidant ability of renal allograft, indicating its potential role as an effective therapeutic agent for CRAD. SIGNIFICANCE: In conclusion, PFD was found to have renoprotective effects on renal injury induced by CRAD, which resulted in the alleviation of inflammation and renal fibrosis, providing novelty for CRAD clinical treatment.

Paeonol reverses promoting effect of the HOTAIR/miR-124/Notch1 axis on renal interstitial fibrosis in a rat model.

Renal interstitial fibrosis (RIF) is a common manifestation of inflammatory and noninflammatory renal diseases, which correlates to renal excretory dysfunction. Recently, the long noncoding RNAs (lncRNAs) have been demonstrated to be involved in the development of various renal diseases. Here, we aim to determine whether paeonol (PAE) affects RIF with involvement of the lncRNA HOX transcript antisense intergenic RNA (HOTAIR)/microRNA-124 (miR-124)/Notch1 axis. RIF rat models were established by performing unilateral ureteral occlusion (UUO), in which interactions between HOTAIR, Notch1, and miR-124 were determined. To identify the roles of PAE and HOTAIR in RIF, rats were injected with HOTAIR or PAE. Subsequently, to further investigate the underlying mechanism of PAE in RIF, epithelial to mesenchymal transition (EMT)- and migration-related genes in NRK-49F cells were measured. Next, rats were further treated with IMR-1 (inhibitor of the Notch1/Jagged1 signaling pathway) to determine how PAE influences the Notch1/Jagged1 signaling pathway. HOTAIR interacted with miR-124, and miR-124 directly targeted Notch1, and HOTAIR was observed to be upregulated in RIF rats. PAE was found to decrease HOTAIR and Notch1 expression but to increase the miR-124 expression in RIF rats. PAE inhibited EMT and migration of NRK-49F cells facilitated by HOTAIR. HOTAIR activated the Notch1/Jagged1 signaling pathway by downregulating miR-124, while PAE reversed these effects of HOTAIR on the Notch1/Jagged1 signaling pathway. Overall, our study demonstrates the contributory effect of lncRNA HOTAIR on RIF by activating the Notch1/Jagged1 signaling pathway via inhibition of miR-124, whereas administration of PAE can alleviate the effects of HOTAIR on RIF.

LncRNA HOTAIR promotes renal interstitial fibrosis by regulating Notch1 pathway via the modulation of miR-124.

AIM: To understand the mechanism of long non-coding RNA (LncRNA) HOTAIR on renal interstitial fibrosis (RIF) by regulating Notch1 pathway via the modulation of miR-124. METHODS: Unilateral ureteral occlusion (UUO) was used to construct the RIF rat model. HK-2 cells induced by TGF-ß1 were used for the in vitro experiment, which were divided into five groups: Vehicle, TGF-ß1, si-HOTAIR+TGF-ß1, miR-124 inhibitor+TGF-ß1, and si-HOTAIR+miR-124 inhibitor+TGF-ß1 groups. Quantitative real-time PCR (qRT-PCR) and Western blot were performed to detect the expression of HOTAIR, miR-124, Notch1- and epithelial-to-mesenchymal transition (EMT)-related proteins. RESULTS: Significant elevated HOTAIR and reduced miR-124 were presented in UUO rats and TGF-ß1-induced HK-2 cells in a time-dependent manner, with the increased Jagged1 (JAG1), Notch1, NICD, α-SMA and FN, as well as the decreased E-cadherin (all P < 0.05). Compared with the TGF-ß1 group, cells in the si-HOTAIR+TGF-ß1 group were remarkably declined in cell proliferation and the protein expressions of JAG1, Notch1, NICD, α-SMA, and FN, but dramatically higher in E-cadherin expression (all P < 0.05). However, in comparison with the si-HOTAIR+TGF-ß1 group, cells in the si-HOTAIR+miR-124 inhibitor+TGF-ß1 group were apparently improved in proliferation and the protein expression of JAG1, Notch1, NICD, α-SMA, and FN, but substantially reduced in the level of E-cadherin protein (all P < 0.05). CONCLUSION: Silencing lncRNA HOTAIR can up-regulate miR-124 to block Notch1 pathway, and thereby alleviating EMT and RIF, indicating HOTAIR as a potential target for RIF treatment.

[Health Risk Assessment of Tunnel Workers Based on the Investigation and Analysis of Occupational Exposure to PM10].

The health risk of tunnel workers' occupational exposure to PM10, was evaluated applying public health exposure evaluation nodel. A questionnaire survey of 250 tunnel workers was conducted in a construction site of Ma-zhu Highway in Hubei Province, and the concentrations of PM10 were monitored. The results showed that the PM10 exposure concentrations of different types of tunnel workers were extremely high. Compared with the limited value, the PM10 exposure concentrations were 83 times, 18 times, 8 times, 9 times Emd 9 times for excavation workers, blasting workers, supporting workers, slag-out workers and secondary-lining workers, respectively. For secondary-lining workers, the average daily exposure time was the longest, which was 11.48 h x d(-1), and the energy metabolism rate was also the highest, which was 1067.43 kj x (m2 x h)(-1). Regarding the inhalation rates, secondary-lining workers could be classified to high-level working intensity, and the other four types of tunnel workers could he classified to middle-level working intensity. The health risk assessment results showed that all tunnel workers had health risk. High exposure concentration of PM10 was the main reason for excavation workers' highest hazard quotient, and it was the same for the blasting workers. The reason for secondary-lining workers' high hazard quotient was that they had higher inhalation rates and longer average daily exposure time. In order to reduce the health risk of tunnel workers, firstly the workers should be equipped with appropriate respiratory protective equipment; secondly an appropriate tunnel working standard should be developed to set a reasonable working-years for reducing the exposure time.