1.MicroRNA-26a-5p protects against podocyte injury via inhibiting TRPC6 expression in diabetic kidney disease
Yan ZHOU ; Haiyan ZHOU ; Nanchi LIU ; Yan XU ; Xingjian ZHANG ; Lin DING ; Ruixia MA
Chinese Journal of Nephrology 2022;38(4):336-343
Objective:To investigate the protective effect and potential mechanisms of microRNA-26a-5p (miR-26a-5p) on podocyte injury in diabetic kidney disease (DKD).Methods:(1) In vivo experiment: Four-week-old db/db mice were divided into db/db group, db/db+agomir-NC group and db/db+miR-26a-5p agomir group according to random number table method, with 10 mice in each group, and 10 db/m mice of the same week-old were set as normal control group. At the age of 10 weeks, pathological changes were observed through light and electron microscopy. Kidney weight/body weight (KW/BW), urinary albumin to creatinine ratio (ACR), fasting blood glucose (FBG) and other biochemical indicators were also detected. The position and expression of miR-26a-5p in kidney tissue were determined through fluorescence in situ hybridization and quantitative real-time PCR, while the expressions of transient receptor potential cation channel-6 (TRPC6) and Nephrin in kidney tissue were determined by Western blotting and immunohistochemistry. (2) In vitro experiment: The immortalized mouse podocytes (MPC5) were divided into 5 groups: normal glucose group, high mannitol group, high glucose group, high glucose+miR-26a-5p mimic group, and high glucose+mimic-NC group. The expressions of miR-26a-5p, TRPC6 and Nephrin were detected. Luciferase reporter assay was conducted to research the relationship of miR-26a-5p and TRPC6. Results:(1) In vivo experiment: Compared with db/m group, db/db mice exhibited lower KW/BW and disrupted conditions of ACR, FBG, total cholesterol, triglycerides and low density lipoprotein cholesterol (all P<0.01). Increased glomeruli volume, more extracellular matrix deposition, thicker basement membrane and more foot process fusion were observed by light and electron microscope. Increased expression of TRPC6 protein as well as decreased expression of Nephrin protein and miR-26a-5p were detected in kidney tissues of db/db mice ( P<0.05). Compared with db/db+agomir-NC group, db/db mice transfected by miR-26a-5p agomir exhibited less albuminuria, with less protein expression of TRPC6 and more Nephrin in kidney tissue (all P<0.05). (2) In vitro experiment: Compared with normal glucose group, high glucose-treated podocytes exhibited increased expression of TRPC6 ( P<0.05), as well as decreased expression of Nephrin ( P<0.05) and miR-26a-5p ( P<0.01). Compared with high glucose+mimic-NC group, lower expression of TRPC6 and higher expression of Nephrin were detected in podocytes transfected by miR-26a-5p mimic (both P<0.05). Luciferase reporter assay confirmed that miR-26a-5p could regulate the expression of TRPC6 precisely. Conclusions:The expression of miR-26a-5p in podocytes is down-regulated in the context of high glucose and miR-26a-5p protects podocytes from injury via inhibiting the expression of TRPC6 in DKD.
2.Research progress on the regulatory mechanism of Hippo signaling pathway in pulmonary fibrosis
Ruixia DING ; Ying SUN ; Xiaohui WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(9):716-720
Pulmonary fibrosis is a difficult to treat fibrotic disease with multiple triggering factors and complex pathogenesis. It is characterized by diffuse inflammatory damage, tissue structure destruction, and persistent fibrosis, resulting in irreversible damage to lung function. The Hippo signaling pathway is involved in regulating various biological processes such as cell proliferation, differentiation, migration, apoptosis, and is closely related to the occurrence of pulmonary fibrosis. In order to further explore the mechanism of pulmonary fibrosis, this paper comprehensively analyzes the Hippo signaling pathway and its cellular and pathological imbalance related to pulmonary fibrosis, revealing the influence of Hippo signaling pathway in pulmonary fibrosis and its possible mechanism of action, which is expected to provide new targets and strategies for the prevention and treatment of pulmonary fibrosis.
3.Research progress on the regulatory mechanism of Hippo signaling pathway in pulmonary fibrosis
Ruixia DING ; Ying SUN ; Xiaohui WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(9):716-720
Pulmonary fibrosis is a difficult to treat fibrotic disease with multiple triggering factors and complex pathogenesis. It is characterized by diffuse inflammatory damage, tissue structure destruction, and persistent fibrosis, resulting in irreversible damage to lung function. The Hippo signaling pathway is involved in regulating various biological processes such as cell proliferation, differentiation, migration, apoptosis, and is closely related to the occurrence of pulmonary fibrosis. In order to further explore the mechanism of pulmonary fibrosis, this paper comprehensively analyzes the Hippo signaling pathway and its cellular and pathological imbalance related to pulmonary fibrosis, revealing the influence of Hippo signaling pathway in pulmonary fibrosis and its possible mechanism of action, which is expected to provide new targets and strategies for the prevention and treatment of pulmonary fibrosis.
4.Study on serum proteomic characteristics of workers with rare earth samarium oxide pneumoconiosis
Xiaohui WANG ; Ruixia DING ; Teng MA ; Yannan BI ; Haijing YIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(11):807-814
Objective:To screen differential proteins in the serum of workers with rare earth samarium oxide pneumoconiosis, in order to provide new ideas for finding its early diagnostic biomarkers.Methods:In April 2019, three male workers diagnosed with samarium oxide pneumoconiosis at a rare earth factory in Baotou City, Inner Mongolia Autonomous Region were selected as the observation group, and three male workers who were not exposed to dust were selected as the control group. The serum was sequenced using the Label-free proteomic method to screen for differentially expressed proteins, followed by cluster of orthologous groups of proteins (COG) annotation, gene ontology (GO) enrichment analysis, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. The interaction gene library retrieval tool database and Cytoscape 3.9.1 software were used to draw protein-protein interaction networks. CytoHubba plugin was used to screen for differentially expressed proteins with high scores, and real-time fluorescence quantitative polymerase chain reaction (RT/q-PCR) was used to validate the proteomic sequencing results.Results:A total of 45 up-regulated differentially expressed proteins and 5 down-regulated differentially expressed proteins were screened out in the serum of workers with rare earth samarium oxide pneumoconiosis. In the COG functional classification, post-translational modifications, protein turnover, and chaperones were the most numerous. GO enrichment included 25 entries for biological processes such as complement activation (classical pathways), 15 entries for cellular components such as extracellular recombinants, and 10 entries for molecular functions such as protein binding. The pathways identified by KEGG enrichment analysis mainly included infectious diseases, immune system, signal transduction, and immune related diseases. The top 10 scoring proteins were haptoglobin, complement C1r subcomponent, complement C1s subcomponent, apolipoprotein C-Ⅲ, apolipoprotein A-Ⅱ, prothrombin, afamin, complement component C8 gamma chain, complement component C6, complement component C7. The RT/q-PCR validation results showed that the mRNA expression levels of haptoglobin, prothrombin and complement C1s subcomponent in the observation group were significantly higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusion:Ten differentially expressed proteins in the serum of workers with rare earth samarium oxide pneumoconiosis are screened, which provides a good idea for the screening of biomarkers for early diagnosis of samarium oxide pneumoconiosis.
5.Study on serum proteomic characteristics of workers with rare earth samarium oxide pneumoconiosis
Xiaohui WANG ; Ruixia DING ; Teng MA ; Yannan BI ; Haijing YIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(11):807-814
Objective:To screen differential proteins in the serum of workers with rare earth samarium oxide pneumoconiosis, in order to provide new ideas for finding its early diagnostic biomarkers.Methods:In April 2019, three male workers diagnosed with samarium oxide pneumoconiosis at a rare earth factory in Baotou City, Inner Mongolia Autonomous Region were selected as the observation group, and three male workers who were not exposed to dust were selected as the control group. The serum was sequenced using the Label-free proteomic method to screen for differentially expressed proteins, followed by cluster of orthologous groups of proteins (COG) annotation, gene ontology (GO) enrichment analysis, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. The interaction gene library retrieval tool database and Cytoscape 3.9.1 software were used to draw protein-protein interaction networks. CytoHubba plugin was used to screen for differentially expressed proteins with high scores, and real-time fluorescence quantitative polymerase chain reaction (RT/q-PCR) was used to validate the proteomic sequencing results.Results:A total of 45 up-regulated differentially expressed proteins and 5 down-regulated differentially expressed proteins were screened out in the serum of workers with rare earth samarium oxide pneumoconiosis. In the COG functional classification, post-translational modifications, protein turnover, and chaperones were the most numerous. GO enrichment included 25 entries for biological processes such as complement activation (classical pathways), 15 entries for cellular components such as extracellular recombinants, and 10 entries for molecular functions such as protein binding. The pathways identified by KEGG enrichment analysis mainly included infectious diseases, immune system, signal transduction, and immune related diseases. The top 10 scoring proteins were haptoglobin, complement C1r subcomponent, complement C1s subcomponent, apolipoprotein C-Ⅲ, apolipoprotein A-Ⅱ, prothrombin, afamin, complement component C8 gamma chain, complement component C6, complement component C7. The RT/q-PCR validation results showed that the mRNA expression levels of haptoglobin, prothrombin and complement C1s subcomponent in the observation group were significantly higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusion:Ten differentially expressed proteins in the serum of workers with rare earth samarium oxide pneumoconiosis are screened, which provides a good idea for the screening of biomarkers for early diagnosis of samarium oxide pneumoconiosis.