Overexpression of miR-155-5p Inhibits the Proliferation and Migration of IL-13-Induced Human Bronchial Smooth Muscle Cells by Suppressing TGF-β-Activated Kinase 1/MAP3K7-Binding Protein 2

Yujia Shi,Xingli Fu,Qi Cao,Zhengdao Mao,Yi Chen,Yun Sun,Zhiguang Liu,Qian Zhang 대한천식알레르기학회 2018 Allergy, Asthma & Immunology Research Vol.10 No.3

Purpose: Molecular mechanisms leading to asthma is still ill-defined. Though the function of microRNAs (miRNAs) in asthma was previously reported, the involvement of miR-155 in important features of this disease remains unknown. The present study was designed to uncover the probable involvement of miR-155-5p in the proliferation and migration of IL-13-induced human bronchial smooth muscle cells (BSMCs) and the intrinsic regulatory mechanism. Methods: The effects of different concentrations of IL-13 on the proliferation and migration of BSMCs as well as the expression of miR-155-5p and its predicted target transforming growth factor (TGF)-β-activated kinase 1/MAP3K7-binding protein 2 (TAB2) were investigated. The effects of miR-155-5p on the proliferation and migration of interleukin (IL)-13-induced BSMCs was determined in vitro using BSMCs transfected with miR-155 mimic/inhibitor and induced by a high concentration of IL-13. The quantitative real-time polymerase chain reaction (qRT-PCR) was employed for determining the expression of miR-155-5p and TAB2. Western blotting was applied to analyze the expression of TAB2 at the protein level. Cell proliferation and migration were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Transwell assays, respectively. Results: The proliferation and migration of BSMCs were dose-dependently increased with IL-13 treatment. Contrariwise, IL-13 dose-dependently inhibited the expression of miR-155-5p in BSMCs. Mechanistic studies showed that inhibition of miR-155-5p further promoted the stimulatory effects of IL-13, whereas overexpression of miR-155 significantly inhibited these effects. In silico studies and luciferase reporter assays indicated that TAB2 was a negatively regulated miR-155-5p target. Conclusions: These results suggested that miR-155-5p-inhibit the IL-13-induced proliferation and migration of BSMCs by targeting TAB2 and that the IL-13/miR-155/TAB2 pathway could serve as a therapeutic target for pulmonary diseases, especially asthma.

Effects of maize straw treated with various levels of CaO and moisture on composition, structure, and digestion by in vitro gas production

Shi Mingjun,Ma Zhanxia,Tian Yujia,Zhang Xuewei,Shan Huiyong 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.12

Objective: The objective of this study was to explore the effects of maize straw treated with calcium oxide (CaO) and various moisture, on the composition and molecular structure of the fiber, and gas production by fermentation in an in vitro rumen environment. Methods: The experiment used 4×3 Factorial treatment. Maize straws were treated with 4 concentrations of CaO (0%, 3%, 5%, and 7% of dry straw weight) and 3 moisture contents (40%, 50%, and 60%). Scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray fluorescence spectroscopy were employed to measure the surface texture, secondary molecular structure of carbohydrate, and calcium (Ca) content of the maize straw, respectively. The correlation of secondary molecular structures and fiber components of maize straw were analyzed by CORR procedure of SAS 9.2. In vitro rumen fermentation was performed for 6, 12, 24, 48, and 72 h to measure gas production. Results: Overall, the moisture factor had no obvious effect on the experimental results. Neutral detergent fiber (NDF), acid detergent fiber, acid detergent lignin, hemicellulose and cellulose contents decreased (p<0.05) with increasing concentrations of CaO treatment. Surface and secondary molecular structure of maize straw were affected by various CaO and moisture treatments. NDF had positive correlation (p<0.01) with Cell-H (H, height), Cell-A (A, area), CHO-2-H. Hemicellulose had positive correlation (p<0.01) with Lignin-H, Lignin-A, Cell-H, Cell-A. Ca content of maize straw increased as the concentration of CaO was increased (p<0.01). Gas production was highest in the group treated with 7% CaO. Conclusion: CaO can adhere to the surface of the maize straw, and then improve the digestibility of the maize straw in ruminants by modifying the structure of lignocellulose and facilitating the maize straw for microbial degradation. Objective: The objective of this study was to explore the effects of maize straw treated with calcium oxide (CaO) and various moisture, on the composition and molecular structure of the fiber, and gas production by fermentation in an in vitro rumen environment.Methods: The experiment used 4×3 Factorial treatment. Maize straws were treated with 4 concentrations of CaO (0%, 3%, 5%, and 7% of dry straw weight) and 3 moisture contents (40%, 50%, and 60%). Scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray fluorescence spectroscopy were employed to measure the surface texture, secondary molecular structure of carbohydrate, and calcium (Ca) content of the maize straw, respectively. The correlation of secondary molecular structures and fiber components of maize straw were analyzed by CORR procedure of SAS 9.2. In vitro rumen fermentation was performed for 6, 12, 24, 48, and 72 h to measure gas production.Results: Overall, the moisture factor had no obvious effect on the experimental results. Neutral detergent fiber (NDF), acid detergent fiber, acid detergent lignin, hemicellulose and cellulose contents decreased (p<0.05) with increasing concentrations of CaO treatment. Surface and secondary molecular structure of maize straw were affected by various CaO and moisture treatments. NDF had positive correlation (p<0.01) with Cell-H (H, height), Cell-A (A, area), CHO-2-H. Hemicellulose had positive correlation (p<0.01) with Lignin-H, Lignin-A, Cell-H, Cell-A. Ca content of maize straw increased as the concentration of CaO was increased (p<0.01). Gas production was highest in the group treated with 7% CaO.Conclusion: CaO can adhere to the surface of the maize straw, and then improve the digestibility of the maize straw in ruminants by modifying the structure of lignocellulose and facilitating the maize straw for microbial degradation.

Genipin-Crosslinked, Immunogen-Reduced Decellularized Porcine Liver Scaffold for Bioengineered Hepatic Tissue

Xiujuan Wu,Yujia Wang,Qiong Wu,Yi Li,Li Li,Jing Tang,Yujun Shi,Hong Bu,Ji Bao,Mingjun Xie 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.6

Liver disease affects millions of patients each year worldwide. Decellularized biologic matrices are plausible biomedical materials for bioengineered replacement hepatic tissue. However, one of the concerns for its safe medical application is the lack of objective assessment of the immunogen within the materials and in vivo immune responses to the matrices. The purpose of this study was to produce immunogen- reduced and biocompatible matrices from porcine liver. Whole porcine livers were perfusion decellularized and cross-linked with glutaraldehyde (GA) or genipin (GP). Proteins were extracted, and the migratory response of human leukocytes toward protein extracts was examined using an in vitro migration chamber. In addition, biopsy specimens of decellularized scaffolds were implanted subcutaneously into rodents to investigate scaffold immunogenicity. Histological staining confirmed cellular clearance from pig livers, with removal of nuclei and cytoskeletal components and widespread preservation of structural extracellular molecules. Polymerase chain reaction analysis showed that galactose-alpha-1,3-galactose-beta-1,4-N-acetylglucosamine (1,3 gal), swine leukocyte antigen, and porcine endogenous retrovirus were completely removed in the matrices. Decellularization significantly reduced the migration of monocytes compared with native porcine tissue. Although the proportion of transmigrating lymphocytes was much lower, repeating the cross-linking procedure reduced the migratory response. After implantation for 4 weeks, the decellularized and native samples were degraded, and the GA-treated group demonstrated a severe inflammatory reaction; however, minimal inflammatory cell infiltration was seen in the GPtreated group during the 8-week investigation period. In conclusion, our study provided evidence that GP crosslinking could significantly reduce the immunogenicity of decellularized liver biomaterials.