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1.
Article in Chinese | WPRIM | ID: wpr-1025342

ABSTRACT

Objective:To observe and verify the changes of transcriptome in hyperoxia-induced acute lung injury (HALI), and to further clarify the changes of pathways in HALI.Methods:Twelve healthy male C57BL/6J mice were randomly divided into normoxia group and HALI group according to the random number table, with 6 mice in each group. The mice in the normoxia group were fed normally in the room, and the mice in the HALI group was exposed to 95% oxygen to reproduce the HALI animal model. After 72 hours of hyperoxia exposure, the lung tissues were taken for transcriptome sequencing, and then Kyoto Encyclopedia of Genes and Genomes database (KEGG) pathway enrichment analysis was performed. The pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to verify the key molecules in the signal pathways closely related to HALI identified by transcriptomics analysis.Results:Transcriptomic analysis showed that hyperoxia induced 537 differentially expressed genes in lung tissue of mice as compared with the normoxia group including 239 up-regulated genes and 298 down-regulated genes. Further KEGG pathway enrichment analysis identified 20 most significantly enriched pathway entries, and the top three pathways were ferroptosis signaling pathway, p53 signaling pathway and glutathione (GSH) metabolism signaling pathway. The related genes in the ferroptosis signaling pathway included the up-regulated gene heme oxygenase-1 (HO-1) and the down-regulated gene solute carrier family 7 member 11 (SLC7A11). The related genes in the p53 signaling pathway included the up-regulated gene tumor suppressor gene p53 and the down-regulated gene murine double minute 2 (MDM2). The related gene in the GSH metabolic signaling pathway was up-regulated gene glutaredoxin 1 (Grx1). The light microscope showed that the pulmonary alveolar structure of the normoxia group was normal. In the HALI group, the pulmonary alveolar septum widened and thickened, and the alveolar cavity shrank or disappeared. RT-RCR and Western blotting confirmed that compared with the normoxia group, the mRNA and protein expressions of HO-1 and p53 in lung tissue of the HALI group were significantly increased [HO-1 mRNA (2 -ΔΔCt): 2.16±0.17 vs. 1.00±0.00, HO-1 protein (HO-1/β-actin): 1.05±0.01 vs. 0.79±0.01, p53 mRNA (2 -ΔΔCt): 2.52±0.13 vs. 1.00±0.00, p53 protein (p53/β-actin): 1.12±0.02 vs. 0.58±0.03, all P < 0.05], and the mRNA and protein expressions of Grx1, MDM2, SLC7A11 were significantly decreased [Grx1 mRNA (2 -ΔΔCt): 0.53±0.05 vs. 1.00±0.00, Grx1 protein (Grx1/β-actin): 0.54±0.03 vs. 0.93±0.01, MDM2 mRNA (2 -ΔΔCt): 0.48±0.03 vs. 1.00±0.00, MDM2 protein (MDM2/β-actin): 0.57±0.02 vs. 1.05±0.01, SLC7A11 mRNA (2 -ΔΔCt): 0.50±0.06 vs. 1.00±0.00, SLC7A11 protein (SLC7A11/β-actin): 0.72±0.03 vs. 0.98±0.01, all P < 0.05]. Conclusions:HALI is closely related to ferroptosis, p53 and GSH metabolism signaling pathways. Targeting the key targets in ferroptosis, p53 and GSH metabolism signaling pathways may be an important strategy for the prevention and treatment of HALI.

2.
Chinese Critical Care Medicine ; (12): 980-983, 2023.
Article in Chinese | WPRIM | ID: wpr-1010895

ABSTRACT

OBJECTIVE@#To investigate the effect of hyperoxia on intestinal metabolomics in mice.@*METHODS@#Sixteen 8-week-old male C57BL/6 mice were randomly divided into hyperoxia group and control group, with 8 mice in each group. The hyperoxia group was exposed to 80% oxygen for 14 days. Mice were anesthetized and euthanized, and cecal contents were collected for untargeted metabolomics analysis by liquid chromatography-mass spectrometry (LC-MS) combined detection. Orthogonal partial least square discriminant analysis (OPLS-DA), volcano plot analysis, heat map analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the effects of hyperoxia on metabolism.@*RESULTS@#(1) OPLS-DA analysis showed that R2Y was 0.967 and Q2 was 0.796, indicating that the model was reliable. (2) Volcano plot and heat map analysis showed significant statistical differences in the expression levels of metabolites between the two groups, with 541 up-regulated metabolites, 64 down-regulated metabolites, and 907 no differences, while the elevated 5-hydroxy-L-lysine was the most significant differential metabolite induced by high oxygen. (3) KEGG pathway enrichment analysis showed that porphyrin and chlorophyll metabolism (P = 0.005), lysine degradation (P = 0.047), and aromatic compound degradation (P = 0.024) were the targets affected by hyperoxia. (4) Differential analysis of metabolic products through KEGG enrichment pathway showed that hyperoxia had a significant impact on the metabolism of porphyrin and chlorophyll, lysine, and aromatic compounds such as benzene and o-cresol.@*CONCLUSIONS@#Hyperoxia significantly induces intestinal metabolic disorders. Hyperoxia enhances the metabolism of porphyrins and chlorophyll, inhibits the degradation of lysine, and delays the degradation of aromatic compounds such as benzene and o-cresol.


Subject(s)
Mice , Male , Animals , Lysine , Hyperoxia , Benzene , Mice, Inbred C57BL , Metabolic Diseases , Oxygen , Chlorophyll , Porphyrins , Biomarkers/metabolism
3.
Chinese Critical Care Medicine ; (12): 344-348, 2021.
Article in Chinese | WPRIM | ID: wpr-883885

ABSTRACT

Objective:To explore whether microRNA-21-5p (miR-21-5p) has the effect of anti-apoptosis of human alveolar typeⅡ epithelial cells (ATⅡ).Methods:ATⅡ cells derived from the human were cultured in vitro and used for experiments when the cells were grown until the presence of lamellar bodies and microvilli were observed by light microscope. The cells were divided into blank control group (direct culture), hydrogen peroxide (H 2O 2) injury group (cultured with 0.5 mmol/L H 2O 2), and miR-21-5p overexpression group (using miR-21-5p with a multiplicity of infection (MOI) of 100 lentiviral overexpression vector with 0.5 mmol/L H 2O 2) and miR-21-5p empty virus control group (miR-21-5p lentiviral blank vector was co-cultured with 0.5 mmol/L H 2O 2). In each group, cell proliferation was detected by cell counting kit-8 (CCK-8) at 0, 12, 24, 36, and 48 hours of cell culture; cell apoptosis was detected by flow cytometry at 24 hours of culture. Results:① Cell proliferation activity test results: with the extension of cell culture time, the cell proliferation activity of the blank control group gradually increased, while the cell proliferation activity gradually decreased after the addition of 0.5 mmol/L H 2O 2. However, the cells proliferation activity in the miR-21-5p overexpression group decreased more slowly than that in the H 2O 2 injury group and the miR-21-5p empty virus control group, and the cell proliferation activity at 48 hours was significantly higher than the H 2O 2 injury group and the miR-21-5pempty virus control group ( A value: 0.295±0.005 vs. 0.184±0.005, 0.169±0.002, both P < 0.05). It showed that both H 2O 2 and lentivirus accelerated cell damage, while miR-21-5p could reduce cell apoptosis. ② Apoptosis rate test results: compared with the blank control group, the apoptosis rate increased significantly after adding 0.5 mmol/L H 2O 2; while the apoptosis rate of the miR-21-5p overexpression group was lower than that of the H 2O 2 injury group and miR-21-5p empty virus control group [early apoptosis rate: (14.31±0.12)% vs. (24.50±0.12)%, (23.41±0.13)%; late apoptosis rate: (8.12±0.13)% vs. (9.71±0.11)%, (10.41±0.15)%; overall apoptosis rate: (22.33±0.12)% vs. (34.21±0.10)%, (33.82±0.14)%; all P < 0.05], which further proved that miR-21-5p had anti-apoptotic effects. Conclusion:miR-21-5p has an anti-apoptotic effect on human ATⅡ.

4.
Chinese Journal of Digestion ; (12): 37-40, 2014.
Article in Chinese | WPRIM | ID: wpr-443474

ABSTRACT

Objective To explore the correlation between hyperlipidemia and colorectal polyps by compare the level of serum lipids in patients with colorectal polyps.Methods The levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein-cholesterol (LDL-C) of 159 patients with colorectal polyps and 138 controls were tested.The serum lipids between colorectal polyps group and control group,of colorectal polyps with different pathological type,of adenomatous polyps with different pathological type,of adenomatous polyps with different location,colorectal polyps of different gender were compared.Chi square test or t test were performed for data analysis.Results The incidence of hyperlipidemia of colorectal polyps group was 41.5% (66/159),which was higher than that of control group (16.7%,23/138) and the difference was statistically significant (x2 =36.56,P<0.01),its levels of TG,TC and LDL-C were all higher than those of the latter ((1.52±0.56) mmol/L vs (1.06 ± 0.42) mmol/L,(5.22±0.86) mmol/L vs (4.52±0.96) mmol/L,(2.85±0.66) mmol/L vs (2.52± 0.35) mmol/L; t=4.23,4.02,3.72,all P<0.01).There were no significant differences in the levels of TC,TG and LDL-C between colorectal polyps with different pathological type (all P> 0.05).The incidence of hyperlipidemia of tubular villous adenoma and villous adenoma (progressive adenomas) was 60.0% (15/25),which was higher than that of tubular adenoma group (33.3%,20/60) and the difference was statistically significant (x2=5.18,P<0.05).The incidence of hyperlipidemia of left colon and rectal polyps group was 46.2% (49/106),which was higher than that of right colon polyps group (28.6 %,12/42) and the difference was statistically significant (x2 =3.87,P<0.05).The incidence of hyperlipidemia of male colorectal polyps group was 47.2% (51/108),which was higher than that of female group (29.4%,15/51) and the difference was statistically significant (x2 =4.53,P<0.05).The level of TG of male colorectal polyps group was higher than that of female group ((1.84 ± 0.73) mmol/L vs (1.55±0.65) mmol/L) and the difference was statistically significant (t=3.98,P<0.05).Age (r=0.766,P=0.009),TG level (r=0.535,P=0.012) and TClevel (r=0.688,P=0.025) were positively correlated with genesis of colorectal polyps.Conclusion There is a significant correlation between hypertriglyceride,hypercholesteremia and colorectal polyps.

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