Transcriptome Analysis of Rat Lungs Exposed to Moxa Smoke after Acute Toxicity Testing

The increasing use of moxibustion has led to a debate concerning the safety of this treatment in human patients. Inhalation of cigarette smoke induces lung inflammation and granulomas, the proliferation of alveolar epithelial cells, and other toxic effects; therefore, it is important to assess the influence of inhaled moxa smoke on the lungs. In the present study, a novel poisoning cabinet was designed and used to assess the acute toxicity of moxa smoke in rats. We evaluated pathological changes in rat lung tissue and analyzed differentially expressed genes (DEGs) using RNA-seq and transcriptomic analyses. Our results show that the maximum tolerable dose of moxa smoke was 290.036 g/m³ and LC50 was 537.65 g/m³. Compared with that of the control group, the degree of inflammatory cell infiltration in the lung tissues of group A rats (all dead group) was increased, while that in group E rats (all live group) remained unchanged. GO and KEGG enrichment analyses showed that the DEGs implicated in cell components, binding, and cancer were significantly enriched in the experimental groups compared with the profile of the control group. The expressions of MAFF, HSPA1B, HSPA1A, AOC1, and MX2 determined using quantitative real-time PCR were similar to those determined using RNA-seq, confirming the reliability of RNA-seq data. Overall, our results provide a basis for future evaluations of moxibustion safety and the development of moxibustion-based technology.

Download Full-text

Maternal factor PABPN1L is essential for maternal mRNA degradation during maternal-to-zygotic transition

10.1101/2020.08.20.258830 ◽

2020 ◽

Author(s):

Ying Wang ◽

Tianhao Feng ◽

Xiaodan Shi ◽

Siyu Liu ◽

Zerui Wang ◽

...

Keyword(s):

Mouse Model ◽

Large Scale ◽

Mrna Degradation ◽

Female Infertility ◽

Control Group ◽

Rna Seq ◽

Cell Stage ◽

Maternal Mrna ◽

Maternal To Zygotic Transition ◽

Group A

AbstractInfertility affects 10% - 15% of families worldwide. However, the pathogenesis of female infertility caused by abnormal early embryonic development is not clear. We constructed a mouse model (Pabpn1l -/-) simulating the splicing abnormality of human PABPN1L and found that the female was sterile and the male was fertile. The Pabpn1l -/- oocytes can be produced, ovulated and fertilized normally, but cannot develop beyond the 2-cell stage. Using RNA-Seq, we found a large-scale upregulation of RNA in Pabpn1l -/- MII oocytes. Of the 2401 transcripts upregulated in Pabpn1l-/- MII oocytes, 1523 transcripts (63.4%) were also upregulated in Btg4 -/- MII oocytes, while only 53 transcripts (2.2%) were upregulated in Ythdf2 -/- MII oocytes. We documented that transcripts in zygotes derived from Pabpn1l -/- oocytes have a longer poly(A) tail than the control group, a phenomenon similar to that in Btg4-/- mice. Surprisingly, the poly(A) tail of these mRNAs was significantly shorter in the Pabpn1l -/- MII oocytes than in the Pabpn1l +/+. These results suggest that PABPN1L is involved in BTG4-mediated maternal mRNA degradation, and may antagonize poly(A) tail shortening in oocytes independently of its involvement in maternal mRNA degradation. Thus, PABPN1L variants could be a genetic marker of female infertility.

Download Full-text

The Biocompatibility and Security of Magnetic Nanoparticles Fe3O4-DNR Used In Hematologic Malignancies Therapy

Blood ◽

10.1182/blood.v116.21.3970.3970 ◽

2010 ◽

Vol 116 (21) ◽

pp. 3970-3970

Author(s):

Bao-An Chen ◽

Wei-wei Wu ◽

Jian Cheng ◽

Jun Wang ◽

Feng Gao ◽

...

Keyword(s):

Magnetic Nanoparticles ◽

Acute Toxicity ◽

Toxicity Testing ◽

Hematologic Malignancies ◽

Control Group ◽

Significant Difference ◽

Magnetic Nanoparticles Fe3o4 ◽

Experimental Group ◽

Fe3o4 Mnps ◽

Acute Toxicity Testing

Abstract Abstract 3970 The objective of this paper is to study the biocompatibility and security of self-prepared magnetic nanoparticles Fe3O4 (Fe3O4-MNPs) loaded with daunorubicin (DNR), which has the potential application in hematologic malignancies therapy. Hemolysis test was carried out to estimate it s blood toxicity; Fe3O4-MNPs loaded with DNR were intra peritoneally injected into mouse to calculate the LD50; micronucleus (MN) assay was reckoned to identify its genotoxicity; acute toxicity testing was evaluated its influence to mouse hepatic and renal functions. The result of hemolysis rate (HR) of Fe3O4-MNPs loaded with DNR was 2.908%, far less than 5%. Therefore, we concluded that self-prepated Fe3O4-MNPs loaded with DNR nanoparticles had no hemolytic reaction, and they consistent with the requirement of hemolytic test of biomaterials. The LD50 of Fe3O4-MNPs loaded with DNR nanoparticles to the mice was 1009.71mg/kg (relative content of DNR was 10mg/kg) and the 95% confidence interval was 769.11∼1262.40mg/kg, it had no significant difference compared with LD50 of using DNR only, which data was 8.51mg/kg and the 95% confidence interval was 6.48∼10.37mg/kg, and it had wide safety value circumscription. In micronucleus assay, compared Fe3O4-MNPs loaded with DNR nanoparticles experimental groups with negative control group, we found that the result had no significant difference (P > 0. 05) in micronucleus formation rate, while compared experimental groups with positive control group, the result had significant difference (P < 0. 05). The result indicated that Fe3O4 magnetic nanoparticles had no cacogenesis and mutagenesis. Acute toxicity testing showed that mice body weigh of control group, Fe3O4-MNPs loaded with DNR nanoparticles experimental group and isodose DNR group had no significant difference in 24h, 48h, and 72h after intra peritoneally injection; they had normal activity, eating and evacuation; toxic reactions such as instability of gait, convulsion, paralysis and respiratory depression were not been found; the alanine transarninase (ALT), blood urea nitrogen (BUN), and creatinine clearance rate (CCr) of Fe3O4-MNPs loaded with DNR nanoparticles experimental group was 66.0±28.55u/L, 9.06±1.05mmol/L, and 18.03±1.84umol/L, respectively, which had no significant difference compared with control group and isodose DNR group. From the results of our experiment, we could consider that self-prepared Fe3O4-MNPs loaded with DNR nanoparticles is a kind of high biocompatibility and security materials and perhaps is suitable for further application in hematologic malignancies therapy. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

The Role Of Epithelial To Mesenchymal Transition In The Remodeling Of Lung Injury Induced By Acute Graft-Versus-Host Disease

Blood ◽

10.1182/blood.v122.21.3257.3257 ◽

2013 ◽

Vol 122 (21) ◽

pp. 3257-3257

Author(s):

Meiqing Wu ◽

Can Liu ◽

Ke Zhao ◽

Xiuli Wu ◽

Yu Zhang ◽

...

Keyword(s):

Public Health ◽

Lung Injury ◽

Natural Science ◽

Research Funding ◽

Science And Technology ◽

Alveolar Epithelial Cells ◽

Control Group ◽

Technology Program ◽

Alveolar Epithelial ◽

National Public Health

Abstract Background Noninfectious lung injury caused by graft-versus-graft disease (GVHD) is a life-threatening complication after allogeneic hematopoietic stem cell transplant (allo-HSCT). Epithelial injury is a central event in the pathogenesis of noninfectious lung injury. Recent studies have shown that alveolar epithelial cells are able to self-renew and reestablish a functional alveolar epithelium. In the inflammatory and fibrotic lung diseases, differentiated epithelia also can acquire a myofibroblast phenotype in the process termed epithelial to mesenchymal transition (EMT), which contributes to aberrant healing and fibrosis. Many factors such as inflammatory cytokine TGF-β have been suggested to induce EMT. However, the role of EMT in the remodeling of acute GVHD (aGVHD) induced lung injury is unclear. Methods BALB/c mice were lethally irradiated and transplanted T cell-deleted (TCD) bone marrow plus whole spleen cells from C57BL/6 mice as aGVHD group, and only transplanted TCD bone marrow cells as control group. Alveolar epithelial cells were isolated from mice of two groups and Ep-CAM expression was measured by flow cytometry. The mRNA expression of cytokines including IFN-γ, TNF-α and TGF-β was detected by RT-PCR. The mRNA and protein expressions of specific markers, including E-cadherin, vimentin, Snail and surfactant proteins (SP)-C in lung tissue, were detected by RT-PCR and western blot. Results All mice in the aGVHD group showed diffuse periluminal infiltrates and parenchymal pneumonitis by histopathology, while the mice in the control group did not show any lung injury evidence. IFN-γ, TNF-α, TGF-β mRNA expressions were markedly up-regulated in the lung injury group as compared to the control group (P=0.045, P=0.032, P=0.025). Alveolar epithelial cells of injured lung expressed higher Ep-CAM (P=0.017) and lower SP-C (P=0.023). RT-PCR and western blot analyses revealed a significant decrease in epithelial marker E-cadherin (P=0.029) and increase in mesenchymal marker vimentin (P=0.026) in the GVHD damaged lung. Snail, a key EMT related transcription factor, was significantly elevated at mRNA and protein level in comparison to control group (P=0.015). Conclusion EMT is involving in the remodeling of lung injury induced by aGVHD. TGF-β is demonstrated to induce EMT. Whether up-regulation of IFN-γ and TNF-α contributes to EMT is deserved to be further explored. Disclosures: Wu: 863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Liu:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Zhao:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Wu:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Zhang:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Fan:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Fan:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Yin:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Zheng:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Yi:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding. Liu:863 Program (No. 2011AA020105) and National Public Health Grand Research Foundation ( No. 201202017): Research Funding; National Natural Science Foundation of China (Grant No.81000231, No.81270647) and Science and Technology Program of Guangzhou of China (11A72121174): Research Funding.

Download Full-text

Transcriptome Sequencing Reveals Gene Expression Differences in the Response of Malignant Melanomas Cells to Hypoxia

10.21203/rs.3.rs-589732/v1 ◽

2021 ◽

Author(s):

Haiting Xu ◽

Huazhen Liu ◽

Zi Li ◽

Qin Xu ◽

Nan Lin ◽

...

Keyword(s):

Tumor Progression ◽

Luciferase Assay ◽

Control Group ◽

Rna Seq ◽

Positive Effects ◽

Cell Hypoxia ◽

Group A ◽

Hypoxic Group ◽

Lower Expression

Abstract BackgroundMalignant melanoma (MM) is the most deadly type of skin cancer, with 5-year survival rate of less than 16%. HIF-1α overexpression is associated with poor prognosis in many cancers including MM. Hence, we characterized differentially expressed genes (DEGs) in the response of MM cells to normal and hypoxia. MethodsWe first successfully constructed cell hypoxia model and then performed RNA-seq to explore the changes of gene transcription in MM cells during hypoxia. The highest expression of the six genes was detected using qRT-PCR and western blot assays. We explored the binding sites between BIRC7 promoter and HIF-1α by dual-luciferase assay. Cellular function assays were used to observe the role of BIRC7 in the effect of hypoxia on tumor progression. ResultsWe found that compared with the transcriptome data of the control group, a total of 2601 DEGs were identified in the hypoxic group. There were 1517 genes with significantly higher expression and 1084 genes with lower expression in the hypoxic group. Among them, OSCAR, BIRC7, HBA1, SFN, GOLT1A, and BEX2 were significantly up-regulated in the hypoxic group. BIRC7 expression was most significantly up-regulated and a downstream factor of HIF-1α. We highlighted that knockdown of BIRC7 reversed the positive effects of HIF-1α on A875 and M14 cells. ConclusionsOur findings demonstrated that BIRC7 was a downstream factor of HIF-1α and reversed the effect of hypoxia on promoting tumor progression of MM cells.

Download Full-text

Time-resolved dual RNA-Seq reveals extensive rewiring of lung epithelial and pneumococcal transcriptomes during early infection

10.1101/048959 ◽

2016 ◽

Author(s):

Rieza Aprianto ◽

Jelle Slager ◽

Siger Holsappel ◽

Jan-Willem Veening

Keyword(s):

Epithelial Cells ◽

Pneumococcal Infection ◽

Alveolar Epithelial Cells ◽

Lung Epithelial Cells ◽

Infection Model ◽

Rna Seq ◽

Time Resolved ◽

Alveolar Epithelial ◽

Lung Epithelial ◽

Host Microbe Interactions

AbstractStreptococcus pneumoniae(pneumococcus) is the main etiological agent of pneumonia. Pneumococcal pneumonia is initiated by bacterial adherence to lung epithelial cells. Infection to the epithelium is a disruptive interspecies interaction involving numerous transcription-mediated processes. Revealing transcriptional changes may provide valuable insights into pneumococcal disease. Dual RNA-Seq allows simultaneous monitoring of the transcriptomes of both host and pathogen. Here, we developed a time-resolved infection model of human lung alveolar epithelial cells byS. pneumoniaeand assessed transcriptome changes by dual RNA-Seq. Our data provide new insights into host-microbe interactions and show that the epithelial glutathione-detoxification pathway is activated by bacterial presence. We observed that adherent pneumococci, not free-floating bacteria, access host-associated carbohydrates and repress innate immune responses. In conclusion, we provide a dynamic dual-transcriptomics overview of early pneumococcal infection with easy online access (http://dualrnaseq.molgenrug.nl). Further database exploration may expand our understanding of epithelial-pneumococcal interaction, leading to novel antimicrobial strategies.Graphical Abstract

Download Full-text

Haemato-biochemical alterations and acute toxicity study of hydro-methanol root extract of Anacardium occidentale in cockerels

Notulae Scientia Biologicae ◽

10.15835/nsb13410727 ◽

2021 ◽

Vol 13 (4) ◽

pp. 10727

Author(s):

Jacinta N. OMEKE ◽

Onyinyechukwu A. AGINA ◽

John I. IHEDIOHA ◽

Aruh O. ANAGA ◽

Wilfred S. EZEMA ◽

...

Keyword(s):

Acute Toxicity ◽

Serum Biochemistry ◽

Control Group ◽

Root Extract ◽

Internal Organs ◽

Serum Total Protein ◽

Biochemical Alterations ◽

Body Weights ◽

Group A ◽

Different Doses

The study evaluated the haemato-biochemical alterations and acute toxicity of hydro-methanol root extract of Anarcaduim occidentale (AO) in cockerels. Forty chickens randomly assigned into four groups (A, B, C and D) of 10 birds each were used for the experiment. Groups A, B, C and D received 3000, 1500 and 500 mg/kg body weight of the extract and distilled water respectively. Relative weights, haematology, serum biochemistry, gross and histopathological features of the lymphoid organs of chickens were studied. No significant (P>0.05) variation was observed in their body weights. The white blood cell count of group A chicken were significantly (P<0.05) lower than that of groups B and C chickens. Serum biochemistry tests revealed significant (P<0.05) variations at different doses for the alanine aminotransferase, alkaline phosphatase, serum total protein, albumin, cholesterol, triacylglycerol, uric acid, creatinine, calcium and phosphorus activities when compared to the control group. Gross and histopathological examinations showed no overt alterations in the tissues examined. It was observed that varied doses of root extract of Anarcaduim occidentale caused significant decrease in haematology and serum biochemistry parameters of chickens and no obvious lesions in internal organs.

Download Full-text

Acute toxicity test of 96% ethanol extract of Syzygium myrtifolium leaves in white mice (Mus musculus)

Pharmacy Education ◽

10.46542/pe.2021.212.201204 ◽

2021 ◽

pp. 201-204

Author(s):

Lusi Indriani ◽

E. Mulyati Effendi ◽

Kevin Christofer Fadillah

Keyword(s):

Acute Toxicity ◽

Oral Administration ◽

Abdominal Cavity ◽

Lethal Dose ◽

Toxicity Testing ◽

Ethanol Extract ◽

Negative Control Group ◽

Negative Control ◽

Toxic Effects ◽

Control Group

Introduction: Acute toxicity effects appear within a short time following the oral administration of either a single dose or repeated doses of toxin within 24 hours. Acute toxicity testing involves the administration of a range of doses across several groups of experimental animals with one dose administered per group, followed by the observation of toxic effects and mortality. Objectives: The purpose of this study was to determine the lethal dose 50 (LD50) and acute toxicity of an ethanol extract of Syzygium myrtifolium leaves in white mice. Methods: Exposed groups consisted of a negative control group (carboxymethylcellulose sodium) and four treatment groups (500, 1000, 2000, and 4000 mg/kg body weight (bw)). Mortality was observed for 14 days following oral administration. Results: The results demonstrated an LD50 of 1995 mg/kg bw, categorised as moderately toxic. Observed toxic effects included white lesions in the lungs, blackened liver, organ swelling, and fluid accumulation in the abdominal cavity and thorax.

Download Full-text

Acute toxicity test ethanol extract of kerehau leaf (Callicarpa longifolia Lamk) using OECD 420 method

Jurnal Ilmiah Farmasi ◽

10.20885/jif.vol16.iss2.art2 ◽

2020 ◽

Vol 16 (2) ◽

pp. 105-111

Author(s):

Aulia Istiqomah ◽

◽

Shintya Safitri ◽

Elis Susilawati

Keyword(s):

Acute Toxicity ◽

Toxicity Test ◽

Toxicity Testing ◽

Ethanol Extract ◽

Liver Weight ◽

Control Group ◽

Test Substance ◽

Acute Toxicity Test ◽

Safety Level ◽

Ld50 Value

Background: Kerehau is empirically used as cooling powder to treat acne. Based on previous research, kerehau leaves have a number of activities, including wound healing, antidiabetic, and anti-inflammatory. Objectives: This study aimed to determine the safety level of a single dose of ethanol extract of kerehau leaves within a 24-hour administration period. Methods: Acute toxicity testing of ethanol extract of kerehau leaves was carried out on female Webster mice. Test animals were divided into 4 treatment groups, consisting of 1 control group and 3 test groups. The testing method referred to OECD (Organization of Economic Cooperation and Development) 420 with modified doses of 2000mg/kgBW, 5000 mg/kgBW, and 8000 mg/kgBW. The observations were made on the behaviour of animals towards toxicity symptoms for 4 hours after administration of the test substance as well as death. The death and weight gain were observed for 14 days. On the 15th day, animals were sacrificed, blood was taken, and biochemical parameters were measured. The heart, kidneys, liver, spleen, and lungs were harvested and weighed. Data was analysed using Oneway ANOVA continued with LSD and Post Hoc. Results: The results showed that ethanol extract of kerehau leaves did not cause death in all of the groups. There were significant differences in liver weight, SGPT, and SGOT levels (p< 0.05) at the dose of 5000 mg/kgBW. No toxicity symptoms and death were found until the end of the experiment. Conclusion: LD50 value of ethanol extract of kerehau leaves was above 8000 mg/kgBW with heart as the most affected organ. Keywords: Ethanol extract of kerehau leaves, acute toxicity test, OECD 420.

Download Full-text

Antiplasmodial Activity of Seed Oil of Moringa oleifera (Lam.)

Journal of Advances in Medical and Pharmaceutical Sciences ◽

10.9734/jamps/2021/v23i130214 ◽

2021 ◽

pp. 24-32

Author(s):

Dada, Ebenezer Oluyemi ◽

Abdulahi, Sikiru Kayode

Keyword(s):

Acute Toxicity ◽

Toxicity Test ◽

Moringa Oleifera ◽

Seed Oil ◽

Treated Group ◽

Positive Control ◽

Antiplasmodial Activity ◽

Control Group ◽

Acute Toxicity Test ◽

Group A

The study was carried out to determine the antiplasmodial activity of seed oil of Moringa oleifera extracted using n-hexane solvent. Acute toxicity test was carried out on twelve albino mice grouped into 3 according to seed oil concentrations (250, 500 and 1000 mg/kg respectively) with each group having four mice. Thirty-five mice were infected with Plasmodium berghei and the mice were randomized into five groups of seven mice each (groups A, B, C, D and E) for antiplasmodial activity. Group A - negative control (non-treated), group B - positive control (chloroquine treated); group C (800 mg/kg), group D (400 mg/kg) and group E (200 mg/kg) of seed oil of Moringa oleifera. All groups were left untreated until after five days when 0.2 mL treatment dose for each group was administered. Treatment was carried out in four days and left for another five days for post treatment effect. The acute toxicity test showed that the seed oil of Moringa oleifera was safe and nontoxic to all mice. There was daily gradual reduction in PCV values; however group with the highest suppression of parasitemia had the highest PCV value after treatment. Group A as expected had the lowest PCV value of 22.23±1.98% which fell short of normal range (40 - 55%) and had its parasitemia load increased by 205% while in other groups, B, C, D and E the parasitemia had decreased by 100%, 97.02%, 90.48% and 67.65%respectively after treatment. Overall, the seed oil of M. oleifera at high concentrations showed a competitive parasite inhibition activity when compared with the result obtained in positive control group; however, few deaths recorded during and after treatment called for further investigation to determine its suitability for the treatment of malaria.

Download Full-text

Transcriptomics, metabolomics and lipidomics of chronically injured alveolar epithelial cells reveals similar features of IPF lung epithelium

10.1101/2020.05.08.084459 ◽

2020 ◽

Author(s):

Willy Roque ◽

Karina Cuevas-Mora ◽

Dominic Sales ◽

Wei Vivian Li ◽

Ivan O. Rosas ◽

...

Keyword(s):

Epithelial Cells ◽

Mitochondrial Dysfunction ◽

Enrichment Analysis ◽

Alveolar Epithelial Cells ◽

Control Group ◽

Lung Epithelium ◽

Chronic Injury ◽

Alveolar Epithelial ◽

Novel Treatments

AbstractThe current hypothesis suggests that Idiopathic pulmonary fibrosis (IPF) arises as a result of chronic injury to alveolar epithelial cells and aberrant activation of multiple signaling pathways. Dysfunctional IPF lung epithelium manifests many hallmarks of aging tissues, including cellular senescence, mitochondrial dysfunction, metabolic dysregulation, and loss of proteostasis. Unfortunately, this disease is often fatal within 3-5 years from diagnosis, and there is no effective treatment. One of the major limitations to the development of novel treatments in IPF is that current models of the disease fail to resemble several features seen in elderly IPF patients. In this study, we sought to develop an in vitro epithelial injury model using repeated low levels of bleomycin to mimic the phenotypic and functional characteristics of the IPF lung epithelium. Consistent with the hallmarks of the aging lung epithelium, we found that chronic-injured epithelial cells exhibited features of senescence cells, including an increase in β-galactosidase staining, induction of p53 and p21, mitochondrial dysfunction, excessive ROS production, and proteostasis alteration. Next, combined RNA sequencing, untargeted metabolomics, and lipidomics were performed to investigate the dynamic transcriptional, metabolic, and lipidomic profiling of our in vitro model. We identified that a total of 8,484 genes with different expression variations between the exposed group and the control group. According to our GO enrichment analysis, the down-regulated genes are involved in multiple biosynthetic and metabolic processes. In contrast, the up-regulated genes in our treated cells are responsible for epithelial cell migration and regulation of epithelial proliferation. Furthermore, metabolomics and lipidomics data revealed that overrepresented pathways were amino acid, fatty acid, and glycosphingolipid metabolism. This result suggests that by using our in vitro model, we were able to mimic the transcriptomic and metabolic alterations of those seen in the lung epithelium of IPF patients. We believe this model will be ideally suited for use in uncovering novel insights into the gene expression and molecular pathways of the IPF lung epithelium and performing screening of pharmaceutical compounds.

Download Full-text