Circular RNA ANKRD36 attends to lipopolysaccharide-aroused MRC-5 cell injury via regulating microRNA-31-3p.

BACKGROUND: Some circular RNAs (circRNAs) are reported to attend to the pathogenesis of pneumonia. This study tested the impact of circRNA ankyrin repeat domain 36 (circANKRD36) on human embryonic lung fibroblast MRC-5 cell injury irritated by lipopolysaccharide (LPS). METHODS: After LPS irritation, viability, apoptosis, ROS, protein, and cytokines, along with circANKRD36 were tested by CCK-8, Annexin V-FITC, DCFH-DA, and ELISA or Western blot. si-circANKRD36 and microRNA-31-3p/5p (miR-31-3p/5p) inhibitor were applied to silence circANKRD36 and miR-31-3p/5p. miR-31-3p/5p mimic was utilized to upregulate miR-31-3p/5p. RT-qPCR was used to detect miRNAs. The relationship between miRNAs and MyD88 or IL-34 was analyzed by luciferase activity reporter assay. RESULTS: LPS aroused a decrease in viability, increases in apoptosis, ROS, and IL-6, IL-8, and TNF-α, along with circANKRD36, and activation of NF-κB pathway. Silencing circANKRD36 weakened the above-mentioned influences of LPS. Moreover, silencing circANKRD36 hoisted miR-31-3p expression. Silencing miR-31-3p mitigated the impacts of circANKRD36 silence on LPS-irritated MRC-5 cells. Besides, MyD88 was a downstream target of miR-31-3p, and 3'UTR of IL-34 mRNA was targeted by miR-31-5p. LPS induced the accumulation of MyD88. Silencing MyD88 was constructive to maintain cell viability, retard apoptosis and inhibit adverse oxidation and inflammation. CONCLUSION: This research verified that silencing circANKRD36 could weaken LPS-irritated MRC-5 cell injury via regulating miR-31/MyD88-mediated repression of NF-κB pathway.

Repression of TSC1/TSC2 mediated by MeCP2 regulates human embryo lung fibroblast cell differentiation and proliferation.

Pulmonary fibrosis (PF) is a severe inflammatory disease with limited effective treatments. It is known that the transdifferentiation of human embryo lung fibroblast (HELF) cells from pulmonary fibroblasts into myofibroblasts, contributes to the progression of pulmonary fibrogenesis. The tuberous sclerosis proteins TSC1 and TSC2 are two key signaling factors which can suppress cell growth and proliferation. However, the roles of TSC1 and TSC2 in lung fibroblast are unclear. Here, we developed a PF model with bleomycin (BLM) in mice and conducted several simulation experiments in HELF cells. Our study shows that the expression of TSC1 and TSC2 in fibrotic mice lung was reduced and stimulation of HELF cells with TGF-ß1 resulted in a down-regulation of TSC1 and TSC2. In addition, overexpression of TSC1 or TSC2 decreased cell proliferation and differentiation. Furthermore, we found that reduced expression of TSC1 and TSC2 caused by TGF-ß1 is associated with the promoter methylation status of TSC1 and TSC2. MeCP2, controls an epigenetic pathway that promotes myofibroblast transdifferentiation and fibrosis. We found that expression of TSC1 and TSC2 can be repressed by MeCP2, which regulates HELF cell differentiation and proliferation as myofibroblasts and lead to PF ultimately.

Protective effects of lemongrass essential oil against benzo(a)pyrene-induced oxidative stress and DNA damage in human embryonic lung fibroblast cells.

Benzo(a)pyrene (BaP) was a well-known environmental pollutant, numerous studies had implicated BaP as a causative agent in human cancer, particularly lung cancer. The lemongrass essential oil (LEO) possessed various pharmacological activities, especially the anti-oxidative stress and cancer prevention. In the current study, human embryonic lung fibroblast (HELF) cells were treated with 25 mM BaP in the absence or presence of 0.5%, 1% or 2.5% LEO and the cell viability and levels of oxidative stress (OS) and DNA damage in the cells were then measured. Nineteen chemical constituents were identified in LEO, with citral being the main component, representing about 68.78%. LEO was able to protect the HELF cells against BaP-induced loss in cell viability, achieving a maximum of 95.58% cell viability at the 0.5% concentration. Treatment of HELF cells with BaP alone significantly increased the level of Malondialdehyde (MDA) and decreased superoxide dismutase (SOD) and catalase (CAT). However, these effects were suppressed when the cells were also treated with LEO, leading to enhanced levels of SOD and CAT activities (2.9- and 2-fold, respectively, compared with BaP treatment only) and reduced the level of MDA in the cells (43% reduction in malondialdehyde level). At the same time, LEO also reduced the level of DNA damage, as shown by a reduced level of 8-hydroxy-deoxyguanosine (8-OHdG). Taken together, the results showed that LEO offered protection against BaP-induced OS and DNA damage, suggesting that LEO could be a promising agent for lung cancer chemoprevention.

Effect of Salvia miltiorrhiza injection on proliferation, apoptosis and expression of amino acid in human embryonic lung fibroblast cells / 实用医学杂志

Objective To investigate the effect of Salvia miltiorrhiza injection on proliferation , apoptosis and expression of hydroxyproline in human embryonic lung fibroblast cells. Methods TGF-β1 was administered to induce the proliferation in human embryonic lung fibroblast cells; the effect of Salvia miltiorrhiza injection on the number of human embryonic lung fibroblast cells was detected by MTT method; ki67 expression by immunocytochemical method;cell apoptosis by flow cytometry and the expression of hydroxyproline by colorimetry method. Results TGF-β1(0, 2.5, 5, 10, 20, 40 μg/L)could up-regulate cell number of human embryonic lung fibroblast cells in a dose-dependent manner , while the OD value of human embryo lung fibroblasts cells declined pretreated with Salvia miltiorrhiza injection in a dose-dependent manner and Salvia miltiorrhiza injection could induce the apoptosis and down regulated hydroxyproline expression in human embryo lung fibroblasts cells. The results of flow cytometry indicated that cell apoptosis increased after treated with Salvia miltiorrhiza injection when compared with TGF-β1 group (P < 0.05). Meanwhile, Salvia miltiorrhiza injection could down regulate the expression of hydroxyproline (P < 0.01). Conclusions Salvia miltiorrhiza injection can target human embryonic lung fibroblast cells , play a potent role in the airway remodeling through the promotion of its apoptosis and down regulate the expression of hydroxyproline.

Effect of Adenovirus Type 3I,7b on Expressions of mRNA and Protein of Transforming Growth Factor-Beta 1 in Human Embryonic Lung Fibroblast Cells / 实用儿科临床杂志

Objective To study the effect of adenovirus type 3I,7b on the expressions of mRNA and protein of transforming growth factor-beta 1(TGF-?1) in human embryonic lung fibroblast cells.Method The expression of mRNA and protein of TGF-?1 were determined in human embryonic lung fibroblast cells before and after being infected by adenovirus type 3I,7b and in normal fibroblast cells with enzyme-linked immunosorbent assay and in situ hybridization.Results The mRNA and protein of TGF-?1 expression in human embryonic lung fibroblast cells increased siginificantly after being infected by adenovirous type 3I,7b compared with those in normal fibroblast cells(Pa0.05).Conclusion Lung fibroblast cells and TGF-?1 may play some roles in pathophysiological processes of viral pneumonia.

Effect of Qingfei Oral Liquid on Transforming Growth Factor-?_1 and Platelet-Derived Growth Factor-BB mRNA Gene Expression of Human Embryonic Lung Fibroblast Cells Infected by Adenovirus Type 3I and 7b / 中国中医药信息杂志

Objective To study the effect of Qingfei oral liquid contained serum on the transforming growth factor-?1 (TGF-?1) and platelet-derived growth factor-BB (PDGF-BB) mRNA gene expression of human embryonic lung fibroblast cells infected by adenovirus (ADV) type 3I and 7b. Method TGF-?1 and PDGF-BB mRNA expression of human embryonic lung fibroblast cells infected by ADV type 3I and 7b were determined by in situ hybridization before and after treated with Qingfei oral liquid contained serum. Results ADV could up-regulate TGF-?1 and PDGF-BB mRNA of human embryonic lung fibroblast cells (P