Inhibition of the ERK1/2-ubiquitous calpains pathway attenuates experimental pulmonary fibrosis in vivo and in vitro.

Idiopathic pulmonary fibrosis (IPF) is a fibrotic lung disease with poor prognosis. Epithelial-mesenchymal transition (EMT) has been reported to play an important role in IPF. The extracellular signal-regulated kinases 1 and 2 (ERK1/2) cascade, which regulates EMT and oncogenesis, has been implicated in the pathogenesis of IPF. Calpains, Ca2+-dependent cysteine proteinases that mediate controlled proteolysis of many specific substrates including epithelial cell marker E-cadherin, participate in organ fibrosis. Calpain-1 and calpain-2 of calpain family are ubiquitous calpains. ERK1/2 signaling stimulates the ubiquitous calpains activity in cancer development, but whether ERK1/2 signaling mediates the ubiquitous calpains activity in pulmonary fibrosis is unknown. Here we investigated whether inhibition of ERK1/2 signaling and the ubiquitous calpains attenuated experimental pulmonary fibrosis and examined the potential mechanism. Our results showed that inhibition of ERK1/2 signaling and the ubiquitous calpains both attenuated bleomycin (BLM)-induced lung fibrosis in mice. Inhibition of ERK1/2 signaling downregulated the expression of calpain-1 and calpain-2 in vivo and in vitro. We detected decreased E-cadherin expression and increased calpain-1 expression in IPF patients. Inhibition of ERK1/2 signaling and the ubiquitous calpains both suppressed the development of EMT in vivo and in vitro. Our study indicated that inhibition of the ERK1/2-ubiquitous calpains pathway protected pulmonary fibrosis from BLM, possibly via inhibition of EMT. Therefore, targeting ubiquitous calpains may be a potential strategy to attenuate IPF.

Two competitive INC-mediated reactions in the gas-phase fragmentation of protonated indolyl benzo[b]carbazoles.

RATIONALE: Ion-neutral complexes (INCs) are intermediates extensively existing in gas-phase ionic reactions. METHODS: Tandem mass spectrometry (MS(n) ) analysis of indolyl benzo[b]carbazoles was performed on an electrospray ionization quadrupole time-of-flight (ESI-Q-TOF) mass spectrometer in positive ion mode. RESULTS: Two competing INC-mediated reactions were obtained in the fragmentation of protonated indolyl benzo[b]-carbazoles. CONCLUSIONS: This study enriches our knowledge of the important roles of INCs in the dissociation of indole compounds in the gas phase. Copyright © 2016 John Wiley & Sons, Ltd.

Two competing ionization processes in electrospray mass spectrometry of indolyl benzo[b]carbazoles: formation of M⁺• versus [M + H]⁺.

RATIONALE: Ionization in electrospray ionization mass spectrometry (ESI-MS) mainly occurs as a result of acid-base reactions or coordination with metal cations. Formation of the radical cation M(+•) in the ESI process has attracted our interest to perform further investigation. METHODS: A series of indolyl benzo[b]carbazoles were investigated using a quadrupole ion trap mass spectrometer equipped with an ESI source or an atmospheric pressure chemical ionization (APCI) source in the positive-ion mode. Theoretical calculations were performed using the density functional theory (DFT) method at the B3LYP/6-31G(d) level. RESULTS: Both the radical ion M(+•) and the protonated molecule [M + H](+) were obtained by ESI-MS analysis of indolyl benzo[b]carbazoles, while only [M + H](+) was observed in the APCI-MS analysis. The relative intensities of M(+•) and [M + H](+) were significantly affected by several ESI operating parameters and the nature of the substituents. CONCLUSIONS: Formation of M(+•) and [M + H](+) was rationalized as two competing ionization processes in the ESI-MS analysis of indolyl benzo[b]carbazoles.

Unveiling the Knowledge Frontier: A Scientometric Analysis of COPD with Sarcopenia.

Objective: Numerous articles and reviews addressing the intersection of Chronic Obstructive Pulmonary Disease (COPD) with sarcopenia have been documented. However, a significant gap exists in the literature concerning scientometric analysis in this field. This study aimed to concentrate on recent research and elucidate emerging research areas through the examination of COPD with sarcopenia. Methods: Articles in the field were systematically retrieved from the Web of Science Core Collections (WoSCC) spanning from 2003 to 2022. The analysis employed scientometric and keyword analyses through specialized software, including VOSviewer, CiteSpace, and Origin. Results: A comprehensive analysis of 758 articles and reviews in the field of COPD with sarcopenia revealed the United States as the leading contributor in terms of publications and overall influence. Maastricht University emerged as the most prolific institution, with Schols Annemie M. W. J. being identified as the most influential scholar in this field. The International Journal of Chronic Obstructive Pulmonary Disease emerged as the most prolific journal. Notably, COPD with sarcopenia exhibits frequent associations with other diseases, underscoring the complexity of the topic and emphasizing the necessity for comprehensive treatment. Mechanistic and diagnostic aspects, particularly computed tomography, are pivotal in this research field. Osteoporosis emerges as a prospective avenue for future research, encompassing both COPD and sarcopenia. Furthermore, nutrition and physical activity are integral components for managing COPD patients with sarcopenia. Conclusion: This study delineates the distribution of fields, the knowledge structure, and the evolution of major research topics related to COPD with sarcopenia. The identification of keyword hotspots enhances the understanding of the occurrence, development, and future study trends associated with the topic.

Exploring influencing factors and predictive analysis of sarcopenia in the Chinese population using the body composition analyzer.

OBJECTIVE: Employing body composition analysis, this study aims to examine the influencing factors and conduct predictive analysis regarding sarcopenia incidence in the middle-aged and elderly population in China. METHODS: This study recruited inpatients from the General Medicine Department of Tongji Medical College Affiliated Union Hospital, Huazhong University of Science and Technology, as the subjects for a single-center retrospective study. Diagnosis was conducted according to the 2019 criteria from the Asian Working Group for Sarcopenia. Binary logistic regression analysis was utilized to identify factors influencing sarcopenia, and predictive modeling for sarcopenia occurrence was performed based on the area under the ROC curve (AUC). RESULTS: This study comprised 1258 hospitalized patients, of whom 340 were diagnosed with sarcopenia and 918 were not, resulting in a prevalence of 27%. The baseline characteristics showed statistically significant differences between the two groups. Binary logistic regression analysis revealed that low protein, low total body water, low minerals, low basal metabolic rate, and age were risk factors for sarcopenia (OR > 1, P < 0.05). Conversely, being male, having a higher BMI, greater fat-free mass index, and a higher InBody score were identified as protective factors against sarcopenia (OR < 1, P < 0.05). The AUC values for predicting sarcopenia occurrence based on low protein, low total body water, low minerals, low basal metabolic rate, and age were 0.871, 0.846, 0.757, 0.645, and 0.649, respectively, indicating their significance as predictive indicators. Combining these five indicators into a new predictive model for sarcopenia yielded an area under the curve (AUC) value of 0.932, demonstrating excellent sensitivity and specificity concurrently. CONCLUSION: The results of body composition analysis indicate that sarcopenia occurrence in the middle-aged and elderly population in China is associated with factors such as low protein, low total body water, low minerals, low basal metabolic rate, age, gender, BMI, fat-free mass index, and InBody score. The combination of specific body composition indicators facilitates the effective prediction of sarcopenia. Clinical practitioners should proactively identify the risk factors influencing sarcopenia, accurately predict.

Diagnostic Value of Nutritional Risk Index and Other Indices for Predicting Sarcopenia in the Middle-Aged and Elderly Population of China Without Cancer: A ROC Curve Analysis.

Background: Emerging evidence suggests that systemic inflammatory and nutritional biomarkers, along with derived indices, could serve as predictors for sarcopenia in cancer population. This study aimed to compare these predictors, focusing on the nutritional risk index (NRI) and evaluate its diagnostic value, for sarcopenic patients without cancer. Methods: This cross-sectional retrospective study included 1674 participants. Sarcopenia is defined by skeletal muscle mass index (SMI). Laboratory data reflected the values of systemic inflammatory and nutritional biomarkers, from which the derived indices were calculated. Multiple logistic regression analysis, ROC curve analysis, and the Youden index were utilized to assess the association between these markers and sarcopenia and determine the cutoff value for predicting sarcopenia. Results: Among all participants (1110 men and 564 women, mean age 61.97 ± 9.83 years), 398 individuals were diagnosed with sarcopenia, indicating a prevalence of 23.78% in China's middle-aged and elderly population without cancer. Logistic regression analysis revealed significant associations between all biomarkers and derived indices with sarcopenia. Following adjustment for potential confounders, lower NRI values were significantly associated with a higher incidence of sarcopenia. For sarcopenia diagnosis, the area under the curve (AUC) for NRI was 0.769 ([95% CI, 0.742, 0.796], P < 0.001), with a cutoff value of 106.016, sensitivity of 75.6% and specificity of 66.1%. NRI demonstrated greater predictive advantage for sarcopenia incidence in men compared to women. Conclusion: A lower NRI value was associated with a higher prevalence of sarcopenia. NRI shows promise for early, rapid, and effective sarcopenia screening, particularly in China's middle-aged and elderly male population without cancer.

Smoking is Associated with Lung Adenocarcinoma and Lung Squamous Cell Carcinoma Progression through Inducing Distinguishing lncRNA Alterations in Different Genders.

BACKGROUND: Smoking participates in pathogenesis of lung cancer. Long non-coding RNAs (lncRNAs) play some specific roles during development of lung cancers. OBJECTIVE: To investigate effects of smoking on lncRNA alterations in lung cancer. METHODS: There are 522 lung adenocarcinoma (LUAD) and 504 lung squamous cell carcinoma (LUSC) participants. Clinical and lncRNA genetic data were downloaded from The Cancer Genome Atlas (TCGA) database. LncRNA alterations were analyzed in lung cancer patients. Smoking category and packs were evaluated. Correlations between smoking and LncRNA alterations were analyzed. Kaplan-Meier analysis was performed to determine overall survival and disease free survival. RESULTS: There are more non-smokers in LUSC than in LUAD. In both LUAD and LUSC, smoking could increase total mutation counts and fraction of copy number alterations. Smoking index positively correlated with total mutations in LUAD, but not in LUSC. Smoking could trigger lncRNA alterations both in LUAD and LUSC. Smoking regulated different lncRNA between male and female. EXOC3-AS1 and LINC00603 alterations were positively correlated with smoking index in male LUAD smokers. In female LUAD smokers, smoking index was positively correlated with SNHG15, TP53TG1 and LINC01600 and negatively with LINC00609 and PTCSC3. In both male and female LUSC patients, smoking increased or decreased several lncRNA alterations. DGCR5 alteration increased in male LUSC than in female LUSC patients. In female LUSC patients, LOH12CR2 alteration was positively correlated with smoking index. CONCLUSION: Smoking promoted LUAD and LUSC development by affecting different lncRNA alterations in different genders.

Latent Transforming Growth Factor-ß Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling.

Despite past extensive studies, the mechanisms underlying pulmonary fibrosis (PF) still remain poorly understood. The aberrantly activated lung myofibroblasts, predominantly emerging through fibroblast-to-myofibroblast differentiation, are considered to be the key cells in PF, resulting in excessive accumulation of extracellular matrix (ECM). Latent transforming growth factor-ß (TGFß) binding protein-2 (LTBP2) has been suggested as playing a critical role in modulating the structural integrity of the ECM. However, its function in PF remains unclear. Here, we demonstrated that lungs originating from different types of patients with PF, including idiopathic PF and rheumatoid arthritis-associated interstitial lung disease, and from mice following bleomycin (BLM)-induced PF were characterized by increased LTBP2 expression in activated lung fibroblasts/myofibroblasts. Moreover, serum LTBP2 was also elevated in patients with COVID-19-related PF. LTBP2 silencing by lentiviral shRNA transfection protected against BLM-induced PF and suppressed fibroblast-to-myofibroblast differentiation in vivo and in vitro. More importantly, LTBP2 overexpression was able to induce differentiation of lung fibroblasts to myofibroblasts in vitro, even in the absence of TGFß1. By further mechanistic analysis, we demonstrated that LTBP2 silencing prevented fibroblast-to-myofibroblast differentiation and subsequent PF by suppressing the phosphorylation and nuclear translocation of NF-κB signaling. LTBP2 overexpression-induced fibroblast-to-myofibroblast differentiation depended on the activation of NF-κB signaling in vitro. Therefore, our data indicate that intervention to silence LTBP2 may represent a promising therapy for PF.

Calpain inhibition attenuates bleomycin-induced pulmonary fibrosis via switching the development of epithelial-mesenchymal transition.

Calpains are intracellular calcium-dependent cysteine proteases, which cleave several substrates proteins, have been proven to play important roles in lung fibrosis. The aim of this study was to investigate the effects of calpain on bleomycin (BLM)-induced pulmonary fibrosis. A lung fibrosis mice model was established successfully by intraperitoneal injection of bleomycin. Calpeptin, a highly selective inhibitor of calpain activation, was administered three times weekly after bleomycin injection. Histological examination was used to assess the fibrosis. Quantitative-PCR and Western blotting were used to assess the development of epithelial-mesenchymal transition (EMT). We found calpeptin treatment decreased the BLM-induced EMT-associated markers, such as muscle actin (α-SMA) and collagen-I, while increased E-cadherin (E-cad). Calpeptin also suppressed the activation of transforming growth factor ß1 (TGFß1)-Smad2/3 signaling pathway, which plays crucial role in lung fibrosis and EMT. Furthermore, we found differentiated embryonic chondrocyte-expressed gene 1 (DEC1), an important transcription factor, was upregulated in both patients with idiopathic pulmonary fibrosis and in bleomycin-induced lung fibrosis. DEC1 was suppressed by calpeptin in bleomycin-induced mice model. Collectively, these findings indicated that calpeptin had a potential anti-fibrosis effect, which focus on the development of EMT.

Interleukin-17 induces human alveolar epithelial to mesenchymal cell transition via the TGF-ß1 mediated Smad2/3 and ERK1/2 activation.

Idiopathic pulmonary fibrosis (IPF) is a chronic and usually progressive lung disease and the epithelial-mesenchymal transition (EMT) may play an important role in the pathogenesis of pulmonary fibrosis. IL-17 is a proinflammatory cytokine which promotes EMT profiles in lung inflammatory diseases. In this study, we investigated the effect of IL-17 on EMT in alveolar epithelial cell line A549 and the role of TGFß1-Smad and ERK signaling pathways in the process. Morphological observation on the cells was performed under inverted microscope. The mRNA and protein expressions of E-cad and α-SMA were detected by quantitative RT-PCR and western blotting. The mRNA and protein expressions of TGF-ß1 were analyzed via quantitative RT-PCR and ELISA. Expressions of Smad2/3, p-Smad2/3, ERK1/2, p-ERK1/2 and p-JNK were examined by western blotting. The results indicated that IL-17 can induce A549 cells to undergo morphological changes and phenotypic markers changes, such as down-regulated E-cad expression and up-regulated α-SMA expression. Additionally, IL-17 enhanced TGF-ß1 expression and stimulated Smad2/3 and ERK1/2 phosphorylation in A549 cells. However, there were no significant differences in the expression of phosphorylated JNK in A549 cells with or without IL-17 treatment. SB431542 or U0126 treated cells showed inhibited morphological changes and phenotypic markers expression, such as up-regulated E-cad expression and down-regulated α-SMA expression. In summary, our results suggest that IL-17 can induce A549 alveolar epithelial cells to undergo EMT via the TGF-ß1 mediated Smad2/3 and ERK1/2 activation.