Effect of allyl isothiocyanate on oxidative stress in COPD via the AhR / CYP1A1 and Nrf2 / NQO1 pathways and the underlying mechanism.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is currently the third leading cause of death globally. Oxidative stress affects various molecular mechanisms and is the main driving factor of COPD. Ally isothiocyanate (AITC) is an effective component of Semen Sinapis Albae, which has favorable effects for the treatment of COPD, but its mechanism has not been fully elucidated. PURPOSE: This study aimed to elucidate the antioxidant effect of AITC on COPD and its molecular mechanism, and preliminarily determine the role of AhR in the progression of COPD. STUDY DESIGN: The COPD rat model was established by smoking combined with intratracheal instillation of lipopolysaccharide. Different doses of AITC, positive control drug acetylcysteine, AhR inhibitor alpha-naphthoflavone, and agonist beta-naphthoflavone were administered by gavage. Human bronchial epithelial cells induced by cigarette smoke extract (CSE) were used in an in vitro model to explore the molecular mechanisms of AITC. METHODS: The effects of AITC on lung function and oxidative stress in rats were evaluated in vivo using the respiratory function test, white blood cell count, enzyme-linked immunosorbent assay, and histological staining. The changes in protein expression in the lung tissue were detected by immunohistochemistry and Western blotting. RT-PCR, western blotting, and immunofluorescence were used to explore the molecular mechanisms of AITC. Enzyme-linked immunosorbent assay, reactive oxygen species probing, and flow cytometry were used to determine the antioxidant effect of AITC. RESULTS: AITC can improve the lung function of rats with COPD, restore lung tissue structure, improve oxidative stress, reduce inflammation, and inhibit lung cell apoptosis. AITC reversed the upregulation of AhR and CYP1A1 and the down-regulation of Nrf2 and NQO1 in the lung tissues of rats with COPD. CSE stimulation can increase the expressions of AhR and CYP1A1 and decrease the expressions of Nrf2 and NQO1 in 16HBE cells, leading to severe oxidative stress and inflammatory response and, ultimately, apoptosis. AITC inhibited AhR and CYP1A1 expressions, induced Nrf2 and NQO1 expressions, promoted Nrf2 nuclear translocation, and improved CSE-induced toxicological effects. CONCLUSION: AITC may improve lung oxidative stress by inhibiting the AhR / CYP1A1 and activating the Nrf2 / NQO1 pathways, thereby delaying the pathological progression of COPD.

Relationship between the stroke mechanism of symptomatic middle cerebral artery atherosclerotic diseases and culprit plaques based on high-resolution vessel wall imaging.

Purpose: For patients with symptomatic middle cerebral artery (MCA) atherosclerotic stenosis, identifying the potential stroke mechanisms may contribute to secondary prevention. The purpose of the study is to explore the relationship between stroke mechanisms and the characteristics of culprit plaques in patients with atherosclerotic ischemic stroke in the M1 segment of the middle cerebral artery (MCA) based on high-resolution vessel wall imaging (HR-VWI). Methods: We recruited 61 patients with acute ischemic stroke due to MCA atherosclerotic stenosis from Shenzhen Bao'an District People's Hospital. According to prespecified criteria based on infarct topography and magnetic resonance angiography, possible stroke mechanisms were divided into parent artery atherosclerosis occluding penetrating artery (P), artery-to-artery embolism (A), hypoperfusion (H), and mixed mechanisms (M). The correlation between the characteristics of MCA M1 culprit plaque and different stroke mechanisms was analyzed using HR-VWI. The indicators included plaque surface irregularity, T1 hyperintensity, location, plaque burden (PB), remodeling index (RI), enhancement rate, and stenosis rate. Results: Parental artery atherosclerosis occluding penetrating artery was the most common mechanism (37.7%). The proposed criteria showed substantial to excellent interrater reproducibility (κ, 0.728; 0.593-0.863). Compared with the P group, the surface irregularity, T1 hyperintensity, and obvious enhancement of the culprit plaque in the A group were more common (p < 0.0125). Compared with the other stroke mechanisms, positive remodeling of culprit plaques was more common (p < 0.0125), the RI was greater (p < 0.05), and the PB was the smallest (p < 0.05) in the P group. The enhancement ratio (ER) was smaller in the P group (p < 0.05). Compared with the A group, T1 hyperintensity of the culprit plaque was more common in the H group (p < 0.0125), and the stenosis rate was greater (p < 0.05). After adjustment for clinical demographic factors in the binary logistic regression analysis, the enhancement level (odds ratio [OR] 0.213, 95% CI (0.05-0.91), p = 0.037) and PB of culprit plaque (OR 0, 95% CI (0-0.477), p = 0.034) were negatively associated with P groups. Conclusion: The culprit plaque characteristics of patients with symptomatic MCA atherosclerotic in different stroke mechanisms may be evaluated using HR-VWI. The plaque characteristics of different stroke mechanisms may have clinical value for the selection of treatment strategies and prevention of stroke recurrence. Clinical trial registration: Identifier: ChiCTR1900028533.

Effects of GNG4 on DNA damage repair and chemosensitivity of ovarian cancer cisplatin-resistant A2780/DDP cells / 肿瘤研究与临床

Objective:To investigate the correlation of GNG4 with DNA damage repair and chemosensitivity of ovarian cancer cisplatin-resistant A2780/DDP cells.Methods:A2780/DDP cells were divided into 500 ng/ml cisplatin group (cDDP group), short hairpin RNA (shRNA)-GNG4 silencing GNG4 expression group (shRNA group), 500 ng/ml cisplatin and shRNA-GNG4 intervention group (shRNA+cDDP group), and non cisplatin and shRNA-GNG4 intervention group (blank control group). Western blot was used to detect the expressions of GNG4 and γH2AX proteins in each group; DNA damage in each group was detected by single cell gel electrophoresis. The focus formation of γH2AX gene at the injury site was detected by immunofluorescence. The ability of cell clone formation was detected by plate clone formation experiment.Results:Compared with the other three groups, the expression level of GNG4 protein in shRNA+cDDP group was the lowest, the expression level of γH2AX protein was the highest, and the differences were statistically significant (all P < 0.01). Single cell gel electrophoresis assay showed that the comet tail DNA% in blank control group, cDDP group, shRNA group and shRNA+cDDP group were (7.7±2.5)%, (12.3±3.6)%, (20.1±2.1)%, (38.6±2.8)%, respectively, and Olive trailing distance were 5.12±1.89, 8.23±2.97, 14.99±3.65, 22.43±3.17, respectively, the comet tail DNA% and Olive tail distance in shRNA+cDDP group were higher than those in the other three groups, and the differences were statistically significant (all P < 0.05). Immunofluorescence assay showed that the focus numbers of γH2AX in each cell of blank control group, cDDP group, shRNA group and shRNA+cDDP group were 4.2±0.7, 5.1±0.5, 26.8±3.3, 71.3±6.2, respectively, the shRNA+cDDP group was higher than the other three groups, and the differences were statistically significant (all P < 0.05). The clone formation rates of blank control group, cDDP group, shRNA group and shRNA+cDDP group were (78.27±5.01)%, (45.67±3.29)%, (26.20±5.76)%, (1.56±0.21)%, respectively, the shRNA+cDDP group was lower than the other three groups, and the differences were statistically significant (all P < 0.001). Conclusions:Down-regulation of GNG4 expression can increase the cisplatin sensitivity of ovarian cancer A2780/DDP cells, which may be achieved by inhibiting the DNA damage repair function induced by cisplatin.

Effects of allyl isothiocyanate on the expression, function, and its mechanism of ABCA1 and ABCG1 in pulmonary of COPD rats.

BACKGROUND AND AIMS: Allyl isothiocyanate(AITC) has been shown to play an important role in the improved symptoms of chronic obstructive pulmonary disease(COPD) and the inhibition of inflammation, but the role in COPD lipid metabolism disorder and the molecular mechanism remains unclear. We aimed to explore whether and how AITC affects COPD by regulating lipid metabolism and inflammatory response. METHODS: The COPD rat model was established by cigarette smoke exposure. Cigarette smoke extract stimulated 16HBE cells to induce a cell model. The effect of AITC treatment was detected by lung function test, H&E staining, Oil red O staining, immunohistochemistry, ELISA, CCK-8, HPLC, fluorescence efflux test, siRNA, RT-PCR, and Western blotting. Biological analysis was performed to analyze the results. Graphpad Prism 8.0 software was used for statistical analysis. RESULTS: AITC can improve lung function and pathological injury in COPD rats. The levels of IL-1 ß and TNF- α in the AITC treatment group were significantly lower than those in the model group(P < 0.05), and the lipid metabolism was also improved (P < 0.05). AITC reverses CSE-induced down-regulation of LXR α, ABCA1, and ABCG1 expression and function in a time-and concentration-dependent manner (P < 0.05). AITC regulates the cholesterol metabolism disorder induced by CSE in NR8383 cells and attenuates macrophage inflammation (P < 0.05). In addition, after silencing LXR α with siRNA, the effect of AITC was also inhibited. CONCLUSION: These results suggest that AITC improves COPD by promoting RCT process and reducing inflammatory response via activating LXR pathways.

Quantitative and Qualitative Analysis of Atherosclerotic Stenosis in the Middle Cerebral Artery Using High-Resolution Magnetic Resonance Imaging.

PURPOSE: We analyzed and compared the imaging characteristics of the vessel wall of the middle cerebral artery (MCA) in symptomatic and asymptomatic patients using a 3.0-T high-resolution magnetic resonance imaging (HR-MRI) protocol, including a 3-dimensional T1-sampling perfection with application-optimized contrasts using different flip angle evolutions sequence. METHODS: Fifty-three patients with atherosclerotic stenosis of the MCA underwent 3.0-T HR-MRI examinations. The characteristics of atherosclerotic plaques in 53 patients (28 symptomatic, 25 asymptomatic) were analyzed, including plaque distribution and signal intensity. Plaque burden (PB), stenosis degree, and the remodeling index were measured and compared between symptomatic and asymptomatic patients. RESULTS: The PB of the symptomatic group was significantly higher than that of the asymptomatic group (P = .006), and moderate-severe stenosis was more common (P = .01). The remodeling index of the symptomatic group was also lower (P = .015) and negative remodeling (NR) was more common (P = .043). Binary logistic regression analysis showed that stenosis degree was a risk factor in symptomatic patients (odds ratio = 135, P = .023). CONCLUSION: There is a trend that some characteristics of plaques and vessels, including the moderate-severe stenosis, larger PB, and NR, were observed more frequently among patients with symptomatic atherosclerotic stenosis of the MCA than among asymptomatic patients.

Effect of junction plakoglobin on radiotherapy-resistant cervical cancer cells / 肿瘤研究与临床

Objective:To explore the effect of junctional plakoglobin (JUP) on the radiotherapy-resistant cervical cancer cells in vitro.Methods:Cervical cancer cell lines SiHa, HeLa and Me-180 were irradiated with 1 Gy of 60Co radioactive rays for 3 times every week to induce the radiotherapy resistance of cells which obtained the radiotherapy-resistant cell lines SiHaIR, HeLaIR and Me-180IR. The corresponding wild-type cell lines were served as control groups. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expressions of JUP mRNA and protein in the 6 groups of cells. The cell scratch test was used to detect the cell migration ability. Results:The relative expressions of JUP mRNA in SiHa and SiHaIR groups, HeLa and HeLaIR groups and Me-180 and Me-180IR groups were the relative expressions of JUP protein were 1.74±0.06 and 0.48±0.02 ( t = 12.327, P < 0.01), 1.77±0.06 and 0.28±0.03 ( t = 14.698, P < 0.01), 2.276±0.061 and 0.780±0.011 ( t = 7.367, P < 0.01); 2.36±0.03 and 0.55±0.02 ( t = 9.245, P < 0.01), 2.13±0.02 and 0.23±0.01 ( t = 15.643, P < 0.01), 1.96±0.05 and 0.73±0.02 ( t = 5.826, P < 0.01). When culturing the cells for 12, 24, and 48 h after scratching, the migration rates in 3 groups of radiotherapy-resistant cell lines were increased compared with the corresponding wild-type cell lines, and the differences were statistically significant (all P < 0.05). Conclusion:The expressions of JUP in radiotherapy-resistant cervical cancer cell lines are lower than those in wild-type cell lines, and the migration ability of radiotherapy-resistant cells is enhanced.

Effect of Oat and Tartary Buckwheat - Based Food on Cholesterol - Lowering and Gut Microbiota in Hypercholesterolemic Hamsters.

The nutritional components in oat and tartary buckwheat had been assessed to have cholesterollowering effects. However, The effect of oat and tartary buckwheat based-food (OF) on cholesterol-lowering and gut microbiota in hypercholesterole hamsters was still limited studied because they are usually consumed in whole gran as well as after being processed. In this study, normal diets, high fat diet (HFD) with/without OF were fed to hamsters for 30 days respectively and growth parameters, metabolic parameters, and gut microbiota were investigated, respectively. It was found that OF significantly decreased plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-cholesterol), lowered liver TC, cholesterol ester (CE), and triglycerides (TG) concentrations, and increased fecal weight and bile acids (BA) concentrations, compared with HFD (p < 0.05). Moreover, the concentrations of acetate, propionate, butyrate and total short-chain fatty acids (SCFAs) were significantly increased in hamsters fed with OF, compared with HFD (p < 0.05). OF changed the overall structure of gut microbiota. The relative abundances of Erysipelotrichaceae, Ruminococcaceae, and Lachnospiraceae were decreased and the relative abundance of Eubacteriaceae was increased, compared with HFD. These results suggested that OF could reduce the concentrations of plasma lipid by inhibiting cholesterol absorption in liver and promoting excretions of fecal lipid and BA. And it also increased SCFAs and modulated the gut microbiota effectively to exert the hypocholesterolemic effects.