Exposure to Heated Tobacco Products Aerosol Causes Acute Stress Responses in the Lung of Mouse.

In the present study, we evaluated the acute response of mice exposed to IQOS aerosol, a brand-name heated tobacco product (HTP), in the lung tissue. First, the thiobarbituric acid-reactive substances (TBA-RS) value was measured as an index to assess oxidative stress, and a significant increase was observed after exposure, followed by a significant increase in the total lung GSH concentration. The stress responses induced by IQOS aerosols was then analyzed by focusing on the changes in Nrf2 and ATF4, which are transcription factors that induce the expression of genes involved in GSH biosynthesis or metabolism. Although Nrf2 activation was not observed, significant accumulation of ATF4 in the nuclear fraction was noted three hours after exposure to IQOS aerosols. Upon an examination of changes in factors in the GSH biosynthetic system, a significant increase in cystine concentration in the lung tissue was measured, and an increase in xCT expression level was observed in the cell membrane fraction three-six hours after IQOS exposure. Furthermore, characteristic changes in HO-1, a stress-response protein regulated by ATF4, was discovered six hours after IQOS exposure. Moreover, analysis of the upstream ATF4 regulatory system revealed that phosphorylation of eIF2α was enhanced in the lung cytoplasmic fraction three hours after exposure to IQOS aerosols. These findings suggest that ER stress might be induced as an early response to IQOS aerosol exposure, accompanied by the activation of the eIF2α-ATF4 axis. These intracellular changes have also been reported after exposure to combustible cigarette smoke. Thus, the acute response found in the lungs of mice in the present study demonstrate that the inhalation of aerosols from IQOS elicits a biological response similar to that of combustible cigarette smoke. In conclusion, our results provide evidence that the biological effects of HTPs, such as IQOS, cannot be ignored in the lungs.

Increased oxidative stress and effects on inflammatory cytokine secretion by heated tobacco products aerosol exposure to mice.

Although the amount of chemicals in heated tobacco products (HTPs) aerosols is reduced compared to conventional combustible cigarette smoke, the association between HTPs and reduced health effects remains unclear. In this study, we hypothesized that exposure to IQOS, an HTP, would increase oxidative stress and affect the secretion of inflammatory cytokines. First, C57BL/6 mice exposed to IQOS aerosols were evaluated to determine the adverse effects of IQOS exposure. IQOS exposure significantly decreased the concentration of GSH in alveolar macrophages in a dose-dependent manner and increased the percentage of GSSG in lung tissues. These results indicate that IQOS exposure increases oxidative stress, and GSH is consumed to remove oxidative stress. In addition, foamy alveolar macrophages were observed in the bronchial alveolar lavage fluid after IQOS exposure. Although the concentration of inflammatory cytokines, IL-6, and GM-CSF, in the plasma increased significantly after IQOS exposure, there were no significant changes in other cytokines. These results indicate that short-term exposure to IQOS aerosols may increase oxidative stress and induce the secretion of inflammatory cytokines. Lastly, the longer-term effects of IQOS aerosols exposure should be evaluated in the future.

Vascular Endothelial Glycocalyx Plays a Role in the Obesity Paradox According to Intravital Observation.

According to the "obesity paradox," for severe conditions, individuals with obesity may be associated with a higher survival rate than those who are lean. However, the physiological basis underlying the mechanism of the obesity paradox remains unknown. We hypothesize that the glycocalyx in obese mice is thicker and more resistant to inflammatory stress than that in non-obese mice. In this study, we employed intravital microscopy to elucidate the differences in the vascular endothelial glycocalyx among three groups of mice fed diets with different fat concentrations. Male C57BL/6N mice were divided into three diet groups: low-fat (fat: 10% kcal), medium-fat (fat: 45% kcal), and high-fat (fat: 60% kcal) diet groups. Mice were fed the respective diet from 3 weeks of age, and a chronic cranial window was installed at 8 weeks of age. At 9 weeks of age, fluorescein isothiocyanate-labeled wheat germ agglutinin was injected to identify the glycocalyx layer, and brain pial microcirculation was observed within the cranial windows. We randomly selected arterioles of diameter 15-45 µm and captured images. The mean index of the endothelial glycocalyx was calculated using image analysis and defined as the glycocalyx index. The glycocalyx indexes of the high-fat and medium-fat diet groups were significantly higher than those of the low-fat diet group (p < 0.05). There was a stronger positive correlation between vessel diameter and glycocalyx indexes in the high-fat and medium-fat diet groups than in the low-fat diet group. The glycocalyx indexes of the non-sepsis model in the obese groups were higher than those in the control group for all vessel diameters, and the positive correlation was also stronger. These findings indicate that the index of the original glycocalyx may play an important role in the obesity paradox.

A Newly Developed Aerosol Exposure Apparatus for Heated Tobacco Products for In Vivo Experiments Can Deliver Both Particles and Gas Phase With High Recovery and Depicts the Time-Dependent Variation in Nicotine Metabolites in Mouse Urine.

INTRODUCTION: There is no standardized aerosol exposure apparatus to deliver heated tobacco products (HTPs) for in vivo experiments. Therefore, we developed a novel HTPs aerosol exposure apparatus for mice and demonstrated that nicotine and other chemicals in HTPs aerosol generated by the apparatus can be delivered to mice which replicate human smoke. AIMS AND METHODS: The amounts of nicotine, tar, and carbon monoxide (CO) in IQOS (Marlboro Regular HeatSticks) aerosol generated by two types of apparatuses were determined. C57BL/6N mice were exposed to IQOS aerosol, followed by determination of the urinary nicotine metabolites. Further, the skin surface temperature of mice was monitored to confirm the vasoconstriction action of nicotine. RESULTS: The amount of chemicals in IQOS aerosol by the novel air push-in inhalation apparatus for HTPs (APIA) was equivalent to that of the analytical vaping machine (LM4E) (1.60 ± 0.08 [APIA] vs. 1.46 ± 0.07 mg/stick [LM4E] in nicotine and 0.55 ± 0.04 [APIA] vs. 0.45 ± 0.01 mg/stick [LM4E] in CO). After mice were exposed to IQOS aerosol by APIA, the urinary nicotine metabolite levels were determined; peak values in cotinine and 3-hydroxycotinine (3-HC) were 6.82 µg/mg creatinine at 1 hour after exposure and 32.9 µg/mg creatinine at 2 hours after exposure, respectively. The skin surface temperature decreased and was lower (33.5°C ± 0.5°C) at 30 minutes than before exposure (37.6°C ± 0.8°C). CONCLUSIONS: The new apparatus for HTPs aerosol exposure to mice showed good performances in terms of both chemical analysis of collected aerosol and fluctuations in the urinary nicotine metabolites. IMPLICATIONS: The APIA reported in this study can expose small animals to HTPs aerosol, including nicotine and other chemical substances as same amounts as LM4E and replicate actual human smoking process by in vivo experiments. Therefore, the experiments using APIA can provide evidence to assess the health risks of HTPs use.

The protective effect of hydroxyethyl starch solution on the glycocalyx layer in an acute hemorrhage mouse model.

PURPOSE: Fluid therapy focused on glycocalyx (GCX) protection in hemorrhagic shock is a current focus of research. Hydroxyethyl starch (HES) solution is commonly used for fluid resuscitation; however, its effects on the GCX remain unclear. The primary aim of this study was to explore the protective effect of HES130 in maintaining GCX thickness and reducing plasma syndecan-1 expression. METHODS: An acute hemorrhage murine model with the dorsal skin chambers was used to measure GCX thickness and to evaluate vascular permeability. Groups of mice were treated with normal saline (NS), albumin (NS-A), HES130 (NS-V), or no exsanguination or infusion (C). We measured syndecan-1 plasma concentrations, performed blood gas analysis, and analyzed the 7-day cumulative mortality. RESULTS: GCX thickness in NS mice was significantly reduced compared to that in group C, but no other groups showed a difference compared to group C. The plasma concentration of syndecan-1 was significantly higher in NS mice than in group C. There were no significant differences in the fluorescence intensity of dextran in the interstitial space. HES70 leakage was suppressed in NS-V mice compared to those in other groups. HES70 was localized to the inner vessel wall in C, NS, and NS-A mice, but not in group NS-V. Blood gas analysis indicated that pH and lactate showed the greatest improvements in NS-V mice. The 7-day cumulative mortality rate was the highest in group NS. CONCLUSION: Resuscitation with HES130 protected the GCX and suppressed vascular permeability of HES70 during early stages of acute massive hemorrhage.