# Exposure to Waterpipe Smoke Disrupts Erythrocyte Homeostasis of BALB/c Mice

## TL;DR

This study shows that waterpipe smoking harms red blood cells in mice by causing inflammation, oxidative stress, and increased cell fragility.

## Contribution

The study provides new evidence on how waterpipe smoke exposure disrupts erythrocyte homeostasis through oxidative stress and eryptosis.

## Key findings

- WPS exposure increased plasma inflammation and oxidative stress markers in mice.
- Erythrocyte counts and hematocrit decreased, while erythrocyte fragility increased in WPS-exposed mice.
- WPS exposure induced eryptosis and hemolysis in erythrocytes in vitro and in vivo.

## Abstract

Although various studies have provided evidence of a link between cardiovascular and hematological disease development and waterpipe smoking (WPS), there are a limited number of studies that have explored the impact of WPS on erythrocyte homeostasis. Here, we show that, compared with the air-exposed group, sub-chronic exposure to WPS in mice induced inflammation and oxidative stress in the plasma, decreased the number of erythrocytes and the hematocrit, and increased erythrocyte fragility. Moreover, erythrocytes in the WPS-exposed group showed increased markers of oxidative stress, ATPase activity, Ca2+ levels, annexin V binding, and calpain activity. In conclusion, our data demonstrate that WPS exposure elevated oxidative stress in the plasma and induced hemolysis in vivo. It also caused alterations in erythrocyte oxidative stress and eryptosis in vitro.

The prevalence of waterpipe tobacco smoking (WPS) is increasing worldwide and is relatively high among youth and young adults. It has been shown, both experimentally and clinically, that WPS exposure adversely affects the cardiovascular and hematological systems through the generation of oxidative stress and inflammation. Our study aimed to evaluate the impact of WPS exposure on erythrocytes, a major component of the hematological system, of BALB/c mice. Here, we assessed the effect of nose-only WPS exposure for four consecutive weeks on erythrocyte inflammation, oxidative stress, and eryptosis. The duration of the session was 30 min/day, 5 days/week. Control mice were exposed to air. Our results showed that the levels of C-reactive protein, lipid peroxidation (LPO), superoxide dismutase, and total nitric oxide (NO) were significantly increased in the plasma of WPS-exposed mice. The number of erythrocytes and the hematocrit were significantly decreased in WPS-exposed mice compared with the control group. Moreover, there was an increase in the erythrocyte fragility in mice exposed to WPS compared with those exposed to air. The levels of lactate dehydrogenase, LPO, reduced glutathione, catalase, and NO were significantly increased in the red blood cells (RBCs) of WPS-exposed mice. In addition, erythrocytes of the WPS-exposed group showed a significant increase in ATPase activity, Ca2+, annexin V binding, and calpain activity. Taken together, our findings suggest that WPS exposure elevated inflammation and oxidative stress in the plasma and induced hemolysis in vivo. It also caused alterations of RBCs oxidative stress and eryptosis in vitro. Our data confirm the detrimental impact of WPS on erythrocyte physiology.