The administration of a high refined carbohydrate diet promoted an increase in pulmonary inflammation and oxidative stress in mice exposed to cigarette smoke

Figures & data

Figure 1 Cumulative food intake of animals fed a standard diet and diet high in refined carbohydrates, and body mass of the experimental groups.

Notes: (A) Food consumption accumulated in grams (g). (B) Cumulative food consumption in kilocalories. (C) Body mass initial (g). (D) Body mass final (g). *Significant difference between the RG compared to CG. ^aSignificant difference between the RG compared to CG and ^bsignificant difference between RG and RCSG compared to CSG. Data are expressed as mean ± standard error of the mean and were analyzed by two- and one-way ANOVA followed by Bonferroni’s post-test (P<0.05). RCSG (n=6).
Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; RG, high refined carbohydrate group; RCSG, high refined carbohydrates diet and cigarette smoke group.

Figure 2 Blood glucose levels of the experimental groups.

Notes: *Significant difference between the RG compared to CG. Data are expressed as mean ± standard error of the mean and were analyzed by two-way ANOVA followed by Bonferroni’s post-test (P<0.05).
Abbreviations: ANOVA, analysis of variance; CG, control group; RG, high refined carbohydrate group (n=6).

Table 1 Cell influx in the bronchoalveolar lavage of the experimental groups

	CG	CSG	RG	RCSG
Leukocytes (×10³/mL)	162.3±6.6	212.0±9.7a	200.8±8.6	270.0±12.2a–c
Macrophages (×10³/mL)	155.3±5.1	181.8±6.9	191.2±6.9	224.7±7.2a,b
Lymphocytes (×10³/mL)	5.3±1.2	4.6±0.5	4.8±0.8	15.8±1.9a–c
Neutrophils (×10³/mL)	1.7±0.3	25.6±2.3a,c	4.8±0.9	29.5±3.1a,c

Notes:

a Significant difference between the groups when compared with the CG,

b significant difference between the groups when compared with the CSG, and

c significant difference between the groups when compared with the RG. Data are expressed as mean ± standard error of the mean and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05).

Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group.

Table 2 Hematological parameters of the experimental groups

	CG	CSG	RG	RCSG
Erythrocyte (×10^6/mm^3)	1.76±0.04	2.07±0.06a,b	1.82±0.04	1.98±0.05a
Hematocrit (%)	36.96±1.47	43.08±1.38a–c	35.41±0.99	37.44±0.70
Hemoglobin (g/dL)	12.36±0.45	14.18±0.26a–c	12.64±0.23	12.96±0.28

Notes:

a Significant difference between the groups when compared with the CG,

b significant difference between the groups when compared with the RG, and

c significant difference between the groups when compared with the RCSG. Data are expressed as mean ± standard error of the mean and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05).

Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group.

Table 3 Biochemical analysis in lung samples from the CG, CSG, RG, and RCSG groups

	CG	CSG	RG	RCSG
TBARS (nmol/mg protein)	0.24±0.05	0.21±0.05	0.28±0.06	0.61±0.12a–c
Protein carbonyl (nmol/mg protein)	12.33±1.37	20.50±3.31a	14.88±1.74	14.78±0.55
SOD (U/mg protein)	27.77±3.60	37.58±2.76	28.36±3.38	48.71±4.15a–c
CAT (U/mg protein)	0.45±0.04	0.86±0.17	0.92±0.11	1.09±0.19a
GSHt (nmol/mL)	20.37±2.33	39.69±2.95a	32.80±2.73	33.06±5.64
GSH/GSSG	0.41±0.86	0.61±0.04a	0.37±0.03a	0.33±0.11a

Notes:

a Significant difference between the groups when compared with the CG,

b significant difference between the groups when compared with the CSG, and

c significant difference between the groups when compared with the RG. Data are expressed as mean ± standard error of the mean and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05).

Abbreviations: ANOVA, analysis of variance; CAT, catalase; CG, control group; CSG, cigarette smoke group; GSH, glutathione reduced; GSHt, glutathione total; GSSG, glutathione oxidized; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group; SOD, superoxide dismutase; TBARS, thiobarbituric acid reactive substances.

Table 4 Levels of TNF-α, IFN-γ, and IL-10 of the pulmonary parenchyma in lung samples of the experimental groups

	CG	CSG	RG	RCSG
TNF-α (pg/mL)	163.1±9.10	179.5±17.40	164.0±10.52	96.81±22.38a,b
IFN-γ (pg/mL)	262.9±32.0	183.5±17.5	177.7±13.4c	127.0±14.2c
IL-10 (pg/mL)	446.8±46.9	399.8±46.9	325.5±19.4	274.2±35.9c

Notes:

a Significant difference between the groups when compared with the CG,

b significant difference between the groups when compared with the CSG, and

c significant difference between the groups when compared with the RG. Data are expressed as mean ± standard error of the mean and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05).

Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; IFN-γ, interferon gamma; IL-10, interleukin-10; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group; TNF-α, tumor necrosis factor alpha.

Table 5 Levels of TNF-α, IFN-γ, and IL-10 of the bronchoalveolar lavage fluid pulmonary parenchyma in lung samples of the experimental groups

	CG	CSG	RG	RCSG
TNF-α (pg/mL)	19.18±1.52	23.05±1.06	18.95±0.81	28.92±1.79a–c
IFN-γ (pg/mL)	30.75±6.56	14.99±1.22	29.27±3.75	39.05±4.58b
IL-10 (pg/mL)	90.17±7.76	112.3±10.69	93.21±5.72	128.5±7.04a,c

Notes:

a Significant difference between the groups when compared with the CG,

b significant difference between the groups when compared with the CSG, and

c significant difference between the groups when compared with the RG. Data are expressed as mean ± standard error of the mean and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05).

Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; IFN-γ, interferon gamma; IL-10, interleukin-10; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group; TNF-α, tumor necrosis factor alpha.

Figure 3 Morphometry of lung tissue.

Notes: (A) Alveolar volume density. (B) Volume density of the alveolar septa. ^aSignificant difference between the CSG and RCSG compared to CG, ^bsignificant difference between the groups when compared with the CSG, and ^csignificant difference between the CSG and RCSG compared to RG. Data are expressed as median, minimum and maximum values and were analyzed by Kruskal–Wallis test followed by Dunn’s post-test (P<0.05). RCSG (n=6).

Abbreviations: CG, control group; CSG, cigarette smoke group; RG, high refined carbohydrate group; RCSG, high refined carbohydrates diet and cigarette smoke group.

Figure 4 Photomicrographs of lung sections stained with hematoxylin and eosin.

Notes: 40× magnification. There was an increase in Vv (alveolar spaces) of the CSG and RCSG when compared to CG and RG and there was a decreased in Vv (alveolar septa) in CSG compared to other groups and in RCSG when compared to CG and RG were observed. Bar =50 μm. RCSG (n=6).
Abbreviations: CG, control group; CSG, cigarette smoke group; RG, high refined carbohydrate group; RCSG, high refined carbohydrates diet and cigarette smoke group; Vv, volume density.

Figure 5 Adipose tissue analysis. (A) Adiposity index in the experimental groups. (B) Area adiposity. (C) Plasma leptin levels.

Notes: ^aSignificant difference between the RG and RCSG compared to CG, ^bsignificant difference between RG and RCSG compared to CSG, ^csignificant difference between the RCSG compared to RG, and ^dsignificant difference between RG compared to RCSG. Data are expressed as mean ± SEM and were analyzed by one-way ANOVA followed by Bonferroni’s post-test (P<0.05). RCSG group size, n=6.

Abbreviations: ANOVA, analysis of variance; CG, control group; CSG, cigarette smoke group; RG, high refined carbohydrate group; RCSG, high refined carbohydrates diet and cigarette smoke group; SEM, standard error of the mean.

Figure 6 Representative photomicrographs of epididymal adipose tissue stained with hematoxylin and eosin.

Notes: 10× magnification. RCSG (n=6). The area of adipocytes was higher in the RG compared to CG, CSG, and RCSG and was higher in RCSG when compared to CG and CSG.
Abbreviations: CG, control group; CSG, cigarette smoke group; RCSG, high refined carbohydrates diet and cigarette smoke group; RG, high refined carbohydrate group.