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Effects of sidestream cigarette smoke exposure on baroreflex components

in spontaneously hypertensive rats
Vitor E. Valentia; Luiz Carlos Abreub; Paulo H. Saldivac; Tatiana D. Carvalhoa; Celso Ferreiraa
a
Departamento de Medicina, Disciplina de Cardiologia, Universidade Federal de São Paulo (UNIFESP),
São Paulo, Brazil b Departamento de Morfologia e Fisiologia, Faculdade de Medicina do ABC, Santo
André, Brazil c Departamento de Patologia, Universidade de São Paulo, São Paulo, Brazil

Online publication date: 14 December 2010

To cite this Article Valenti, Vitor E. , Abreu, Luiz Carlos , Saldiva, Paulo H. , Carvalho, Tatiana D. and Ferreira,
Celso(2010) 'Effects of sidestream cigarette smoke exposure on baroreflex components in spontaneously hypertensive
rats', International Journal of Environmental Health Research, 20: 6, 431 — 437
To link to this Article: DOI: 10.1080/09603123.2010.491852
URL: http://dx.doi.org/10.1080/09603123.2010.491852

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 International Journal of Environmental Health Research
Vol. 20, No. 6, December 2010, 431–437

Effects of sidestream cigarette smoke exposure on baroreflex

components in spontaneously hypertensive rats
Vitor E. Valentia*, Luiz Carlos Abreub, Paulo H. Saldivac, Tatiana D. Carvalhoa
and Celso Ferreiraa
a
Departamento de Medicina, Disciplina de Cardiologia, Universidade Federal de São Paulo
(UNIFESP), São Paulo, Brazil; bDepartamento de Morfologia e Fisiologia, Faculdade de
Medicina do ABC, Santo Andre´, Brazil; cDepartamento de Patologia, Universidade de São
Paulo, São Paulo, Brazil
(Received 4 September 2009; final version received 4 April 2010)
Downloaded By: [Valenti, Vitor E.] At: 01:23 15 December 2010

We evaluated short-term effects of sidestream cigarette smoke (SSCS) exposure

on baroreflex function in spontaneously hypertensive rats (SHR) and Wistar
Kyoto (WKY) normotensive rats. Rats were exposed to SSCS during three weeks,
180 min, five days per week, in a concentration of carbon monoxide (CO)
between 100 and 300 ppm. We observed that SSCS exposure increased
tachycardic peak and heart rate range while it attenuated bradycardic reflex in
WKY. In respect to SHR, SSCS also increased tachycardic peak. Taken together,
our data suggests that three weeks of exposure to SSCS affects the sympathetic
and parasympathetic component of the baroreflex in normotensive WKY while it
tended to affect the sympathetic component in SHR.
Keywords: tobacco; baroreflex; air pollutants; smoke

Introduction
There is growing evidence that long-term exposure to fine particulate matter air
pollution contributes to the risk of all-cause and cardiovascular mortality (Pope
et al. 2002). Moreover, the inspired dose of fine particles from ambient pollution is
extremely small compared to that from sidestream cigarette smoke (SSCS) (Gamble
and Nicolich 2000; Stinn et al. 2005; Tricker et al. 2009). A study of ambient fine
particulate pollution and secondhand cigarette smoke estimates that its cardiovas-
cular mortality effect is much higher than would be expected on the basis of effects
extrapolations of active cigarette smoking that assumes a linear dose-response
relationship that goes through the origin (Howard and Thun 1999).
Baroreflex is one of the body’s homeostatic mechanisms to maintain blood
pressure (Valenti et al. 2009a). Besides cardiac hypertrophy observed in stress models
and in spontaneous hypertensive rats (SHR) (Cenko et al. 2004; Meneghini et al.
2008; Daud et al. 2009; Ferreira et al. 2009; Meneghini et al. 2009), baroreflex is also
impaired in this strain (Valenti et al. 2009a). Hence, this strain is already at a higher
risk due to their baroreflex impairment (Valenti et al. 2009a).

*Corresponding author. Email: valenti@unifesp.br

ISSN 0960-3123 print/ISSN 1369-1619 online

Ó 2010 Taylor & Francis
DOI: 10.1080/09603123.2010.491852
http://www.informaworld.com
 432 V.E. Valenti et al.

Although it is documented in the literature that long-term (years) exposure to

both SSCS and mainstream cigarette smoke impairs baroreflex in human
(Shinozaki et al. 2008), the short-term effects of SSCS on baroreflex function
in SHR have not yet been demonstrated. Thus, in this study we investigated the
effects of SSCS exposure on baroreflex in SHR and Wistar Kyoto (WKY)
normotensive rats.

Method
We used WKY and SHR (15–17 weeks old). They were kept under a 12 h light/dark
cycle (lights on at 07:00 h) and had free access to food and water. The Institution’s
Animal Ethics Committee authorized housing conditions and experimental
procedures. Rats were placed in the transparent chamber, with a volume of
approximately 95 6 80 6 65 cm3, where four rats remained. The concentration of
smoke carbon monoxide (CO) was maintained between 100 and 300 ppm
Downloaded By: [Valenti, Vitor E.] At: 01:23 15 December 2010

(23 + 18C, 50–60%) (Gairola et al. 2001). Rats were exposed to SSCS (1.1 mg of
nicotine, 14 mg of tar and 15 mg of carbon monoxide) during 180 min, five days/
week, for three weeks between 08:00 and 11:00 h. Control animals were maintained
at the same place and the same conditions as the SSCS group but exposed to fresh air
not contaminated by smoke CO.
Rats were anesthetized one day before the experiments with ketamine (50 mg/kg
i.p.) and xylazine (50 mg/kg i.m.) and a catheter of PE-10 polyethylene was inserted
into the abdominal aorta through the femoral artery for blood pressure and heart
rate recording. The catheters were tunneled under the skin and exteriorized at the
animal’s dorsum. One day after surgery, animals were allowed 20 min to adapt to
the conditions of the experimental room such as sound and illumination before
starting blood pressure and heart rate recording. Pulsatile arterial pressure (PAP) of
freely moving animals was recorded using an HP-7754A preamplifier (Hewlett
Packard, USA) and an acquisition board (MP100A, Biopac Systems Inc, USA)
connected to a computer. Mean arterial pressure (MAP) and heart rate (HR) values
were derived from the PAP recordings and processed on-line (Valenti et al. 2009a,
2009b).
The baroreflex was tested with a pressor dose of phenylephrine (PE-bolus-8
mg/kg IV; Sigma Chemical) and depressor doses of sodium nitroprusside (NaNP-
bolus-50 mg/kg IV; RBI). The baroreflex was calculated as the derivation of HR
in function of the MAP variation (DHR/DMAP). There was an interval of at
least 15 min between the infusions to allow the recovery of basal values. We also
evaluated bradycardic and tachycardic peak and HR range, the difference
between bradycardic and tachycardic peak. Variables calculation was performed
by using the Biopac system program (MP100A, Biopac Systems Inc, USA). The
delay in reflex HR responses was about 1.2 s because of the time for baroreflex
synapse processing, which is 700 ms according to Su et al. (1992), and the
integration factor of the recording system, which was about 500 ms (Cisternas
et al. 2010; Valenti et al. 2010).
The results were reported as means + standard error of means (SEM). In order
to verify if SSCS exposure effects were different between two strains (WKY and
SHR) we applied two-way analysis of variance (ANOVA) (strain vs. SSCS) followed
by post hoc Tukey test. Differences were considered significant when the probability
of a Type I error was less than 5% (p 5 0.05).
 International Journal of Environmental Health Research 433

Results
SSCS exposure during three weeks affected tachycardic peak and HR range in WKY
rats, while in SHR the tachycardic peak and HR range were altered by SSCS
exposure (Table 1).
In relation to bradycardic responses to intravenous PHE, WKY control rats
presented the highest response, which was higher than WS, SC and SS groups. Thus,
it leads us to suggest that SSCS exposure attenuated this component of the
baroreflex in WKY rats (Figure 1). NaNP-induced decrease in MAP was reduced in
WKY groups compared to SHR groups and tachycardic reflex responses to
intravenous NaNP were enhanced in WKY groups compared to the SHR control
group, but were not different in the SHR exposed to SSCS (Figure 2).

Discussion
Downloaded By: [Valenti, Vitor E.] At: 01:23 15 December 2010

Due the toxic effects of cigarette components (Yamasaki et al. 2009) and other
pollutant agents on the cardiovascular system (Solomon et al. 2003; Fukuda-
Matsuda et al. 2007) presented in the literature, we evaluated baroreflex function in
WKY and SHR exposed to SSCS during three weeks. Our exposure protocol showed
that SSCS affected bradycardic reflex, tachycardic peak and HR range in WKY rats,
whereas it influenced the tachycardic peak and HR range in SHR.
Maximal parasympathetic activity were not attenuated or elevated in WKY
and SHR exposed to SSCS compared to age-matched control groups, while the
maximal sympathetic activity was significantly increased in SHR and WKY rats
exposed to SSCS. Our data support the hypothesis that less than 30 days of
cigarette exposure is sufficient to change the sympathetic component of the
baroreflex without change basal arterial pressure (Paiva et al. 2003). This method

Table 1. Baseline level of mean arterial pressure (MAP) and heart rate (HR), bradycardic
and tachycardic peak, HR range and baroreflex gain (BG) in WKY (n ¼ 15) and SHR
(n ¼ 16) exposed to SSCS and to ambient air (WKY: n ¼ 22; SHR: n ¼ 18). Mean þ SEM,
maximum and minimum.

WKY exposed SHR exposed

Variable WKY control to SSCS SHR control to SSCS
MAP (mmHg) 115.6 + 2.2 108.3 + 2.3 166.3 + 15.2 168.9 + 2
(101–124) (96–121) (139–194)* (160–192)*
HR (bpm) 338.2 + 8.7 320.3 + 7.9 344.8 + 6.5 350.7 + 7.5
(239–378) (280–363) (318–402) (297–446)
Bradycardic peak (bpm) 208.2 + 7 226 + 8.7 284.2 + 10 281.3 + 9.5
(160–260) (155–266) (211–342)* (214–352)*
Tachycardic peak (bpm) 456.7 + 9 490.1 + 6.9 443 + 7.8 487.5 + 7.6
(352–500) (461–545)* (400–480) (444–521)*$
HR range (bpm) 248.5 + 12 265.8 + 6 158.8 + 7 202.6 + 9.8
(149–316) (229–323)*$ (100–189)* (148–236)$
BG (bpm 6 mmHg71) 72.2 + 0.15 71.78 + 0.15 70.77 + 0.11 71.03 + 0.19
PHE (71.07–3.01) (71.14–3.03) (70.28–1.83)*# (70.24–2.73)*#
BG (bpm 6 mmHg71) 72.8 + 0.15 73.01 + 0.15 71.52 + 0.14 71.43 + 0.14
NaNP (71.29–3.9) (72.2–3.93) (70.52–2.61)*# (70.37–2.32)*#

*p 5 0.05: Different from control WKY. #p 5 0.05: Different from WKY exposed to SSCS. $p 5 0.05:
Different from control SHR.
 434 V.E. Valenti et al.
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Figure 1. Increase in mean arterial pressure (MAP, mmHg) and decrease in heart rate (HR,
bpm) in response to phenylephrine (PHE, 8 mg/kg i.v.) in WKY and SHR exposed to SSCS
(WS, n ¼ 15; SS, n ¼ 16, respectively) and to ambient air (WC, n ¼ 22; SC, n ¼ 18,
respectively). *p 5 0.05: Different from control WKY; #p 5 0.05: Different from WKY
exposed to SSCS.

to activate baroreflex is well accepted by the literature (Valenti et al. 2009a,

2009b; Cisternas et al. 2010). PHE increases arterial pressure and activates the
parasympathetic component of the baroreflex, the bradycardic reflex and then
calculates bradycardic baroreflex gain. NaNP reduces blood pressure and
activates the sympathetic component of the baroceptor reflex (Valenti et al.
2009a, 2009b; Cisternas et al. 2010).
To the best of our knowledge, no study has specifically investigated the impact
of a short period of exposure to SSCS, the component usually inhaled by non-
smokers exposed to passive smoking, on baroreflex components in SHR. Apart
from nicotine, cigarette smoke contains thousands of other chemical substances,
including carbon monoxide, hydrogen cyanide, nitrogen oxides, aldehydes, N-
nitrosamines, polyaromatic hydrocarbons (Cenko et al. 2004; Pouliou et al. 2008).
SSCS has a higher concentration of toxic substances compared to mainstream
smoke due to a lower temperature of combustion as well as lack of filtering. For
instance, there is five times more acrolein in SSCS compared to mainstream smoke
(Talbot et al. 1998).
 International Journal of Environmental Health Research 435
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Figure 2. Decrease in mean arterial pressure (MAP, mmHg) and decrease in heart rate (HR,
bpm) in response to sodium nitroprussied (NaNP, 50 mg/kg i.v.) in WKY and SHR exposed to
SSCS (WS, n ¼ 15; SS, n ¼ 16, respectively) and to ambient air (WC, n ¼ 22; SC, n ¼ 18,
respectively). *p 5 0.05: Different from control SHR; #p 5 0.05: Different from SHR
exposed to SSCS.

Our results from ANOVA suggest that while in normotensive WKY rats SSCS
exposure affects parasympathetic and sympathetic components of baroceptor reflex,
in SHR it influences only the sympathetic component before change arterial
pressure. Therefore, short-term SSCS exposure affects different components of
baroreflex in WKY and SHR.
In conclusion, sidestream cigarette smoke exposure over three weeks affected the
sympathetic and parasympathetic component of the baroreflex in WKY while it
affected the sympathetic component of the baroreflex in SHR. We believe that
secondhand smoke affects the cardiovascular system firstly by changing the
autonomic function.

Acknowledgments
We thank Mr Jason Saltsgiver for critically evaluating the English grammar. This
research was supported by grants from Fundação de Amparo à Pesquisa do Estado
de São Paulo (FAPESP) (003/08).
 436 V.E. Valenti et al.

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