Effect of Empagliflozin on the Reactivity of Mesenteric Arteries and Skin Microvessels in Rats Treated with Doxorubicin

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Abstract

The study assessed the potential protective effect of empagliflozin (EMPA) on the functional state of various types of vessels in Wistar rats that received a single injection of the anthracycline antibiotic doxorubicin (DOX), used clinically as a chemotherapeutic agent for cancer. The rats were divided into 3 groups, 15 animals in each group. Rats in the DOX group were administered DOX (4 mg/kg) once, while animals in the DOX+EMPA group after a single administration of DOX (4 mg/kg) received EMPA (1 mg/kg) daily through a tube for 5 weeks. The control group consisted of intact animals. After 4 weeks of the experiment, the rats were examined for the initial skin microcirculation indices and their changes after iontophoresis of acetylcholine (ACh) and sodium nitroprusside (NP) using Laser Doppler Flowmetry (LDF). One week after LDF, the mesenteric artery dilation value was analyzed by assessing the changes in vessel diameter before and after the action of ACh and NP, without blockers and under conditions of preliminary incubation of vessels with the NO synthase blocker L-NAME. In the control group rats, ACh iontophoresis caused an increase in perfusion intensity by 78.5%, in the DOX group the change was less pronounced (by 55.2%). EMPA prevented a decrease in the skin microvessel response to ACh, the perfusion index in rats of the DOX+EMPA group increased by 82.8%. The increase in the microcirculation index after NP iontophoresis in the DOX+EMPA group did not differ from the control, and in the DOX group it was significantly lower. ACh-induced dilation of the mesenteric arteries of the DOX group was 24.3% lower than in the control rats. The use of EMPA in rats that received DOX improved arterial reactivity. Compared with the reactivity of vessels without blockers, incubation of vessels with L-NAME reduced the amplitude of dilation under the action of ACh in all groups, but a less pronounced change was observed in the DOX group (45.6%). When using EMPA, the differences in the relaxation amplitude before and after NO synthase blockade increased (54.4%), but did not reach the control (64.1%). Conclusion. DOX leads to a decrease in the reactivity of various types of vessels to the action of vasodilators, in particular, ACh and NP. The use of EMPA has a protective effect in animals after the introduction of DOX, improving the dilation of the mesenteric arteries and vessels of the skin microcirculatory bed. It is possible that the effect of EMPA is associated with an improvement in the efficiency of NO-dependent vasorelaxation pathways, the disruption of which is observed upon the introduction of DOX.

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G. T. Ivanova

Pavlov Institute of Physiology of the RAS

Author for correspondence.
Email: ivanovagt@infran.ru
Russian Federation, Saint Petersburg

O. N. Beresneva

First Pavlov Saint Petersburg State Medical University

Email: ivanovagt@infran.ru
Russian Federation, Saint Petersburg

S. V. Okovity

Saint Petersburg State Chemical and Pharmaceutical University

Email: ivanovagt@infran.ru
Russian Federation, Saint Petersburg

A. N. Kulikov

First Pavlov Saint Petersburg State Medical University

Email: ivanovagt@infran.ru
Russian Federation, Saint Petersburg

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2. Fig. 1. Effect of empagliflozin on left ventricular mass index (IMLV) and myocardial mass index (IMM) in rats after doxorubicin administration. Experimental groups: control - control group, DOX - animals that received a single 4 mg/kg doxorubicin, DOX + EMPA - animals that received a single 4 mg/kg doxorubicin and daily 1 mg/kg empagliflozin. Data are presented as mean ± standard error. p - significance of differences between groups.

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3. Fig. 2. Initial microcirculation index in the skin of rats of experimental groups. On the ordinate axis: microcirculation index, perfusion units (PU). Experimental groups: control - control group, DOX - animals that received once 4 mg/kg doxorubicin, DOX + EMPA - animals that received once 4 mg/kg doxorubicin and daily 1 mg/kg empagliflozin. Data are presented as mean ± standard error.

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4. Fig. 3. Calculated value of endothelial (ET), neurogenic (NT) and myogenic (MT) vascular tone of the rat skin microcirculatory channel. On the ordinate axis - tonus value, units. Experimental groups: control - control group, DOX - animals that received 4 mg/kg doxorubicin once, DOX + EMPA - animals that received 4 mg/kg doxorubicin once and 1 mg/kg empagliflozin daily. Data are presented as mean ± standard error. p - significance of differences between groups.

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5. Fig. 4. Mean microcirculation index in rat skin after ionophoresis of acetylcholine solution (ACh) and sodium nitroprusside (NP). On the ordinate axis: microcirculation index, perfusion units (PU). Experimental groups: control - control group, DOX - rats that received doxorubicin, DOX + EMPA - rats that received doxorubicin and empagliflozin treatment.

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6. Fig. 5. Relaxation of rat mesenteric arteries under the action of sodium nitroprusside. On the axis of ordinates - amplitude of dilatation of mesenteric arteries precontracted by phenylephrine (1 × 10-6 mol/l) during sodium nitroprusside (NP, 1 × 10-6 mol/l) administration, expressed as % of the amplitude of constriction on phenylephrine. Experimental groups: control - control group, DOX - rats that received doxorubicin, DOX + EMPA - rats that received doxorubicin and empagliflozin treatment. Data are presented as mean value ± standard error. p - significance of differences between groups.

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7. Fig. 6. Acetylcholine-induced dilatation of rat mesenteric arteries precontracted by phenylephrine, cumulative effect. (a) - amplitude of dilatation of mesenteric arteries during acetylcholine action (ACh): abscissa axis - logarithm of acetylcholine concentration, ordinate axis - amplitude of relaxation expressed as a percentage of the contraction amplitude to phenylephrine (1 x 10-6 mol/L); (b) - area under the curve for acetylcholine: on the ordinate axis is the area under the curve of acetylcholine concentration-dependent dilatation amplitude, units. units. Experimental groups: control - control group, DOX - rats that received doxorubicin, DOX + EMPA - rats that received doxorubicin and empagliflozin treatment. Data are presented as mean value ± standard error. p - significance of differences between groups.

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8. Fig. 7. Acetylcholine-induced dilatation of phenylephrine-pretreated rat mesenteric arteries in the absence of blockers and after incubation of vessels with L-NAME (1 × 10-4 mol/L). Abscissa axis: amplitude of dilatation of mesenteric arteries precontracted by phenylephrine (1 × 10-6 mol/L) under the action of acetylcholine (1 × 10-5 mol/L) expressed as a percentage of the amplitude of constriction to phenylephrine. Experimental groups: control - control group, DOX - rats that received doxorubicin, DOX + EMPA - rats that received doxorubicin and empagliflozin treatment. Data are presented as mean value ± standard error. p - significance of differences between groups.

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