Intravenous Nicotinamide Riboside Administration Has a Cardioprotective Effect in Chronic Doxorubicin-Induced Cardiomyopathy

Abstract

Doxorubicin, which is widely used to treat a broad spectrum of malignancies, has pronounced dose-dependent side effects leading to chronic heart failure development. Nicotinamide riboside (NR) is one of the promising candidates for leveling the cardiotoxic effect. In the present work, we performed a comparative study of the cardioprotective and therapeutic actions of various intravenous NR administration modes in chronic doxorubicin-induced cardiomyopathy in Wistar rats. The study used 60 mature male SPF Wistar rats. The animals were randomized into four groups (a control group and three experimental groups) which determined the doxorubicin (intraperitoneally) and NR (intravenous) doses as well as the specific modes of NR administration (combined, preventive). We demonstrated the protective effect of NR on the cardiovascular system both with combined and preventive intravenous drug administration, which was reflected in a fibrous tissue formation decrease, reduced fractional-shortening decrease, and better antioxidant system performance. At the same time, it is important to note that the preventive administration of NR had a more significant protective effect on the animal organism as a whole. This was confirmed by better physical activity parameters and vascular bed conditions. Thus, the data obtained during the study can be used for further investigation into chronic doxorubicin-induced cardiomyopathy prevention and treatment approaches.

Keywords: NAD+; anthracyclines; doxorubicin-induced cardiomyopathy; fibrosis; intravenous administration; nicotinamide riboside; sirtuins.

Figure 1

Body−weight dynamics. 1—drug administration endpoint for control, DOX, and NR+DOX groups; 2—drug administration endpoint for NR/NR+DOX group. The first weighing was carried out before the start of each manipulation. During the drug administration, the animals were weighed every 2 days on the day of the substance administration (9 times). At the end of the administration in NR/NR+DOX group, the weight and clinical condition of the animals were monitored once a week. */*—p < 0.05, **—p < 0.01, and ***—p < 0.001 which shows significant difference compared to control group (*—DOX; *—NR+DOX). Data are presented as median [25%; 75%]. Control represents control group (n = 15); DOX represents doxorubicin group (n = 15); NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode (n = 15); and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode (n = 15).

Figure 2

Treadmill activity duration. */*—p < 0.05 compared to the initial value (start of the experiment) (*—DOX; *—NR+DOX). Data are presented as median [25%; 75%]. Control represents control group (n = 15); DOX represents doxorubicin group (n = 15); NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode (n = 15); and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode (n = 15).

Figure 3

Left-ventricular parameters according to ECHO. (A)—left-ventricular end-systolic internal diameter, or LVIDs; (B)—left-ventricular end-diastolic internal diameter, or LVIDd; (C)—fractional shortening, or FS; (D)—anterior wall thickness, or IVS; and (E)—posterior wall thickness, or LVPW. */*/*—significant difference compared to the start of the experiment (*—DOX; *—NR+DOX; and *—NR/NR+DOX); #/#/#—significant difference in FS between DOX, NR+DOX, and NR/NR+DOX groups compared to control group; and #/#/#—significant difference in LVIDs and LVIDd between control, DOX, and NR+DOX groups compared to NR/NR+DOX group (#—DOX; #—NR+DOX; and #—NR/NR+DOX). (*/*/*—p < 0.05, **/**/**—p < 0.01, and ***/***/***—p < 0.001; #/#/#—p < 0.05, ##/##—p < 0.01, and ###—p < 0.001.) Data are presented as median [25%; 75%]. Control represents control group (n = 15); DOX represents doxorubicin group (n = 15); NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode (n = 15); NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode (n = 15).

Figure 4

Representative ECHO photographs of control, DOX, NR+DOX, and NR/NR+DOX groups at the start of the experiment and two months after doxorubicin administration completion. (A,B)—control group; (C,D)—DOX group; (E,F)—NR+DOX group; and (G,H)—NR/NR+DOX group. DOX represents doxorubicin group; NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode; and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode.

Figure 5

Cardiac muscle tissue. (A)—control group, normal structure. (B)—DOX group, dystrophic changes in cardiomyocytes: loss of striation (1,2 arrows) and vacuolization of the cytoplasm (stars). (C)—NR+DOX group, minimal alterative changes. (D)—NR/NR+DOX group, minimal alterative changes. Stained with hematoxylin–eosin, ×50. DOX represents doxorubicin group; NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode; and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode.

Figure 6

Morphometric values (% area of collagen fibers) in cardiac muscle. (A,B)—interstitial fibrosis; (C,D)—perivascular fibrosis; (E,F)—epicardial fibrosis. Collagen fibers are blue. Staining by Mallory, × 100. *—p < 0.05, **—p < 0.01, and ***—p < 0.001. Data are presented as median [25%; 75%]. Control represents control group (n = 15); DOX represents doxorubicin group (n = 15); NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode (n = 15); and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode (n = 15).

Figure 7

Mesenteric artery functional activity. (A)—dose-dependent response curves to phenylephrine (PE); (B)—dose−dependent response curves to acetylcholine (ACh); (C)—area under the curves (AUC) for PE-dependent response; and (D)—AUC for ACh-dependent response. *—p < 0.05, and ***—p < 0.001, significant difference compared to control group. Values represent mean ± SEM mean from 9 arteries of 3 rats in every group (n = 36). Control represents control group; DOX represents doxorubicin group; NR+DOX represents nicotinamide riboside + doxorubicin group—combined mode; and NR/NR+DOX represents nicotinamide riboside/nicotinamide riboside + doxorubicin group—preventive mode.

Figure 8

Experimental design.