CN113662955A

CN113662955A - Application of nicotinamide adenine dinucleotide in treatment of adriamycin-induced cardiotoxicity

Info

Publication number: CN113662955A
Application number: CN202111085332.0A
Authority: CN
Inventors: 孙爱军; 葛均波; 章金延; 蒋昊
Original assignee: Zhongshan Hospital Fudan University
Current assignee: Zhongshan Hospital Fudan University
Priority date: 2021-09-16
Filing date: 2021-09-16
Publication date: 2021-11-19
Also published as: WO2023040716A1

Abstract

The invention discloses application of nicotinamide adenine dinucleotide in treating adriamycin-induced cardiotoxicity, and belongs to the technical field of biological medicines. Animal experiments prove that the heart function of the DOX + NAD group is obviously improved compared with that of the DOX + Control group; cell experiments prove that the ratio of dead cells of the DOX + NAD group is obviously reduced compared with the DOX + PBS group; the invention firstly provides the application of nicotinamide adenine dinucleotide in treating adriamycin-induced cardiotoxicity, can reduce myocardial cell death caused by adriamycin by adopting NAD, relieve the cardiotoxicity caused by adriamycin, resist side effects generated by adriamycin, enlarge the application range of adriamycin and improve the prognosis of tumor patients clinically using adriamycin.

Description

Application of nicotinamide adenine dinucleotide in treatment of adriamycin-induced cardiotoxicity

Technical Field

The invention relates to application of nicotinamide adenine dinucleotide in treating adriamycin-induced cardiotoxicity, and belongs to the technical field of biological medicines.

Background

Doxorabicin (DOX) is one of the most widely used anti-tumor drugs in clinical use at present, and the main mechanism is to achieve the effect of killing tumor cells by inhibiting the synthesis of DNA and RNA. However, cardiotoxicity caused by doxorubicin often limits its clinical use. Although the current techniques for reducing cardiotoxicity by liposome-encapsulated adriamycin have been greatly developed, it has not been possible to completely prevent myocardial damage caused by adriamycin. Therefore, there is a need to discover new targets for doxorubicin-induced cardiotoxicity therapy from a new perspective.

Abnormal myocardial energy metabolism is an important pathological basis for doxorubicin-induced myocardial injury, and mitochondria play a key role therein as a major site for ATP supply. It has been reported that doxorubicin causes increased apoptosis of the mitochondrial pathway of cardiomyocytes, dysregulation of mitochondrial autophagy homeostasis, metabolic dysfunction such as the tricarboxylic acid cycle, etc. Therefore, the mitochondria function is improved, and the protein can be used as a potential target for preventing myocardial damage induced by adriamycin.

Nicotinamide adenine dinucleotide (NAD +) is a coenzyme necessary for the maintenance of mitochondrial metabolism and function. First, NAD + can act as an electron acceptor, transferring energy in different metabolic pathways, such as fatty acid oxidation and the tricarboxylic acid cycle. Second, NAD + is also required for mitochondrial oxidative phosphorylation, and NAD + deficiency will result in mitochondrial failure to produce ATP. Studies have shown that abnormalities in mitochondrial function are often accompanied by a decrease in NAD +. And supplementation with NAD + can significantly improve mitochondrial function. Currently, there are several clinical studies in which NAD + levels are increased by replenishing NAD + precursors to treat metabolic diseases. But the therapeutic advantages of NAD + on doxorubicin-induced cardiotoxicity have not been explored at present.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: how to improve doxorubicin-induced cardiotoxicity.

In order to solve the technical problems, the invention provides application of nicotinamide adenine dinucleotide in preparing a medicine for treating adriamycin-induced cardiotoxicity.

Preferably, the medicament comprises a pharmaceutically acceptable carrier and an effective amount of an active ingredient which is nicotinamide adenine dinucleotide.

Compared with the prior art, the invention has the beneficial effects that:

1. animal experiments prove that the heart function of the DOX + NAD group is obviously improved compared with that of the DOX + Control group; cell experiments prove that the ratio of dead cells of the DOX + NAD group is obviously reduced compared with the DOX + PBS group;

2. the invention firstly provides the application of nicotinamide adenine dinucleotide in treating adriamycin-induced cardiotoxicity, and the NAD can reduce myocardial cell death caused by adriamycin, relieve cardiotoxicity caused by adriamycin, resist side effects caused by adriamycin, enlarge the application range of adriamycin and improve the prognosis of tumor patients clinically using adriamycin.

Drawings

FIG. 1 shows the results of Left Ventricular Ejection Fraction (LVEF) measurements in a mouse model;

FIG. 2 shows the results of measuring the left ventricular minor axis shrinkage (LVFS) of a mouse model;

FIG. 3 shows the result of dead and alive staining of cardiomyocytes; wherein green represents live cells (shown as dots in the figure) and red represents dead cells (shown as circles in the figure);

FIG. 4 shows the ratio of dead cells in each group;

wherein, in the figure, "NS": no significant match (no statistical difference); *: p < 0.05; **: represents p < 0.01; ***: representing p < 0.001.

Detailed Description

In order to make the invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings.

Examples

This example demonstrates the use of nicotinamide adenine dinucleotide in the treatment of doxorubicin-induced cardiotoxicity by in vivo animal experiments:

1) grouping experimental animals and preparing animal models: SPF-grade male C57BL/6 mice (purchased from Shanghai Jie Si Jie laboratory animals Co., Ltd.) were selected and aged 8-10 weeks and weighed 20-25g, and divided into four groups (Sham + Control group; Sham + NAD group; DOX + Control group; DOX + NAD group), Sham + NAD group and DOX + NAD group were intraperitoneally injected for 7 consecutive days with NAD (50mg/kg/D, Sereck, S2518), Sham + Control group and DOX + Control group with an equal amount of saline, DOX + Control group for 8 days, DOX + NAD group was intraperitoneally injected with DOX (15mg/kg, Sigma, D1515), and then Sham + NAD group and DOX + NAD group were intraperitoneally injected daily with NAD (50mg/kg/D, Sereck, S2518), Sham + Control group and DOX + Control group, and mice were examined for an equal amount of cardiac function after 7 days.

2) Echocardiography assessment of mouse cardiac function: echocardiography was examined at 4 weeks with a probe frequency of 30 MHz. Specifically isoflurane animals, M-mode images were recorded while the heart rate of the mice was maintained at 450-. A B-Mode image of a long axis section beside a sternum and a four-cavity section at the apex of the heart is collected. And taking a short left ventricular axis beside the sternum, and performing 2D ultrasonic left ventricular short axis section, and applying M-type ultrasonic to record the motion condition of the left ventricle at the papillary muscle level. The functional indicators include: left Ventricular Ejection Fraction (LVEF), left ventricular minor axis contraction (LVFS). The heart morphology and function changes of the mice in each group are compared. All measurements were averaged over 5 consecutive cardiac cycles and were performed by 3 experienced technicians. The results show that: the heart function index of the mice in the DOX group is obviously lower than that of the Sham group, the heart function of the mice in the DOX group is improved after nicotinamide adenine dinucleotide is injected, and the EF percent of the mice in the DOX + NAD group is 10.24 percent higher than that of the mice in the DOX + Control group (P is less than 0.05); the FS% of the mice in the DOX + NAD group is 8.987% higher than that of the mice in the DOX + Control group (P < 0.05); the mice in the Sham group injected with NAD showed no difference in EF and FS compared with the mice in the normal saline control group, as shown in fig. 1 and 2.

3) Myocardial cell death and survival staining: SPF-grade male C57BL/6 mice (purchased from Shanghai Jie Si laboratory animals Co., Ltd.) with week age of 8-10 weeks and body weight of 20-25g were selected, and primary cardiomyocytes of adult mice were extracted and divided into 4 groups (PBS group; NAD group; DOX + PBS group; DOX + NAD group) under the following intervention conditions: cells were subjected to dead-live staining using a Calcein/PI cell activity and cytotoxicity detection kit (beyond, C2015S) after 24 hours of intervention in a DOX + NAD group (DOX 1uM + NAD 500uM), a DOX + PBS group (DOX 1uM + PBS equal amount), a NAD group (NAD 500uM), a PBS group (PBS equal amount), and a significant increase in dead cell ratio after DOX intervention (P <0.001), which was less than that in the DOX + NAD group (P <0.001), and a no difference in dead cell ratio between the PBS group and the NAD group, as shown in fig. 3 and 4.

The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way and substantially, it should be noted that those skilled in the art may make several modifications and additions without departing from the scope of the present invention, which should also be construed as a protection scope of the present invention.

Claims

1. Application of nicotinamide adenine dinucleotide in preparation of medicine for treating adriamycin-induced cardiotoxicity.

2. The use of claim 1, wherein the medicament comprises a pharmaceutically acceptable carrier and an effective amount of an active ingredient which is nicotinamide adenine dinucleotide.