Application of beta-hydroxybutyric acid and ketogenic diet in malaria prevention and treatment

Abstract

The embodiment of the invention discloses application of beta-hydroxybutyric acid and ketogenic diet in malaria prevention and treatment, and belongs to the technical field of malaria prevention and treatment, the ketogenic diet can obviously increase the content of beta-hydroxybutyric acid generated in the body, the beta-hydroxybutyric acid has an obvious inhibiting effect on development and reproduction of plasmodium in the body, and the effect of preventing and treating malaria is achieved. Beta-hydroxybutyric acid can completely resist infection of plasmodium, and beta-hydroxybutyric acid as a normal metabolite has no obvious side effect on the organism, so that ketogenic diet or use of beta-hydroxybutyric acid can endow the organism with the capability of resisting malaria infection, and the propagation path of malaria is cut off while malaria is treated; the method is simple, convenient and economical, and a new method is provided for malaria prevention and control in a mode of replacing therapy with food, namely metabolism therapy.

Description

Technical Field

[0001] The embodiments of the present invention relate to the technical field of malaria prevention and treatment, and specifically to an application of β-hydroxybutyric acid and a ketogenic diet in malaria prevention and treatment. Background Art

[0002] Malaria has always been a major infectious disease that endangers human health. Every year, about 250 million people are infected with the disease worldwide, and about 3 billion people are threatened by malaria. About 800,000 people die from falciparum malaria infection, most of whom are children under the age of 5. At present, the prevention and control of malaria mostly rely on vaccines and drugs. However, due to the continuous mutation of the malaria-causing virus and the development of population mobility and international trade, drug resistance outbreaks from different sources have occurred many times, posing a huge challenge to global prevention and control work.

[0003] In recent years, due to the continuous emergence and spread of artemisinin-resistant strains, the number of malaria infections and deaths has gradually increased, and malaria prevention and control faces huge challenges. The slow development of new antimalarial drugs has led to a decrease in the number of effective drugs available in clinical practice, and the shortage of antimalarial drugs has become a serious problem. In order to better control the spread and harm of malaria, it is urgent to develop new malaria treatment strategies. Summary of the invention

[0004] To this end, an embodiment of the present invention provides an application of β-hydroxybutyrate and a ketogenic diet in the prevention and treatment of malaria, so as to solve the problem in the prior art that the spread of malaria cannot be well controlled due to the lack of treatment methods for the continuous emergence and spread of artemisinin-resistant strains.

[0005] In order to achieve the above purpose, the embodiment of the present invention provides the following technical solutions:

[0006] According to a first aspect of an embodiment of the present invention, the present invention provides use of β-hydroxybutyrate (βOHB) in preparing a drug for preventing and treating malaria.

[0007] Furthermore, the β-hydroxybutyric acid provides energy for the body and is also involved in metabolism and immune regulation.

[0008] According to a second aspect of an embodiment of the present invention, the present invention provides a use of a ketogenic diet in preventing and treating malaria, wherein the ketogenic diet exerts its effect by increasing β-hydroxybutyrate in the body.

[0009] Furthermore, the ketogenic diet consists of high fat content and low protein content, wherein the fat content is 90% cal and the protein content is 10% cal, and is purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd. using the formula of XTKD01.

[0010] Further, the ketogenic diet is fed before and during malaria infection.

[0011] The embodiments of the present invention have the following advantages:

[0012] 1. The present invention aims to solve the above-mentioned problems of drug resistance and drug shortage of malarial parasites, and provides a new strategy for the prevention and treatment of malaria. β-hydroxybutyric acid, as one of the main components of ketone bodies in the human body, can not only provide energy for the body, but also participate in the regulation of metabolism and immunity.

[0013] 2. The ketogenic diet can significantly increase the production of β-hydroxybutyric acid in the body. β-hydroxybutyric acid has a significant inhibitory effect on the development and reproduction of malarial parasites in the body, and can completely resist the infection of malarial parasites. Moreover, β-hydroxybutyric acid, as a normal metabolite, has no obvious side effects on the body. Therefore, the ketogenic diet or the use of β-hydroxybutyric acid can give the body the ability to resist malarial infection, while treating malaria and cutting off its transmission route;

[0014] 3. The present invention is simple, economical, and provides a new method for malaria prevention and control by using the method of "food instead of therapy" or "metabolic therapy". BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the implementation methods of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the implementation methods or the description of the prior art. Obviously, the drawings in the following description are only exemplary, and for ordinary technicians in this field, other implementation drawings can be derived from the provided drawings without creative work.

[0016] Figure 1 The survival curves of two groups of mice provided in Example 1 of the present invention;

[0017] Figure 2 The statistical graph of the worm infection rate of RD+PbA and KD+PbA mice 30 days after attacking worms provided in Example 1 of the present invention;

[0018] Figure 3 This is a graph of serum βOHB concentrations in four groups of mice, RD, RD+PbA, KD, and KD+PbA, seven days after being challenged with parasites, provided in Example 2 of the present invention;

[0019] Figure 4 A line graph of the infection rate and median lethal concentration determined by βOHB provided in Example 3 of the present invention;

[0020] Figure 5 This is a statistical chart of the weight changes of mice in Example 4 of the present invention when they were fed with conventional feed and ketogenic feed for 30 days;

[0021] Figure 6This is a statistical chart of the routine blood test of mice in Example 4 of the present invention after being fed with conventional feed and ketogenic feed for 30 days. DETAILED DESCRIPTION

[0022] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] βOHB (S1031) (β-Hydroxybutyric acid) was purchased from Selleck; ketogenic diet XTKD01 was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.; β-Hydroxybutyrate Colorimetric Assay Kit was purchased from Elabscience; 2,2,2-Tribromoethanol was purchased from Aladdin.

[0024] Example 1

[0025] The preventive and therapeutic effects of ketogenic diet on Plasmodium infection in mice

[0026] Step 1: Take 20-23g female C57BL / 6J mice, and keep them in an environment with 12 hours of light, 12 hours of darkness, moderate temperature and humidity, with free access to food and water, and adaptive feeding for one week;

[0027] Step 2: The mice were randomly divided into two groups: a regular diet (RD) + parasite group and a ketogenic diet (KD) + parasite group; the malarial parasites infected with the mice were Plasmodium bergheiANKA (PbA); the mice in the RD + parasite group were fed with a regular diet, and the mice in the KD + parasite group were fed with a ketogenic diet (XTKD01, Synergy Biotech);

[0028] Step 3: After two weeks of pre-feeding, mice in the RD and KD groups were intraperitoneally injected with 2×10 Plasmodium berghei. 5 Worms / mouse;

[0029] Step 4: After the third day of infection, blood was collected from the tail tips of the two groups of mice every 2-3 days to prepare thin blood smears, which were then stained with Giemsa to calculate the infection rate of each group;

[0030] Step 5: Record the survival status of the two groups of mice and draw a survival curve.

[0031] Depend on Figure 1 The results showed that feeding a ketogenic diet could significantly prolong the survival of the mice in the infected group. Figure 2 The results showed that after testing each group of mice after worm attack, it was found that compared with the mice in the conventional feed group, whose worm infection rate continued to rise, the worm infection rate in the ketogenic group appeared later and disappeared quickly. Only a few worms were detected on the 10th day, which was consistent with the survival curve.

[0032] Example 2

[0033] Increased concentration of circulating ketone bodies in mice fed a ketogenic diet

[0034] Step 1: Take 20-23g female C57BL / 6J mice, and keep them in an environment with 12 hours of light, 12 hours of darkness, moderate temperature and humidity, with free access to food and water, and adaptive feeding for one week;

[0035] Step 2: The mice were randomly divided into four groups: RD group, RD+PbA group, KD group, and KD+PbA group. The mice in the RD group and RD+PbA group were fed with conventional feed, and the mice in the KD group and KD+PbA group were fed with ketogenic feed (XTKD01, Synergy Biotech);

[0036] Step 3: After two weeks of pre-feeding, mice in the RD+PbA group and the KD+PbA group were intraperitoneally injected with 2×10 Plasmodium berghei. 5 Parasites / mouse, the malarial parasite that infects mice is Plasmodium berghei ANKA (PbA);

[0037] Step 4: 7 days after the attack, mice were anesthetized by intraperitoneal injection of avertin (2,2,2-Tribromoethanol, Aladdin 75-80-9), and blood was collected from the inferior vena cava;

[0038] Step 5: After the blood was allowed to stand at 4°C for 30 min, it was centrifuged at 3000 rpm for 15 min;

[0039] Step 6: Use a pipette to aspirate the upper layer of serum, and use the ketone body detection kit β-Hydroxybutyrate (KetoneBody) Colorimetric Assay Kit (Elabscience, E-BC-K785-M) to detect the serum ketone body content.

[0040] Depend on Figure 2 The results showed that the concentration of circulating ketone body βOHB in mice was significantly increased.

[0041] Example 3

[0042] Inhibitory effect of β-hydroxybutyrate on the development and reproduction of Plasmodium falciparum 3D7 strain that infects humans only in vitro

[0043] Step 1: Collect Plasmodium falciparum 3D7 parasites at the ring stage and adjust the percentage of infected parasite cells to 0.7%;

[0044] Step 2, culturing the worms in vitro with culture medium containing 0, 1, 2, 4, 8, 10, 12, 14, 16, 20, and 25 mM βOHB (Selleck, S1031);

[0045] Step 3, after the parasites have developed for two cycles, the infection rate of the falciparum malaria 3D7 strain cultured with ketone bodies of different concentrations is detected by flow cytometry;

[0046] Step 4, plot and calculate the median lethality of ketone body βOHB against the falciparum malaria 3D7 strain.

[0047] In vitro experiments have shown that ketone body βOHB also has a strong inhibitory effect on Plasmodium falciparum infection in humans and its median lethal concentration was measured to be 7.268 mM ( Figure 4 ), which is consistent with the effect in mice.

[0048] Example 4

[0049] Determination of toxicological effects of β-hydroxybutyrate on mice

[0050] Step 1: Take 20-23g female C57BL / 6J mice, and keep them in an environment with 12 hours of light, 12 hours of darkness, moderate temperature and humidity, with free access to food and water, and adaptive feeding for one week;

[0051] Step 2: One week later, the mice were fed with regular feed (RD) and ketogenic feed (KD) and given normal drinking water;

[0052] Step 3: Weigh the mice every three days;

[0053] Step 4: After the 30th day of feeding, the mice were anesthetized with Avertin and blood was collected from the inferior vena cava for blood index detection;

[0054] Step 5: Use the Tecon veterinary fully automatic blood cell analyzer (TEK-VET3) to perform a routine blood test.

[0055] The results showed that a one-month health monitoring of the ketogenic diet mice revealed no significant difference in body weight compared with the mice in the conventional feed group ( Figure 5 ), and no significant differences were found in the indicators of the two groups of mice in the subsequent blood routine tests ( Figure 6), indicating that the ketogenic diet has no obvious toxic effect on the body.

[0056] The above results show that ketone body βOHB has a significant inhibitory effect on the development and reproduction of malarial parasites both in vivo and in vitro. A ketogenic diet can increase the circulating concentration of ketone bodies in the body to achieve the effect of killing malarial parasites without obvious toxic effects on the body.

[0057] Although the present invention has been described in detail above by general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. Application of β-hydroxybutyric acid in the preparation of drugs for preventing and treating malaria.

2. The use according to claim 1, characterized in that: The β-hydroxybutyric acid provides energy for the body and is also involved in metabolism and immune regulation.

3. A use of a ketogenic diet in preventing and treating malaria, characterized in that: The ketogenic diet works by increasing beta-hydroxybutyrate in the body.

4. The use according to claim 3, characterized in that: The ketogenic diet was administered prior to and during malaria infection.

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