Application of phycocyanin in the preparation of drugs for alleviating nickel toxicity

Abstract

The present invention discloses the use of phycocyanin in the preparation of drugs for alleviating nickel toxicity, and belongs to the field of biomedicine technology. Phycocyanin is easy to obtain, has no biological toxicity, and has stable properties. The present invention has confirmed through experiments that phycocyanin can improve the renal glomerular enlargement and tubular epithelial cell vacuoles caused by nickel, and significantly restore blood uric acid levels. It can also restore the levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and total cholesterol in mice, and alleviate the metabolic disorders caused by nickel. It can also restore the renal damage gene Cndp2, Fga, and Ubc The expression level of the protein provides a reference for solving the environmental problem of heavy metal nickel pollution.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to the application of phycocyanin in the preparation of a drug for alleviating nickel toxicity. Background Art

[0002] Nickel is a metal pollutant widely distributed in the environment. The International Agency for Research on Cancer has classified nickel compounds and metallic nickel as Class I and Class IIB carcinogens, respectively. 2+ It exists in the form of chlorinated nickel and can cause damage to the liver, lungs, kidneys and immune system of organisms through water and air. Studies have shown that the main toxic mechanisms of nickel are: (1) Leading to pulmonary toxicity: Epithelial-mesenchymal transition is a key factor in the pathogenesis of nickel exposure-related lung diseases. (2) Leading to immunotoxicity: High doses of nickel can induce excessive apoptosis of cells in immune organs such as the thymus and spleen, and G0 / G1 phase arrest of thymic and bursal cells. At the same time, NiCl2 can also reduce the content of antioxidant enzymes in immune organs such as the thymus and spleen. Antioxidant function damage further leads to the accumulation of reactive oxygen species (ROS), which subsequently induces DNA damage and cell apoptosis. (3) Leading to neurotoxicity: The neurotoxicity of nickel includes the destruction of trace element homeostasis, neurotransmitter imbalance and destruction, and energy metabolism disorders. (4) Leading to renal toxicity: NiCl2 can induce autophagy and apoptosis of TCMK-1 cells through the oxidative stress-dependent AMPK / AKT-mTOR pathway. Nickel is widely used in human life, but excessive exposure to nickel is extremely harmful to human health, and accumulation in the human body may cause serious organ damage. Therefore, how to reduce the damage caused by nickel is crucial. Summary of the invention

[0003] The purpose of the present invention is to provide the use of phycocyanin in the preparation of a drug for alleviating nickel toxicity, so as to solve the problems existing in the above-mentioned prior art.

[0004] To achieve the above object, the present invention provides the following solutions:

[0005] One of the technical solutions of the present invention is the use of phycocyanin in the preparation of drugs for alleviating nickel toxicity.

[0006] The second technical solution of the present invention is a drug for alleviating nickel toxicity, comprising phycocyanin.

[0007] The third technical solution of the present invention is a pharmaceutical preparation for alleviating nickel toxicity, comprising phycocyanin or the drug described in the second technical solution of the present invention, and a pharmaceutically acceptable carrier.

[0008] Based on the above technical solution, the present invention has the following technical effects:

[0009] The present invention provides the use of phycocyanin in the preparation of a drug for alleviating nickel toxicity. Phycocyanin is easy to obtain, has no biological toxicity, and has stable properties. The present invention has confirmed through experiments that phycocyanin can improve the renal glomerular enlargement and tubular epithelial cell vacuoles caused by nickel, and significantly restore blood uric acid levels. It can also restore the levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and total cholesterol in mice, and alleviate the metabolic disorders caused by nickel. It can also restore the renal damage gene Cndp2, Fga, Ubc The expression level of the protein provides a reference for solving the environmental problem of heavy metal nickel pollution. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0011] Figure 1 The effect of phycocyanin on the kidney of nickel-treated mice. A is the HE staining result of the kidney tissue of the NaCl control group; B is the HE staining result of the kidney tissue of the NiCl2-treated group; C is the HE staining result of the kidney tissue of the PC group.

[0012] Figure 2 Effect of phycocyanin on blood uric acid levels in nickel-treated mice.

[0013] Figure 3 The effect of phycocyanin on blood lipids in nickel-treated mice. A is the content of high-density lipoprotein cholesterol; B is the content of low-density lipoprotein cholesterol; C is the content of serum total cholesterol.

[0014] Figure 4 Verification of key gene expression in renal injury. DETAILED DESCRIPTION

[0015] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0016] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0017] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0018] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0019] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0020] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0021] The embodiment of the present invention provides the use of phycocyanin in preparing a drug for alleviating nickel toxicity.

[0022] In some specific embodiments, phycocyanin alleviates nickel-induced renal damage, reduces nickel-induced low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and total cholesterol levels, and regulates renal damage genes. Cndp2, Fga, Ubc expression level, alleviating nickel toxicity.

[0023] An embodiment of the present invention further provides a drug for alleviating nickel toxicity, comprising phycocyanin.

[0024] An embodiment of the present invention further provides a pharmaceutical preparation for alleviating nickel toxicity, comprising phycocyanin or the drug described in the second technical solution of the present invention, and a pharmaceutically acceptable carrier.

[0025] In some specific embodiments, the pharmaceutically acceptable carrier is suitable for liquid dosage forms, solid dosage forms or paste dosage forms.

[0026] Phycocyanin is a high-value protein with antioxidant, neuromodulatory and immunomodulatory effects, and plays an important protective role against organ damage caused by heavy metals. In order to explore the effect of phycocyanin on nickel-induced renal damage in mice, the present invention used NiCl2 solution to stress mice for 28 days, and intervened with phycocyanin at the same time, exploring the control of nickel pollution and the resource utilization of phycocyanin. The present invention explored whether phycocyanin can effectively alleviate the kidney damage in mice caused by nickel, and found that phycocyanin can effectively alleviate the kidney damage in mice caused by nickel, providing a reference for solving the environmental problems of heavy metal nickel pollution.

[0027] The present invention obtained 6-week-old ICR male mice from Peking University Health Science Center, raised them for one week to adapt to the environment, stressed the mice with 10 mg / kg NiCl2 solution for 28 days, and treated them with phycocyanin. Through the detection of blood uric acid (UA) content, blood lipid content and the expression of key genes for kidney damage, it was found that phycocyanin can alleviate the damage of mouse kidneys caused by nickel. Example 1

[0028] Thirty male ICR mice were randomly divided into three groups: NaCl control group (10 mice), 10 10 mg / kg NiCl2 treatment group (10 mice), and 25 mg / kg phycocyanin (PC) group (10 mice), with 5 mice randomly divided into cages in each group. The specific administration methods were as follows: NaCl control group: 0.9% saline intraperitoneal injection (ip) + PBS gavage (ig); NiCl2 treatment group: 10 mg / kg NiCl2 (ip) + PBS (ig); PC group: 10 mg / kg NiCl2 (ip) + 25 mg / kg PC (ig); once a day, the experimental period was 28 days in total.

[0029] 1. Construction of renal injury model: After 28 days of nickel treatment, mice (5 mice in each group) were anesthetized and killed by cervical dislocation. The kidney tissue of each mouse was isolated and immediately rinsed with 1× PBS, and then fixed with 4% paraformaldehyde for 24 h. The kidney specimens were dehydrated, embedded in paraffin, and cut into 4 μm thick sections. HE staining was performed using a hematoxylin and eosin (HE) staining kit. The results are shown in Figure 2. Figure 1 As shown in the figure, there was no significant change in the morphology of glomeruli and tubules in mice treated with NaCl; the mice treated with NiCl2 showed glomerular enlargement and interstitial shrinkage, accompanied by vacuoles in tubular epithelial cells, epithelial cell shedding and cell fragmentation; after treatment with phycocyanin, the glomerular enlargement, tubular epithelial cell shedding and cell fragmentation in mice in PC group were improved.

[0030] 2. Determination of blood uric acid content: After 28 days of nickel treatment, the mice were anesthetized and blood was collected from the eyeballs. After standing in a 4°C refrigerator for 30 min, the serum was separated using a low-temperature high-speed centrifuge. The serum was collected and immediately tested for UA content using a blood uric acid detection kit to assess the degree of renal damage. Figure 2 As shown, the blood uric acid level of mice was significantly restored after phycocyanin perfusion, indicating that phycocyanin can alleviate nickel-induced renal damage in mice.

[0031] 3. Determination of blood lipid content in mice: After 28 days of nickel treatment, the mice were anesthetized and blood was collected from the eyeballs. After standing in a refrigerator at 4°C for 30 min, the serum was separated using a low-temperature high-speed centrifuge. The collected serum was immediately used to detect the blood lipid content of the mice using a total cholesterol (T-CHO) detection kit, a high-density lipoprotein cholesterol (HDL-C) detection kit, and a low-density lipoprotein cholesterol (LDL-C) detection kit. Figure 3 As shown, phycocyanin significantly restored the levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and total cholesterol in mice, indicating that phycocyanin alleviated the metabolic disorders caused by nickel to some extent.

[0032] 4. Determination of key genes for kidney damage in mice:

[0033] After 28 days of nickel treatment, mice (5 mice in each group) were anesthetized and killed by cervical dislocation. Kidney tissues were collected and placed in sterile cryopreservation tubes, quickly frozen in liquid nitrogen, and stored in a -80°C refrigerator. Total RNA from mouse kidneys was extracted using an RNA extraction kit and PrimeScript ® cDNA was synthesized using the RT Reagent Kit With gDNA Eraser (Perfect Real Time) and gene expression was detected using the TB Green TM Premix Ex Taq TM II Kit. Figure 4 As shown, after phycocyanin treatment, the renal injury gene Cndp2, Fga, Ubc The expression levels of phycocyanin were significantly restored, proving that phycocyanin can effectively alleviate nickel-induced kidney damage in mice.

[0034] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. The use of phycocyanin in the preparation of a drug for alleviating nickel toxicity, characterized in that: Phycocyanin alleviates nickel-induced renal damage, restores nickel-induced LDL-C, HDL-C and total cholesterol levels, and modulates renal damage genes Cndp2, Fga, and Ubc expression level, alleviating nickel toxicity.

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