Application of Glycyrrhizinol in the Preparation of Drugs for Preventing and / or Treating Acute Kidney Injury

Abstract

The present invention discloses the use of liquiritol in the preparation of a drug for preventing and / or treating acute kidney injury. Experimental results show that liquiritol can reduce the levels of creatinine and blood urea nitrogen in the serum of mice, improve the pathological characteristics of folic acid-induced acute kidney injury, restore the activity of GSH-Px, and inhibit the molecular mechanism of the increase in iron content induced by folic acid, thereby having the effect of alleviating acute kidney injury; at the same time, liquiritol is derived from the medicinal and edible homologous plant - licorice, with low toxicity and side effects, and is expected to be further developed into a safe and reliable therapeutic drug.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of glycyrol in the preparation of drugs for preventing and / or treating acute kidney injury. Background Art

[0002] Licorice is a plant with both medicinal and edible uses announced by the Ministry of Health of China. It belongs to the Leguminosae family and is a perennial herb. The effects of licorice include treating various liver diseases, anti-inflammatory, antitussive and expectorant, anti-cancer, etc. It is reported that the three main types of active ingredients of licorice are glycyrrhizin, licorice flavonoids and coumarin compounds. Glycyrol (GC) is a unique bioactive ingredient in licorice roots and belongs to one of the coumarin compounds in licorice. It has biological activities such as antibacterial, anti-inflammatory, immunosuppressive, and anti-tumor.

[0003]

[0004] Acute kidney injury (AKI) is a common and costly critical illness in clinical practice. Its incidence and mortality rates are both increasing year by year, and it is a global public health problem. AKI is mainly manifested as a sudden and continuous rapid decline in renal function and the continuous accumulation of metabolites. Some patients develop chronic kidney disease as the disease progresses. Even if the renal function of the patient partially or completely recovers, there is still a risk of developing end-stage renal disease.

[0005] Currently, it is not clear whether glycyrol has an inhibitory and therapeutic effect on acute kidney injury. Summary of the Invention

[0006] The purpose of the present invention is to provide a new medical use of glycyrol.

[0007] The new medical use of glycyrol provided by the present invention is: the application of glycyrol in the preparation of drugs for preventing and / or treating acute kidney injury.

[0008] The acute kidney injury may specifically be drug-induced acute kidney injury, and more specifically may be folic acid-induced acute kidney injury.

[0009] Specifically, the application of glycyrol in the preparation of drugs with the following functions:

[0010] 1) Reducing the content of creatinine and blood urea nitrogen in the serum,

[0011] 2) Restoring the activity of GSH-Px to inhibit ferroptosis.

[0012] The drug takes glycyrol as the only active ingredient; it may also further include other active ingredients, and those skilled in the art can determine other active ingredients according to the drug effect.

[0013] The dosage form of the drug can be one of tablets, ointments, creams, capsules, sustained-release tablets, controlled-release tablets, oral liquids, syrups, injection dosage forms, dripping pills, and lyophilized powder injection dosage forms.

[0014] Experimental findings of the present invention show that liquiritol can reduce the levels of creatinine and blood urea nitrogen in the serum of mice, improve the pathological characteristics of folic acid-induced acute kidney injury, restore the activity of GSH-Px, and inhibit the molecular mechanism of the increase in iron content induced by folic acid, thereby having the effect of alleviating acute kidney injury. At the same time, liquiritol is derived from the medicinal and edible homologous plant - licorice, with low toxicity and side effects, and is expected to be further developed into a safe and reliable therapeutic drug. Description of the Drawings

[0015] Figure 1 It is the flow chart of the animal experiment in Example 1 of the present invention.

[0016] Figure 2 It is the content of BUN in the serum of mice in each group in Example 1 of the present invention.

[0017] Figure 3 It is the content of CRE in the serum of mice in each group in Example 1 of the present invention.

[0018] Figure 4 It is the HE staining of mice in each group in Example 1 of the present invention.

[0019] Figure 5 It is the change results of MDA, GSH-Px activity and iron content in the kidneys of mice in each group in Example 2 of the present invention, as well as the change results of ferroptosis-related proteins. (A) MDA levels in the kidneys of mice in each group; (B) GSH-Px activities in the kidneys of mice in each group; (C) Iron contents in the kidneys of mice in each group; (D) Expression levels of Tfr1, FPN, and HO-1 in the kidneys of mice in each group. Detailed Embodiments

[0020] The present invention will be further described in detail below in combination with specific embodiments. The provided embodiments are only for clarifying the present invention, rather than limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and do not limit the present invention in any way.

[0021] The experimental methods in the following embodiments are all conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following embodiments can be obtained from commercial sources unless otherwise specified.

[0022] The present invention provides a new medical use of liquiritol.

[0023] The new medical use of liquiritol provided by the present invention is: the application of liquiritol in the preparation of drugs for preventing and / or treating acute kidney injury.

[0024] The acute kidney injury may specifically be drug-induced acute kidney injury, and more specifically be folic acid-induced acute kidney injury.

[0025] Example 1: Protection of liquiritol against folic acid-induced acute kidney injury in mice

[0026] Test method: 24 male Balb / c mice, 6 - 8 weeks old. All mice were randomly divided into 4 groups, with 6 mice in each group: control group (NaHCO 3 ), folic acid group (FA), liquiritol group (GC), and folic acid + liquiritol group (FA + GC). The treatment conditions for each group are shown in the following table. The dose of GC is 10 mg / kg bw, and the dose of FA is 250 mg / kg bw. GC was prepared with PBS containing 5% Tween-80, and FA was prepared with 0.3 M NaHCO 3 aqueous solution. GC was administered continuously 4 times, and the mice were sacrificed 48 h after FA administration. Blood and kidneys of the mice were collected. The blood samples of the mice were allowed to stand at room temperature for 1 h and then centrifuged at 1200 g for 10 min at 4°C. The supernatant was aspirated to obtain serum. Then, the determination was carried out according to the instructions for the determination of blood urea nitrogen (BUN) and creatinine (CRE) contents. The fresh kidney tissues were washed with pre-cooled physiological saline, blotted dry with filter paper, fixed overnight with 4% paraformaldehyde, and then dehydrated, cleared, embedded in paraffin and sectioned. The section thickness was 4 μm, and the staining was carried out according to the instructions for hematoxylin and eosin (H&E) staining.

[0027] Animal grouping and treatment

[0028] Grouping Treatment Control <![CDATA[Intraperitoneal injection, equal volume of 0.3M NaHCO 3 aqueous solution, equal volume of PBS with 5% Tween-80]]> FA Intraperitoneal injection, FA dose is 250 mg / kgbw, PBS with an equal amount of 5% Tween-80 GC <![CDATA[Intraperitoneal injection, equal volume of 0.3M NaHCO 3 aqueous solution, GC dose is 10 mg / kgbw]]> FA + GC Intraperitoneal injection, FA dose is 250 mg / kgbw, GC dose is 10 mg / kgbw

[0029] Experimental results ( Figure 2 , Figure 3 ) showed that the contents of BUN and CRE in the serum of mice were significantly increased 48 h after folic acid injection, and the contents of BUN and CRE in the serum of the FA + GC group were significantly decreased. It can be seen from Figure 4 that in group C (normal control group), the glomerular morphological structure was normal, and the renal tubular epithelial cells were arranged tightly; there was no obvious abnormality in the kidneys of the GC group; a large number of renal tubules were dilated and there was infiltration of inflammatory cells in the renal interstitium in the FA group (folic acid group); administration of GC could significantly improve the pathological characteristics of acute kidney injury in the kidneys of mice in the FA group.

[0030] Example 2: Liquiritol alleviates acute renal injury by inhibiting ferroptosis

[0031] Experimental method: 24 male Balb / c mice, 6 - 8 weeks old. All mice were randomly divided into 4 groups, with 6 mice in each group: control group (NaHCO 3 ), folic acid group (FA), glycyrrhols group (GC), and folic acid + glycyrrhols group (FA + GC). The treatment conditions for each group were the same as in Example 1. The dose of GC was 10 mg / kg bw, and the dose of FA was 250 mg / kg bw. GC was prepared with PBS containing 5% Tween - 80, and FA was prepared with 0.3 M NaHCO 3 aqueous solution. GC was administered continuously 4 times. The mice were sacrificed 48 h after FA administration, and the blood and kidneys of the mice were collected. The animal grouping, treatment, and experimental procedures were the same as in Example 1. The kidney tissues were weighed, and pre - cooled normal saline was added at a ratio of 1:9 (m / v), and then ground with a tissue homogenizer on ice to prepare 10% tissue homogenate for the determination of Fe, MDA, and GSH - Px. The above - mentioned indexes were measured according to the relevant instructions. The kidney tissues were weighed, and an appropriate amount of pre - cooled RIPA lysis buffer containing 1% protease inhibitor was added to lyse the cells or tissues. Ultrasonic treatment was performed on ice, and then centrifuged. The supernatant was taken to measure the protein concentration and the concentrations of all protein samples were adjusted to be the same. 6× protein loading buffer was added, boiled for 5 min, and stored at - 20 °C or - 80 °C for later use. Western blotting was used to detect the changes in related proteins.

[0032] Experimental results Figure 5 Figure A shows that MDA in the kidneys of mice increased significantly 48 hours after folic acid injection, and the FA + GC group significantly reduced the MDA level in the kidneys of mice. As can be seen from Figure 5 Figure B, administration of GC could improve the decrease in the activity of GSH - Px in the kidneys of FA - group mice. As can be seen from Figure 5 Figures C and D, compared with the FA group, the FA + GC group reduced the iron content in the kidneys, probably by reducing the expression of Tfr1 and HO - 1 in the kidneys to affect iron metabolism and inhibit ferroptosis to alleviate acute kidney injury.

[0033] The above has detailed the present invention. For those skilled in the art, without departing from the purpose and scope of the present invention and without unnecessary experiments, the present invention can be implemented within a relatively wide range under equivalent parameters, concentrations, and conditions. Although specific embodiments of the present invention are given, it should be understood that the present invention can be further improved. In short, according to the principle of the present invention, this application intends to include any changes, uses, or improvements to the present invention, including changes made with conventional techniques known in the art that are outside the scope disclosed in this application.

Claims

1. Use of liquiritol in the preparation of a medicament for preventing and / or treating acute kidney injury.

2. The use according to claim 1, wherein: the acute kidney injury is drug-induced acute kidney injury.

3. The use according to claim 2, wherein: the acute kidney injury is folic acid-induced acute kidney injury.

4. The use according to claim 1, wherein: 1) Liquiritol reduces the contents of creatinine and blood urea nitrogen in serum; 2) Liquiritol restores the activity of GSH-Px to inhibit ferroptosis.

5. The use according to any one of claims 1-4, wherein: the medicament takes liquiritol as the sole active ingredient.

6. The use according to any one of claims 1-4, wherein: the medicament contains other active ingredients in addition to liquiritol.

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