A GSH-Fe 3-X Se3 nanomaterial, its preparation method and application

Abstract

This application provides a GSH-Fe 3‑X Se3 nanomaterial, its preparation method and application, which are applicable to the technical field of nanomaterials. The GSH-Fe 3‑X Se3 nanomaterial comprises the following raw materials: 0.175 - 0.32 g of glutathione, 0.033 - 0.066 g of Fe(ClO4)2⋅xH2O, 0.045 - 0.09 g of Na2SeO 3 0, and 35 - 45 mL of dimethyl sulfoxide. The GSH-Fe 3‑X Se3 nanomaterial provided by this application has higher synergistic anti-inflammatory and antioxidant activities compared with glutathione, can inhibit the generation of excessive ROS and inflammatory storms. At the same time, the synergistic treatment using the glutathione-coated Fe 3‑X Se3 nanomaterial can effectively improve the therapeutic effect on cisplatin-induced acute kidney injury.

Description

Technical Field

[0001] This application relates to the technical field of nanomaterials. Specifically, it relates to a GSH-Fe 3-X Se3 nanomaterial and its preparation method and application. Background Art

[0002] Acute Kidney Injury (AKI) is one of the common complications in clinical practice. Its clinical manifestations are complex, seriously affecting the prognosis of patients. According to the analysis of 154 studies, the incidence of AKI in inpatients is 20.0 - 31.7%, and the mortality rate is 23%. In addition, even after renal function recovery, AKI often develops into fibrosis and chronic kidney disease. In the past few decades, the global death burden related to AKI has exceeded that of heart failure, diabetes, or breast cancer. However, except for supportive treatment, there is still no effective treatment method for diagnosed AKI. Therefore, it is urgent to explore advanced treatment strategies to improve the clinical outcomes of AKI.

[0003] The two common major causes of AKI are ischemic injury and nephrotoxic drugs. Cisplatin (Cis), as one of the most commonly used chemotherapeutic drugs in clinical practice, is used to treat tumors of various solid organs. Its anti-cancer activity is proportional to the dosage. As the dosage increases, the toxicity of cisplatin becomes more obvious. Among them, cisplatin nephrotoxicity is one of the main side effects. Some studies have shown that the incidence of cisplatin-induced kidney damage is 20% - 30%. It is statistically shown that in the anti-cancer chemotherapy treatment regimens in China, those mainly based on cisplatin or with cisplatin in combination account for 70% - 80% of all chemotherapy regimens.

[0004] Current research has found that AKI can cause an increase in reactive oxygen species (ROS) and inflammatory factors, thereby triggering apoptosis and necrosis of cells. In many studies, glutathione (GSH), as a common antioxidant, has a certain application in the treatment of acute kidney injury. However, during its treatment process, a large dosage is often required, and the treatment effect is generally average. Selenium-related nanodrugs, as one of the most widely studied nanomaterials currently, have advantages such as high biocompatibility, high bioavailability, and low toxicity. In addition, the anti-inflammatory and antioxidant activities of selenium-related nanodrugs have been used in various anti-inflammatory and antioxidant disease models. However, there are relatively few current studies on the treatment of acute kidney injury with iron selenide-related nanodrugs. Summary of the Invention

[0005] To solve one of the above technical deficiencies, this application provides a GSH-Fe 3-X Se3 nanomaterial and its preparation method and application.

[0006] According to the first aspect of this application, there is provided a GSH-Fe 3-XSe3 nanomaterials, comprising the following raw materials: 0.175 - 0.32 g of GSH, 0.033 - 0.066 g of Fe(ClO4)2·xH2O, 0.045 - 0.09 g of Na2SeO3, and 35 - 45 mL of dimethyl sulfoxide.

[0007] Preferably, the GSH-Fe 3-X Se3 nanomaterials are GSH-coated Fe 3-X Se3 nanomaterials.

[0008] Preferably, the GSH-Fe 3-X Se3 nanomaterials include GSH-Fe3Se3 nanomaterials and GSH-Fe2Se3 nanomaterials.

[0009] Preferably, the GSH is reduced glutathione.

[0010] Preferably, the particle size of the GSH-Fe 3-X Se3 nanomaterials is 150 nm.

[0011] According to the second aspect of the present application, a method for preparing GSH-Fe 3-X Se3 nanomaterials is provided, wherein the GSH-Fe 3-X Se3 nanomaterials are the GSH-Fe 3-X Se3 nanomaterials described in any of the above, and the preparation method includes the following steps:

[0012] S1. Add GSH to a three-necked flask containing dimethyl sulfoxide, and stir evenly at a rotation speed of 700 - 900 r / min to obtain reaction system A;

[0013] S2. Add Fe(ClO4)2·xH2O to reaction system A, and stir evenly at a rotation speed of 700 - 900 r / min to obtain reaction system B;

[0014] S3. Under an argon atmosphere, heat the stirred reaction system B to 135 - 145 °C, and stir at a rotation speed of 700 - 900 r / min for 10 min. Then, add Na2SeO3 to the heated reaction system B and continuously stir for 16 - 22 h to obtain reaction system C;

[0015] S4. Cool reaction system C to 25 °C, and then centrifuge and wash the cooled reaction system C with isopropanol to obtain GSH-Fe 3-X Se3 nanomaterials;

[0016] S5. Lyophilize and collect the obtained GSH-Fe 3-X Se3 nanomaterials and store them at room temperature.

[0017] Preferably, in the step S4, the volume ratio of isopropanol to the cooled reaction system C is 8:1.

[0018] Preferably, in the step S4, the centrifugation speed is 15000 - 18000 r / min, the centrifugation time is 10 - 30 min, and the centrifugation temperature is 4 - 25 °C; in the step S5, the freeze-drying time is 24 h.

[0019] Preferably, in the step S5, the mass of the obtained GSH-Fe 3-X Se3 nanomaterial is 40 mg.

[0020] According to the third aspect of the present application, there is provided the use of the GSH-Fe 3-X Se3 nanomaterial described in any of the above in the preparation of a drug for treating acute kidney injury.

[0021] The beneficial effects of the present application are as follows: In the GSH-Fe 3-X Se3 nanomaterial provided by the present application, there is a synergistic effect between GSH and Fe 3-X Se3. Therefore, the GSH-Fe 3-X Se3 nanomaterial has higher anti-inflammatory and antioxidant activities compared with GSH, can inhibit the generation of excessive ROS and cytokine storms. At the same time, the GSH-Fe 3-X Se3 nanomaterial has good stability and dispersibility in aqueous solution, and the GSH-Fe 3-X Se3 nanomaterial also has good biocompatibility and has low irritation in vivo. In addition, the synergistic treatment using the GSH-coated Fe 3-X Se3 nanomaterial can effectively improve the therapeutic effect on cisplatin-induced acute kidney injury. Description of the Drawings

[0022] Figure 1 It is the scanning electron microscope image of the GSH-Fe 3-X Se3 nanomaterial provided by Example 1 of the present application;

[0023] Figure 2 It is the infrared spectrum of the GSH-Fe 3-X Se3 nanomaterial provided by Example 1 of the present application;

[0024] Figure 3 It is the XPS diagram of the GSH-Fe 3-X Se3 nanomaterial provided by Example 1 of the present application; among them, A is the full XPS spectrum of the GSH-Fe 3-X Se3 nanomaterial, B is the fine XPS spectrum of Se, and C is the fine XPS spectrum of Fe;

[0025] Figure 4 The stability characterization diagram of the GSH-Fe 3-X Se3 nanomaterial provided in Example 1 of this application;

[0026] Figure 5 The GSH-Fe 3-X Fluorescence effect diagram of the GSH-Fe Se3 nanomaterial provided in Example 1 of this application for eliminating ROS in RAW264.7 cells;

[0027] Figure 6 The GSH-Fe 3-X Quantitative characterization diagram of the GSH-Fe Se3 nanomaterial provided in Example 1 of this application for eliminating ROS in RAW264.7 cells;

[0028] Figure 7 The GSH-Fe 3-X Bar graph of the effect of the GSH-Fe Se3 nanomaterial provided in Example 1 of this application on the expression level of inflammatory factors in RAW264.7 cells;

[0029] Figure 8 Cell viability diagram of RAW264.7 cells treated with different concentrations of the GSH-Fe 3-X Se3 nanomaterial provided in Example 1 of this application;

[0030] Figure 9 The experimental diagram of the effect of the GSH-Fe 3-X Se3 nanomaterial provided in this application on acute kidney injury markers; where A is the H&E section diagram of the mouse kidney, and B is the Tunel staining diagram of the mouse kidney;

[0031] Figure 10 The GSH-Fe 3-X Blood compatibility detection diagram of the Se3 nanomaterial; where A is the blank comparison diagram of the GSH-Fe 3-X Se3 nanomaterial, and B is the hemolytic activity diagram of the GSH-Fe 3-X Se3 nanomaterial. Detailed implementation manners

[0032] In order to make the technical solutions and advantages in the embodiments of this application clearer and more understandable, the exemplary embodiments of this application are further described in detail below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, rather than an exhaustive list of all embodiments. It should be noted that, without conflict, the embodiments in this application and the features in the embodiments can be combined with each other.

[0033] The reagents and materials in the embodiments can be conventionally purchased from the market without special instructions; the test methods in the following embodiments are all conventional methods in this field without special instructions.

[0034] RAW264.7 cells (Wuhan Punosai Life Science Co., Ltd.); C57BL / 6 mice (Beijing Huafukang Biotechnology Co., Ltd.); The infrared spectrometer model is NICOLET iS 50; The XPS instrument model is EXCALAB 250 XI; The dynamic light scattering instrument model is Malvern Zetasizer Nano ZS90; Flow cytometer (BD); Scanning electron microscope (ApreoC); NO detection kit (Beyotime); The inverted fluorescence microscope model is ECLIPSE Ts2; The fluorescence microplate reader model is Synergy NEO; The microplate reader model is synergy lx; Enzyme-linked immunosorbent assay kit (elabscience).

[0035] Example 1

[0036] This application provides a GSH-Fe 3-X Se3 nanomaterial, including the following raw materials: 0.175 - 0.32 g of GSH, 0.033 - 0.066 g of Fe(ClO4)2·xH2O, 0.045 - 0.09 g of Na2SeO3, and 35 - 45 mL of dimethyl sulfoxide.

[0037] Specifically, when implemented, the GSH-Fe 3-X Se3 nanomaterial includes the following raw materials: 0.32 g of GSH, 0.066 g of Fe(ClO4)2·xH2O, 0.045 g of Na2SeO3, and 40 mL of dimethyl sulfoxide.

[0038] Further, the GSH-Fe 3-X Se3 nanomaterial is a GSH-coated Fe 3-X Se3 nanomaterial.

[0039] Further, the GSH-Fe 3-X Se3 nanomaterial includes GSH-Fe3Se3 nanomaterial and GSH-Fe2Se3 nanomaterial. Among them, the valence state of Fe element in GSH-Fe3Se3 nanomaterial is +2, and the valence state of Se element is -2; The valence state of Fe element in GSH-Fe2Se3 nanomaterial is +3, and the valence state of Se element is -2.

[0040] Further, the GSH is reduced glutathione, which is used to reduce the +4 valence selenium in Na2SeO3 to -2 valence selenium.

[0041] Further, the particle size of the GSH-Fe 3-X Se3 nanomaterial is 150 nm.

[0042] The present application also provides a preparation method of GSH-Fe 3-X Se3 nanomaterials, which specifically includes the following steps:

[0043] S1. Add 0.32 g of GSH into a three-necked flask containing 40 mL of dimethyl sulfoxide, and stir evenly at a rotation speed of 900 r / min to obtain reaction system A;

[0044] S2. Add 0.066 g of Fe(ClO4)2·xH2O into reaction system A, and stir evenly at a rotation speed of 900 r / min to obtain reaction system B;

[0045] S3. Under an argon atmosphere, heat the stirred reaction system B to 140 °C, and stir at a rotation speed of 900 r / min for 10 min. Then, add 0.045 g of Na2SeO3 into the heated reaction system B and continuously stir for 18 h to obtain reaction system C;

[0046] S4. Cool reaction system C to 25 °C, and then perform centrifugal washing on the cooled reaction system C with isopropanol. Centrifugal washing can purify and precipitate the cooled reaction system C, and finally obtain a brownish-black precipitate, namely GSH-Fe 3-X Se3 nanomaterials;

[0047] S5. Place the obtained GSH-Fe 3-X Se3 nanomaterials in a freeze dryer for 24 h, collect them, and store them at room temperature for subsequent use.

[0048] Further, in S4, the volume ratio of isopropanol to reaction system C is 8:1.

[0049] Further, in S4, the centrifugal rotation speed is 15000 - 18000 r / min, the centrifugal time is 10 - 30 min, and the centrifugal temperature is 4 - 25 °C. Specifically in implementation, the centrifugal rotation speed is 15000 r / min, the centrifugal time is 20 min, and the centrifugal temperature is 25 °C.

[0050] Further, in S5, the mass of the obtained GSH-Fe 3-X Se3 nanomaterials is 40 mg.

[0051] To better understand the essence of the present application, relevant characterizations were carried out on the GSH-Fe 3-X Se3 nanomaterials prepared in this example.

[0052] The present application characterized the GSH-Fe 3-X Se3 nanomaterials prepared in this example by scanning electron microscopy (SEM). As Figure 1As shown, GSH-Fe 3-X The particle size of Se3 nanomaterials is 150 nm.

[0053] In this application, an infrared spectrometer was used to characterize the GSH-Fe 3-X Se3 nanomaterials prepared in this example. As Figure 2 shown, by comparing the infrared spectra of GSH and GSH-Fe 3-X Se3 nanomaterials, the conjugation between GSH-Fe 3-X Se3 nanomaterials and GSH was confirmed. Among them, in the infrared spectrum of GSH-Fe 3-X Se3 nanomaterials, the S─H stretching vibration band at 2524 cm ﹣1 disappeared by forming an Fe─S coordination bond, which means that GSH molecules are bound to FeSe through thiol bonds. In addition, the absorption band at 1716 cm ﹣1 corresponds to the >C═O stretching mode of the carboxyl group of GSH.

[0054] In this application, an XPS instrument was used to characterize the GSH-Fe 3-X Se3 nanomaterials prepared in this example. As Figure 3 shown, it can be concluded from the figure that the selenium element in GSH-Fe 3-X Se3 nanomaterials is divalent negative, and the iron element is divalent positive and trivalent positive, indicating that the divalent positive and trivalent positive of the iron element coexist, further proving that GSH-Fe 3-X Se3 nanomaterials include GSH-Fe3Se3 nanomaterials and GSH-Fe2Se3 nanomaterials.

[0055] From the above characterization results, it can be seen that GSH-Fe 3-X Se3 nanomaterials were prepared in this application.

[0056] In addition, this application also used a dynamic light scattering instrument to characterize the aqueous solution stability of the GSH-Fe 3-X Se3 nanomaterials prepared in this example. As Figure 4 shown, the particle size and polydispersity coefficient of GSH-Fe 3-X Se3 nanomaterials remained basically stable within one week. It can be concluded that GSH-Fe 3-X Se3 nanomaterials can maintain good stability within one week.

[0057] Example 2

[0058] GSH-Fe 3-X Se3 nanomaterials scavenging reactive oxygen species in RAW264.7 cells experiment.

[0059] To detect GSH-Fe3-X The ability of Se3 nanomaterials to eliminate reactive oxygen species in activated macrophages was examined. The fluorescence intensity of reactive oxygen species in RAW264.7 cells after various treatments was detected by inverted fluorescence microscopy and flow cytometry.

[0060] Specifically, the detection method using inverted fluorescence microscopy was as follows: Activated RAW264.7 cells were seeded in a 6-well plate at a density of 1×10 6 and incubated for 24 h. Then, lipopolysaccharide (LPS, 1 μg / mL), LPS (1 μg / mL) + GSH (200 μg / mL), and LPS (1 μg / mL) + GSH-Fe 3-X Se3 nanomaterials (200 μg / mL) were added respectively. A blank control group was also set up. After the cells and drugs were incubated for 24 h, the cells were washed three times with phosphate buffer saline (PBS) and stained with a reactive oxygen species probe (DCFH-DA) for 30 min. The cell samples were washed once more with PBS and observed under an inverted fluorescence microscope. The excitation wavelength and emission wavelength of the inverted fluorescence microscope were 488 nm and 525 nm respectively. The results are as Figure 5 shown.

[0061] The detection method using flow cytometry was as follows: Activated RAW264.7 cells were seeded in a 6-well plate at a density of 1×10 per well 6 and cultured in an incubator at 37 °C and 5% CO2 for 24 h. Then, LPS (1 μg / mL), LPS (1 μg / mL) + GSH (200 μg / mL), and LPS (1 μg / mL) + GSH-Fe 3-X Se3 nanomaterials (200 μg / mL) were added respectively. A blank control group was also set up. After the cells and drugs were incubated for 24 h, the cells were washed three times with PBS and stained with DCFH-DA for 30 min. The cell samples were washed once more with PBS, and the cell samples were collected and detected by flow cytometry. The results are as Figure 6 shown.

[0062] It can be seen from Figure 5 that after incubating with LPS-induced RAW264.7 cells for 24 h, the fluorescence intensity of reactive oxygen species in the model group (LPS) was the strongest. In each experimental group with the addition of nanomaterials, the fluorescence intensity of reactive oxygen species in the cells decreased to varying degrees. It was also found that the efficiency of GSH-Fe 3-X Se3 nanomaterials in eliminating reactive oxygen species was significantly stronger than that of GSH alone. This also proved that GSH-Fe 3-X Se3 nanomaterials had a stronger ability to eliminate reactive oxygen species. The detection by flow cytometry also showed the same trend, as Figure 6As shown, the detection results indicate that the fluorescence intensity of reactive oxygen species is the strongest in the model group (LPS), while the fluorescence intensity of intracellular reactive oxygen species in each experimental group with the addition of nanomaterials shows varying degrees of decrease, and it can be found that GSH-Fe 3-X The efficiency of Se3 nanomaterials in eliminating reactive oxygen species is significantly stronger than that of GSH alone, which also proves that GSH-Fe 3-X Se3 nanomaterials have stronger reactive oxygen species elimination ability. In summary, GSH-Fe 3-X Se3 nanomaterials have good ability to eliminate intracellular reactive oxygen species in macrophages.

[0063] Example 3

[0064] Effect of GSH-Fe 3-X Se3 nanomaterials on the expression levels of inflammatory factors in RAW264.7 cells. The inflammatory factors include NO, tumor necrosis factor α (TNF-α), and interleukin-6 (IL-6).

[0065] Specifically, a NO detection kit was used to measure the concentration of NO. More specifically, RAW264.7 cells were seeded in a 96-well plate at a density of 4×10 4 , incubated overnight for 24 h, and then LPS (1 μg / mL), LPS (1 μg / mL) + GSH (200 μg / mL), LPS (1 μg / mL) + GSH-Fe 3-X Se3 nanomaterials (200 μg / mL) were added in the form of medium replacement. Meanwhile, a blank control group was set up. After the cells and drugs were incubated for 24 h respectively, the supernatant was collected by centrifugation (12,000 g, 5 min, 4 °C). Subsequently, the concentration of NO was detected using a NO detection kit, and the OD value was measured at 540 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0066] In addition, enzyme-linked immunosorbent assay (ELISA) kits were used to measure the concentrations of TNF-α and IL-6. Specifically, RAW264.7 cells were seeded in a 96-well plate at a density of 4×10 4 , and cultured in an incubator for 24 h. Then, LPS (1 μg / mL), LPS (1 μg / mL) + GSH (200 μg / mL), LPS (1 μg / mL) + GSH-Fe 3-X Se3 nanomaterials (200 μg / mL) were added in the form of medium replacement. Meanwhile, a blank control group was set up. After the cells and drugs were incubated for 24 h respectively, the supernatant was collected. Subsequently, the corresponding inflammatory factor kits were used to detect TNF-α and IL-6, and the OD value was measured at 450 nm using an ELISA reader.

[0067] As Figure 7As shown, after incubating with LPS-induced RAW264.7 cells for 24 h, the amount of inflammatory factors in the model group (LPS) was the highest, while the intracellular inflammatory intensity in each experimental group treated with the drug showed varying degrees of decrease, indicating that both GSH and GSH-Fe 3-X Se3 nanomaterials could inhibit the release of inflammatory factors by macrophages. By comparison, it was also found that the efficiency of GSH-Fe 3-X Se3 nanomaterials in eliminating inflammation was significantly stronger than that of GSH alone, thus proving that GSH-Fe 3-X Se3 nanomaterials had stronger anti-inflammatory ability.

[0068] Example 4

[0069] Toxicity evaluation of GSH-Fe 3-X Se3 nanomaterials.

[0070] Specifically, RAW264.7 cells at a density of 5×10 4 were seeded in a 96-well plate and cultured in an incubator at 37°C and 5% CO2 for 24 h. Then, different concentrations of GSH-Fe 3-X Se3 nanomaterials (0, 12.5, 25, 50, 100, 200, 400 μg / mL) were added for co-culture, and three replicates were set for each concentration; the 96-well plate was placed in an incubator at 37°C and 5% CO2 and continued to be statically cultured for 24 h; after incubation, the supernatant was aspirated, fresh medium containing CCK-8 was added, and it was continued to be placed in the incubator for 20 min. The absorbance of all samples at 450 nm was measured using an enzyme-linked immunosorbent assay (ELISA) reader. The results are as Figure 8 shown. As can be seen from the figure, the viability of RAW264.7 cells was still greater than 80% at a concentration of 200 μg / mL, indicating that the GSH-Fe 3-X Se3 nanomaterials prepared in this application had low toxicity.

[0071] Example 5

[0072] Effect of GSH-Fe 3-X Se3 nanomaterials on a mouse model of acute kidney injury.

[0073] The specific test method is as follows: A mouse model of acute kidney injury was established using cisplatin. After 72 h of the experiment, the mice were sacrificed by cervical dislocation, and the kidneys were collected. The organ tissues were fixed with 4% paraformaldehyde fixative for subsequent histological pathological section staining. In addition, Tunnel staining was performed on the kidneys of mice in different treatment groups to observe the apoptosis of renal lymphocytes.

[0074] As Figure 9 shown, compared with the Control group, the renal epithelial cells in the cisplatin-induced model group (Cis group) were shed after injecting GSH-Fe 3-XAfter the Se3 nanomaterial, compared with GSH treatment, in the mice treated with GSH-Fe 3-X Se3 nanomaterial, the shedding of renal epithelial cells decreased and the apoptosis of lymphocytes was more inhibited. It can also be seen that as the administration concentration increased (200, 400, 800 mg / kg), the treatment effects were not significantly different. This proves that GSH-Fe 3-X The appropriate administration concentration of Se3 nanomaterial in mice is 200 mg / kg. It has excellent synergistic anti-inflammatory and antioxidant activities, can scavenge ROS in the body of mice with acute kidney injury and reduce the expression level of inflammatory factors, and effectively alleviates the organ damage caused by acute kidney injury.

[0075] Example 6

[0076] GSH-Fe 3-X The blood compatibility of Se3 nanomaterial.

[0077] The specific test method is as follows: First, collect fresh blood by removing the eyeballs of healthy mice, collect the obtained fresh blood in an anticoagulation tube, shake well and then centrifuge (1500r / min, 20 min) to collect red blood cells. Second, resuspend the obtained red blood cells with PBS (the volume ratio of PBS to red blood cells is 49:1), centrifuge and wash (3000r / min, 10min) until the supernatant has no obvious red color. Then, add 20µL of red blood cells to 980µL of PBS solution containing different concentrations of GSH-Fe 3-X Se3 nanomaterial (200, 100, 50, 25, 12.50µg / mL). Finally, use ultrapure water and PBS as the positive control and negative control respectively, and use PBS solutions containing different concentrations of GSH-Fe 3-X Se3 nanomaterial (200, 100, 50, 25, 12.50µg / mL) as the blank control. Place it at room temperature for 2h. After the incubation is over, centrifuge at 15000r / min for 3min, take pictures, take 100µL of the supernatant and put it in a 96-well plate, and use an enzyme-linked immunosorbent assay instrument to detect the OD value at 540nm. Calculate its hemolysis rate according to the following formula:

[0078]

[0079] As Figure 10 shown, the red blood cells dissolved in ultrapure water are used as the positive control group, the red blood cells dissolved in PBS are used as the negative control group, the PBS solutions containing different concentrations of GSH-Fe 3-X Se3 nanomaterial are used as the blank control group, and the corresponding concentrations of GSH-Fe 3-XThe red blood cells dissolved in the PBS solution of the GSH-Fe Se3 nanomaterial were used as the experimental group. As can be seen from the figure, the blood cells in the positive control group were completely lysed, while those in the negative control group did not lyse. By subtracting the absorbance value of the blank control group from the absorbance values of different concentration groups of the material (12.5, 25, 50, 100, 200 μg / mL respectively), the hemolysis rate was calculated, and it was found that GSH-Fe 3-X When the concentration of the GSH-Fe Se3 nanomaterial was 200 μg / mL, the hemolysis rate of the blood cells did not exceed 5%, indicating that GSH-Fe 3-X The Se3 nanomaterial has good blood compatibility.

[0080] The GSH-Fe Se3 nanomaterials in Examples 2 to 6 are all the GSH-Fe Se3 nanomaterials prepared in Example 1. 3-X The incubators in Examples 2 to 4 can be constant temperature incubators. 3- X There is a synergistic effect between GSH and Fe Se3 in the GSH-Fe Se3 nanomaterial provided in this application. Therefore, compared with GSH, the GSH-Fe Se3 nanomaterial has higher anti-inflammatory and antioxidant activities, can inhibit the generation of excessive ROS and inflammatory storms, and the synergistic treatment using the GSH-coated Fe Se3 nanomaterial can effectively improve the therapeutic effect on cisplatin-induced acute kidney injury. At the same time, the GSH-Fe Se3 nanomaterial has good stability and dispersibility in aqueous solution, and the GSH-Fe Se3 nanomaterial also has good biocompatibility and low irritation in vivo. In addition, the product system of the GSH-Fe Se3 nanomaterial provided in this application is simple, the product can be directly stored and used, and the preparation method is simple.

[0081] Although the preferred embodiments of this application have been described, those skilled in the art can make additional changes and modifications once they know the basic creative concept. Therefore, the appended claims are intended to be construed to include the preferred embodiments and all changes and modifications falling within the scope of this application. 3-X 3-X 3-X 3-X 3-X 3-X 3-X

[0082]

[0083] Obviously, those skilled in the art can make various changes and modifications to this application without departing from the spirit and scope of this application. Thus, if these modifications and variations of this application fall within the scope of the claims of this application and their equivalent technologies, this application is also intended to include these modifications and variations.​​​​​​​​

Claims

1. A GSH-Fe 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: The GSH-Fe 3-X Se3 nanomaterials include the following raw materials: glutathione 0.175-0.32 g, Fe(ClO4)2·xH2O 0.033-0.066 g, Na2SeO3 0.045-0.09 g; The GSH-Fe 3-X Se3 nanomaterials include GSH-Fe3Se3 nanomaterials and GSH-Fe2Se3 nanomaterials.

2. GSH-Fe according to claim 1 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: The GSH-Fe 3-X Se3 nanomaterials are glutathione-coated Fe 3-X Se3 nanomaterials.

3. GSH-Fe according to claim 1 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: The glutathione is reduced glutathione.

4. GSH-Fe according to claim 1 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: The GSH-Fe 3-X The particle size of Se3 nanomaterial is 150nm.

5. GSH-Fe according to claim 1 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: The GSH-Fe 3-X The preparation method of Se3 nanomaterials comprises the following steps: S1. Add glutathione into a three-necked flask containing dimethyl sulfoxide, and stir evenly at a speed of 700-900 r / min to obtain a reaction system A; S2, adding Fe(ClO4)2·xH2O to reaction system A, stirring evenly at a speed of 700-900 r / min to obtain reaction system B; S3. In an argon atmosphere, the stirred reaction system B is heated to 135-145° C. and stirred at a speed of 700-900 r / min for 10 min. Thereafter, Na2SeO3 is added to the heated reaction system B and stirred continuously for 16-22 h to obtain a reaction system C. S4, cooling the reaction system C to 25°C, and then centrifugally washing the cooled reaction system C with isopropanol to obtain GSH-Fe 3-X Se3 nanomaterials; S5. The obtained GSH-Fe 3-X The Se3 nanomaterials were collected by freeze-drying and stored at room temperature.

6. GSH-Fe according to claim 5 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: In S4, the volume ratio of isopropanol to the cooled reaction system C is 8:

1.

7. GSH-Fe according to claim 5 3-X The application of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized by: In S4, the centrifugal speed is 15000-18000 r / min, the centrifugal time is 10-30 min, and the centrifugal temperature is 4-25° C.; In S5, the freeze-drying time is 24 hours.

8. GSH-Fe according to claim 5 3-X The use of Se3 nanomaterials in the preparation of drugs for treating acute kidney injury is characterized in that: In S5, the obtained GSH-Fe 3-X The mass of Se3 nanomaterial is 40 mg.

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