Application of DHGC in preparation of medicine for treating SIRT3 / NRF2 axis mediated acute kidney injury ferroptosis

Abstract

The invention belongs to the technical field of medicine preparation, and particularly relates to application of DHGC in preparation of a medicine for treating acute kidney injury ferroptosis mediated by an SIRT3 / NRF2 axis. Experiments prove that DHGC can activate SIRT3 in a targeted manner, up-regulate the expression level of SIRT3, promote NRF2 deacetylation and drive NRF2 nuclear translocation, and further up-regulate the expression level of GPX4, so that the effect of resisting ferroptosis caused by acute kidney injury is achieved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine preparation, and particularly relates to an application of DHGC in preparation of a medicine for treating SIRT3 / NRF2 axis mediated acute kidney injury ferroptosis. BACKGROUND

[0002] Acute kidney injury (AKI) is one of the serious complications of sepsis, characterized by inflammation, oxidative stress and ferroptosis. More than 50% of sepsis patients develop AKI, which seriously threatens the survival and death risk of patients. Survivors may progress to chronic kidney disease, and even end-stage renal disease. Dahuang Gancao Decoction (DHGC) is a traditional prescription derived from Shanghan Zajiabianlun, composed of Dahuang and Gancao, with the effects of clearing heat and resolving toxicity, promoting blood circulation and removing blood stasis, and has a long history in clinical application.

[0003] The key bioactive components of DHGC are emodin and glycyrrhizin, which can improve AKI by inhibiting inflammatory cascade and enhancing antioxidant defense, showing therapeutic potential in the treatment of AKI, but its exact mechanism and therapeutic target still need further research. In the literature “Exploring anti-acute kidney injury mechanism of Dahuang-Gancao decoction by network pharmacology and experimental validation”, the above research screened out 161 potential targets of DHGC against AKI through network pharmacology analysis, among which SIRT3 was determined as a key target. Subsequent animal experiments used LPS to induce AKI mouse models, which showed that DHGC could significantly reduce blood urea nitrogen, creatinine, and reduce renal tissue pathological damage, inhibit cell apoptosis and inflammatory factors (IL-1β, TNF-α, IL-6), confirming that DHGC plays a role in anti-apoptosis and protection of mitochondrial function through activation of the SIRT3 / NRF2 / HO-1 signaling pathway.

[0004] Ferroptosis and apoptosis are two essentially different cell death pathways with different characteristics and key proteins, which can independently or synergistically exacerbate kidney injury. In the process of ferroptosis, mitochondria are not only the main site of lipid peroxidation, but also the central regulator of intracellular iron homeostasis. The destruction of mitochondrial iron balance directly increases the susceptibility of cells to ferroptosis, so maintaining mitochondrial stability becomes a key therapeutic target. SIRT3 is a mitochondria-targeted NAD + There is no literature report on whether DHGC can reduce ferroptosis in AKI through SIRT3. SUMMARY

[0005] The application aims at providing a DHGC application in preparing a SIRT3 / NRF2 axis-mediated acute kidney injury ferroptosis drug to solve the above problems in the prior art.

[0006] To achieve the above object, the technical scheme of the application is as follows:

[0007] The application provides a DHGC application in preparing a SIRT3 / NRF2 axis-mediated acute kidney injury ferroptosis drug.

[0008] Specifically, the mechanism of action of the DHGC is as follows: targeting activation of SIRT3, up-regulation of the expression level of SIRT3, promotion of NRF2 deacetylation and driving of NRF2 nuclear translocation, and then up-regulation of the expression level of GPX4.

[0009] Compared with the prior art, the application has the following beneficial effects:

[0010] The application verifies through experiments that the Dahuang Gancao Decoction can target activation of SIRT3, up-regulation of the expression level of SIRT3, promotion of NRF2 deacetylation and driving of NRF2 nuclear translocation, and then up-regulation of the expression level of GPX4, thereby playing a role in acute kidney injury anti-ferroptosis, and thus can be used for preparing an acute kidney injury anti-ferroptosis drug. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 LC-MS / MS analysis results of the water extract of the Dahuang Gancao Decoction.

[0012] Figure 2 LPS-induced SA-AKI mouse ferroptosis-related index detection results.

[0013] Figure 3 Erastin-induced HK-2 cell ferroptosis-related index detection results.

[0014] Figure 4 The figure shows the physical and chemical parameter changes of the SA-AKI mice before and after the intervention of the Dahuang Gancao Decoction.

[0015] Figure 5 The figure shows the kidney tissue morphological changes of the SA-AKI mice before and after the intervention of the Dahuang Gancao Decoction.

[0016] Figure 6 The figure shows the ferroptosis-related index changes of the SA-AKI mice before and after the intervention of the Dahuang Gancao Decoction.

[0017] Figure 7Changes of SIRT3, GPX4, nuclear NRF2, ac-NRF2 in SA-AKI mice before and after intervention of Dahuang Gancao Decoction.

[0018] Figure 8 Changes of SIRT3, GPX4, nuclear NRF2, ac-NRF2 in HK-2 cells induced by Erastin before and after intervention of Dahuang Gancao Decoction.

[0019] Figure 9 Changes of physical and chemical parameters of SA-AKI mice before and after SIRT3 knockout.

[0020] Figure 10 Changes of renal histomorphology of SA-AKI mice before and after SIRT3 knockout.

[0021] Figure 11 Changes of SIRT3, GPX4, nuclear NRF2, ac-NRF2 in SA-AKI mice before and after SIRT3 knockout.

[0022] Figure 12 Changes of indicators of ferroptosis in HK-2 cells induced by Erastin before and after SIRT3 knockout.

[0023] Figure 13 Changes of SIRT3, GPX4, nuclear NRF2, ac-NRF2 in HK-2 cells induced by Erastin before and after SIRT3 knockout.

[0024] Figure 14 Mechanism of Dahuang Gancao Decoction. DETAILED DESCRIPTION

[0025] In order to further illustrate the technology for achieving the purpose of the present application, the detailed methods, techniques, processes and features related to determining the application of DHGC in preparing drugs for treating SIRT3 / NRF2 axis-mediated ferroptosis in acute kidney injury are described below. The following cases are used to describe but not limit the application of the present application.

[0026] 1. Materials and methods

[0027] 1.1 Reagents

[0028] Lipopolysaccharide (LPS) was derived from Solarbio (L8880-10, Beijing, China). Erastin gene (E424821) was derived from Aladdin (Shanghai, China). Ferrostatin-1 (abbreviated as Fer-1, RM02804) was provided by Abmabio Biotech Co., Ltd. (Wuhan, China). Primary antibody SIRT3 (GB11354-100), NRF2 (GB113808-100), GPX4 (GB115275-100), Histone H3 (GB11102), and β-actin (GB15003) were purchased from Servicebio (Wuhan, China). Anti-acetylated lysine antibody (ab21623) was purchased from Abeam (Cambridge, UK). Chromatography, mass spectrometry, and methanol chromatography purity were obtained from Thermo Fisher Scientific (Shanghai, China). Formic acid chromatography purity was obtained from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China).

[0029] 1.2 Preparation of Dahuanggancao Decoction Water Extract and Identification of Components

[0030] According to the ratio of medicine to solvent being 1:8, i.e. 12g of Dahuang (derived from Hubei Chenmei Pharmaceutical Co., Ltd., Huanggang City, China) and 3g of Gancao (derived from Hubei Tianji Pharmaceutical Co., Ltd., Wuhan, China) were added to 240mL boiling water in a mass ratio of 4:1. After 10min of extraction, it was concentrated to 150mL to obtain the crude Dahuanggancao Decoction with a concentration of 200mg / mL, which was then placed in a vacuum drying machine for drying. The ratio of Dahuanggancao Decoction powder to crude medicine was calculated to be 9.53%. Therefore, 2g / kg of crude Dahuanggancao Decoction in this example was equivalent to 190.6mg / kg of powder. In the in vivo study, the dosage was within the concentration range of the extract (100-200mg / kg).

[0031] 2 Animal Experiments

[0032] Forward validation: Male C57BL / 6 mice (6-8 weeks old, body weight (20±2) g, from Henan Skope Biotechnology Co., Ltd. (Anyang, China). After a 7-day acclimation period under controlled environmental conditions (12 h light / dark cycle, 23±2°C, 55%±5% humidity), the mice were randomly divided into 5 experimental groups of 6 each: control group (Ctrl), LPS group, and 3 DHGC water extract treatment groups (low, medium, and high groups). The low, medium, and high dose groups of Dahuang Gancao Decoction water extract (1 g / kg, 2 g / kg, and 4 g / kg) were given Dahuang Gancao Decoction water extract 1 g / kg, 2 g / kg, and 4 g / kg, respectively, 3 days before intraperitoneal injection of LPS. Subsequently, the LPS group, low dose group, medium dose group, and high dose group were intraperitoneally injected with LPS (15 mg / kg), and the Ctrl group was intraperitoneally injected with an equal amount of normal saline. At 24 h after modeling, all mice were euthanized with 2% sodium pentobarbital (50 mg / kg) in accordance with the requirements of the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals to minimize animal suffering. The kidneys and serum were collected. The left kidney was fixed with 4% paraformaldehyde, dehydrated with ethanol, and embedded in paraffin; the remaining samples were stored at -80°C for subsequent analysis.

[0033] Reverse validation: Wild-type (WT) C57BL / 6 mice were randomly divided into Ctrl, LPS, and LPS+DHGC groups, each with 6 mice. Similarly, SIRT3 gene knockout (SIRT3 - / - ) mice were randomly divided into SIRT3 - / - , SIRT3 - / - +LPS, and SIRT3 - / - +LPS+DHGC groups, each with 6 mice. The same modeling and dosing regimen as in the forward validation was used, and the subsequent experimental procedures were repeated.

[0034] All animal experiments were conducted in accordance with the guidelines approved by the Hubei University of Chinese Medicine Ethics Committee (HUCMS61616355).

[0035] 3 Experimental methods

[0036] 3.1 Assessment of kidney function and biochemical indicators

[0037] The E-BC-K183-M and E-BC-K188-M kits were used to detect serum urea nitrogen (BUN) and creatinine (SCr) levels, respectively, to assess kidney function. Oxidative stress markers, including glutathione (E-BC-K030-M), superoxide dismutase (E-BC-K020-M), and malondialdehyde (E-BC-K025-M), were obtained from Elabscience (Wuhan, China). These oxidative stress markers were quantified in mouse serum and cell suspensions using the corresponding detection kits. Fe 2+Kidney Fe content was detected by a kit (E-BC-K773-M, Elabscience, Wuhan, China). 2+ All operations were strictly in accordance with the manufacturer's operating procedures.

[0038] 3.2 Histological analysis

[0039] Renal tissues were first fixed with 4% paraformaldehyde, then dehydrated and embedded in paraffin. These tissues were then cut into 5 pm-thick sections. After deparaffinization with xylene and ethanol, hematoxylin-eosin (HE) staining (G1004-500ML, Servicebio, Inc, Wuhan, China) and TUNEL staining kit (G1507-50T, Servicebio, Inc, Wuhan, China) were used to quantitatively analyze cell apoptosis.

[0040] 3.3 Cell transfection

[0041] SIRT3 expression plasmid was custom synthesized by GenePharma (Shanghai, China). 2 pg of expression plasmid or negative control vector was transfected into HK-2 cells using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). In addition, specific siRNA or control siRNA (GenePharma) was transfected under the same conditions using Lipofectamine 3000 for 48 hours. siRNA sequence: SIRT3, forward 5'-GAAACUACAAGCCCAACGUTT-3', reverse 5'-acguuggcuuguaguuctt-3'.

[0042] 3.4 Statistical analysis

[0043] Data were expressed as mean ± standard deviation. The differences between two groups were analyzed by unpaired t test. Comparison among multiple groups was performed by one-way ANOVA. P < 0.05 was considered statistically significant. Regular analysis was performed using GraphPad Prism (Version 8.0.1, GraphPad Software Inc, CA, USA) and SPSS 25.0 (IBM Corp, USA).

[0044] 4 Experimental results

[0045] 4.1 LC-MS / MS identification of Dahuang Gancao Decoction aqueous extract

[0046] LC-MS / MS was used to identify the Dahuang Gancao Decoction aqueous extract, and the results are shown in Figure 1 Figure 1 ​Figures A and B in the middle represent the mass spectra of the rhubarb and licorice decoction aqueous extract obtained in negative ion mode and positive ion mode in LC-MS / MS analysis, respectively. Based on the peak areas in negative and positive ion modes, representative compounds include citric acid, rhein, 18-β-glycyrrhetinic acid, and aloe-emodin.

[0047] 4.2 LPS induces ferroptosis in SA-AKI mice

[0048] See Figure 2 As shown in Figure A, C57BL / 6 mice were acclimatized for 7 days, followed by intraperitoneal injection of LPS (15 mg / kg) to establish an SA-AKI model. Figure 2 As shown in the BC plot, compared with the Ctrl group, the LPS group mice had significantly higher levels of BUN and SCr; from Figure 2 The DF plot shows that the expression of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α was upregulated in the LPS group mice; from Figure 2 The G-plot shows an elevated renal tubular injury score; from Figure 2 As shown in the H-image, kidney histopathology (HE staining 20×) reveals renal tubular dilation with epithelial cell edema and interstitial inflammatory infiltration; immunohistochemistry (GPX4 staining 20×) shows decreased GPX4 expression; simultaneously, transmission electron microscopy (TEM detection 12000×) confirms mitochondrial cristae breakage and outer membrane swelling. Figure 2 As can be seen from the IL plot, LPS treatment also reduced the activity of SOD and GSH, while increasing the activity of MDA and Fe. 2+ The level.

[0049] 4.3 Erastin induces ferroptosis in HK-2 cells

[0050] The cell viability of HK-2 cells treated with different concentrations of Erastin was detected using the CCK-8 assay. The results of the cell viability experiment are as follows: Figure 3 As shown in Figure AB, HK-2 cell viability was inhibited after 24 hours of treatment with 0.4 μmol / L Erastin, exhibiting a dose-response curve. Therefore, subsequent experiments used 0.4 μM Erastin for 24 hours to induce HK-2 cell damage. Similarly, based on the cell viability experiment results, 5 μmol / L Fer-1 ferroptosis inhibitor was used as the intervention dose. Figure 3 As shown in the CH diagram, Erastin induction inhibited cell viability, GSH and SOD activity in HK-2 cells, increased MDA content, and significantly elevated ROS levels. The Fer-1 ferroptosis inhibitor reversed these changes.

[0051] 4.4 DHGC improves physicochemical parameters and reduces ferroptosis in SA-AKI mice

[0052] The schematic diagram of animal experiment is shown in Figure 4 Figure A. After 3 days of DHGC intervention, SA-AKI model was established by intraperitoneal injection of LPS (15 mg / kg). The experimental results are shown in Figure 4 Figures B-G, the medium dose DHGC group (2 g / kg) and the high dose DHGC group (4 g / kg) significantly inhibited the increase of serum BUN and SCr levels induced by LPS, significantly reduced the expression of inflammatory factors IL-6, IL-1β and TNF-α, and the renal tubular injury score. See Figure 5 From the observation of HE and PAS staining, compared with the LPS group, the kidney pathology of SA-AKI mice was significantly improved after intervention with different doses of DHGC, including relief of renal tubular dilation, reduction of epithelial cell edema and reduction of inflammatory cell infiltration, immunohistochemistry showed that GPX4 expression was increased, at the same time, transmission electron microscopy showed that the mitochondrial cristae rupture and outer membrane swelling caused by LPS were improved to different degrees after intervention with DHGC. The above results showed that compared with the LPS group, DHGC dose-dependently improved the pathological changes of SA-AKI mice.

[0053] To investigate the anti-ferroptosis effect of DHGC on SA-AKI mice, SOD activity, GSH content, MDA and Fe 2+ levels were analyzed. The results are shown in Figure 6 Figures I-L, compared with the Ctrl group, the ferroptosis level (MDA and Fe2 + level) of the LPS group was increased, and the antioxidant stress level (SOD activity and GSH content) was reduced. And DHGC also reversed the changes of the above indexes in a dose-dependent manner.

[0054] To investigate the mechanism of renal protection of DHGC in SA-AKI, RNA-seq analysis was performed. RNA-seq analysis was performed by comparing the gene expression in the kidneys of mice before and after DHGC treatment to determine the differentially expressed mRNAs. After normalization and filtering of the RNA-seq data, 5867 differentially expressed mRNAs were identified, as shown in Figure 6 Figure M, the volcano plot showed that 2637 mRNAs were up-regulated (red indicates up-regulation) and 3230 mRNAs were down-regulated (blue indicates down-regulation). As shown in Figure 6 Figure N, principal component analysis (PCA) showed that there were different clusters between the treatment group and the model group, which revealed the difference in mRNA expression between the two experimental groups. As shown in Figure 7As shown in the figure, the heatmap shows that rhubarb and licorice decoction upregulates SIRT3 and NRF2 (red indicates upregulation, blue indicates downregulation). Western blot analysis of mouse kidney tissue yielded the following results: Figure 7 As shown in the PS figure, the rhubarb and licorice decoction upregulated the expression of SIRT3 and GPX4 in SA-AKI mice, increased nuclear NRF2 levels, and decreased Ac-NRF2 levels. This indicates that NRF2 acetylation inhibits its nuclear translocation and promotes ferroptosis, while the rhubarb and licorice decoction alleviates ferroptosis and kidney injury by activating SIRT3-mediated NRF2 deacetylation, thereby promoting NRF2 nuclear translocation and GPX4 expression.

[0055] 4.5 Rhubarb and Licorice Decoction Restores Erastin-Induced Decline in HK-2 Cell Viability and Alleviates Ferrapholysis

[0056] like Figure 8 As shown in Figure A, Erastin-induced HK-2 cell viability was significantly reduced, while medium and high doses of rhubarb and licorice decoction could reverse Erastin-induced cytotoxicity and restore cell viability. Figure 8 As shown in the BF diagram, Erastin-induced oxidative stress imbalance in HK-2 cells led to increased MDA and ROS levels and decreased SOD and GSH levels. These pathological changes were effectively reversed by Da Huang Gan Cao Tang (rhubarb and licorice decoction). Figure 8 As shown in the GJ figure, Western blot analysis of SIRT3, GPX4, nuclear NRF2, and ac-NRF2 in HK-2 cells revealed that rhubarb and licorice decoction dose-dependently upregulated the expression of SIRT3 and GPX4, significantly increased the nuclear level of NRF2, and reduced the acetylated form of NRF2. Notably, the anti-ferroptosis effect of rhubarb and licorice decoction was comparable to that of the Fer-1 ferroptosis inhibitor. These results indicate that rhubarb and licorice decoction promotes the nuclear translocation of NRF2 through SIRT3-mediated deacetylation, activates downstream target genes of GPX4, and ultimately inhibits the ferroptosis pathway.

[0057] 4.6 Rhubarb and licorice decoction did not improve the physicochemical parameters or reduce ferroptosis in SIRT3 knockout SA-AKI mice.

[0058] To investigate whether rhubarb and licorice decoction plays an important role in protecting against ferroptosis in SA-AKI mice by targeting SIRT3, subsequent experiments were conducted using the CRISPR / Cas9 system to knock out SIRT3 in C57BL / 6 mice. Figure 9 As shown in the middle AE diagram, SIRT3 - / - The serum levels of BUN and SCr, as well as the expression of inflammatory factors IL-6, IL-1β, and TNF-α in the group of mice, were compared with those of SIRT3. + / +There was no significant difference between the groups of mice. At the same time, as shown in Figs. Figure 9 Figs. Figure 11 K, SIRT3 - / - oxidative stress and ferroptosis indicators GSH, SOD, MDA and Fe 2+ compared with SIRT3 + / + There was no significant difference between the groups of mice. As shown in Fig. Figure 10 I, histological analysis showed that SIRT3 - / - group of mice, no mitochondrial ultrastructure damage was observed, immunohistochemistry showed no change in GPX4 expression in both groups, and transmission electron microscopy showed no significant difference between the two groups. As shown in Figs. Figure 11 L-Q, compared with SIRT3 + / + group of mice, SIRT3 - / - group of mice, SIRT3 protein expression was absent, while there was no statistically significant difference in GPX4, nuclear NRF2 and ac-NRF2 protein expression between the two groups, as shown in Fig. Figure 11 J, there was no significant difference in tubular damage score between the two groups. The above evidence proves that SIRT3 knockout does not affect the physicochemical parameters of the kidney and the occurrence of ferroptosis.

[0059] Compared with the LPS group, the LPS+DHGC group significantly improved the above indicators. See Figs. Figure 9 A-H, Figure 10 I, Figure 11 K-J, however, in the SIRT3 - / - +LPS+DHGC group, the deteriorating physicochemical parameters of the kidney, including renal function, inflammation, oxidative stress and ferroptosis indicators, were not improved. Histological analysis showed that DHGC could not improve the SIRT3 - / - +SA-AKI group of mice, the renal tubular structure was destroyed, abnormal glycogen deposition and mitochondrial ultrastructure damage occurred. In addition, see Figs. Figure 11 L-Q, DHGC treatment could not significantly restore the SIRT3 - / - +SA-AKI mice GPX4 and nuclear NRF2 protein expression, nor could it reduce ac-NRF2 protein expression. The above results suggest that DHGC cannot improve the ferroptosis of SIRT3 knockout SA-AKI mice.

[0060] 4.7 DHGC cannot improve Erastin-induced ferroptosis in SIRT3 knockout HK-2 cells

[0061] To study whether DHGC can inhibit Erastin-induced ferroptosis in SIRT3 knockout HK-2 cells, SIRT3 siRNA was transiently transfected into HK-2 cells to silence SIRT3 gene expression. See Figs. Figure 12Fig. 2A, there was no significant difference in cell viability between the si-SIRT3-Ctrl group and the Ctrl group. See Figure 12 Figs. 2B-2D, there was no significant difference in the levels of GSH, MDA and SOD between the si-SIRT3-Ctrl group and the Ctrl group. Meanwhile, see Figure 12 Figs. 2E-2I, there was no significant difference in ROS fluorescence intensity, GPX4 protein expression, nuclear NRF2 protein expression and ac-NRF2 protein expression between the si-SIRT3-Ctrl group and the Ctrl group, but the SIRT3 protein expression in the si-SIRT3-Ctrl group was significantly inhibited compared with the Ctrl group. See Figure 13 Fig. 2J, immunofluorescence detection of NRF2 nuclear translocation, similar to the Ctrl group, only a small amount of NRF2 was detected in the cytoplasm of the si-SIRT3-Ctrl group. The above results show that silencing SIRT3 gene alone has no damage effect on HK-2 cells.

[0062] See Figure 12 Figs. 2A-2E, Figure 13 Fig. 2F, compared with the Erastin group, the cell viability, SOD and GSH levels of the Erastin+DHGC group were increased, and the MDA level and ROS fluorescence intensity were decreased, and see Figure 13 Figs. 2G-2I, DHGC treatment significantly restored GPX4 and nuclear NRF2 protein expression, and reduced ac-NRF2 levels. See Figure 13 Fig. 2J, NRF2 in the Erastin group mainly accumulated in the cytoplasm, and after DHGC treatment, NRF2 was obviously translocated into the nucleus. In the SIRT3-knockdown cells, the improvement effect of DHGC on the above indicators was almost completely eliminated. The above results prove that the deacetylation of NRF2 by DHGC targeting SIRT3 is the key molecular mechanism for inhibiting ferroptosis.

[0063] In summary, see Figure 14 , the present application induces a SA-AKI mouse model by injecting 15 mg / kg LPS in vivo, and induces HK-2 cell ferroptosis by using 0.4 μM Erastin in vitro. It is observed that the levels of BUN and SCr significantly increase after modeling, the levels of SOD and GSH significantly decrease, the levels of MDA and Fe 2+The content is significantly increased, SIRT3, GPX4 and nuclear NRF2 level is significantly inhibited, ac-NRF2 level is significantly increased, and the gradient dose intervention of Dahuang Lianqiao Decoction reverses the content change of the above indexes, however, the protection of Dahuang Lianqiao Decoction disappears through SIRT3 gene knockout or silencing. The above results verify that Dahuang Lianqiao Decoction plays an anti-ferroptosis role by targeting SIRT3-mediated deacetylation of NRF2, promoting nuclear translocation of NRF2. In summary, the Dahuang Lianqiao Decoction has ideal anti-ferroptosis effect, and can be used for preparing acute kidney injury anti-ferroptosis related drugs.

Claims

1. Use of the DHGC in the preparation of a drug for treating acute kidney injury ferroptosis mediated by the SIRT3 / NRF2 axis.

2. Use according to claim 1, characterized in that: The mechanism of action of the DHGC is: targeting activation of SIRT3, up-regulating the expression level of SIRT3, promoting deacetylation of NRF2 and driving nuclear translocation of NRF2, and then up-regulating the expression level of GPX4.

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