Application of paeoniflorin as an aryl hydrocarbon receptor agonist

Abstract

The present invention relates to the field of medical technology, and provides the application of paeoniflorin as an aryl hydrocarbon receptor agonist. The aryl hydrocarbon receptor agonist is paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin. Through the scheme of the present invention, paeoniflorin as an aryl hydrocarbon receptor agonist can regulate the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor, thereby producing an effect of resisting oxidative stress damage to the central nervous system, thereby improving central nervous system toxicity. Moreover, as an aryl hydrocarbon receptor agonist, paeoniflorin can activate multiple signaling pathways regulated by the aryl hydrocarbon receptor, target and improve central nervous system toxicity, and has no cytotoxicity. Therefore, paeoniflorin as an aryl hydrocarbon receptor agonist is used in the preparation of drugs for improving central nervous system toxicity and has potential application value.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to the application of paeoniflorin as an aryl hydrocarbon receptor agonist. Background Art

[0002] Central nervous system toxicity can cause serious interference with the normal functioning of the nervous system, and its causes are wide-ranging, including drug side effects, chemical exposure, infection, autoimmune reactions, etc. In the current treatment of central nervous system toxicity, drug therapy is one of the main means, but there are still some problems with existing drug treatments. Some traditional central nervous system toxicity treatment drugs have therapeutic effects, but at the same time they are toxic to normal nerve cells and other tissue cells. In addition, the central nervous system has a complex structure and function, and different brain regions and nerve cells have different sensitivities and responses to drugs. Existing drug treatments often lack precise targeting and are difficult to accurately act on damaged nerve cells or lesions.

[0003] The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcriptional protein widely distributed in various tissues and cells of humans and animals. Its protein structure is highly conserved across different organisms, and it plays a role in detoxification, immunity, and protection against invasion by microorganisms such as viruses and bacteria. Currently, aryl hydrocarbon receptor ligands are primarily divided into two categories: exogenous ligands such as dioxins, and endogenous ligands, primarily tryptophan metabolites. Albiflorin, a monoterpene glycoside found in high concentrations in the commonly used traditional Chinese medicine white peony root, exhibits a wide range of pharmacological activities and is used to treat rheumatoid arthritis, bacillary dysentery, enteritis, viral hepatitis, and geriatric diseases. However, the use of albiflorin as an aryl hydrocarbon receptor agonist in ameliorating central nervous system toxicity has not been reported. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention provides the use of paeoniflorin as an aryl hydrocarbon receptor agonist, which aims to solve the problems mentioned in the background technology.

[0005] In one aspect, the present invention provides an aryl hydrocarbon receptor agonist, wherein the aryl hydrocarbon receptor agonist is paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin.

[0006] Furthermore, the structural formula of the paeoniflorin is as follows:

[0007] .

[0008] On the other hand, the present invention also provides the use of paeoniflorin as an aryl hydrocarbon receptor agonist in the preparation of a drug for improving central nervous system toxicity.

[0009] Furthermore, the drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor and improving central nervous system toxicity.

[0010] Furthermore, the drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aromatic hydrocarbon receptor in hCMEC / D3 cells, thereby improving central nervous system toxicity.

[0011] Furthermore, the drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aromatic hydrocarbon receptor in hCMEC / D3 cells, downregulating the expression of PI3K, AKT, VEGFA, 4EBP-1, and HIF-1α, and upregulating the expression of p70S6k, mTOR, and Bcl-2 / Bax, thereby improving central nervous system toxicity.

[0012] Furthermore, the drug includes paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin, and one or more pharmaceutically acceptable excipients, carriers or adjuvants.

[0013] Furthermore, the drug is prepared into clinically acceptable tablets, pills, capsules, liquids, suspensions, gels, dispersions, solutions, emulsions, ointments or lotions.

[0014] The present invention has the following technical effects:

[0015] (1) Bifidobacterium lactone has an activating effect on the aryl hydrocarbon receptor, and bifidobacterium lactone can form multiple intermolecular hydrogen bonds with the active site of the aryl hydrocarbon receptor protein molecule. Therefore, bifidobacterium lactone can be used as an aryl hydrocarbon receptor agonist.

[0016] (2) As an aryl hydrocarbon receptor agonist, paeoniflorin can regulate the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor, thereby producing an anti-oxidative stress injury to the central nervous system and improving central nervous system toxicity. Moreover, as an aryl hydrocarbon receptor agonist, paeoniflorin can activate multiple signaling pathways regulated by the aryl hydrocarbon receptor, target and improve central nervous system toxicity, and has no cytotoxicity. Therefore, paeoniflorin as an aryl hydrocarbon receptor agonist has potential application value in the preparation of drugs to improve central nervous system toxicity. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] A more complete understanding of exemplary embodiments of the present invention may be obtained by referring to the following drawings:

[0018] Figure 1 This is a graph showing the detection results of the activation effect of paeoniflorin on the aryl hydrocarbon receptor according to Example 1 of the present invention; compared with the control group, * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001.

[0019] Figure 2 This is the optimal binding mode and interaction diagram between paeoniflorin and aromatic hydrocarbon receptor in Example 2 of the present invention.

[0020] Figure 3 This is a graph showing the qRT-PCR test results of Example 4 of the present invention; compared with the control group, * indicates p < 0.05; compared with the aconitine group, # indicates p < 0.05.

[0021] Figure 4 This is a graph showing the qRT-PCR detection results of Example 5 of the present invention; compared with the control group, * indicates p < 0.05.

[0022] Figure 5 : This is a graph showing the qRT-PCR test results of Example 6 of the present invention; compared with the control group, * indicates p < 0.05; compared with the paeoniflorin group, △ indicates p < 0.05; wherein:

[0023] Figure 5 A in the figure is the expression result of Bcl-2 / Bax gene mRNA in hCMEC / D3 of transfected and normal group, biloba lactone group and biloba lactone group;

[0024] Figure 5 B in the figure is the expression result of mTOR gene mRNA in hCMEC / D3 of transfected and normal groups, biloba lactone group and biloba lactone group. DETAILED DESCRIPTION

[0025] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; the terms used herein are only for the purpose of describing specific embodiments rather than limiting the present invention.

[0027] In some embodiments, the present invention provides an aryl hydrocarbon receptor agonist, which is abiogenin or a pharmaceutically acceptable salt of abiogenin.

[0028] Specifically, the structural formula of paeoniflorin is as follows:

[0029] .

[0030] In some embodiments, the present invention provides the use of paeoniflorin as an aryl hydrocarbon receptor agonist in the preparation of a medicament for improving central nervous system toxicity.

[0031] Specifically, the drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor and improving central nervous system toxicity.

[0032] Specifically, the drug regulates the PI3K / AKT / HIF-1 signaling pathway and improves central nervous system toxicity by stimulating the aromatic hydrocarbon receptor in hCMEC / D3 cells.

[0033] Specifically, the drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aromatic hydrocarbon receptor in hCMEC / D3 cells, downregulating the expression of PI3K, AKT, VEGFA, 4EBP-1, and HIF-1α, and upregulating the expression of p70S6k, mTOR, and Bcl-2 / Bax, thereby improving central nervous system toxicity.

[0034] Specifically, the drug includes paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin, and one or more pharmaceutically acceptable excipients, carriers or adjuvants.

[0035] Specifically, the drug is formulated into clinically acceptable tablets, pills, capsules, liquids, suspensions, gels, dispersions, solutions, emulsions, ointments or lotions.

[0036] Experimental Materials:

[0037] (1) Cell line: hCMEC / D3 cells (human cerebral vascular endothelial cells / D3 cell line) were purchased from Shanghai Fuheng Biotechnology Co., Ltd.;

[0038] (2) Drugs: Paeoniflorin (purity > 98%) was purchased from Chengdu Zhibiao Huachun Biotechnology Co., Ltd.

[0039] (3) Reagents: ECM culture medium (endothelial cell culture medium) was purchased from Beijing Solaibao Technology Co., Ltd.; pPRO-RB-Report-ABCB1-WT1 plasmid was constructed by Guangzhou Ruibo Biotechnology Co., Ltd.; CV702-AhR overexpression plasmid was provided by Shanghai Jikai Gene Technology Co., Ltd.; dual-luciferase reporter gene detection kit was purchased from Shanghai Biyuntian Biotechnology Co., Ltd.; OptiMEMI reduced serum culture medium and HiLiTran transfection reagent were purchased from Aorui Dongyuan Biotechnology Co., Ltd.; polymerase chain reaction (qPCR) primers were purchased from Shanghai Shenggong Biotechnology Co., Ltd.; total RNA extraction kit, reverse transcription kit and qPCR kit were purchased from Beijing Jumei Biotechnology Co., Ltd.

[0040] Example 1: Dual luciferase reporter gene assay to demonstrate the activation effect of paeoniflorin on aryl hydrocarbon receptors

[0041] (1) Cell culture and transient transfection: hCMEC / D3 cells were inoculated into cell culture flasks (T-25) and cultured in a cell culture incubator. hCMEC / D3 cells in the logarithmic growth phase were washed twice with 3 mL of Hanks' balanced salt solution, digested with 1 mL of 0.25% trypsin for 1 min, and terminated with 5 mL of ECM culture medium. The cells were centrifuged, the supernatant was discarded, and the cells were resuspended with ECM culture medium. The cells were mixed evenly and diluted to a cell concentration of 2 × 10 5 per well at 1×10 5 Cells were seeded in 24-well plates, with six replicates per group, and cultured to a cell density of 80%. Plasmids were diluted in OptiMEMI reduced serum medium. Co-transfection groups were set up with the pPRO-RB-ReportABCB1 plasmid (a promoter plasmid constructed by inserting the ABCB1 gene promoter into the upstream multiple cloning site of the Renilla luciferase gene using the pPro-RB-Report vector) and the CV702-AhR overexpression plasmid (CMVenhancer-MCS-3FLAG-SV40-Puromycin). A blank control group and a CV702-AhR empty vector group were also set up. Transfection was performed by adding HiLiTran transfection reagent to each group at a 1:50 ratio. After incubation for 20 minutes, 100 μL / well of the transfected cells were added to hCMEC / D3 cells. The cells were incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0042] (2) Drug intervention: 24 hours after co-transfection of hCMEC / D3 cells with pPRO-RB-ReportABCB1 plasmid and CV702-AhR overexpression plasmid, the cells were replaced with ECM culture medium containing drugs. The following were set: positive drug TCDD group: 2 nmol / L tetrachlorodiphenyl-p-dioxin group ECM culture medium; low-dose peoniflorin group: 4 μmol / L peoniflorin group ECM culture medium; medium-dose peoniflorin group: 10 μmol / L peoniflorin group ECM culture medium; high-dose peoniflorin group: 20 μmol / L peoniflorin group ECM culture medium; and a control group: ECM culture medium without drugs. 24 hours after drug intervention, dual luciferase activity was detected using a dual-luciferase activity assay kit, repeated 6 times.

[0043] (3) Dual luciferase activity detection: The culture medium was discarded from each group, and the hCMEC / D3 cells were washed once with phosphate buffer. 100 μL / well reporter gene lysis buffer was added, and the cells were shaken at room temperature for 15 min. After centrifugation at 12000 r / min for 5 min, 20 μL of supernatant was mixed with 100 μL firefly luciferase detection buffer, and the activity of firefly luciferase was detected using a multifunctional microplate reader. Then 100 μL Renilla luciferase luciferase detection buffer was added to detect the activity of Renilla luciferase. The ratio of Renilla luciferase activity to firefly luciferase activity was calculated, which was the corrected luciferase activity. The ratio of the luciferase activity of the drug to the luciferase activity of the control group was the induction fold.

[0044] The results of the detection of the activation effect of paeoniflorin on aryl hydrocarbon receptors are as follows Figure 1 As shown in the figure, the results showed that treatment of hCMEC / D3 cells with 4µmol / L, 10µmol / L, and 20µmol / L of paeoniflorin all activated the aryl hydrocarbon receptor in a dose-dependent manner. With the increase of the dose of paeoniflorin, the activation effect of paeoniflorin on the aryl hydrocarbon receptor was enhanced.

[0045] Example 2: Investigation of protein binding sites and binding free energy of peony lactone glycosides to aryl hydrocarbon receptor

[0046] Molecular docking studies were performed using AutoDock Vina software. The protein X-ray crystal structure (Protein Data Bank, code: 5V0L) was input. Water molecules were removed from the protein X-ray crystal structure and hydrogen atoms were added. Energy minimization was performed using the Tripos force field and Pullman charges. The structure of paeoniflorin was drawn using ChemDraw version 19.0 and optimized using ChemDraw3D. The ligand structure was optimized using AutoDock Vina software, dispersing the charge within the ligand residues, determining the root site of the ligand, and selecting torsional bonds within it. AutoDock Vina software was then loaded, followed by the optimized protein and energy-optimized baicalein. Grid energies were calculated using Autogrid, and the Lamarckian genetic algorithm was used to evaluate the binding between the ligand and the receptor using the free energy method. The docking results were visualized using PyMOL software and displayed in different modes.

[0047] The optimal binding mode and interaction between paeoniflorin and aryl hydrocarbon receptors Figure 2As shown in the figure, the binding energy of biloba lactone to the aryl hydrocarbon receptor docking is -8.1 kcal / mol, and the binding between them is primarily through hydrogen bonding and hydrophobic interactions. The amino acid that forms a hydrogen bond between the aryl hydrocarbon receptor and biloba lactone is Arg143, with hydrogen bond lengths of 2.0 and 2.4 Å. The hydrophobic amino acid surrounding biloba lactone (5 Å) is Arg143.

[0048] The results of Example 1 and Example 2 show that paeoniflorin is an aryl hydrocarbon receptor agonist.

[0049] Example 3: Protective effect of paeoniflorin on oxidative stress damage induced by aconitine in hCMEC / D3 cells

[0050] hCMEC / D3 cells were collected at 1×10 4 The cells were seeded at a density of cells / mL in a 6-well culture plate, 2 mL per well, and incubated in an incubator for 24 hours; the groups were set as follows: blank control group: no drug treatment; aconitine group: treated with 2 μg / ml aconitine; aconitine-peoniflorin low-dose combination group: treated with 2 μg / ml aconitine and 4 μg / ml paeoniflorin; aconitine-peoniflorin medium-dose combination group: treated with 2 μg / ml aconitine and 10 μg / ml paeoniflorin; aconitine-peoniflorin high-dose combination group: treated with 2 μg / ml aconitine and 20 μg / ml paeoniflorin; after the cells in each group were treated with drugs, the old culture medium was carefully aspirated, 500 μL of 0.25% trypsin was added, 1 mL of complete culture medium was added to terminate the digestion, and the cells were blown off the bottom of the plate with a pipette. Transfer the hCMEC / D3 cell suspension to a 5mL centrifuge tube and centrifuge at 1000 rpm for 10 minutes. Aspirate the supernatant, add 1mL of phosphate buffered saline (PBS), and rinse again by pipetting. Centrifuge again, aspirate the supernatant, and add 500μL of phosphate buffered saline to the hCMEC / D3 cell pellet. Prepare an ice-water bath and sonicate the cells using a cell ultrasonic disruptor set to 300W. Ultrasonicate once every 3-5 seconds, with 30 seconds between each sonication. Prepare hCMEC / D3 cell homogenates and measure the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the hCMEC / D3 cells.

[0051] The results of SOD and MDA content detection in hCMEC / D3 cells are shown in Table 1. The results show that aconitine has significant oxidative stress toxicity to hCMEC / D3 cells, and that paeoniflorin can reverse the oxidative stress toxicity of aconitine in a dose-dependent manner. This indicates that paeoniflorin has a protective effect against aconitine-induced oxidative stress damage in hCMEC / D3 cells.

[0052] Table 1 SOD and MDA contents in hCMEC / D3 cells

[0053]

[0054] In the table, * indicates that compared with the blank control group, p < 0.05; # indicates that compared with the aconitine group, p < 0.05.

[0055] In summary, aconitine exerts oxidative stress toxicity on hCMEC / D3 cells, leading to central nervous system toxicity. Biflorin can reverse the oxidative stress toxicity of aconitine. Furthermore, as shown in the test results in Table 1, biflorin ameliorates central nervous system toxicity by producing an effect that protects against oxidative stress damage to the central nervous system.

[0056] It is known in the prior art that the aryl hydrocarbon receptor, as a ligand-activated transcription factor, regulates the PI3K / AKT / HIF-1 signaling pathway and its downstream related genes PI3K, AKT, VEGFA, 4EBP-1, p70S6k, mTOR, Bcl-2 / Bax, and HIF-1α. Among them:

[0057] PI3K / AKT / HIF-1 signaling pathway: phosphatidylinositol 3-kinase / protein kinase B / hypoxia-inducible factor-1 signaling pathway;

[0058] PI3K: Phosphatidylinositol 3-kinase;

[0059] AKT: Protein Kinase B, protein kinase B;

[0060] VEGFA; Vascular Endothelial Growth Factor A, vascular endothelial growth factor A;

[0061] 4EBP-1: Eukaryotic Translation Initiation Factor 4E-Binding Protein 1;

[0062] p70S6k: p70 Ribosomal Protein S6 Kinase, p70 ribosomal protein S6 kinase;

[0063] mTOR: Mechanistic Target of Rapamycin, rapamycin target protein;

[0064] Bcl-2 / Bax: B-cell Lymphoma-2 / Bcl-2-associated X protein, B-cell lymphoma-2 / Bcl-2-associated X protein;

[0065] HIF-1α: Hypoxia-Inducible Factor-1α, hypoxia-inducible factor-1α.

[0066] Example 4: Regulatory Effects of Bifidobacterium on the mRNA Expression of PI3K, AKT, VEGFA, 4EBP-1, p70S6k, mTOR, Bcl-2 / Bax, and HIF-1α Genes in hCMEC / D3 Cells

[0067] (1) Cell culture: hCMEC / D3 cells were inoculated into cell culture flasks (T-25) and cultured in a cell culture incubator. hCMEC / D3 cells in the logarithmic growth phase were washed twice with 3 mL of Hanks solution, digested with 1 mL of 0.25% trypsin for 1 min, and then digested with 5 mL of ECM culture medium. The cells were centrifuged, the supernatant was discarded, and the cells were resuspended with ECM culture medium. The cells were mixed evenly and the cell concentration was diluted to 1 × 10 5 per well at 2×10 5 The cells were seeded in 6-well plates with 3 replicates per group and cultured until the cell density was about 80%.

[0068] (2) Drug intervention: The normal group was set up: no drug treatment; the aconitine group was treated with 2 μmol / L aconitine; the aconitine-peoniflorin low-dose combination group was treated with 2 μmol / L aconitine and 4 μmol / L paeoniflorin; the aconitine-peoniflorin medium-dose combination group was treated with 2 μmol / L aconitine and 10 μmol / L paeoniflorin; the aconitine-peoniflorin high-dose combination group was treated with 2 μmol / L aconitine and 20 μmol / L paeoniflorin. After 24 hours of drug intervention, the expression of PI3K, AKT, VEGFA, 4EBP-1, p70S6k, mTOR, Bcl-2 / Bax, and HIF-1α gene mRNA in hCMEC / D3 cells in each group was detected by qRT-PCR (real-time fluorescence quantitative polymerase chain reaction).

[0069] Activation of the PI3K / AKT pathway plays a dual role in ischemia-reperfusion injury. Early on, it may play a protective role by inhibiting apoptosis, but excessive activation may lead to inflammation and cell death. Upregulation of VEGFA and HIF-1α expression indicates cerebral ischemia. Downregulation of Bcl-2 / Bax and mTOR is often associated with mechanisms such as oxidative stress, disrupted energy metabolism, inflammatory responses, and aging. These changes are particularly prominent in neurodegenerative diseases, cerebral ischemia, and traumatic brain injury, ultimately leading to neuronal apoptosis and decreased brain function.

[0070] The results of qRT-PCR were as follows Figure 3 The results showed that compared with the normal group, the aconitine group increased the expression of PI3K, AKT, 4EBP-1, VEGFA, and HIF-1α genes in hCMEC / D3 cells, and decreased the expression of p70S6k, mTOR, and Bcl-2 / Bax genes, indicating that aconitine can cause hypoxia, inflammation, metabolic disorders, and apoptosis in hCMEC / D3 cells, resulting in central nervous system toxicity. When aconitine was combined with low, medium, and high doses of paeoniflorin, the expression of PI3K, AKT, 4EBP-1, VEGFA, and HIF-1α genes in hCMEC / D3 cells was downregulated, and the expression of p70S6k, mTOR, and Bcl-2 / Bax genes was upregulated, indicating that paeoniflorin can reduce the central nervous system toxicity of aconitine.

[0071] In summary, it is suggested that the mechanism by which paeoniflorin alleviates the neurotoxicity of aconitine may be related to hypoxia-induced activation of HIF-lα, improvement of vascular damage, and promotion of neurological function recovery.

[0072] Example 5: Verification of hAhR-siRNA Plasmid Inhibiting AhR Gene Expression

[0073] hCMEC / D3 cells were seeded in cell culture flasks (T-25) and cultured in a cell culture incubator. hCMEC / D3 cells in the logarithmic growth phase were washed twice with 3 mL of Hanks solution, digested with 1 mL of 0.25% trypsin for 1 min, and then digested with 5 mL of ECM medium. The cells were centrifuged, the supernatant was discarded, and the cells were resuspended in ECM medium and mixed evenly to dilute the cell concentration to 1×10 5 per well at 2×10 5Cells were seeded in 6-well plates, with triplicate wells per group, and cultured to a cell density of approximately 80%. hAhR siRNA groups were established: hCMEC / D3 cells were transfected with hAhR-siRNA plasmids. A control group (hCMEC / D3 cells were not transfected) and an NC siRNA group (hCMEC / D3 cells were transfected with NC-siRNA plasmids, which do not interfere with the AhR gene) were also established. Each group was transfected with plasmids according to the instructions for HiLiTran transfection reagent. Twenty-four hours after transfection, the culture medium was replaced with fresh medium, and the cells were cultured in a 37°C, 5% CO2 incubator for an additional 24 hours. qRT-PCR was used to examine AhR mRNA expression in hCMEC / D3 cells in the hAhR siRNA, control, and NC siRNA groups.

[0074] The results of qRT-PCR were as follows Figure 4 As shown, the results showed that there was no significant difference in the expression of AhR gene mRNA in hCMEC / D3 cells between the NC siRNA group and the control group, proving that the reagents and operations used in transfection did not affect the expression of the AhR gene; compared with the control group, the expression of AhR gene mRNA in hCMEC / D3 cells in the hAhR siRNA group was reduced, and the inhibition rate was greater than 70%.

[0075] In summary, hCMEC / D3 cells transfected with hAhR-siRNA plasmid inhibited AhR gene expression in hCMEC / D3 cells.

[0076] Example 6: Activation of aryl hydrocarbon receptor by paeoniflorin affects the induction of Bcl-2 / Bax and mTOR by paeoniflorin

[0077] hCMEC / D3 cells were transfected with hAhR-siRNA plasmids. 24 hours later, the cells were replaced with ECM culture medium containing 10 µmol / L paeoniflorin. These cells were designated as the transfection and paeoniflorin groups. A control group (hCMEC / D3 cells were not transfected or treated with the drug) and a paeoniflorin group (hCMEC / D3 cells were not transfected and treated with 10 µmol / L paeoniflorin) were also established. After 24 hours of drug treatment, the expression of Bcl-2 / Bax and mTOR gene mRNA in the cells was assessed by qRT-PCR.

[0078] The results of qRT-PCR were as follows Figure 5As shown in A and B in Figure 3, the results showed that compared with the normal group, the expressions of Bcl-2 / Bax and mTOR in hCMEC / D3 cells in the bifennilide group were significantly increased, indicating that bifennilide can induce the expression of Bcl-2 / Bax and mTOR in hCMEC / D3 cells; compared with the bifennilide group, the expressions of Bcl-2 / Bax and mTOR in hCMEC / D3 cells in the transfection and bifennilide groups were significantly decreased, indicating that inhibiting the expression of AhR can reduce the induction effect of bifennilide on Bcl-2 / Bax and mTOR, that is, the induction effect of bifennilide on Bcl-2 / Bax and mTOR is related to the activation of aryl hydrocarbon receptor by bifennilide.

[0079] Based on the analysis of the results of Examples 1 to 6, the present invention reaches the following conclusions:

[0080] (1) Bifidobacterium lactone has an activating effect on the aryl hydrocarbon receptor, and bifidobacterium lactone can form multiple intermolecular hydrogen bonds with the active site of the aryl hydrocarbon receptor protein molecule. Therefore, bifidobacterium lactone can be used as an aryl hydrocarbon receptor agonist.

[0081] (2) As an aryl hydrocarbon receptor agonist, paeoniflorin can regulate the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor, thereby producing an anti-oxidative stress injury to the central nervous system and improving central nervous system toxicity. Moreover, as an aryl hydrocarbon receptor agonist, paeoniflorin can activate multiple signaling pathways regulated by the aryl hydrocarbon receptor, target and improve central nervous system toxicity, and has no cytotoxicity. Therefore, paeoniflorin as an aryl hydrocarbon receptor agonist has potential application value in the preparation of drugs to improve central nervous system toxicity.

[0082] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Use of paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin as an aryl hydrocarbon receptor agonist in the preparation of a medicament for improving central nervous system toxicity caused by aconitine.

2. The use according to claim 1, characterized in that: The drug regulates the PI3K / AKT / HIF-1 signaling pathway by stimulating the aryl hydrocarbon receptor, thereby improving the central nervous system toxicity caused by aconitine.

3. The use according to claim 2, characterized in that: The drug comprises paeoniflorin or a pharmaceutically acceptable salt of paeoniflorin, and one or more pharmaceutically acceptable excipients, carriers or adjuvants.

4. The use according to claim 3, characterized in that: The drug is prepared into clinically acceptable tablets, pills, capsules, suspensions, gels, solutions, emulsions, ointments or lotions.

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