Application of rutin composition in preparing medicine for relieving cytotoxicity of porcine hepatocytes

Abstract

The present invention discloses the application of a rutin composition in the preparation of a drug for alleviating porcine hepatocyte toxicity, belonging to the field of pharmaceutical technology. The rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B. According to the verification of the embodiments of the present invention, it can be obtained that the rutin composition of the present invention can alleviate porcine hepatocyte toxicity by increasing the viability of IPLs cells, restoring the inhibition of IPLs cell proliferation by zearalenone, and restoring the increase in the ROS production in IPLs cells by zearalenone.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to the application of rutin composition in the preparation of a drug for alleviating porcine hepatocyte toxicity. Background Art

[0002] In modern animal husbandry, the scale of pig farming has been continuously expanding, and the accompanying disease problems have become increasingly prominent. Among them, hepatocyte toxicity is one of the important factors affecting pig health. After the hepatocytes are damaged, it will not only affect the growth and development and production performance of pigs, but may also lead to death in severe cases, causing economic losses to farmers. Therefore, developing effective methods to alleviate or treat porcine hepatocyte toxicity has become one of the key research directions. At present, there are some products for animal liver protection on the market, but most of them have limited effects or high costs, restricting their wide use in practical applications.

[0003] In recent years, with the progress of natural product chemistry and pharmacology research, it has been found that many plant extracts have good biological activities, including antioxidant, anti-inflammatory, and hepatoprotective effects, etc. Rutin is a flavonoid compound widely present in various plants, and it has been proven to have strong free radical scavenging ability and anti-lipid peroxidation ability. Based on these characteristics, scientists have begun to explore the potential role of rutin and its derivatives as potential drug ingredients in preventing and treating liver damage caused by various reasons. However, the application of rutin in the form of a composition in the preparation of a drug specifically for alleviating porcine hepatocyte toxicity has not been reported. Summary of the Invention

[0004] The purpose of the present invention is to provide the application of rutin composition in the preparation of a drug for alleviating porcine hepatocyte toxicity to solve the problems existing in the above-mentioned prior art. The rutin composition provided by the present invention can alleviate porcine hepatocyte toxicity by increasing the viability of IPLs cells.

[0005] To achieve the above purpose, the present invention provides the following solutions:

[0006] Technical Solution 1: The application of rutin composition in the preparation of a drug for alleviating porcine hepatocyte toxicity, wherein the rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B.

[0007] Further, the concentration of rutin is 60 μmol / L, the concentration of vitamin C is 2 μmol / L, the concentration of vitamin E is 20 μmol / L, the concentration of tanshinone IIA is 10 μmol / L, the concentration of puerarin is 40 μmol / L, and the concentration of licorinchalcone B is 20 μmol / L.

[0008] Further, the porcine hepatocyte toxicity is caused by zearalenone.

[0009] Further, the alleviation of porcine hepatocyte toxicity includes enhancing the viability of IPLs cells.

[0010] Further, the alleviation of porcine hepatocyte toxicity includes restoring the inhibition of IPLs cell proliferation by zearalenone.

[0011] Further, the alleviation of porcine hepatocyte toxicity includes restoring the increase in ROS production in IPLs cells by zearalenone.

[0012] Technical solution two: A drug capable of alleviating porcine hepatocyte toxicity, the active ingredient includes a rutin composition; the rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B.

[0013] Further, the concentration of rutin is 60 μmol / L, the concentration of vitamin C is 2 μmol / L, the concentration of vitamin E is 20 μmol / L, the concentration of tanshinone IIA is 10 μmol / L, the concentration of puerarin is 40 μmol / L, and the concentration of licorinchalcone B is 20 μmol / L.

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

[0015] The present invention discloses the application of a rutin composition in the preparation of a drug for alleviating porcine hepatocyte toxicity. The rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B. According to the verification of the embodiments of the present invention, it can be obtained that the rutin composition of the present invention can alleviate porcine hepatocyte toxicity by enhancing the viability of IPLs cells, restoring the inhibition of IPLs cell proliferation by zearalenone, and restoring the increase in ROS production in IPLs cells by zearalenone. Description of the Drawings

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0017] Figure 1Effects of zearalenone, vitamin C (A), vitamin E (B), tanshinone IIA (C), puerarin (D), licorinchalcone B (E), and rutin (F) on the viability of porcine hepatocytes (n = 6); where CON: control; ZEA: zearalenone; VC: vitamin C; VE: vitamin E; Tan IIA: tanshinone IIA; PUE: puerarin; LCB: licorinchalcone B; RUT: rutin; ZEA×VC: zearalenone×vitamin C; ZEA×VE: zearalenone×vitamin E; ZEA×Tan IIA: zearalenone×tanshinone IIA; ZEA×PUE: zearalenone×puerarin; ZEA×LCB: zearalenone×licorinchalcone B; ZEA×RUT: zearalenone×rutin; a, b, c indicate significant differences (P < 0.05);

[0018] Figure 2 Effects of each treatment group on the viability of porcine hepatocytes (n = 6); where CON: control; ZEA: zearalenone; VC: vitamin C; VE: vitamin E; RUT: rutin; VC×VE×ZEA: vitamin C×vitamin E×zearalenone; VC×RUT×ZEA: vitamin C×rutin×zearalenone; VE×RUT×ZEA: vitamin E×rutin×zearalenone; VC×VE×RUT×ZEA: vitamin C×vitamin E×rutin×zearalenone; VC×VE×Tan IIA×ZEA: vitamin C×vitamin E×tanshinone IIA×zearalenone; VC×VE×PUE×ZEA: vitamin C×vitamin E×puerarin×zearalenone; VC×VE×LCB×ZEA: vitamin C×vitamin E×licorinchalcone B×zearalenone; VC×VE×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×tanshinone IIA×puerarin×licorinchalcone B×zearalenone; VC×VE×RUT×Tan IIA×ZEA: vitamin C×vitamin E×rutin×tanshinone IIA×zearalenone; VC×VE×RUT×PUE×ZEA: vitamin C×vitamin E×rutin×puerarin×zearalenone; VC×VE×RUT×LCB×ZEA: vitamin C×vitamin E×rutin×licorinchalcone B×zearalenone; VC×VE×RUT×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×rutin×zearalenone×tanshinone IIA×puerarin×licorinchalcone B; a, b, c indicate significant differences (P < 0.05);

[0019] Figure 3Effect of each treatment group on the proliferation of porcine hepatocytes under fluorescence microscopy (n = 6), where CON: control; ZEA: zearalenone; VC×VE×RUT×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×rutin×zearalenone×tanshinone IIA×puerarin×licochalcone B; a, b, c indicate significant differences (P < 0.05), and the magnification ratio is 200 μm;

[0020] Figure 4 Statistics of the effect of the zearalenone and six - drug combination treatment group on the proliferation of porcine hepatocytes (n = 6), where CON: control; ZEA: zearalenone; VC×VE×RUT×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×rutin×zearalenone×tanshinone IIA×puerarin×licochalcone B; a, b, c indicate significant differences (P < 0.05);

[0021] Figure 5 Effect of the zearalenone and six - drug combination treatment group on the production of reactive oxygen species in porcine hepatocytes under fluorescence microscopy (n = 6), where CON: control; ZEA: zearalenone; VC×VE×RUT×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×rutin×zearalenone×tanshinone IIA×puerarin×licochalcone B; a, b, c indicate significant differences (P < 0.05), and the magnification ratio is 200 μm;

[0022] Figure 6 Statistics of the effect of the zearalenone and six - drug combination treatment group on the production of reactive oxygen species in porcine hepatocytes (n = 6), where CON: control; ZEA: zearalenone; VC×VE×RUT×Tan IIA×PUE×LCB×ZEA: vitamin C×vitamin E×rutin×zearalenone×tanshinone IIA×puerarin×licochalcone B; a, b, c indicate significant differences (P < 0.05). Detailed implementation manners

[0023] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0024] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0025] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0026] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the specification of the present invention, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0027] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0028] Example 1

[0029] 1 Test method

[0030] 1.1 Cell resuscitation

[0031] The IPLs cells were donated by the College of Animal Science and Technology (College of Veterinary Medicine), Shandong Agricultural University. Take out the cryopreservation tube of cells in the frozen state from the liquid nitrogen tank and place it in a 37 °C water bath. After melting, take it out. Take a 15 mL centrifuge tube and add 5 mL of complete cell culture medium (89% Dulbecco’s Modified Eagle’s Medium (DMEM)-high glucose medium (SH30022.01, Hyclone, USA) + 10% fetal bovine serum (FBS, 10099141, Gibco, USA) + 1% penicillin-streptomycin mixture (PS, 15140122, Gibco, USA)). Transfer the melted cells to the centrifuge tube using a pipette, gently pipette and mix well, centrifuge at 1000 r / min for 3 min, discard the supernatant, add 1 mL of complete cell culture medium, gently pipette and mix well 40 - 50 times to resuspend the cells. Take a 100 mm culture dish, add 9 mL of complete cell culture medium using a Pasteur pipette, and evenly inoculate the mixed cell suspension into the culture dish. Gently shake it in a cross shape on the plane. Observe the cell status under the microscope. If there is no abnormality, it can be placed in a 37 °C, 5% CO2 cell culture incubator for culture.

[0032] 1.2 Cell passage

[0033] When the cells are cultured to approximately 80% of the entire culture dish area, passage can be carried out. Use a Pasteur pipette to suck out and discard the culture medium in the culture dish, add 2 mL of PBS using a pipette to rinse, suck out and discard it, add 2 mL of 0.25% trypsin (15400054, Gibco, USA), gently shake to make it evenly distributed, place it in a 37 °C, 5% CO2 cell culture incubator for digestion for 2 min. Observe the cell status under the microscope. After most cells become round and detached, add 4 mL of complete cell culture medium to terminate digestion, gently pipette and mix well, transfer the cells to a 15 mL centrifuge tube, centrifuge at 1000 r / min for 3 min, discard the supernatant, add an appropriate amount of complete cell culture medium, gently pipette and mix well 40 - 50 times to resuspend the cells, and evenly inoculate them into a culture dish containing complete cell culture medium. Observe the cell status under the microscope. If there is no abnormality, it can be placed in a 37 °C, 5% CO2 cell culture incubator for culture.

[0034] 1.3 Cell cryopreservation

[0035] When the cells are cultured to approximately 80% of the entire culture dish area, the cell collection procedure is the same as in 1.2. Add cell cryopreservation solution (80% complete cell culture medium + 10% FBS + 10% dimethyl sulfoxide (DMSO) (VWRC0231, Amresco, USA)), gently pipette and mix 40 - 50 times to resuspend the cells, aliquot them into sterile cryotubes (labeled with passage number, date, name, and cell name), place them at 4°C for 0.5 h, transfer to -20°C for 0.5 h, then transfer to -80°C for 24 h, and subsequently transfer them to a liquid nitrogen tank for long-term storage.

[0036] 1.4 Cell treatment

[0037] 1.4.1 Toxin and drug preparation

[0038] ZEA: Zearalenone; VC: Vitamin C; VE: Vitamin E; Tan IIA: Tanshinone IIA; PUE: Puerarin; LCB: Licoricidin B; RUT: Rutin. The powder of ZEA (Fermentek, Israel, purity ≥98%) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored in the dark at -20 °C for preparing ZEA with appropriate concentrations. Previous studies in this invention found that the appropriate concentration of ZEA acting on IPLs was 60 μmol / L for 24 h treatment. Therefore, the same concentration and treatment time were also selected in this invention. The powder of Vitamin C (VC, Solarbio, China, purity ≥98%) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing VC with different concentrations. The powder of Vitamin E (VE, Solarbio, China, purity ≥98%) was dissolved in diethyl pyrocarbonate (DEPC) water (R0021, Beyotime, China) and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing VE with different concentrations. The powder of eutectic rutin (RUT, Eutectic Technology, China, rutin content 75%, the remaining 25% is L-proline) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing RUT with different concentrations. The powder of tanshinone IIA (Tan IIA, Huacui Biotechnology, China, purity ≥99%) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing Tan IIA with different concentrations. The powder of puerarin (PUE, Anhui Medical University, China, purity ≥98%) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing PUE with different concentrations. The powder of licoricidin B (LCB, Herbest Biotech, China, purity ≥98%) was dissolved in DMSO and prepared into a stock solution with a concentration of 20 mmol / L, stored at 4 °C for preparing LCB with different concentrations.

[0039] 1.4.2 Screening the Concentrations of Toxins and Drugs for Treating IPLs

[0040] Add 60 μL of 1 mg / mL polylysine solution to each well of a 96-well plate, place it in an incubator for 10 min, then aspirate and discard; add 100 μL of PBS to each well, place it in an incubator for 10 min, aspirate and discard, wash twice with PBS, and air dry. When the cells are cultured to cover about 80% of the entire culture dish area, resuspend the cells with trypsin, following the same steps as in 1.2. Use a counting plate to count the cells, dilute the cells with complete cell medium, and inoculate 10 4IPLs cells were seeded in 96-well plates. When the cells grew to 70% of the bottom area of the culture dish, the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × VC (0.5, 1, 2, 4, and 6 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × VC for treating IPLs cells; the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × VE (15, 20, 25, 30, and 35 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × VE for treating IPLs cells; the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × RUT (20, 40, 50, 60, 80 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × RUT for treating IPLs cells; the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × Tan IIA (2, 5, 8, 10, 15 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × Tan IIA for treating IPLs cells; the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × PUE (10, 20, 40, 60, 80 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × PUE for treating IPLs cells; the IPLs cells were treated with 0, 60 μmol / L ZEA, 60 μmol / L ZEA × LCB (10, 20, 40, 60, 80 μmol / L) for 24 h to detect cell viability and screen out the final concentration of ZEN × LCB for treating IPLs cells; using the concentrations of VC, VE, Tan IIA, PUE, LCB, and RUT screened above, twelve groups of drug treatment groups were prepared, namely 0, 60 μmol / L ZEA, VC × VE × ZEA, VC × RUT × ZEA, VE × RUT × ZEA, VC × VE × RUT × ZEA, VC × VE × Tan IIA × ZEA, VC × VE × PUE × ZEA, VC × VE × LCB × ZEA, VC × VE × Tan IIA × PUE × LCB × ZEA, VC × VE × RUT × Tan IIA × ZEA, VC × VE × RUT × PUE × ZEA, VC × VE × RUT × LCB × ZEA, and VC × VE × RUT × Tan IIA × PUE × LCB × ZEA, to detect cell viability and screen out the optimal treatment group.

[0041] 1.4.5 Cell viability detection

[0042] Cell seeding, toxin, and drug treatments were the same as in 1.4.2. The CCK-8 kit (C0038, Beyotime, Shanghai, China) was used to detect the viability of IPLs cells. 10 μL of CCK reagent was added to each well and incubated in a cell culture incubator for 2 h. The absorbance was measured at a wavelength of 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the cell viability was calculated.

[0043] 1.4.6 Cell proliferation detection

[0044] Cell seeding, toxin, and drug treatments were the same as in 1.4.2. The EdU cell proliferation detection kit (C10310-1, Beyotime, Shanghai, China) was used to detect the proliferation of IPLs cells. After the cells were treated with toxins and drugs for 22 h, 100 μL of complete cell medium containing 50 μmol / L EdU was added to each well and incubated in a cell culture incubator for 2 h. The supernatant was aspirated and discarded. The cells were washed twice with PBS, and 50 μL of cell fixative (PBS containing 4% paraformaldehyde) was added and incubated at room temperature for 30 min. The supernatant was aspirated and discarded. 50 μL of 2 mg / L glycine was added and incubated for 5 min. The supernatant was aspirated and discarded. The cells were washed with PBS, and 100 μL of permeabilizing agent (PBS containing 0.5% Triton X-100) was added and incubated for 10 min. The supernatant was aspirated and discarded. The cells were washed with PBS, and 100 μL of Apollo staining reaction solution was added and incubated in the dark at room temperature for 30 min. The supernatant was aspirated and discarded. 100 μL of Hoechst 33342 reaction solution was added and incubated in the dark at room temperature for 30 min. The supernatant was aspirated and discarded. After washing with PBS, the cells were observed under a 40× fluorescence microscope.

[0045] 1.4.7 Detection of intracellular reactive oxygen species (ROS) production

[0046] Cell seeding, toxin, and drug treatments were the same as in 1.4.2. A positive control well was set up separately. The ROS detection kit (S0033M, Beyotime, Shanghai, China) was used to detect the production of ROS in IPLs cells. After the cells were treated with toxins and drugs for 23.5 h, the ROS positive control reagent (Rosup, 50 mg / mL) was added to the positive control well and incubated in the incubator for an additional 0.5 h. Then, a reactive oxygen fluorescence probe with a final concentration of 10 μmol / L was added. After incubating in the incubator for 20 min, the cells were washed three times and observed under a 40× fluorescence microscope. The fluorescence intensity was scanned using ImageJ software.

[0047] 2 Results and analysis

[0048] 2.1 Cell viability detection

[0049] The effects of ZEA 60 μmol / L × VC (0.5, 1, 2, 4, and 6 μmol / L), ZEA 60 μmol / L × VE (15, 20, 25, 30, and 35 μmol / L), ZEA 60 μmol / L × Tan IIA (2, 5, 8, 10, and 15 μmol / L), ZEA 60 μmol / L × PUE (10, 20, 40, 60, and 80 μmol / L), ZEA 60 μmol / L × LCB (10, 20, 40, 60, and 80 μmol / L), and ZEA 60 μmol / L × RUT (20, 40, 50, 60, 80, and 100 μmol / L) on the viability of IPLs cells after 24 h of treatment are shown in Figure 1 . As can be seen from the figure, compared with the control group, 60 μmol / L ZEA can significantly reduce the viability of IPLs cells (P < 0.05); 2 μmol / L VC, 20 μmol / L VE, 10 μmol / L Tan IIA, 40 μmol / L PUE, 20 μmol / L LCB, and 60 μmol / L RUT can significantly increase the viability of IPLs cells (P < 0.05). Therefore, ZEA 60 μmol / L × VC 2 μmol / L, ZEA 60 μmol / L × VE 20 μmol / L, ZEA 60 μmol / L × Tan IIA 10 μmol / L, ZEA 60 μmol / L × PUE 40 μmol / L, ZEA 60 μmol / L × LCB 40 μmol / L, and ZEA 60 μmol / L × RUT 60 μmol / L were selected for subsequent experiments.

[0050] The effects of VC × VE × ZEA, VC × RUT × ZEA, VE × RUT × ZEA, VC × VE × RUT × ZEA, VC × VE × TanIIA × ZEA, VC × VE × PUE × ZEA, VC × VE × LCB × ZEA, VC × VE × Tan IIA × PUE × LCB × ZEA, VC × VE × RUT × Tan IIA × ZEA, VC × VE × RUT × PUE × ZEA, VC × VE × RUT × LCB × ZEA, and VC × VE × RUT × Tan IIA × PUE × LCB × ZEA on the viability of IPLs cells after 24 h of treatment with each treatment group are shown in Figure 2 . As Figure 2 can be seen, each treatment group can significantly increase the viability of IPLs cells (P < 0.05). Among them, the viability of IPLs cells in the VC × VE × RUT × Tan IIA × PUE × LCB × ZEA treatment group increased most significantly. Therefore, this treatment group was selected for subsequent experiments.

[0051] [[ID=

[0052] The effects of ZEA and the co - treatment of VC×VE×RUT×Tan IIA×PUE×LCB×ZEA for 24 h on the proliferation of IPLs cells are shown in Figure 3 and 4 . As can be seen from the figure, compared with the control group, 60 μmol / L ZEA can significantly inhibit the proliferation of IPLs cells (P < 0.05); the co - treatment group of VC×VE×RUT×Tan IIA×PUE×LCB×ZEA can significantly reverse the inhibition of IPLs cell proliferation by ZEA (P < 0.05).

[0053] 2.3 Detection of cellular reactive oxygen species production

[0054] The effects of ZEA and the co - treatment of VC×VE×RUT×Tan IIA×PUE×LCB×ZEA for 24 h on the proliferation of IPLs cells are shown in Figure 5 and 6 . As can be seen from the figure, compared with the control group, 60 μmol / L ZEA can significantly increase the production of ROS in IPLs cells (P < 0.05); the co - treatment group of VC×VE×RUT×Tan IIA×PUE×LCB×ZEA can significantly reverse the increase in ROS production in IPLs cells by ZEA (P < 0.05).

[0055] The embodiments described above are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of a rutin composition in the preparation of a drug for relieving cytotoxicity of porcine hepatocytes, characterized in that, The rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B; the porcine hepatocyte toxicity is induced by zearalenone.

2. The application according to claim 1, wherein The alleviation of porcine hepatocyte toxicity includes increasing the viability of IPLs cells.

3. The application according to claim 1, characterized in that, The alleviation of porcine hepatocyte toxicity includes restoring the inhibition of IPLs cell proliferation by zearalenone.

4. The application according to claim 1, characterized in that The alleviation of porcine hepatocyte toxicity includes restoring the increase in ROS production in IPLs cells by zearalenone.

5. A drug capable of alleviating the cytotoxicity of porcine hepatocytes, characterized in that, The active ingredient includes the rutin composition; the rutin composition includes rutin, vitamin C, vitamin E, tanshinone IIA, puerarin, and licorinchalcone B.

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