Application of active component of humifuse euphorbia herb in preparation of medicine for treating lipotoxicity inflammation

Abstract

The invention provides an application of an active ingredient of humifuse euphorbia herb in preparation of a medicine for treating lipotoxicity inflammation. The active ingredient of the humifuse euphorbia herb is a composition formed by gallic acid, quercetin, kaempferol and umbelliferone. The invention also provides an application of the active component of the humifuse euphorbia herb in preparation of a medicine for treating or improving ulcerative colitis induced by high fat combined with DSS. The composition of four main active ingredients in the humifuse euphorbia herb extract can effectively improve lipotoxicity inflammation.

Description

Technical Field

[0001] The present invention relates to the fields of Chinese medicine chemistry, and in particular to a compatibility pharmaceutical composition of an active ingredient group of Herba Euphorbiae Herba Extract, i.e., a combination of effective monomer components contained therein, and a use thereof for improving the effect of high fat promoting dextran sulfate sodium (DSS)-induced ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is a chronic inflammatory disease of the intestine, which belongs to inflammatory bowel disease (IBD). Currently, more than 7 million people suffer from IBD worldwide. UC is common in the rectum, with diffuse inflammation, leading to increased permeability of the intestinal mucosa, edema, ulcers, bleeding, and electrolyte loss. Epidemiological and experimental studies of UC have found that diet is one of the factors leading to the onset of ulcerative colitis. Excessive intake of sugar, animal fat, linoleic acid, etc. can promote the occurrence of intestinal inflammation.

[0003] Inflammasomes are recognized as important players in innate immunity, among which NLRP3 inflammasomes are one of the most famous members. NLRP3 inflammasomes are mainly present in immune and inflammatory cells activated by inflammatory stimuli. Studies have shown that cellular NLRP3 protein levels are essential for the assembly and activation of NLRP3 inflammasomes. Inflammasome NLRP3 plays an important role in the process of colitis and inflammation-related colitis, and studies have shown that the expression of NLRP3 can be indirectly inhibited through the NF-κB pathway in colonic inflammation, attenuating DSS-induced colitis. The propagation of cytokine-driven stress / inflammatory responses depends on the activity of transcription factors, which in turn activate multiple downstream genes.

[0004] Transcription factor E74 factor 3 (ELF3) is one of the epithelial-restricted members of the Ets transcription factor family. ELF3 is an inflammatory mediator that is normally expressed in epithelial tissues. After being induced by inflammatory stimuli, it induces the expression of downstream inflammatory-related genes such as nitric oxide synthase 2 (NOS2), cyclooxygenase (COX2), and lipocalin 2 (LCN2). At the same time, the high-affinity NF-κB binding site on the proximal ELF3 promoter can bind to NF-κB (p65 / p50) in a dependent manner. ELF3 can also accelerate the apoptosis of intestinal epithelial cells in ulcerative colitis by accelerating the activation of NF-κB. At the same time, studies have also found that EMT in colorectal cancer cells requires the expression of NLRP3 protein. Studies have shown that ELF3 can participate in the inflammatory response of the endothelium under high glucose conditions, mediating the activation of the NLRP3 inflammasome by inducing upregulation of ELF3 expression.

[0005] Euphorbia euphorbiae is the dried whole herb of Euphorbia euphorbiae or Euphorbia euphorbiae, which has the effects of clearing heat, detoxifying, diuresis, hemostasis, and lactation. It is mainly used for dysentery, diarrhea, hematuria, bloody stool, hemoptysis, metrorrhagia, damp-heat jaundice, etc. Summary of the invention

[0006] Based on the above research background, this study aims to explore the compatibility of the active ingredient group of Herba Euphorbiae Iris extract, that is, the combination of effective monomer components contained therein and observe their intervention effects on the lipotoxic enteritis cell model and the high-fat diet combined with dextran sulfate sodium (DSS)-induced ulcerative colitis in mice.

[0007] Disclosed is an application of an active ingredient of Herba Euphorbiae in preparing a medicine for treating or improving lipotoxic inflammation. The active ingredient of Herba Euphorbiae is a composition formed by gallic acid, quercetin, kaempferol and umbelliferone.

[0008] The active ingredients of the Herba Euphorbiae Herba are used in reducing the expression of ELF3, NLRP3 and the pro-inflammatory factor TNF-α to prepare a drug for treating or improving lipotoxic inflammation.

[0009] The active ingredients of the herb are a composition formed by gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 3-8:0.5-2:0.5-2:0.5-2.

[0010] In some preferred embodiments, the active ingredient of the Herba Euphorbiae Ichthyophthirii is a composition formed by gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 4:1:1:1.

[0011] The four main active ingredients in the Herba Euphorbiae extract of the present invention can effectively improve lipotoxic inflammation and reduce inflammatory response. The four main active ingredients in the Herba Euphorbiae extract of the present invention can effectively protect the damage of the intestinal mucosa in the colorectal mucosa caused by high fat combined with DSS, and the mechanism is related to reducing the level of inflammatory cytokines in colorectal tissue and reducing inflammatory response. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 The expression of related inflammatory factors and ELF3 in the lipotoxic inflammatory cell model, including: a. The result of the change of TNFα, NLRP3, and ELF3 protein expression in cells; b. The statistical chart of the expression of TNFα, NLRP3, and ELF3 proteins in cells. Note: One-way ANOVA, n=4; compared with the normal group: * P <0.05, ** P <0.01, *** P <0.001.

[0013] Figure 2 This is the screening of the combination of four monomer compounds in the extract of Euphorbia pulegiae (combination group). Note: One-way ANOVA, n=4; compared with the normal group: * P <0.05, ** P <0.01, *** P <0.001; compared with the model group: # P <0.05, ## P <0.01, ### P <0.001.

[0014] Figure 3 The protective effect of the monomer compound compound (combination group) in the extract of Herba Euphorbiae on the lipotoxic enteritis cell model. a. The change levels of ELF3, NLRP3, and TNFα mRNA were detected in the Herba Euphorbiae monomer combination group after intervention in the lipotoxic enteritis model; b. The expression of ELF3, NLRP3, and TNFα proteins were detected in the Herba Euphorbiae monomer combination group after intervention in the lipotoxic enteritis model; c. The statistical chart of the expression of ELF3, NLRP3, and TNFα proteins in the Herba Euphorbiae monomer combination group after intervention in the lipotoxic enteritis model; Note: One-way ANOVA, n=4; Compared with the normal group: * P <0.05, ** P <0.01, *** P <0.001; compared with the model group: # P <0.05, ## P <0.01, ### P <0.001.

[0015] Figure 4 The gross morphology of the mouse colon and the detection of colorectal tissue morphology, a. Mouse weight change and DAI score; b. Mouse colorectal length and statistical chart of mouse colorectal length; c. H&E staining of mouse colon tissue; Note: Normal: normal control group mice, DSS+HFD: high-fat diet + 2.5% DSS model group mice, Drug-L: low-dose group of Herba Dichondrium intervened in high-fat diet + 2.5% DSS group mice, Drug-H: high-dose group of Herba Dichondrium intervened in high-fat diet + 2.5% DSS group mice; One-way ANOVA, n=6; Compared with the normal group: # P <0.05,## P<0.01; compared with the model group: * P <0.05,** P <0.01,*** P <0.001.

[0016] Figure 5 Figure 2 is the expression of colon mucosal barrier-related genes in each group of mice.

[0017] Figure 6 The expression of related inflammatory factors in the colon tissue of each group of mice. a. Expression of related inflammatory factors mRNA in the colorectal tissue of mice; Note: Normal: normal control group mice, DSS+HFD: high-fat diet + 2.5% DSS model group mice, Drug-L: low-dose group of Herba Dichondrium intervening with high-fat diet + 2.5% DSS group mice, Drug-H: high-dose group of Herba Dichondrium intervening with high-fat diet + 2.5% DSS group mice; One-way ANOVA, n=6; Compared with the normal group: # P <0.05, ## P <0.01, ### P <0.001; compared with the model group: * P <0.05,** P <0.01,*** P <0.001. DETAILED DESCRIPTION

[0018] Example 1 Monomer compound standards such as gallic acid, quercetin, kaempferol, and umbelliferone were purchased from Chengdu Plant Standard Pure Biotechnology Co., Ltd. DSS was produced by Yisheng Biotechnology Co., Ltd. Lipopolysaccharide (LPS) was produced by Sigma, Germany. Palmitic acid was produced by GENE TECH, Spain. HRP-labeled goat anti-rabbit IgG: produced by Wuhan Sewell Biotechnology Co., Ltd., batch number GB23303. ELF3 antibody was produced by Abcam, UK, batch number ab133621. NLRP3 antibody was produced by Wuhan Abotek Biotechnology Co., Ltd., batch number A21906. TNF-α antibody was produced by Santa Cruz, USA, batch number SC-52746. 31) β-actin antibody was produced by Wuhan Abotek Biotechnology Co., Ltd., batch number AC026. 30). The primers used for qRT-PCR were synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0019] Experimental cells Mouse colon cancer cells MC38 were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China Collection Culture Center, and cultured in RPMI 1640 medium containing 10% FBS in a 37°C temperature-controlled CO2 incubator, with medium replacement every other day. All experiments used cells with a confluence of 80-90%.

[0020] Western blot assay RIPA lysis buffer was used to prepare the original protein lysate of colon tissue and cells. The total protein concentration was determined using the BCA protein quantification kit; 25 μg of protein samples were separated using a 10% SDS-PAGE gel. Then, the proteins on the gel were transferred to a PVDF membrane. Subsequently, nonspecific antigens were blocked with 5% skim milk in TBST for 1 h, incubated overnight at 4° in the prepared primary antibody solution, and then incubated with the secondary antibody at room temperature for 1 h. Finally, the PVDF membrane was soaked in the prepared ECL developing working solution. The ChemiScope mini chemiluminescent imaging system was used for development. The protein level was quantified using ImageJ software.

[0021] Real-time RT-PCR assay Real-time RT-PCR was performed according to the manufacturer's method as follows: Total RNA was extracted from mouse colon tissue or cells using Trizol solution. Total RNA samples were reverse transcribed into cDNA and quantitative PCR was performed using SYBR Green (Vazyme, China) and evaluated using the QuantStudio™ Design and Analysis Real-Time PCR System (Applied Biosystems, Carlsbad, CA, USA). β-actin or U6 RNA was used as an internal reference for mRNA and miRNA levels, respectively. The primer sequences are shown in Table 2 Table 2 Primer sequence list

[0022] Statistical analysis All data are expressed as mean ± SD. The results were analyzed using GraphPad Prism 5 software. One-way ANOVA and independent sample t-test were used, and statistical differences were determined when p < 0.05.

[0023] Establishment of lipotoxic inflammatory cell model In order to explore the intervention effect of Herba Euphorbiae Herba Extract on intestinal inflammatory response at the in vitro level, the present invention first used LPS and PA to treat MC38 cells together to induce lipotoxic inflammatory response in cells, which was used to simulate the in vitro model of ulcerative colitis in which lipotoxicity aggravates enteritis. After MC38 cells were treated with LPS:PA (1 μg / mL: 20 μmol / mL) at a concentration of 1 μg / mL, the protein expression of related inflammatory factors was detected. The results are as follows: Figure 1 As shown in the figure, compared with the control group, the protein levels of inflammatory factors TNFα and NLRP3 were significantly upregulated after LPS and PA combined treatment (P < 0.05). Subsequently, the combination of LPS∶PA (1 μg / mL: 20 μmol / mL) was tested to see whether it could induce high expression of ELF3. The results are shown in the figure. Compared with the control group, both PA and LPS alone could induce high expression of ELF3, and the expression level of ELF3 in the LPS∶PA group was more significant than that in the PA and LPS treatment groups alone (P < 0.05). This suggests that LPS∶PA combined treatment of MC38 cells can replicate the in vitro lipotoxic enteritis model, so this concentration was selected for subsequent experiments.

[0024] Screening of monomer compound combination (combination group) in Herba Euphorbiae Herba Extracts According to the literature search, the monomer components contained in the extract of Herba Euphorbiae Herba were obtained. In the preliminary experiment, 7 monomer compounds were screened, and 4 monomer components were further obtained to have a protective effect on the lipotoxic inflammatory cell model, namely gallic acid, quercetin, kaempferol, and umbelliferone, and the protective effect was dose-dependent (P < 0.05). Figure 2 shown.

[0025]

[0026] Protective effect of the combination of monomeric compounds in Herba Euphorbiae Herba Extracts (combination group) on inflammatory cell models Orthogonal experimental optimization design of monomer composition: Drug concentration design: Unit (μg / mL) Gallic acid (A) Umbelliferone (B) Kaempferol (C) Quercetin (D) High (1) 2.5 2.5 2.5 2.5 Medium(2) 1.25 1.25 1.25 1.25 Low (3) 0.625 0.625 0.625 0.625 Orthogonal array According to the design scheme in the table below, the present invention combines the above four compounds and adds them to the lipotoxic inflammatory cell model respectively. After the treatment, the cells are harvested and the total cell RNA is extracted according to the instructions. Subsequently, the relative expression level of the NLRP3 gene is detected by RT-qPCR technology.

[0027]

[0028] In the above, NO refers to the serial number, and 1, 2, and 3 refer to the high (1), medium (2), and low (3) content of the components of each raw material in the above-mentioned agents, respectively.

[0029] The ability to inhibit inflammatory response is measured by the relative expression of the NLRP3 gene. Specifically, the lower the expression of the NLRP3 gene, the stronger the ability of the monomer composition to inhibit inflammation. Our experimental results show that some monomer compositions have the potential to inhibit inflammation. Among these compositions, 3# and 7# have the most significant inhibitory effects on inflammation, while 4# and 8# have the second highest inhibitory abilities. Combining various data, we can derive the following inhibitory ability ranking: 3>7>4>8>9>6>2>5>1.

[0030] Further analysis of the factors that affect the ability to inhibit inflammation found that the degree of influence of each factor was D>A>B>C, which indicates that quercetin (factor D) plays the most critical role in inhibiting inflammation. Based on this, we determined that the concentration of each monomer in the 3# ratio relationship is the best ratio, that is, A1:B3:C3:D3, or 4:1:1:1, which can achieve the maximum inhibitory activity.

[0031] In the above orthogonal experiment, the four monomer components were subjected to orthogonal experiments, and the combination of gallic acid: quercetin: kaempferol: umbelliferone in a ratio of 4:1:1:1 (2.5 μg / mL: 0.625 μg / mL: 0.625 μg / mL: 0.625 μg / mL) was applied to the cell model of lipotoxic enteritis, and the intervention effect was the most significant. The results are as follows Figure 3 As shown, compared with the control group, the expression of ELF3 and NLRP3 in the model group was significantly upregulated (P < 0.01). Compared with the model group, after the intervention of the monomer combined group, the expression of ELF3 and NLRP3 was significantly downregulated, and it was dose-dependent (P < 0.05). At the same time, the present invention also detected the expression of the pro-inflammatory factor TNF-α in each group, and its expression trend was the same as that of ELF3 and NLRP3. Compared with the control group, the expression of the model group was significantly upregulated, and after the intervention of the Herba Euphorbiae Monomer Combined Group, its expression was significantly decreased.

[0032] Example 2 The improvement effect of Herba Euphorbiae Herba Extract on ulcerative colitis in mice, that is, the combination of different extracts in Herba Euphorbiae Herba Extract, specifically, in Example 1, gallic acid: quercetin: kaempferol: umbelliferone were mixed in a ratio of 4:1:1:1 (2.5 μg / mL: 0.625 μg / mL: 0.625 μg / mL: 0.625 μg / mL), and then 1.75 mg / mL was used as the Herba Euphorbiae Herba Extract low-dose group and 5.25 mg / mL was used as the Herba Euphorbiae Herba Extract high-dose group for the following test.

[0033] Establishment and testing of ulcerative colitis mouse model Experimental animals and feed The present invention uses 20 SPF grade 6-week-old C57 male and female mice, 20 ± 2 g, all purchased from the Experimental Animal Center of Three Gorges University (animal production license is SCXK (E) 2017-0012), and the mice are raised in the Experimental Animal Center of Three Gorges University. The mouse breeding conditions are: relative humidity 40-70%, ambient temperature 22 ± 2 ° C, light and dark: 12 h each, mice are free to drink water, drink 2.5% DSS solution, and the bedding is changed regularly every week. The ordinary feed consumed by the mice was purchased from the Experimental Animal Center of Three Gorges University, and the high-fat feed was homemade feed, with a specific weight ratio of: 53.75% ordinary feed, 20% lard, 20% fructose, 5% cholesterol, 1% edible salt and 0.25% sodium cholate. The nutritional components of the ordinary feed are shown in Table 1, and the nutritional components of the high-fat feed are shown in Table 2. All animal experiments in this study were carried out in accordance with animal husbandry requirements and operations and were approved by the Animal Ethics Committee of China Three Gorges University (approval number: 4201020005879).

[0034] Table 1 Nutrient composition of common feed (per kg feed)

[0035] Table 2 Nutritional composition of high-fat feed (per kg feed)

[0036] Construction of experimental animal models, specimen collection and preservation According to the experimental data of the previous research group, it was found that drinking 2.5% DSS solution in a high-fat environment can quickly induce ulcerative colitis in mice, and the colon inflammation of mice is more significant, so this condition was used to model mice. 20 male and female C57 mice of SPF grade were completely randomly divided into 8 groups: normal group, model group, low-dose group of Herba Euphorbiae Herba Extract, and high-dose group of Herba Euphorbiae Herba Extract (male and female C57 mice were divided into 4 groups according to this grouping). After all mice were fed with ordinary feed in the first week, the ordinary feed of the model group, low-dose group of Herba Euphorbiae Herba Extract, and high-dose group of Herba Euphorbiae Herba Extract was replaced with high-fat feed in the second week. The mice in the normal group were fed with ordinary feed. At the same time, the drug group was given different doses of Herba Dijincao extract (80 mg / mL, 240 mg / mL) by gavage according to the weight of each mouse every day, and the normal group and model group were given PBS by gavage. From the fifth week, mice in all groups except the normal group were given free drinking of 2.5% DSS solution, and the DSS solution was replaced with a fresh 2.5% DSS solution every two days. The weight of the mice was recorded every day, and after the administration of 2.5% DSS solution, the redness and swelling of the anus of the mice, whether there was bleeding, etc., as well as the shape, color, and blood in the stool of the mice were observed. After a total of 5 weeks of feeding, the mice were taken out of the animal room, euthanized, and the residual excrement in the colorectal tissue of the mice was carefully irrigated with saline, and the excess saline on the surface of the colorectal tissue was absorbed with filter paper. The length of the colorectum of the mice was measured and photographed. Cut 0.5 cm of mouse colorectal tissue and place it in an embedding clip, then place it in 4% paraformaldehyde. The remaining tissue is placed in a sterile 1.5 mL EP tube and stored at -80° for long-term storage for subsequent experiments. Note: Colorectal tissue is a digestive organoid and should be collected quickly.

[0037] H&E staining After the colon tissue was fixed with 4% paraformaldehyde solution, the length and shape of the colon tissue were trimmed. Then it was dehydrated in a dehydrator. After dehydration, it was embedded in a paraffin embedding machine. After the wax block was cooled, it was sliced ​​with a slicer to a thickness of 4 μm. After fully unfolding in warm water, it was attached to a slide and spread at 60°C for at least 4 hours before H&E staining. The steps of H&E staining are as follows: Xylene I immersion time: 5 min; Xylene II immersion time: 10 min; Anhydrous ethanol I immersion time: 5 min; Anhydrous ethanol II immersion time: 10 min; 95% ethanol immersion time: 3-5 min; 80% ethanol immersion time: 3-5 min; 70% ethanol immersion time: 3-5 min; ddH2O immersion time: 3 min; Hematoxylin staining time: 3 min; ddH2O soaking time: 30 sec; Eosin staining time: 45 sec; 70% ethanol immersion time: 1 sec; 80% ethanol immersion time: 1 sec; 95% ethanol I immersion time: 1 sec; 95% ethanol II immersion time: 1 sec; Anhydrous ethanol I immersion time: 5 min; Soaking time in anhydrous ethanol II is 10 min; Xylene I immersion time 5 min; Xylene II immersion time 10 min; After staining, drop neutral resin next to the tissue and seal it with a coverslip. It is important to avoid creating bubbles as much as possible during the sealing process. Place it in a ventilated and cool place and store it at room temperature for a long time after it is completely solidified, or observe and take pictures under an optical microscope.

[0038] Extract of Herba Euphorbiae Herba improves body weight and DAI score in mice with ulcerative colitis According to the experimental data of the previous research group, feeding mice with a 2.5% DSS solution combined with a high-fat diet can accelerate the occurrence of inflammatory response in mice. This condition was used to model the animal experiment of this project. The weight changes of mice were recorded every day, and the fecal characteristics and blood in the stool of mice were observed after the mice drank the 2.5% DSS solution, which was combined into the DAI score. The results showed that compared with the normal control group, the DAI score of the mice in the model group increased and the weight loss was obvious ( P <0.01). After the mice were gavaged with Herba Euphorbiae Herba Extract, the weight loss of the mice was reduced compared with the model group mice, and the DAI scores were reduced to varying degrees, such as Figure 4 As shown in a.

[0039] Changes in colorectal tissue morphology in mice After the mice were killed by cervical dislocation, their colons were removed and irrigated with saline to measure the length of the colon. Compared with the normal control group, the colon length of the mice in the model group was significantly shortened ( P <0.01). The colon length of the group treated with Dijincao medicine was shortened compared with the model group ( P <0.05), such as Figure 4As shown in b. Part of the colon tissue at the lesion site was cut, dehydrated, embedded, sliced ​​and stained with H&E. It can be seen that the colon tissue mucosa of the normal control group was intact, with regular cell structure and glands in normal position, showing a complete and healthy intestinal barrier. However, in the model group, epithelial cells were damaged, the colon mucosa had abnormal cell structure and infiltration of inflammatory cells that destroyed glandular tissue, while the colon of mice in the group intervened by Dijincao showed mild infiltration and damage of inflammatory cells, as shown in Figure 2. Figure 4 As shown in c.

[0040] Expression of intestinal mucosal barrier-related genes in mouse colorectal tissue Real-time PCR was used to detect the expression of intestinal tight junction proteins and mucus secretion proteins ZO-1 and Connexin-43 in the colorectal tissues of each group of mice. The results showed that the expression levels of ZO-1 and Connexin-43 in the model group were significantly lower than those in the normal control group ( P <0.01), while the expression levels of ZO-1 and Connexin-43 in the mice in the Euphorbia pulex drug intervention group increased compared with those in the model group, and gradually approached the level of the normal control group ( P <0.05), such as Figure 5 shown.

[0041] Expression of inflammatory-related factors in mouse colorectal tissue Real time PCR was used to detect the expression of inflammatory factors TNFα, TLR4, and IL-1β mRNA in the colorectal tissues of each group of mice. The results showed that compared with the normal control group, the expression of inflammatory factors in the model group was significantly increased ( P >0.05), while the expression of inflammatory-related factors in mice in the Herba Euphorbiae Herba Extract intervention group was significantly reduced compared with the model group ( P >0.001), such as Figure 6 shown.

Claims

1. An application of an active ingredient of Herba Euphorbiae in the preparation of a drug for treating or improving lipotoxic inflammation, characterized in that: The active ingredients of the herb are a composition formed by gallic acid, quercetin, kaempferol and umbelliferone.

2. The use according to claim 1, characterized in that: The active ingredients of the Herba Euphorbiae Herba are used in reducing the expression of ELF3, NLRP3 and the pro-inflammatory factor TNF-α to prepare a drug for treating or improving lipotoxic inflammation.

3. The use according to claim 1 or 2, characterized in that: The active ingredients of the herb are a composition formed by gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 3-8:0.5-2:0.5-2:0.5-2.

4. The use according to claim 3, characterized in that: The active ingredients of the herb are a composition of gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 4:1:1:

1.

5. Use of an active ingredient of Herba Euphorbiae Ichthyophthirius in the preparation of a drug for treating or improving ulcerative colitis induced by high fat and DSS.

6. The use according to claim 5, characterized in that: The active ingredients of the Herba Euphorbiae Herba are used in reducing the expression of inflammatory factors TNFα, TLR4, and IL-1β mRNA to prepare a drug for treating or improving lipotoxic inflammation.

7. The use according to claim 6, characterized in that: The active ingredients of the Herba Euphorbiae Herba are used in increasing the expression of ZO-1 and Connexin-43 to prepare a drug for treating or improving lipotoxic inflammation.

8. The use according to any one of claims 5 to 7, characterized in that: The active ingredients of the herb are a composition formed by gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 3-8:0.5-2:0.5-2:0.5-2.

9. The use according to claim 8, characterized in that: The active ingredients of the herb are a composition of gallic acid, quercetin, kaempferol and umbelliferone in a mass ratio of 4:1:1:1.

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