Use of safflower oil bodies in the preparation of a medicament for preventing or treating intestinal inflammatory diseases

Abstract

The present application relates to a kind of safflower oil body in the preparation of preventing or treating intestinal inflammatory disease drug application, belong to the technical field of new use of material.The present application provides the new use of safflower oil body in medicine, i.e.safflower oil body is applied in the new drug of preventing and treating colitis and other inflammatory bowel disease.The present application is committed to the experimental research of safflower oil body preventing and treating colitis and other inflammatory bowel disease, and it is proved by mouse in-vivo experiment that safflower oil body has significant prevention and treatment effect on colitis and other inflammatory bowel disease.The present application has explored new medical use for safflower oil body, and developed a new natural medicine research field.Safflower oil body of the present application can be used as drug in the prevention and treatment of colitis, with good physical and chemical stability, no toxic side effects, and good anti-inflammatory capacity and other advantages.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of new use of substances, and particularly relates to an application of safflower oil body in preparation of a medicine for preventing and treating intestinal inflammation. BACKGROUND

[0002] At present, the medicines for treating ulcerative colitis in clinical application mainly include amino salicylic acid drugs, immunosuppressants, glucocorticoid drugs, traditional Chinese medicine preparations, and microecological preparations, and although they have certain therapeutic effects on ulcerative colitis, there are still certain controversies and potential safety problems due to the unsatisfactory therapeutic effects and the existence of side effects.

[0003] The safflower oil body is a kind of organelle with a spherical structure with a diameter of 0.5-2.5 μm extracted from the seeds of the Chinese herbal medicine safflower (Carthamus tinctorius L.), which is composed of a central triacylglycerol and a phospholipid monolayer wrapped outside, and oil body proteins embedded on the surface. The core component triacylglycerol is a mixture of various unsaturated fatty acid esters, including oleic acid, linoleic acid, linolenic acid, etc. The special structure of the safflower oil body can produce resistance and electrostatic repulsion, so that it can be uniformly dispersed in the solution and has good stability.

[0004] The main uses of the safflower oil body in the prior art are as follows:

[0005] The oleosin-hEGF fusion protein with biological activity expressed by the safflower oil body protein system has better transdermal absorption effect, has the effect of promoting skin wound healing and improving healing quality, and can better help the tissue repair of the wound site (Qiang Weidong, Jilin Agricultural University, 2019: 61).

[0006] As a natural emulsifier, the safflower oil body is an effective component of the recombinant collagen polypeptide oil body skin care emulsion, which can effectively delay or inhibit the aging of the skin and restore it, achieving the effect of skin care (Yang Jing, Jilin Agricultural University, 2015: 114).

[0007] The fibroblast growth factor recombinant fusion protein expressed by the safflower oil body has the effect of promoting hair regeneration (Cai Jingbo, Chinese Journal of Chinese Medicine, 2018: 208).

[0008] Although the safflower oil body has a wide range of applications, there is no report that the safflower oil body and its main components can prevent and treat ulcerative colitis, as well as treat different types of intestinal inflammation diseases such as viral enteritis, bacterial enteritis, fungal enteritis, parasitic enteritis, tuberculous enteritis, small intestine inflammation, colitis, and rectal inflammation. SUMMARY

[0009] In view of this, the purpose of this invention is to provide an application of safflower oil in the preparation of drugs for the prevention or treatment of enteritis, safflower oil having significant preventive and therapeutic effects on inflammatory bowel diseases such as colitis.

[0010] To achieve the above objectives, the technical solution of the present invention is as follows:

[0011] Application of a safflower oil in the preparation of drugs for the prevention or treatment of enteritis.

[0012] Furthermore, the application of the safflower oil in the preparation of drugs for the prevention or treatment of ulcerative colitis.

[0013] Furthermore, the safflower oil is dissolved in a sterile single-distilled aqueous solution to obtain a mass concentration of 1.2 μg / μL.

[0014] Furthermore, the safflower oil is used in the preparation of drugs for treating viral enteritis, bacterial enteritis, fungal enteritis, parasitic enteritis, tuberculous enteritis, enteritis, colitis, or proctitis.

[0015] The beneficial effects of this invention are:

[0016] This invention provides a new medicinal application for safflower oil, specifically its use in novel drugs for the prevention and treatment of inflammatory bowel diseases such as colitis. This invention focuses on experimental research into the prevention and treatment of inflammatory bowel diseases such as colitis using safflower oil. In vivo experiments on mice have confirmed that safflower oil has significant preventive and therapeutic effects on these diseases. This invention unlocks new medical applications for safflower oil and opens up a new field of natural drug research.

[0017] The safflower oil of the present invention can be used as a medicine in the prevention and treatment of colitis. It has the advantages of good physicochemical stability, no toxic side effects, and good anti-inflammatory ability. Attached Figure Description

[0018] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0019] Figure 1 The images show the microstructure of safflower oil bodies under visible light and fluorescence conditions, where A and C are the original safflower oil bodies, and B and D are the safflower oil bodies after treatment with artificial gastrointestinal fluid.

[0020] Figure 2 A schematic diagram of colon length in each group for the prevention of colitis using safflower oil.

[0021] Figure 3 A schematic diagram of colon length in different groups of patients treated with safflower oil for colitis.

[0022] Figure 4HE staining image of safflower oil for treating colitis.

[0023] Figure 5 The expression of inflammatory factors in safflower oil for the treatment of colitis. Detailed Implementation

[0024] This invention provides a new medicinal application for safflower oil, specifically its use in novel drugs for the prevention and treatment of inflammatory bowel diseases such as colitis. This invention focuses on experimental research into the prevention and treatment of inflammatory bowel diseases such as colitis using safflower oil. In vivo experiments on mice have confirmed that safflower oil has significant preventive and therapeutic effects on these diseases. This invention unlocks new medical applications for safflower oil and opens up a new field of natural drug research.

[0025] In the prevention and treatment of ulcerative colitis, safflower oil is obtained using conventional safflower oil extraction processes (such as the extraction process disclosed in patent number: CN201710793460.8).

[0026] The liquid used in the safflower oil group in the following examples was obtained by dissolving safflower oil in sterile single-distilled water, with a mass concentration of 1.2 μg / μL.

[0027] Example 1: Safflower oil for the prevention of ulcerative colitis

[0028] Laboratory animals:

[0029] Nine female C57BL / 6J mice, weighing 18-20g, were purchased from Yisi Biotechnology Co., Ltd. and raised at the Small Animal Experiment Center of Jilin Agricultural University.

[0030] Experimental grouping and methods:

[0031] Mice were randomly divided into three groups: a control group, a DSS model group (DSS), and a safflower oil body group (OB; 0.25 mg / kg); each group consisted of three mice. The other two groups, except for the safflower oil body group, underwent a one-week environmental acclimatization period before the experiment, housed in an SPF environment with a 12-hour day / night cycle and free access to water. For seven days prior to the establishment of the enteritis model, each mouse in the safflower oil body group received 90 μL of liquid orally daily. After this period, except for the control group, the other two groups began using 2% DSS solution to replace drinking water to establish an acute enteritis model on day 1 of the experiment, switching to sterile single-distilled water on day 6. The control mice continued to drink sterile single-distilled water. In addition, from day 1 of the experiment, the control and model groups received sterile single-distilled water orally once daily; the safflower oil body group received safflower oil body (0.25 mg / kg; dissolved in sterile single-distilled water) orally once daily, with each mouse receiving 90 μL of liquid orally. The weight changes of mice were recorded at the same time every day. On the 8th day, the mice were euthanized after anesthesia. Colon tissue was taken and the colon length of the safflower oil group and the model group was measured and observed to compare with the colon length of the normal group using the same ruler. The results were used in subsequent experiments.

[0032] Statistical analysis:

[0033] Graphpad Prism 8.0 was used for analysis; one-way ANOVA and Tukey's method were used for subsequent comparisons of multiple groups; P < 0.05 was considered statistically significant.

[0034] All data from this experiment were processed using Excel software.

[0035] Figure 1 The dispersibility of safflower oil bodies under normal conditions and simulated gastrointestinal fluid indicates that safflower oil bodies can exist stably under gastrointestinal fluid. Table 1 shows the Disease Activity Index (DAI) scores of mice 7 days after ulcerative colitis prevention.

[0036] The DAI score includes changes in mouse weight, rectal bleeding, and shortening of colon length. Figure 2 No weight change, no rectal bleeding, normal stool, and no colon shortening: DAI score is 0. Weight loss of 1%-5% with loose stool and 2%-10% colon shortening: score is 1. Weight loss of 5%-10% with sticky stool and 10%-20% colon shortening: score is 2. Weight loss of 10%-20% or more with visible rectal bleeding and significant colon shortening: score is 3 or 4.

[0037] As shown in Table 1, safflower oil has a good preventive effect, indicating that safflower oil can effectively prevent ulcerative colitis.

[0038] Table 1. Preventive effects of safflower oil on ulcerative colitis.

[0039] Group Control DSS OB Bleeding condition Normal Macroscopic bleeding Mucous Mouse weight (g) 21±5 17.8±0.7 20.4±2.2 Colon length (cm) 9.1±0.2 6.2±0.44 7.9±0.21 DAI score 0 4 1

[0040] Example 2: Safflower oil used to treat ulcerative colitis

[0041] Laboratory animals:

[0042] Twelve female C57BL / 6J mice, weighing 18-20g, were purchased from Yisi Biotechnology Co., Ltd. and raised at the Small Animal Experiment Center of Jilin Agricultural University.

[0043] Experimental grouping and methods:

[0044] All mice underwent a one-week environmental acclimatization period before the experiment, housed in an SPF environment with a 12-hour day / night cycle and free access to water. One day prior to the experiment, mice were randomly assigned to four groups: a control group, a DSS model group (DSS), a methotrexate-methylxyltransferase (MTX) group (DSS + MTX; 5 mg / kg), and a safflower oil body group (DSS + OB; 1 mg / kg); with three mice in each group. Except for the control group, all other groups received a 2% DSS solution as a substitute for drinking water to induce acute enteritis starting on day 1. This was replaced with sterile single-distilled water on day 6, while the control mice continued to drink sterile single-distilled water. In addition, starting on day 1, the control and model groups received sterile single-distilled water orally once daily; the MTX group received MTX solution (5 mg / kg; dissolved in sterile single-distilled water) orally once daily; and the safflower oil body group received safflower oil body (1 mg / kg; dissolved in sterile single-distilled water) orally once daily. Each mouse received 90 μL of liquid orally each time. Mouse weight changes were recorded at the same time every day. On day 8, mice were euthanized after anesthesia. Colon tissue was harvested, and the colon lengths of the drug-treated group and model group were measured and compared with those of the normal group using the same ruler. These measurements were then used in subsequent experiments.

[0045] Detection of intestinal IL-1β, IL-6, and TNF-α inflammatory factors:

[0046] Mouse colon tissue was collected in centrifuge tubes and allowed to stand at 0-4℃ for 1-2 hours to coagulate and separate into layers. The tissue was centrifuged at 3000 rpm for 5 min, and the supernatant was transferred to a new centrifuge tube. It was then centrifuged again at 12000 rpm at 4℃ for 10 min, and the supernatant was collected again. The tissue was then flash-frozen in liquid nitrogen for 15 min and stored at -80℃. The levels of intestinal IL-1β, IL-6, and TNF-α inflammatory factors were detected using an ELISA kit, strictly following the manufacturer's instructions.

[0047] HE staining: HE staining is one of the most commonly used staining methods for tissue sections. (1) Tissue is embedded and dehydrated with ethanol, (2) cleared, (3) impregnated with paraffin, (4) the paraffin surrounding the embedded tissue is kept at an appropriate level so that the sections can be stained. The paraffin sections are stained with hematoxylin for 0.5-1 min, rinsed with tap water, differentiated with 1% hydrochloric acid alcohol for a few seconds, rinsed with tap water, then blued with 1% ammonia water solution for 1 min, rinsed with running water for a few seconds, and then stained with eosin staining solution for a few seconds, and rinsed with running water. The cell nucleus is blue and the cytoplasm is red, which means HE staining is performed.

[0048] Statistical analysis:

[0049] Graphpad Prism 8.0 was used for analysis; one-way ANOVA and Tukey's method were used for subsequent comparisons of multiple groups; P < 0.05 was considered statistically significant.

[0050] Table 2 shows the histological scores of mice 7 days after treatment for ulcerative colitis, including mouse weight, colon length, and DAI score (including changes in mouse weight, fecal bleeding, and shortening of colon length). Figure 3 No weight change, no rectal bleeding, normal stool, and no colon shortening: DAI score is 0. Weight loss of 1%-5% with loose stool and 2%-10% colon shortening: score is 1. Weight loss of 5%-10% with sticky stool and 10%-20% colon shortening: score is 2. Weight loss of 10%-20% or more with visible rectal bleeding and significant colon shortening: score is 3 or 4.

[0051] As shown in Table 2, safflower oil has a good therapeutic effect, indicating that safflower oil can effectively treat ulcerative colitis.

[0052] Table 2. Therapeutic effects of safflower oil on ulcerative colitis.

[0053]

[0054]

[0055] This invention's experimental research found that safflower oil can effectively prevent and treat ulcerative colitis. Specifically, a 2% DSS colitis model was established, with a normal group, a 2% DSS group, and a safflower oil group (0.25 mg / kg). The safflower oil group was given 0.25 mg / kg of safflower oil (dissolved in sterile single-distilled water) orally once daily during the 7-day normal feeding period before modeling and after modeling. Each mouse was given 90 μL of the liquid orally each time. On the 8th day, the mice were sacrificed after anesthesia. Clinical scores were obtained: normal group 0 points, DSS group 4 points, safflower oil group 1 point; mouse weight: normal group 21±5 g, DSS group 17.8±0.7 g, safflower oil group 20.4±2.2 g; colon length (cm): normal group 9.1±0.2 cm, DSS group 6.2±0.44 cm, safflower oil group 7.9±0.21 cm. This effectively prevented colitis. In the treatment experiment, to simultaneously compare the efficacy of the drugs, three mice were included in each group: a normal control group, a DSS group, a safflower oil group (1 mg / kg), and a methotrexate (MTX) group (5 mg / kg). Except for the normal control group, all other groups of mice underwent acute enteritis modeling by replacing their drinking water with a 2% DSS solution starting on day 1 of the experiment. The MTX group received oral MTX solution (5 mg / kg; dissolved in sterile single-distilled water) once daily; the safflower oil group received oral safflower oil (1 mg / kg; dissolved in sterile single-distilled water) once daily. Each mouse received 90 μL of the liquid orally each time. Clinical scores were obtained as follows: normal group 0 points, DSS group 4 points, safflower oil group 1 point, MTX group 2 points; mouse weight: normal group 22±3g, DSS group 18.5±1.7g, safflower oil group 216±2g, MTX group 20±2.1g; colon length (cm): normal group 9.8±0.8cm, DSS group 6.3±0.11cm, safflower oil group 8.3±0.15cm, MTX group 6.9±0.3cm; and Figure 5 and Figure 4 The verification of inflammatory factors and the results of HE section analysis both indicate that safflower oil can effectively prevent and treat ulcerative colitis.

[0056] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. Use of safflower oil body in the preparation of a drug for preventing or treating intestinal inflammatory disease, wherein the intestinal inflammatory disease is ulcerative colitis.

2. Use according to claim 1, characterized in that, The mass concentration of the safflower oil body is 1.2 μg / μL.

3. Use according to claim 1, characterized in that, The safflower oil body is dissolved in sterilized single distilled water to obtain a mass concentration of 1.2 μg / μL. The safflower oil body is dissolved in sterilized single distilled water to obtain a mass concentration of 1.2 μg / μL.

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