An OPO-structured lipid for preventing or improving intestinal epithelial barrier injury and its application

Abstract

The present invention relates to the use of OPO structured lipids in the preparation of a drug for preventing or improving intestinal epithelial barrier damage. The present invention discovers that OPO structured lipids can improve colonic epithelial barrier damage by promoting the proliferation of intestinal stem progenitor cells and the secretion of goblet cell mucin, increasing the protein expression of tight junction proteins ZO-1, Occludin, and Claudin-1, and inhibiting apoptosis of colonic epithelial cells.

Description

Technical Field

[0001] The present invention relates to the field of drugs, and particularly relates to the use of OPO structured lipids in preventing or improving intestinal epithelial barrier damage. Background Art

[0002] At present, the intestinal health status of the people is not optimistic. According to incomplete statistics, 90% of people in our country have varying degrees of intestinal health problems. As the "gatekeeper" of intestinal health, the intestinal epithelial barrier is an important barrier for maintaining the homeostasis of the internal environment of the body. Once the barrier function is damaged, toxins, harmful bacteria and harmful substances in the intestine will enter the blood circulation system, triggering various diseases such as autoimmune diseases and even the central nervous system. Therefore, improving the function of the intestinal epithelial barrier is crucial for promoting intestinal health and the overall health level.

[0003] The structure of triglycerides has a significant impact on intestinal health. Sn-2 palmitate, as a functional structured lipid, is rich in breast milk fat and animal fats such as lard. Research shows that milk fat with high sn-2 palmitate can increase beneficial intestinal bacteria, relieve colonic pathological damage, and improve intestinal inflammation. Sn-2 palmitate exists in various forms, among which 1,3-dioleoyl-2-palmitoyl glycerol (OPO structured lipid) is the main type, and it has physiological functions such as promoting the absorption of fatty acids and calcium in infants, promoting bone development, improving the intestinal flora, and improving constipation. At present, there are no relevant reports on the use of OPO structured lipids in preventing or improving intestinal epithelial barrier damage. Summary of the Invention

[0004] To solve the above technical problems, the present invention includes the following aspects:

[0005] The first aspect of the present invention provides the application of OPO structured lipids in the preparation of drugs for preventing or improving intestinal epithelial barrier damage.

[0006] Preferably, the intestinal epithelial barrier damage is colonic epithelial barrier damage.

[0007] The second aspect of the present invention provides the application of OPO structured lipids in the preparation of health products for preventing or improving intestinal epithelial barrier damage.

[0008] Preferably, the intestinal epithelial barrier damage is colonic epithelial barrier damage.

[0009] The third aspect of the present invention provides the application of OPO structured lipids in the preparation of functional foods for preventing or improving intestinal epithelial barrier damage.

[0010] Preferably, the intestinal epithelial barrier damage is colonic epithelial barrier damage.

[0011] Preferably, the functional food is dairy products.

[0012] More preferably, the dairy product is milk powder.

[0013] Even more preferably, the milk powder is infant milk powder.

[0014] The fourth aspect of the present invention provides the use of OPO structured lipid in the preparation of a drug, a health product or a functional food having the following effects, and the effects are selected from one or more of (1) promoting the proliferation of intestinal stem progenitor cells and the secretion of goblet cell mucin, (2) increasing the expression of tight junction proteins, and (3) inhibiting the apoptosis of colonic epithelial cells.

[0015] Preferably, the present invention provides the use of OPO structured lipid in the preparation of a drug, a health product or a functional food having the following effects simultaneously, and the effects are (1) promoting the proliferation of intestinal stem progenitor cells and the secretion of goblet cell mucin, (2) increasing the expression of tight junction proteins, and (3) inhibiting the apoptosis of colonic epithelial cells.

[0016] Preferably, the tight junction proteins are selected from one or more of ZO-1 protein, Occludin protein and Claudin-1 protein.

[0017] Preferably, the increase in the expression of tight junction proteins is to increase the expression of tight junction proteins ZO-1, Occludin and Claudin-1 simultaneously.

[0018] Preferably, the functional food is a dairy product.

[0019] More preferably, the dairy product is milk powder.

[0020] Even more preferably, the milk powder is infant milk powder.

[0021] Technical effects produced by the present invention:

[0022] The present invention unexpectedly discovers that OPO structured lipid has the effect of improving colonic epithelial barrier injury, and can improve colonic epithelial barrier injury by promoting the proliferation of intestinal stem progenitor cells and the secretion of goblet cell mucin, increasing the expression of tight junction proteins ZO-1, Occludin and Claudin-1 proteins, and inhibiting the apoptosis of colonic epithelial cells, and can exert a protective effect even at a low dose of 400 mg / kg concentration. Description of the Drawings

[0023] Figure 1 is the design diagram of the animal experiment dosing scheme in Test Example 1;

[0024] Figure 2 is the test result of the effect of OPO structured lipid on alleviating colonic epithelial barrier injury symptoms in DSS-induced mice, wherein Figure 2A is the percentage change in body weight, Figure 2 B is the colon length, Figure 2 C is the disease activity index, Figure 2 D is the H&E staining of colon tissue;

[0025] Figure 3 are the test results of OPO-structured lipid in improving intestinal epithelial barrier injury, where Figure 3 A is the result of OPO-structured lipid promoting the proliferation of intestinal stem progenitor cells; Figure 3 B is the result of OPO-structured lipid promoting the expression of goblet cell MUC2; Figure 3 C is the result of OPO-structured lipid inhibiting the apoptosis of colon epithelial cells;

[0026] Figure 4 are the test results of OPO-structured lipid in increasing the expression of colon tight junction proteins, where Figure 4 A is the result of the expression level of ZO-1 protein, Figure 4 B is the result of the expression level of Occludin protein, Figure 4 C is the result of the expression level of Claudin-1 protein. Detailed implementation manners

[0027] Test Example 1: Effect of OPO-structured lipid on improving intestinal epithelial barrier injury in model mice

[0028] 1. Test method

[0029] The animal experiment design diagram is as shown in Figure 1 7-week-old male C57BL / 6J mice were acclimated in a SPF barrier facility for one week, given free access to water and sufficient diet. The mice were randomly divided into 4 groups, with 10 mice in each group. The control group and the model group were respectively gavaged with 0.5% sodium carboxymethylcellulose solution (0.5% CMC-Na), the low-dose OPO group was gavaged with 400 mg / kg, and the high-dose OPO group was gavaged with 800 mg / kg, which continued until the end of the experiment. Seven days after gavage of the mice, the control group was fed with drinking water without dextran sulfate sodium (DSS, molecular weight: 36000 - 50000 Da); the model group and the OPO treatment group were fed with drinking water containing 3% DSS to induce an intestinal barrier injury model in mice. On the 5th day of modeling, the 3% DSS drinking water was removed and replaced with normal drinking water for 3 days, and then the mice were sacrificed by cervical dislocation. During the modeling process, the body weight change and fecal occult blood of the mice were measured every day.

[0030] The specific measurement indexes and methods are as follows:

[0031] (1) Body weight change: The body weight of the mice was weighed using an electronic balance, and the percentage change in body weight was calculated.

[0032] (2) Fecal occult blood: Measured using a fecal occult blood test kit.

[0033] (3) Disease activity index: It is evaluated and scored from three aspects: percentage of weight loss, fecal viscosity, and fecal occult blood.

[0034] (4) Colon length: The mouse colon segment is taken out and its length is measured with a ruler.

[0035] (5) Histopathological score: The mouse colon tissue is stained with H&E, and the morphology of the colon tissue is observed under a microscope. The damage degrees of epithelial cell morphology, crypt structure, mucosal thickness, etc. are scored.

[0036] (6) Tight junction protein expression: The protein expression levels of ZO-1, Occludin, and Claudin-1 are determined by immunohistochemical staining.

[0037] (7) Intestinal stem progenitor cell proliferation: The protein expression level of Ki67 is determined by immunohistochemical staining.

[0038] (8) Goblet cell differentiation: The protein expression level of MUC2 is determined by immunohistochemical staining.

[0039] (9) Intestinal epithelial cell apoptosis: TUNEL staining method.

[0040] 2. Test results

[0041] As Figure 2 shown, in the mice with a DSS-induced colon barrier injury model, their body weights decreased significantly, the colon lengths shortened significantly, and the disease activity index increased significantly. H&E staining showed that in the DSS model group, the mice had colon pathologies such as crypt loss and tissue destruction in the colon. Both the high-dose OPO group (HOPO: 800 mg / kg) and the low-dose OPO group (LOPO: 400 mg / kg) could delay the weight loss of the mice with a colon barrier injury model, reduce the disease activity index, increase the colon length, and improve the pathological damage of the colon tissue.

[0042] The integrity of the intestinal epithelial barrier depends on the proliferation, differentiation of intestinal stem progenitor cells, and the expression of tight junction proteins. As Figure 3 and 4As shown, in the mice treated with DSS, the proliferation (Ki67 protein expression) and differentiation (MUC2 protein expression) levels of intestinal stem progenitor cells were significantly decreased, the apoptosis level was significantly increased, and the expression of tight junction proteins was significantly decreased. While OPO could promote the proliferation of intestinal stem progenitor cells and the secretion of mucin by goblet cells, inhibit the apoptosis of colon epithelial cells, increase the expression of tight junction proteins ZO-1, Occludin and Claudin-1 proteins, and improve the damage of the colon epithelial barrier. The above experimental results indicate that OPO structured lipids have the function of maintaining the integrity of the intestinal epithelial barrier, have a significant effect on preventing or improving the damage of the intestinal epithelial barrier, indicating that OPO structured lipids can play a role as an active ingredient in drugs, health products or functional foods in the field of preventing or improving the damage of the intestinal epithelial barrier.

[0043] Although specific embodiments of the invention have been described, those skilled in the art should recognize that various changes and modifications can be made to the invention without departing from the scope or spirit of the invention. Thus, the invention is intended to cover all such changes and modifications that fall within the scope of the appended claims and their equivalents.

Claims

1. The use of OPO structured lipid as the sole active ingredient in the preparation of a drug for preventing or improving intestinal epithelial barrier damage, characterized in that: The intestinal epithelial barrier damage is colonic epithelial barrier damage.

2. Use of OPO structured lipid as the sole active ingredient in the preparation of a health product for preventing or improving intestinal epithelial barrier damage, characterized in that: The intestinal epithelial barrier damage is colonic epithelial barrier damage.

3. Use of OPO structured lipid as the sole active ingredient in the preparation of functional food for preventing or improving intestinal epithelial barrier damage, characterized in that: The intestinal epithelial barrier damage is colonic epithelial barrier damage.

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