Tea saponin derivative liposome and application thereof

Abstract

The invention belongs to the technical field of cultivation, and discloses a theasaponin derivative liposome which comprises a liposome capsule material and a theasaponin derivative wrapped in the liposome capsule material. The theasaponin derivative lipidosome is coated with a theasaponin derivative, through lipidosome coating, the distribution and permeation of the theasaponin derivative in the intestinal tract of Qingyuan chicken can be effectively improved, the antibacterial and antioxidant activity functions of the theasaponin derivative can be fully exerted, and the theasaponin derivative lipidosome can at least partially replace antibiotics. Meanwhile, the invention also provides application of the liposome.

Description

Technical Field

[0001] The invention relates to the field of breeding, and in particular to a tea saponin derivative liposome and uses thereof. Background Art

[0002] Our unit cooperates with South China Agricultural University in industry-university-research cooperation to conduct research on the resource utilization of tea oil cake and the breeding of Qingyuan chickens.

[0003] Professor Ye Yong is the core researcher of this project. Through Professor Ye Yong’s previous research on camellia oleifera meal, it is hoped that camellia oleifera meal can be used as a resource in the breeding of Qingyuan chickens.

[0004] The technical problem to be solved by this scheme is: how to use tea-leaf meal or its derivatives to replace antibiotics in the breeding of Qingyuan chickens to achieve good application. Summary of the invention

[0005] The object of the present invention is to provide a tea saponin derivative liposome, which is coated with a tea saponin derivative. The liposome coating can effectively improve the distribution and penetration of the tea saponin derivative in the intestine of Qingyuan chicken, so that the antibacterial and antioxidant activity functions of the tea saponin derivative can be fully exerted, and it can at least partially replace antibiotics.

[0006] Meanwhile, the invention also provides the use of the liposome.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] A tea saponin derivative liposome, comprising a liposome capsule material and a tea saponin derivative encapsulated in the liposome capsule material;

[0009] The structure of the tea saponin derivative is shown in Formula 1 below;

[0010]

[0011] R is one of glucose, fructose, arabinose or maltose.

[0012] Tea saponin derivatives can be found in CN102030803B A tea saponin derivative and its preparation method and application, applicant: South China University of Technology. Different sugars are used as substituents in Examples 1 to 5.

[0013] This scheme demonstrates its use as a surface active.

[0014] In subsequent studies, we found that the above-mentioned derivatives have certain antibacterial effects. At the same time, when they are encapsulated in liposomes, they can be effectively dispersed in water and feed, and can be sustained-released in the intestinal wall, which can play a good antibacterial role and improve intestinal health.

[0015] In the above-mentioned liposome, the capsule material of the liposome is composed of phospholipids and cholesterol.

[0016] In the above liposomes, the phospholipids are one or more combinations of phosphatidylcholine, phosphatidylcholine and phosphatidylethanolamine.

[0017] In the above-mentioned liposomes, the mass ratio of phospholipid to cholesterol is 4:1-3.

[0018] In the above liposomes, the mass ratio of the tea saponin derivative to the capsule material is 1:200-300.

[0019] In the above-mentioned liposome, the preparation method of the liposome is:

[0020] Step 1: Add phospholipids, cholesterol and tea saponin derivatives into the oil phase and mix well;

[0021] Step 2: drying the mixture in step 1 by rotary evaporation to form a lipid film on the inner wall of the rotary evaporation container;

[0022] Step 3: adding an aqueous solution containing a surfactant into a rotary evaporation container for hydration;

[0023] Step 4: The hydration product of step 3 is homogenized to obtain a liposome-containing solution.

[0024] In the above liposomes, the surfactant is Tween 80.

[0025] Meanwhile, the present invention discloses the use of any of the above liposomes as a poultry feed additive.

[0026] At the same time, the present invention discloses the use of any of the above liposomes as a drinking water additive for poultry.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] After in-depth research on tea saponin derivatives, the present invention found that the above-mentioned derivatives have certain antibacterial effects. At the same time, when they are encapsulated in liposomes, they can be effectively dispersed in water and feed, can be slowly released on the inner wall of the intestine, and can play a good antibacterial and intestinal health improvement function. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the antibacterial effect test result diagram of Escherichia coli antibacterial circle;

[0030] Figure 2 This is a graph showing the antibacterial effect test results on Staphylococcus aureus. DETAILED DESCRIPTION

[0031] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] Source of raw materials:

[0033] Tea saponin derivatives: provided by Professor Ye Yong's research and development team, with the same structure as Example 1 of CN102030803B, a tea saponin derivative and its preparation method and application;

[0034] Lecithin, cholesterol, and Tween 80 are all commercially available products, and this embodiment does not impose too many restrictions.

[0035] Part I: In vitro antibacterial experiments

[0036] Culture medium: Beef extract peptone solid culture medium. This product is a prepared powder made of 10g peptone, 5g sodium chloride, 3g beef extract powder and 15g agar powder, with a neutral pH.

[0037] The method of use is: take a certain amount of powder, add distilled water in the ratio of powder to distilled water 33:1000, heat and boil with constant stirring until completely dissolved, and when there is no obvious granular matter in the solution, take it out and cool it.

[0038] Activate Escherichia coli and Staphylococcus aureus; streak the frozen bacteria onto the plate culture medium and culture at 37°C for 24 hours; pick a single colony and inoculate it into 100 mL of liquid culture medium, culture it at 37°C and 200 r / min in a shaking incubator overnight, and keep the bacterial solution for later use.

[0039] Inoculation: Add the prepared culture medium into the culture dish, let it stand horizontally, and inoculate 0.1 ml of bacterial solution.

[0040] Use tweezers to soak the filter paper in the sample, one sample is a camellia saponin solution, and the other sample is a liposome solution; the concentration of camellia saponin in the camellia saponin solution is 100 ppm; the liposome solution contains 25 ppm of a camellia saponin derivative (Example 1), the mass ratio of the camellia saponin derivative to the capsule material is 1:300, the capsule material is composed of phospholipids and cholesterol, and the mass ratio of phospholipids to cholesterol is 4:1-3.

[0041] The results can be found in Figure 1 and Figure 2 , Figure 1 This is the test result of the E. coli inhibition zone. Figure 2 It was the test result of Staphylococcus aureus.

[0042] It can be seen from the above experiments that the liposomes of the present invention have a strong antibacterial function.

[0043] Part II: Farming Experiment

[0044] Sample preparation

[0045] 1. Preparation of liposomes:

[0046] Step 1: Add phospholipids, cholesterol and tea saponin derivatives into the oil phase and mix well;

[0047] Step 2: drying the mixture in step 1 by rotary evaporation to form a lipid film on the inner wall of the rotary evaporation container;

[0048] Step 3: adding an aqueous solution containing a surfactant Tween 80 (with a weight ratio of 1:300 to water) into a rotary evaporation container for hydration;

[0049] Step 4: The hydration product of step 3 is homogenized to obtain a liposome-containing solution.

[0050] The above liposome preparation process is a conventional liposome hydration method in the art. The innovation of this embodiment lies in the selection and dosage of the tea saponin derivatives.

[0051] The dosage of different cases is as follows:

[0052] Table 1 Formula table Unit: g

[0053]

[0054] Performance Testing

[0055] Breed: Qingyuan Ma chicken hens, a total of 3600, divided into 6 groups, 6 replicates in each group, 100 in each replicate;

[0056] The 5 groups were divided into experimental group A, experimental group B, experimental group C, experimental group D, experimental group E and blank group;

[0057] Experimental group A used the sample of Example 1, experimental group B used the sample of Example 2, experimental group C used the sample of Example 3, experimental group D used the sample of Comparative Example 1, and experimental group D used the sample of Comparative Example 2;

[0058] Farming method: Ground farming + online farming

[0059] The breeding conditions are:

[0060] Feed the whole process until 20 weeks of age, and market them when the weight is above 1.6kg. Feed 610 small chicken feed from 1-28 days of age, 611 medium chicken feed from 29-119 days of age, and 612 large chicken feed from 120 days of age to market. The transition period for feed change is 1-3 days. During the feeding process, drinking water is controlled before immunization, and drinking water is free at other times.

[0061] The test group conducted a full test. During the Qingyuan silkie chicken breeding process, drinking water medication was performed according to the requirements of the above embodiments and comparative examples. Other drug use schemes and vaccine immunization were in accordance with the original field practice, and various different drugs were added at different times. During the test, the sample addition amount in the drinking water of each group was 20kg / 1000kg drinking water. No drugs were added to the blank group. The remaining breeding conditions were controlled according to normal breeding conditions.

[0062] Experimental Results

[0063] During the entire experiment, the growth status of the chickens was recorded, mainly recording the following performance indicators: 1. Feed-to-weight ratio; 2. Market weight; 3. Intestinal health status, including but not limited to: diarrhea, indigestion, and feed feces. By counting the status of the chickens' excrement every day, when diarrhea, indigestion, and feed feces in the excrement exceeded the normal level, it was defined as a day when an intestinal disease had obviously occurred; 4. Mortality rate.

[0064] Relevant experimental data can be found in Table 2 below;

[0065] Table 2 Experimental results statistics

[0066]

[0067] Result analysis:

[0068] 1. The data of experimental group A, experimental group B, and experimental group C show that the use of liposomes as drinking water additives can effectively increase the slaughter weight and reduce the feed-to-meat ratio; by comparing experimental group D and the blank group, it can be seen that there is no substantial difference between the two; by comparing experimental group E and the blank group, it can be seen that the use of tea saponin derivatives can increase the slaughter weight and reduce the feed-to-meat ratio to a certain extent.

[0069] 2. The data of experimental groups A, B and C show that when liposomes are used as drinking water additives, the chickens almost do not suffer from intestinal diseases, and their mortality rate is the lowest among all groups; it has obvious advantages over experimental group D.

[0070] We believe that the reason is that tea saponin derivatives themselves have certain antibacterial and anti-inflammatory abilities. At the same time, in the preliminary experiments of this program, we found that this substance also has good antioxidant ability, which can increase the activity of T-AOC and SOD enzymes. The antibacterial and anti-inflammatory abilities can inhibit the growth of harmful intestinal bacteria, improve intestinal immunity, and reduce the probability of intestinal diseases. Its antioxidant ability can improve its digestion and absorption of feed, improve the body's immunity, and ultimately manifest as a reduction in mortality.

[0071] 3. In the experimental group E of the present invention, it has a certain effect, but the effect is not significant. We believe that the possible reason is that the tea saponin derivatives are digested and decomposed in advance in the digestive tract, causing the modified groups to separate from the tea saponin, reducing the biological performance of the derivatives. At the same time, through the form of liposomes, its water solubility can be improved, and the derivatives can be directly delivered to the intestines for action.

[0072] In the experimental group D of the present invention, the effect of using camellia saponin is not obvious, which is related to the weak antibacterial ability of camellia saponin itself.

[0073] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

Claims

1. A tea saponin derivative liposome, characterized in that: It includes liposome capsule material and tea saponin derivatives encapsulated in the liposome capsule material; The structure of the tea saponin derivative is shown in Formula 1 below; R is one of glucose, fructose, arabinose or maltose.

2. The liposome according to claim 1, characterized in that The capsule material of the liposome is composed of phospholipids and cholesterol.

3. The liposome according to claim 1, characterized in that The phospholipid is one or more combinations of phosphatidylcholine, phosphatidylcholine and phosphatidylethanolamine.

4. The liposome according to claim 1, characterized in that The mass ratio of the phospholipid to cholesterol is 4:1-3.

5. The liposome according to claim 1, characterized in that The mass ratio of the tea saponin derivative to the capsule material is 1:200-300.

6. The liposome according to claim 1, characterized in that The preparation method of the liposome is: Step 1: Add phospholipids, cholesterol and tea saponin derivatives into the oil phase and mix well; Step 2: drying the mixture in step 1 by rotary evaporation to form a lipid film on the inner wall of the rotary evaporation container; Step 3: adding an aqueous solution containing a surfactant into a rotary evaporation container for hydration; Step 4: The hydration product of step 3 is homogenized to obtain a liposome-containing solution.

7. The liposome according to claim 6, characterized in that The surfactant is Tween 80.

8. Use of the liposome as claimed in any one of claims 1 to 7 as a poultry feed additive.

9. Use of the liposome as any one of claims 1 to 7 as a drinking water additive for poultry.

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