CN115040492A - Metformin-loaded milk exosome nano preparation, preparation method and application - Google Patents
Metformin-loaded milk exosome nano preparation, preparation method and application Download PDFInfo
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Abstract
The invention discloses a metformin-loaded milk exosome nano preparation, wherein a milk exosome is used as a carrier to load metformin, and the preparation method of the metformin-loaded milk exosome nano preparation comprises the following steps: (1) extracting fresh cow milk to obtain milk exosome; (2) and ultrasonically mixing the milk exosome with metformin in an ice bath, and performing refrigerated centrifugation to prepare the metformin-loaded milk exosome nano preparation. The invention utilizes the milk exosome as the drug delivery carrier of the metformin, can potentially enhance the distribution of the metformin drug in the liver and brain, and improve the efficacy of the drug, thereby improving the treatment effect of the metformin in alcoholic liver and brain injury.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a milk exosome nano preparation loaded with metformin, a preparation method and application thereof.
Background
Excessive drinking and drinking can cause multiple organ and system damages to the human body, of which liver and brain damage is the most serious. The alcoholic liver injury is chronic liver disease caused by long-term heavy drinking, is usually manifested as fatty liver at the initial stage, can further develop into alcoholic hepatitis, alcoholic hepatic fibrosis and alcoholic cirrhosis, and can induce extensive hepatocyte necrosis and even liver failure when being seriously drunk. Alcohol, as a lipophilic central nervous system inhibitor, also has strong affinity with the central nervous system, and acute excessive intake can cause irreversible permanent damage to the central nervous system. Alcohol and its metabolite acetaldehyde can pass through blood brain barrier, combine with brain protein to cause nerve cell injury, directly influence central nervous system and brain normal function. Alcohol exposure also activates the expression of inflammatory factors, affecting the activation of glial cells in the nervous system, leading to aberrant signaling associated with neuroinflammation and causing cell damage. At present, no effective medicine for treating liver and brain injury caused by alcoholism exists, so that the research of effective treatment means is a hot spot of the current research.
Metformin is a biguanide compound extracted from goat beans (Galega officinalis L.), and a large number of studies show that metformin has anti-inflammatory, anti-tumor and anti-apoptosis effects, in addition to being used as a first-line therapeutic agent for type 2 diabetes. For example, metformin protects the liver by anti-oxidative and anti-inflammatory effects in liver ischemia models, reduces lipopolysaccharide-induced hepatitis by inhibiting the levels of inflammatory factors, and also improves cognitive function in patients with alzheimer's and parkinson's disease.
Exosomes are nano-sized extracellular vesicles (30-150 nm in diameter) secreted by cells as a result of environmental stimuli or self-activation. Exosomes are extremely important medium systems among cells, are rich in proteins, lipids, complex polysaccharides, nucleic acids, cell adhesion molecules and the like, and can realize material exchange and information exchange among cells. In recent years, research on drug carrier delivery systems in which exosomes are loaded with small-molecule chemical drugs, proteins, peptides, nucleic acid drugs, and the like has been attracting attention. However, the collection efficiency of the exosomes from the cells is low, the cost is high, and certain toxicity risks exist. The milk is used as a natural beverage for supplementing nutrition in daily life, contains a large amount of exosomes, and a double-layer membrane of the milk exosomes can be completely maintained in vitro for a long time, so that the milk is a potential excellent drug carrier.
Chinese patent document with publication number CN112791092A discloses a milk exosome-loaded icariin nano-preparation, and the preparation method comprises: firstly, preparing a milk exosome suspension by taking fresh cow milk as a raw material, and then mixing the raw material and the milk exosome suspension at room temperature in a volume ratio of 1: 9 mixing icariin with the exosome suspension; the exosome load icariin nanometer preparation prepared by freeze drying. The exosome-loaded icariin nano preparation has the capability of enhancing osteogenic repair.
Chinese patent publication No. CN112656836A discloses the use of a transdermal peptide-modified radix Puerariae exosome nano-preparation in the preparation of anti-skin aging products, the invention firstly prepares a radix Puerariae exosome suspension, and then mixes the suspension with palmitic acid-modified transdermal peptide to prepare the transdermal peptide-modified radix Puerariae exosome nano-preparation, which can effectively penetrate the stratum corneum of the skin and has a significant anti-skin (cell) aging function.
Disclosure of Invention
The invention provides a metformin-loaded milk exosome nano preparation, wherein the milk exosome is used as a carrier to load metformin, the brain targeting property of the milk exosome can improve the curative effect of the metformin on treating alcoholic liver and brain injury, and the metformin-loaded milk exosome nano preparation is low in cost, easy to obtain, and high in biocompatibility and safety.
The technical scheme is as follows:
a milk exosome nano preparation loaded with metformin comprises a milk exosome and metformin, wherein the milk exosome is used as a carrier to load the metformin, and the mass ratio of the milk exosome to the metformin is 1: 1-3.
The milk exosome has stable property, good biocompatibility, low cost, small toxicity and brain targeting property, and can potentially enhance the distribution of the metformin medicament in the liver and brain and improve the medicament effect by using the milk exosome as the medicament delivery carrier of the metformin so as to improve the treatment effect of the metformin in the liver and brain injury.
The invention also provides a preparation method of the metformin-loaded milk exosome nano preparation, which comprises the following steps:
(1) extracting fresh milk to obtain milk exosome;
(2) and ultrasonically mixing the milk exosome with metformin in an ice bath, and freezing and centrifuging to prepare the milk exosome nano preparation loaded with metformin.
Preferably, the preparation method of the milk exosome comprises the following steps:
(1) centrifuging fresh cow milk at 0-10 deg.C to obtain middle layer whey under the centrifugation condition of 3000g at 2000-20 min;
(2) mixing the middle layer whey with sodium citrate solution, centrifuging at 0-10 deg.C, and collecting supernatant to obtain defatted whey; the centrifugation condition is 10000-12000g for 60-90 min;
(3) at the temperature of 0-10 ℃, after centrifugation of 35000-40000g of defatted whey for 60-90min, the supernatant is subsequently centrifuged at 70000-80000g for 60-90min, and then the precipitate is obtained after centrifugation of 100000-135000g for 120-180min to obtain the milk exosome.
Further preferably, the preparation method of the milk exosome comprises the following steps:
(1) centrifuging fresh cow milk at 4 deg.C to obtain middle layer whey, wherein the centrifugation condition is 3000g for 15 min;
(2) uniformly mixing the middle layer whey with 2 wt% sodium citrate solution with the same volume, centrifuging at 4 ℃, and taking supernatant to obtain defatted whey; centrifuging at 12000g for 60 min;
(3) centrifuging 35000g of defatted whey for 60min at 4 deg.C, collecting supernatant, centrifuging 70000g for 60min, and collecting supernatant; centrifuging at 100000g for 60min and at 135000g for 120min, and collecting precipitate to obtain milk exosome precipitate.
Preferably, the average particle size of the milk exosomes is 100-200 nm. The milk exosome prepared by the method has a proper particle size range, and is suitable for loading metformin to prepare a nasally-administered nano preparation.
In the step (2), the milk exosome and the metformin are mixed in a PBS buffer solution according to the mass ratio of 1: ultrasonic mixing in ice bath 1-3.
Preferably, in the step (2), the milk exosome and the metformin are mixed in the PBS buffer solution according to the mass ratio of 1: 1, mixing by ultrasonic treatment for 5-10min under the power of 90-110W in ice bath.
Preferably, in step (2), the centrifugation is performed at 0-10 ℃ for 100000-135000g and 60-180 min.
The invention also provides application of the metformin-loaded milk exosome nano preparation in treating liver oxidative damage and/or brain oxidative damage, in particular application in treating alcoholic liver and brain damage.
The administration mode of the metformin-loaded milk exosome nano preparation is nasal delivery. Nasal delivery may enhance the bioavailability of the metformin-loaded milk exosome nano-formulation.
In an alcoholic hepatic and cerebral injury animal model, the milk exosome nano preparation loaded with the metformin is delivered through the nose to act on the animal model, the hepatic index, the glutamic-pyruvic transaminase (ALT), the glutamic-oxalacetic transaminase (AST) and the Triglyceride (TG) level in serum are verified to obtain the milk exosome nano preparation loaded with the metformin, the damage degree of hepatic cells can be reduced, and meanwhile, the total superoxide dismutase (SOD) and Malondialdehyde (MDA) level in hepatic and cerebral tissues are verified to obtain the milk exosome nano preparation loaded with the metformin, which has a protection effect on oxidative damage.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention utilizes the milk exosome as the drug delivery carrier of the metformin, can potentially enhance the distribution of the metformin drug in the liver and brain, and improve the efficacy of the drug, thereby improving the treatment effect of the metformin in alcoholic liver and brain injuries.
(2) The metformin-loaded milk exosome nano preparation prepared by the method has good biocompatibility, low cost, low toxicity and high yield, can be produced in a large scale, can be used for treating liver oxidative damage and/or brain oxidative damage, and has wide application prospect in the aspect of treating alcoholic liver and brain damage.
Drawings
Fig. 1 is a transmission electron microscope image of the metformin-loaded milk exosome nano-preparation prepared in example 2.
Fig. 2 shows the therapeutic results of the metformin-loaded milk exosome nano-preparation prepared in example 2 on a liver-brain injury model after nasal administration, wherein a is a healthy rat control group, B is an alcoholic liver-brain injury rat model group, and C is a metformin-milk exosome treatment group.
Detailed Description
The invention is further elucidated with reference to the figures and the examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
In the embodiment of the invention, the fresh cow milk is from the Ministry laboratory pasture of the milk science research institute of Zhejiang university, and the metformin is from Shanghai Michelin Biotechnology, Inc.
Example 1
Extraction of milk exosomes
(1) Centrifuging fresh cow milk at 4 deg.C for 15min at 3000g, and collecting middle layer whey;
(2) mixing the middle layer whey with 2 wt% sodium citrate solution of the same volume, centrifuging at 4 deg.C for 60min at 12000g, carefully taking supernatant, and discarding precipitate to obtain defatted whey;
(3) centrifuging about 200mL of defatted whey 35000g at 4 deg.C for 60min, and collecting the supernatant;
(4) centrifuging the supernatant obtained in the step (3) at 4 ℃ at 70000g for 60min, and taking the supernatant again;
(5) centrifuging the supernatant of the step (4) at 4 ℃ at 100000g for 60min, increasing the speed, continuing to centrifuge at 135000g for 120min, discarding the supernatant, and taking the precipitate to obtain the milk exosome.
The size, structure and morphology of the milk exosome prepared in this example were observed by transmission electron microscope, and the result showed that the average particle size of the milk exosome prepared in this example was 100-200nm, and the milk exosome had a typical cup-shaped structure of exosome.
Example 2
Synthesis of metformin-loaded milk exosome nano preparation
In a PBS buffer, in a mass ratio of 1: 1, mixing the milk exosome prepared in the example 1 and metformin, performing ultrasonic treatment on the mixture for 5min by using a cell disruptor probe under the ice bath at the power of 105W, centrifuging the mixture for 60min at 100000g under the ice bath at the temperature of 4 ℃, removing supernatant, and re-dissolving the precipitate by using 1mL of PBS buffer solution to obtain the milk exosome nano preparation loaded with the metformin.
The shape, size and structure of the metformin-loaded milk exosome nano preparation prepared in the embodiment are observed and analyzed by a transmission electron microscope, and as a result, as shown in fig. 1, a plurality of cup-shaped vesicles with the particle size of about 100nm can be seen in a visual field, the boundary is clear, the metformin-loaded milk exosome nano preparation has a stereoscopic impression, and the structure is relatively complete.
Example 3
In a PBS buffer, in a mass ratio of 1: 3, mixing the milk exosome prepared in the example 1 with the metformin, performing ultrasonic treatment on the mixture for 5min by using a cell disruptor probe under ice bath at the power of 110W, then performing centrifugal treatment on the mixture for 100min at the temperature of 4 ℃ under ice bath at the power of 120000g, removing supernatant, and re-dissolving and precipitating the mixture by using 1mL of PBS buffer solution to obtain the milk exosome nano preparation loaded with the metformin.
Application example 1
The metformin-loaded milk exosome nano preparation prepared in example 2 is nasally delivered to a rat model for alcoholic liver and brain injury, blood is taken after the last administration, the rat model is kept still for 30min, then is centrifuged at 3000rpm and 4 ℃ for 10min, serum on the upper layer is absorbed and placed in a centrifuge tube, and the contents of alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and Triglyceride (TG) in the serum are measured. Taking isolated liver and brain tissues, weighing, homogenizing the tissues, centrifuging for 30min at 4 ℃ and 12000rpm, taking supernatant, and determining the content of total superoxide dismutase (SOD) and Malondialdehyde (MDA) in the homogenate according to the instruction of the kit. The damage degree of the liver cells is examined according to the liver index and ALT, AST and TG levels in serum, and the protective effect of the liver cells on oxidative damage is examined according to SOD and MDA levels in liver and brain tissue homogenate. The result proves that the metformin-loaded milk exosome nano preparation can reduce the damage degree of liver cells and has a protection effect on oxidative damage of liver and brain tissues.
Meanwhile, the therapeutic result of the metformin-loaded milk exosome nano preparation on a liver and brain injury model is investigated by HE (human immunodeficiency virus) dyeing, the result is shown in figure 2, and the results in the figure show that the liver tissues of a healthy rat control group are arranged regularly, the streak structure is full and clear, and the structure of liver cells is clear. No significant dilation of the hepatic sinus was seen (a in fig. 2, right side is left enlarged view); in contrast, the alcoholic hepatic brain injury rat model group had disordered liver tissue arrangement, a cord structure was shrunk, fat droplets with different sizes were observed in hepatocytes, and most of the hepatocytes showed edema (fig. 2, B, right side is an enlarged view of the left side); the liver tissues of rats in the metformin milk exosome treatment group are arranged neatly, and the liver cell edema alleviating and fat drop vacuolation phenomena are reduced compared with those of a model group (C in figure 2, the right side is an enlarged view on the left side), so that the metformin-loaded milk exosome nano preparation prepared by the invention has an obvious treatment effect on alcoholic hepatic and cerebral injuries.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. The metformin-loaded milk exosome nano preparation is characterized by comprising a milk exosome and metformin, wherein the milk exosome is used as a carrier to load the metformin, and the mass ratio of the milk exosome to the metformin is 1: 1-3.
2. The method for preparing the metformin-loaded milk exosome nano-formulation according to claim 1, comprising the steps of:
(1) extracting fresh milk to obtain milk exosome;
(2) and ultrasonically mixing the milk exosome with metformin in an ice bath, and freezing and centrifuging to prepare the milk exosome nano preparation loaded with metformin.
3. The preparation method of the metformin-loaded milk exosome nano-formulation according to claim 2, wherein the preparation method of the milk exosome comprises the following steps:
(i) centrifuging fresh cow milk at 0-10 deg.C to obtain middle layer whey under the centrifugation condition of 3000g at 2000-20 min;
(ii) mixing the middle layer whey with sodium citrate solution, centrifuging at 0-10 deg.C, and collecting supernatant to obtain defatted whey; the centrifugation condition is 10000-12000g, 60-90 min;
(iii) at the temperature of 0-10 ℃, after centrifugation of 35000-40000g of defatted whey for 60-90min, the supernatant is subsequently centrifuged at 70000-80000g for 60-90min, and then the precipitate is obtained after centrifugation of 100000-135000g for 120-180min to obtain the milk exosome.
4. The method for preparing the metformin-loaded milk exosome nano-formulation according to claim 2, wherein the average particle size of the milk exosomes is 100-200 nm.
5. The preparation method of the metformin-loaded milk exosome nano-formulation according to claim 2, wherein in the step (2), the milk exosome and metformin are mixed in a PBS buffer solution in a mass ratio of 1: ultrasonic mixing in ice bath 1-3.
6. The preparation method of the metformin-loaded milk exosome nano-formulation according to claim 5, wherein in the step (2), the milk exosome and metformin are mixed in PBS buffer solution in a mass ratio of 1: 1, ultrasonically mixing for 5-10min under the power of 90-110W in ice bath.
7. The method for preparing the metformin-loaded milk exosome nano-formulation according to claim 2, wherein in the step (2), 100000-135000g is frozen and centrifuged at 0-10 ℃ for 60-180 min.
8. Use of the metformin-loaded milk exosome nano-formulation according to claim 1 in treating oxidative damage to the liver and/or oxidative damage to the brain.
9. The use of the metformin-loaded milk exosome nano-formulation according to claim 8 in the treatment of alcoholic liver and brain injury.
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| Title |
|---|
| 周东方等: "二甲双胍在大鼠酒精性肝损伤中的保护作用及其机制", 《中华肝脏病杂志》 * |
| 周东方等: "二甲双胍在大鼠酒精性肝损伤中的保护作用及其机制", 《中华肝脏病杂志》, 31 December 2012 (2012-12-31) * |
| 蒋怡冰: "血浆外泌体作为药物载体的制备及其对脑缺血再灌注损伤保护机制研究", 《中国优秀硕士学位论文全文数据库》 * |
| 蒋怡冰: "血浆外泌体作为药物载体的制备及其对脑缺血再灌注损伤保护机制研究", 《中国优秀硕士学位论文全文数据库》, 15 January 2022 (2022-01-15), pages 68 * |
| 郝海宁等: "2种不同的牛乳外泌体提取方法的比较研究", 《食品安全质量检测学报》 * |
| 郝海宁等: "2种不同的牛乳外泌体提取方法的比较研究", 《食品安全质量检测学报》, vol. 11, no. 8, 30 April 2020 (2020-04-30), pages 2569 - 2574 * |
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