32 results about "Cerebral microvessel" patented technology

In one embodiment, the present invention is a method of creating a fully-human blood-brain barrier (BBB) model, comprising the steps of (a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs), wherein the neural cells and BMECs that comprise the mixture were produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs of step (a); and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of pericytes, astrocytes and differentiated neural progenitor cells (NPCs), wherein a blood brain barrier model is created.

The invention discloses an application of 3,5,4'-trihydroxy-stilbyl-3'-O-glucoside in the preparation of microcirculatory disturbance improvement medicines. The microcirculatory disturbance improvement medicines comprise 3,5,4'-trihydroxy-stilbyl-3'-O-glucoside as an effective active component, and one or more pharmaceutically acceptable auxiliary agents, especially gastrodin and / or water-soluble cyclodextrin derivatives and PEG400 and mannitol, wherein the structural formula of 3,5,4'-trihydroxy-stilbyl-3'-O-glucoside is shown in the specification. The microcirculatory disturbance improvement medicines treat 3,5,4'-trihydroxy-stilbyl-3'-O-glucoside as the effective active component, and are used for improving disturbances caused by cerebral microvascular damages and / or disturbances caused by ischemia reperfusion damages and / or disturbances caused by leukocyte adhesion damages, and / or disturbances caused by peroxides. Normal mice of Kunming species and artificially induced microcirculatory disturbance model mice undergo tail vein injection treatment, and results of the observation of arteriole and venule systole and capillary network under a microcirculatory microscope show that 3,5,4'-trihydroxy-stilbyl-3'-O-glucoside has an obvious expansion effect on the blood vessels of the normal mice, can obviously resist the reduction of the arteriole and venule systole and capillary network opening number of the microcirculatory disturbance model mice and can obviously resist the reduction of the blood flow speed.

The invention discloses application of TTR (transthyretin) to angiogenesis inhibition, and belongs to the fields of biochemistry, molecular biology and medicine. After the TTR is given to mammals, inSTZ induced diabetes rat and mouse models, the new vessels of eyes are obviously reduced. Tumor new vessel, diabetes new vessel, eye new vessel and brain new vessel in vitro experiment models verify that the TTR has the effect of inhibiting the angiogenesis; the result shows that the TTR can act on an in vitro cell model in anoxic environment of brain and can continuously and efficiently inhibit the generation of new vessels. When the TTR acts on endothelial cells of umbilical veins, the formation of 90 percent of vessels can be obviously inhibited; when the TTR acts on endothelial cells of cerebral microvessel, the formation of 95 percent of vessels can be effectively inhibited; when the TTR acts on the endothelial cells of eye microvessel, the formation of 90 percent of vessels can be effectively inhibited; when the TTR acts on rat and mouse models, the as high as generation of 90 percent of new vessels can be effectively inhibited.

The invention relates to a medical prescription of catalpol freezing-drying powder for injection and a preparation method thereof. The invention is characterized in that the medical prescription of the catalpol freezing-drying powder for injection comprises 1 to 1000 mg of catalpol, 50mg to 2000mg of mannite, 100mg to 4000mg of glycine, 25mg to 1000mg of fucose, and 10% of sodium hydroxide solution right amount (pH 6.8-7.0). The invention also contains the medicinal application of the catalpol freezing-drying powder for injection which is used for treating clinical ischemia cerebral disease. The invention also comprises accessories used in the prescription which are used for preparing the catalpol and the freezing-drying powder for injection of the composition which is prepared by the catalop and other medicines according to different mixing ratios and is used for treating the ischemia cerebral disease. The invention aims at providing the traditional Chinese medicine active ingredient medicinal preparation. The preparation can permeate the blood brain barrier, alleviates the injury degree of the cerebral ischemia, increases the tolerance capability of the ischemia and oxygen-poor of brain tissues, promotes the newborn and the mature of brain microvascular, protects neurovascular units, promotes the neuron axon and dendrite growth, strengthens the synapse reconstruction, improves the back neutral behaviors of the cerebral ischemia, increases the neutral function score, and promotes the motor function recovery of affected limbs after the cerebral ischemia.

The invention discloses a construction method of an adeno-associated virus cross-blood brain barrier model. The construction method comprises the following steps: inoculating prepared astrocytes to the upper layer of a chip; inervsely inoculating prepared human brain microvascular endothelial cells to the lower layer of the chip; placing the chip in a matched precise shaking table so as to simulate the fluid shear force in the blood vessel of a human body; and performing continuous culture on the double-layer co-culture chip so that brain microvascular endothelial cells and astrocytes grow to form a blood brain barrier structure. According to the technical scheme provided by the invention, the in-vitro blood-brain barrier model is successfully constructed, the characteristics of the in-vivo blood-brain barrier are reflected more accurately and more closely, and the method can be used for related research of AAV crossing the blood-brain barrier. According to the invention, fluid shear force is added, so that a real in-vivo environment is better met, high throughput can be realized, and consumption of cells and reagents is reduced.

The invention discloses a preparation method and an application of a prodrug-supported metastatic encephaloma targeted drug-delivery system for inhibiting Mfsd2a. A nano drug-delivery system is constructed by adopting a biodegradable high polymer material as a basic carrier, functional molecules for inhibiting Mfsd2a and a metastatic encephaloma enzyme specificity prodrug are loaded in the nano drug-delivery system, and the nano drug-delivery system is modified with BMEC (brain microvascular endothelial cell) targeted ligands and metastatic encephaloma targeted ligands on the surface; the nanodrug-delivery system can efficiently penetrate blood brain barrier under the action of the BMEC targeted ligands and the Mfsd2a inhibiting functional molecules; after crossing the blood brain barrier, the nano drug-delivery system can enter tumor cells in a targeted manner under guidance of the metastatic encephaloma targeted ligands; the prodrug in the nano drug-delivery system can further specifically act on the tumor cells on the basis of targeted delivery and reduces side effects on normal brain tissue and surrounding tissue, and the method is simple and has operability and economic benefit.

The present invention relates to nucleic acids and polypeptides encoded thereby, whose expression is modulated in brain microvascular endothelial cells undergoing early dynamic inflammation-induced changes in blood-brain bather functionality. Such polypeptides are referred to as lipopolysaccharide-sensitive (LPSS) polypeptides. These nucleic acids and polypeptides may be useful in methods for controlling blood-brain bather properties in mammals in need of such biological effects. This includes the diagnosis and treatment of disturbances in the blood-brain / retina barrier, brain (including the eye) disorders, as well as peripheral vascular disorders. Additionally, the invention relates to the use of anti-LPSS polypeptide antibodies or ligands as diagnostic probes, as blood-brain barrier targeting agents or as therapeutic agents as well as the use of ligands or modulators of expression, activation or bioactivity of LPSS polypeptides as diagnostic probes, therapeutic agents or drug delivery enhancers.

The invention discloses a traditional Chinese medicine composition, fermented medicine liquor prepared from the traditional Chinese medicine composition, a preparation method and application. The traditional Chinese medicine composition is prepared from, by weight, 160-480 parts of kudzuvine roots, 80-320 parts of radix salvia miltiorrhiza, 120-400 parts of radix paeoniae alba, 120-400 parts of milkvetch roots, 40-240 parts of rhizoma curcumae longae, 60-280 parts of rhizoma atractylodis macrocephalae, 40-240 parts of semen brassicae, 60-280 parts of fructus crataegi, 60-280 parts of radix bupleuri, 60-280 parts of flos chrysanthemi, 40-240 parts of rhizoma gastrodiae, 60-280 parts of rhizome panacis japonica, 60-280 parts of rhizoma cyperi and 40-240 parts of herba salviae chinensis. Every 1,000 parts by weight of the fermented medicine liquor is prepared by adding and fermenting, by weight, 10-50 parts of saccharomyces cerevisiae, 10-50 parts of bacillus aceticus, 10-50 parts of lactobacillus and the balance traditional Chinese medicine decoction. Experiments show that the fermented medicine liquor has the obvious effect on oxidative stress injuries of cerebral microvascular endothelial cells and can significantly inhibit cerebral microvascular endothelial cell apoptosis and aging which are induced by hydrogen peroxide. Clinical experiments show that the fermented medicine liquor prepared from the traditional Chinese medicine composition can effectively treat ischemic encephalopathy, is safe and effective in treatment, has no adverse influence on the liver and kidney functions of a patient, can be taken for a long term and is worthy of clinical application and popularization.

The invention discloses a brain microvascular endothelial cell extraction and purification method, and relates to the technical field of cell purification. The extraction and purification method comprises the following steps: S1, coarsely extracting brain microvascular endothelial cells in brain tissues of a mouse; S2, carrying out primary CD31 immunomagnetic bead sorting on a brain microvascular endothelial cell mixture treated in the step S1 for purification; and S3, carrying out secondary CD31 immunomagnetic bead sorting on brain microvascular endothelial cells treated in the step S2 for purification so as to obtain the purified brain microvascular endothelial cells. According to the method, a mode of carrying out CD31 magnetic bead sorting for twice is adopted, damage to the activity of the brain microvascular endothelial cells is reduced, the separation and purification efficiency is high, the operation steps are relatively simple, and the brain microvascular endothelial cells with high purity, high yield and high activity can be obtained.

The invention relates to a medical prescription and a preparation method of catalpol-puerarin freeze-dried powder injection. The invention is characterized in that the medical prescription of the catalpol-puerarin freeze-dried powder injection comprises 1 to 1000mg of catalpol, 1 to 3000mg of puerarin, 100 to 8000mg of mannite, 50 to 4000mg of fucose as well as proper amount of 50% propanediol. The freeze-dried powder injection prepared by catalpol and puerarin has medical application in the treatment of clinic cerebral ischemic diseases. The invention also comprises the adjuvant used in the medical prescription for preparing the freeze-dried powder injection of the composition prepared by catalpol, puerarin and other drugs according to different mixture ratio to cure the ischemic diseases. The invention aims at providing the compound drug preparation of the active components of Chinese herbs for curing the ischemic diseases. The preparation can penetrate blood brain barriers, lighten the injury degree of cerebral ischemia, improve the durable capacity of ischemia and oxygen deficiency of brain tissues, promote the rebirthing and maturity of brain microvascular, protect the neurovascular units, promote the growth of neuron axon and neurodendrite, enhance the synaptic reorganization, improve the neurological behavior after cerebral ischemia, increase the neurological scores and promote the motor function resuming of the tired limbs after the cerebral ischemia.

A method for detection or monitoring status of silent brain ischemia (SBI) and cerebrovascular health. The assay reagents and methods described herein provide a specific indicator of cerebral microvascular disease, enabling clinicians to identify patients at risk for the development of SBI. A method of treating a subject having silent brain ischemia and / or metabolic syndrome comprises administering to the subject aspirin therapy, blood pressure therapy, body weight management, and / or a program of diet and exercise when levels of two or more SBI markers are elevated. Described herein are molecules that are produced by cerebral endothelial cells exposed to chronic vascular risk factors including obesity, hyperlipidemia, hypertension, and glucose intolerance. These stress molecules produced by cerebral endothelial cells are detectable in the serum and serve as diagnostic indicators of brain-specific endothelial cell damage and correlate with MRI indicators of silent stroke and impaired cognitive function.

The invention discloses an application and preparation method of a metastatic encephaloma targeted nano drug delivery system with effects of LRP1 up-regulation and targeting. By a biologically-degradable high polymer material serving as a basic carrier, internally-loaded functional small molecules capable of up-regulating low-density lipoprotein receptor related protein 1 (LRP1), and an LRP1 targeting ligand for surface modification, the nano drug delivery system is capable of realizing targeted delivery of the internally-loaded LRP1 up-regulation molecules under the action of the surface ligand so as to specifically up-regulate the surface LRP1 level of cerebral microvascular endothelial cells and metastatic encephaloma cells, accumulation of the LRP1 targeting ligand modified nano drug delivery system in metastatic encephaloma is promoted, and accordingly the nano drug delivery system forms a self-promoted drug delivery system. The nano drug delivery system which loads antitumor drugs realizes delivery of drugs in therapeutic concentration, and the preparation method is simple and high in operability and economic benefit.

The invention discloses application of medioresinol in preparation of a drug for preventing or treating the cerebral injury and a pharmaceutical composition thereof. The invention provides applicationof medioresinol in preparation of a drug for preventing or treating the cerebral injury for the first time; the medioresinol has an obvious effect on prevention or treatment of the cerebral injury, especially the cerebral injury caused by the cerebral arterial thrombosis; and the cerebral injury caused by the cerebral arterial thrombosis can be obviously reduced. The experiment discovers that themedioresinol has an effect of inhibiting brain microvascular endothelial cell damage caused by oxygen-glucose deprivation and is capable of obviously reducing the permeability of a blood brain barrier under oxygen-glucose deprivation. The medioresinol can reduce the volume of cerebral infarction under middle cerebral artery occlusion remarkably. The pharmaceutical composition containing the medioresinol can become a novel drug for preventing and / or treating the cerebral injury and is used for treating various cerebrovascular diseases including cerebral arterial thrombosis.

The invention provides a bionic object testing platform of a human brain microvascular system blood flow. The bionic object testing platform comprises the components of a container; a bracket assemblywhich is mounted in the container; an accommodating chamber which is mounted at the bracket assembly and is accommodated in the container; wherein the accommodating chamber comprises a first sub-chamber and a second sub-chamber which are separately arranged, and the first sub-chamber is above the second sub-chamber; the first sub-chamber and the second sub-chamber respectively extend outwards forforming a pipe; the first sub-chamber and the second sub-chamber are internally provided with blocking members; and a probe mounting member which is arranged at top of the first sub-chamber. Throughthe container, the bracket assembly, the accommodating chamber and the components of the probe mounting member, a bionic object of the human brain microvascular system blood flow is formed, and a technical gap in the technical solution is filled.

The invention belongs to the technical field of tissue engineering simulation, and particularly relates to a blood-brain barrier in-vitro model and a preparation method thereof. The blood-brain barrier in-vitro model comprises a quartz tube, and a first ceramic conductive gel layer and a second ceramic conductive gel layer which are respectively positioned at two ends of the quartz tube, and a brain microvascular endothelial cell monolayer formed under the condition of an ultrasonic pressure composite field is arranged in the quartz tube. Cells in the brain microvascular endothelial cell monolayer are connected through DNA chains, the brain microvascular endothelial cell monolayer and the inner wall of the quartz tube are connected through DNA chains, and astrocytes and microglia grow on the upper surface of the brain microvascular endothelial cell monolayer in an adhesion mode. The blood-brain barrier in-vitro model can simulate a blood-brain barrier structure more truly, and direct co-culture contact among different cells can reflect actual conditions more truly, so that the blood-brain barrier in-vitro model can be beneficial to drug development and screening of a central nervous system and has a good application prospect.

The invention discloses an in vitro construction method for simulating the blood-brain barrier of human brain microvascular formation, which comprises the following steps: preparing human brain microvascular endothelial cell suspension and human brain astrocyte suspension, and preparing fibrinogen mother liquor and thrombin mother liquor ; Mix endothelial cell suspension, astrocyte suspension, DMEM medium, fibrinogen mother solution and thrombin mother solution to prepare a mixed cell gel solution; inject the mixed cell gel solution into the microfluidic chip After incubating at a constant temperature until gelation, adding endothelial cell growth medium to the microfluidic chip to construct a 3D cell culture chip; continuously culturing the 3D cell culture chip so that the endothelial cells and astrocytes The plasmocytes grow to form the microvascular network structure of the brain, which corresponds to the generation of a simulated blood-brain barrier. The technical solution provided by the invention has successfully established an in vitro model of the blood-brain barrier, which more clearly and accurately reflects the characteristics of the blood-brain barrier.

The invention discloses a pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application. The pharmaceutical composition is composed of the following components at final concentrations: 1-10 μg / mL verbascoside, 0.1-1 μg / mL formononetin, 0.5-5 μg / mL7-2'-dihydroxy-3',4 '‑Dimethoxyisoflavane. In the present invention, by optimizing the proportion of the three effective active ingredients in Buyang Huanwu Decoction, the anti-cerebral ischemia-reperfusion injury effect of the pharmaceutical composition is significantly enhanced, and the cell apoptosis caused by cerebral ischemia-reperfusion injury can be alleviated. , improve the survival rate of brain microvascular endothelial cells, and can increase the expression levels of vascular endothelial growth factor, fibroblast growth factor, lactate dehydrogenase and superoxide dismutase in brain microvascular endothelial cells, and help restore brain function, It can be used to prepare medicines or preparations for preventing or treating ischemic cerebral apoplexy, and has broad application prospects.

Methods for generating functional brain microvascular endothelial cells (BMECs) under chemically defined, serum-free conditions are provided. In particular, efficient and cost-effective methods for generating functional BMECs under chemically defined culture conditions are provided. BMECs obtained according to the methods provided herein are suitable for in vitro blood brain barrier (BBB) formation.

The invention discloses application of TIMP-1 in preparation of a medicine for preventing or treating traumatic brain injury. The invention discloses application of TIMP-1 in preparation of a medicinefor treating blood-brain barrier dysfunction of traumatic brain injury. The medicine can be in the form of injection powder, water aqua, lipidosome and nano materials. The protein TIMP-1 can prolongthe incubation period of a mouse with cerebral trauma in a rotarod test; the motion balance capability of the mouse with cerebral trauma on a balance beam is improved; and the Evans blue permeabilityof brain tissues is reduced. In brain microvascular endothelial cells HBMEC, the TIMP-1 can participate in maintaining intercellular tight junction integrity, up-regulating tight junction complex expression, reducing luciferase leakage and increasing intercellular resistance. The invention also provides application of the protein TIMP-1 as a CD63-Integrin[beta]1 ligand in preparation of a medicinefor treating central nervous system diseases caused by blood-brain barrier dysfunction. The TIMP-1 plays a role by combining with a cell membrane complex CD63-Integrin[beta]1 to regulate and controlthe assembly of important molecules F-actin of cytoskeleton so as to maintain the intercellular tight junction integrity. The protein TIMP-1 has a good application prospect in treating the blood-brainbarrier dysfunction of thetraumatic brain injury.

The invention aims at disclosing polyester cotton for cranial nerve microvascular decompression and application of the polyester cotton, the polyester cotton with an unspecific three-dimensional shape is developed, the polyester cotton is of a cotton-fiber-shaped structure and comprises a plurality of wavy polyester fibers, the fineness d of the polyester fibers ranges from 100 D to 200 D, and the fineness d of the polyester fibers ranges from 100 D to 200 D. The number f of filaments of the polyester fibers is equal to (0.2-2) * d, the density of the polyester cotton is 0.003-0.01 g / cm < 3 >, and the filaments of the polyester fibers are in a separated state, the polyester cotton has the advantages that the polyester cotton is soft and fluffy, has certain elasticity, can be shaped at will in the clinical process, can be used in combination with the polyester cotton of different sizes, and has the advantages of being simple in structure and convenient to use. Compared with the prior art, the polyester cotton with the proper size and shape can be quickly selected according to the space structure between the cranial nerve and the cerebral microvessel, the shaped polyester cotton is placed between the cranial nerve and the cerebral microvessel, isolation of the cranial nerve and the cerebral microvessel is achieved, and cranial neuropathy of a patient is treated.

The invention discloses a method for separating and purifying brain microvessels, which comprises the following steps: (1) taking fresh whole brain tissue of mice, and grinding it up and down for 12-15 times with a Dounce homogenizer to prepare tissue homogenate; (2) homogenizing the tissue The slurry was double-filtered with sterile glass magnetic beads and a 70 μm nylon cell filter to obtain brain blood vessels and microvascular tissues. The method of the invention can not only effectively improve the integrity, quantity and purity of the isolated cerebral microvessels, but also not destroy the content of each target protein in the cerebral microvessels.

The invention discloses a use of a single targeted reduction-responsive vesicle nano-drug in preparation of a drug for treating brain tumors. Through reduction-responsive reversibly crosslinked vesicles of block polymers such as PEG-P(TMC-DTC), PEG-P(LA-DTC), PEG-P(TMC-DTC)-PEI, PEG-P(LA-DTC)-PEI, PEG-P(TMC-DTC)-Sp and PEG-P(LA-DTC)-Sp and a targeting polymer using ApoE as a targeting molecule, small-molecule chemotherapeutic drugs, protein drugs and gene drugs sensitive to brain glioma cells are efficiently encapsulated. The drug-carrying targeting vesicles can efficiently target multiple receptors (including LRP-1, LRP-2 and LDLR) that are highly expressed on the surfaces of brain microvascular endothelial cells in tumor regions thereby efficiently penetrating the blood-brain barrier andbeing efficiently enriched in the brain tumor region. ApoE-targeting related receptors are also highly expressed on the surfaces of glioma cells so that the drug-carrying targeting vesicles can be efficiently endocytosed by glioma cells and fast release drugs to induce apoptosis.

Disclosed is a method for producing an in vitro model for blood-brain barrier, including (a) a culturing conditionally immortalized astrocytes on one surface of a porous membrane and culturing conditionally immortalized brain pericytes on the other surface of the porous membrane, until both of the cells become a sheet; (b) culturing conditionally immortalized brain microvascular endothelial cells in a culture vessel, until the cells become a sheet; (c) peeling off the sheet of conditionally immortalized brain microvascular endothelial cells; (d) allowing the sheet of conditionally immortalized brain microvascular endothelial cells to come into contact with the sheet of conditionally immortalized brain pericytes, so that the sheets are arranged in layers; and (e) co-culturing a cell culture comprising three layers consisting of the sheet of conditionally immortalized brain microvascular endothelial cells, the sheet of conditionally immortalized brain pericytes, and the sheet of conditionally immortalized astrocytes.

The invention discloses a preparation method and application of a prodrug-loaded brain metastases targeting drug delivery system that inhibits Mfsd2a. The nano drug delivery system is constructed with a biodegradable polymer material as the basic carrier, and functional molecules are loaded inside it to Inhibition of Mfsd2a and brain metastases intracellular enzyme-specific prodrugs, surface modification of brain microvascular endothelial cell targeting ligands and brain metastases targeting ligands; the nano drug delivery system can target ligands in brain microvascular endothelial cells and Mfsd2a inhibitory functional molecules efficiently penetrate the blood-brain barrier; after crossing the blood-brain barrier, the nano-drug delivery system can target and enter tumor cells under the guidance of the brain metastases targeting ligand; On the basis of targeted delivery, the body drug can further specifically act on tumor cells and reduce side effects on normal brain tissue and surrounding tissues. The above method is simple, operable and economical.

The invention provides a method for detection or monitoring status of silent brain ischemia (SBI) and cerebrovascular health. The assay reagents and methods described herein provide a specific indicator of cerebral microvascular disease, enabling clinicians to identify patients at risk for the development of SBI. A method of treating a subject having silent brain ischemia and / or metabolic syndromecomprises administering to the subject aspirin therapy, blood pressure therapy, body weight management, and / or a program of diet and exercise when levels of two or more SBI markers are elevated. Described herein are molecules that are produced by cerebral endothelial cells exposed to chronic vascular risk factors including obesity, hyperlipidemia, hypertension, and glucose intolerance. These stress molecules produced by cerebral endothelial cells are detectable in the serum and serve as diagnostic indicators of brain-specific endothelial cell damage and correlate with MRI indicators of silentstroke and impaired cognitive function.

In one embodiment, the present invention is a method of creating a fully-human blood-brain barrier (BBB) model, comprising the steps of (a) obtaining a mixture of neural cells and brain microvascular endothelial cells (BMECs), wherein the neural cells and BMECs that comprise the mixture were produced from the differentiation of human pluripotent stem cells (hPSCs); (b) purifying BMECs from the mixture of neural cells and BMECs of step (a); and (c) co-culturing the purified BMECs with a cell type selected from the group consisting of pericytes, astrocytes and differentiated neural progenitor cells (NPCs), wherein a blood brain barrier model is created.

The invention relates to application of epalrestat in preparation of a medicine for preventing and treating cerebral arterial thrombosis (cerebral infarction), and belongs to the technical field of medicines. The invention provides application of epalrestat in preparation of the medicine for preventing and treating cerebral arterial thrombosis for cerebral arterial thrombosis patients who have missed a treatment time window and cannot be treated by thrombolysis in the early stage of cerebral arterial thrombosis. Epalrestat is used for treatment after cerebral arterial thrombosis, expression of aldose reductase can be inhibited, tight junction protein of cerebral microvascular endothelial cells can be up-regulated, endothelial cell apoptosis can be relieved, neurological dysfunction after cerebral arterial thrombosis can be improved, and integrity of blood-brain barrier after cerebral arterial thrombosis can be maintained; in addition, epalrestat can effectively reduce infiltration of peripheral inflammatory cells and relieve inflammatory response in ischemic brain tissue. The invention provides a new effective way for preventing and treating cerebral arterial thrombosis, and provides a theoretical basis for treating clinical cerebral arterial thrombosis by epalrestat.

Disclosed is a method for producing an in vitro model for blood-brain barrier, including (a) a culturing conditionally immortalized astrocytes on one surface of a porous membrane and culturing conditionally immortalized brain pericytes on the other surface of the porous membrane, until both of the cells become a sheet; (b) culturing conditionally immortalized brain microvascular endothelial cells in a culture vessel, until the cells become a sheet; (c) peeling off the sheet of conditionally immortalized brain microvascular endothelial cells; (d) allowing the sheet of conditionally immortalized brain microvascular endothelial cells to come into contact with the sheet of conditionally immortalized brain pericytes, so that the sheets are arranged in layers; and (e) co-culturing a cell culture comprising three layers consisting of the sheet of conditionally immortalized brain microvascular endothelial cells, the sheet of conditionally immortalized brain pericytes, and the sheet of conditionally immortalized astrocytes.

A liver-blood-brain barrier system based on a microfluidic chip for simulating metabolic processes in vivo. The microfluidic chip is mainly composed of a top chip, a porous filter membrane, and a bottom chip. The top chip is inoculated with liver cells, which can simulate the liver metabolism process of drugs in vivo; the bottom chip is mainly connected by the left main channel and the right main channel through the collagen channel, and the brain glial cell layer and the brain microvascular endothelial cell layer are seeded on the collagen interface Can simulate blood-brain barrier function. The invention integrates the construction and characterization of the in vitro blood-brain barrier model and the evaluation of the barrier permeability of the drug through the liver metabolism process, and can be used for the in vitro simulation of the blood-brain barrier model and the application of drug evaluation. The invention solves the problem of secondary inoculation and long time consumption of the cell co-culture model, adds flow conditions, is more in line with the real environment in the body, and significantly reduces the consumption of cells and reagents, and can obtain multiple experimental related parameters at the same time.