CN110179799A - The application of tetrandrine and the extracting method of tetrandrine - Google Patents
The application of tetrandrine and the extracting method of tetrandrine Download PDFInfo
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Abstract
The invention discloses a kind of applications of tetrandrine, and the extracting method of tetrandrine, specifically application and its extracting method of the tetrandrine in treatment rat microwave radiation cerebral injury, rat escape latency can be significantly reduced in tetrandrine, rat Spatial memory ability is improved, rat spirit of exploration and autonomic activities behavior are enhanced.
Description
Technical field
The present invention relates to chemical fields.It is more particularly related to a kind of application of tetrandrine and tetrandrine
Extracting method.
Background technique
Tetrandrine, also known as tetrandrine are also known as tetrandrine, and tetrandrine A prime is colorless needle crystals, main to use
In analgesia, flesh pine, antiallergy, anti-arrhythmia, the antitumor decompression anticoagulant of antibacterial.Tetrandrine is mainly mentioned from Fourstamen Stephania Root
It takes.Fourstamen Stephania Root is the dry root of Menispermaceae plant powder Radix Stephaniae Tetrandrae, has effects that inducing diuresis for removing edema, wind-expelling pain-stopping, modern pharmacology experiment
Show that Fourstamen Stephania Root total alkaloid has analgesia, decompression, myorelaxant effects.Alkaloid is higher in Fourstamen Stephania Root, general reachable
1.5~2.0%, wherein mainly tetrandrine, followed by fangchinoline.
It is more about the patent and document of the extraction of tetrandrine at present, mainly use alcohol extraction and acid precipitation method, supercritical fluid
There are many deficiencies in extraction or sour water percolation etc..And tetrandrine is methoxyl group, fangchinoline is phenolic hydroxyl group, powder
The polarity of menispermine is smaller, can be dissolved in cold benzene, and fangchinoline polarity is larger, is insoluble in cold benzene, therefore generally using cold benzene point
From tetrandrine and fangchinoline, but cold benzene is more toxic, and the influence of health and environment to operator is larger and mentions
It takes that rate is low, waste liquid amount is big, pollutes environment.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
It is an object of the present invention to provide a kind of applications of tetrandrine, and specifically tetrandrine is in treatment rat
The extracting method of application and tetrandrine in microwave radiation cerebral injury, tetrandrine, which can be significantly reduced rat and escape, hides
Phase improves rat Spatial memory ability, enhances rat spirit of exploration and autonomic activities behavior.
In order to realize purposes and other advantages according to the present invention, a kind of application of tetrandrine, tetrandrine are provided
Application in treatment rat microwave radiation cerebral injury.
The present invention also provides a kind of for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, comprising:
S1, it takes the Fourstamen Stephania Root of 100~120 parts by weight to clean, dry, crush, obtains Fourstamen Stephania Root powder, then to Fourstamen Stephania Root powder
The water of 300~500 parts by weight and the solid sodium chloride of 30~50 parts by weight are added in end, then is placed in the ultrasonic wave of 30~50kHz
20~30min of middle processing, obtains the first mixed liquor, is then placed in the first mixed liquor in the electric field that electric current is 4~8mA, is passed through CO2
Gas, CO2The flow velocity that is passed through of gas is 20mL/min, and being passed through the time is 15~25min, obtains the second mixed liquor；
S2, the second mixed liquor is placed in a centrifuge, 20~30min, removal is centrifuged with the revolving speed of 5000~8000r/min
Sediment takes supernatant, then supernatant is dissolved in the first diethyl ether solution of 30~50 parts by weight, obtains third mixed liquor, then
Third mixed liquor is placed in 40~50min of water-bath heating and refluxing extraction at 70~80 DEG C, filters, obtains the first solids, wherein the
The volume ratio of ether and water is 2:1 in one diethyl ether solution；
S3, the ether of 30~50 parts by weight and the water of 50~80 parts by weight are added into the first solids and is uniformly mixed,
The quick lime of 20~30 parts by weight is added, is filtered after stirring 20~30min, obtains the second solids, then the second solids is molten
In the second diethyl ether solution of 4 DEG C of 30~50 parts by weight, diethyl ether solution is then heated to by no ether with the speed of 5 DEG C/min
Until smell escapes, filtering, drying obtain tetrandrine, wherein the volume ratio of ether and water is 1:3 in the second diethyl ether solution.
Preferably, the extracting method of the tetrandrine for treating rat microwave radiation cerebral injury, step S1
The partial size of middle Fourstamen Stephania Root powder is 80~100nm.
Preferably, the extracting method of the tetrandrine for treating rat microwave radiation cerebral injury, step S3
In when quick lime is added, N is passed through with the flow velocity of 10mL/min2Gas, N2The time that is passed through of gas is 25~35min.
Preferably, the extracting method of the tetrandrine for treating rat microwave radiation cerebral injury, step S3
In the first solids need to be purified before ether and water is added, purification process are as follows: the first solids is dissolved in 10~15 weights
The ether of part is measured, then will be dissolved with the ether cation exchange resin exchange adsorption of the first solids, then use diethylether-ethanol
Solution elutes the cation exchange resin column of adsorption saturation, flow velocity 5mL/min, until outflow eluent inanimate object alkali reaction is
Only, the volume ratio of ether and ethyl alcohol is 1:2 in diethylether-ethanol solution.
Preferably, the extracting method of the tetrandrine for treating rat microwave radiation cerebral injury, step S3
Middle drying is cryogenic vacuum spray drying, and the vacuum degree of cryogenic vacuum spray drying is 6~8Pa, and temperature is 60 DEG C.
The present invention is include at least the following beneficial effects:
1, tetrandrine is applied to treatment rat microwave radiation cerebral injury by the present invention, and it is latent to significantly reduce rat average escape
Fu Qi improves Spatial memory ability, enhances rat spirit of exploration and increases autonomic activities behavior.
2, the present invention uses and is passed through CO into the first mixed liquor2The method of gas, CO2Gas be dissolved in the first mixed liquor with
Water reaction generates carbonic acid, and the first mixed liquor is made to have certain acidity, in order to extract the alkaloid in Fourstamen Stephania Root, and use to
CO is passed through in first mixed liquor2Gas changes the method for the first pH of mixed degree, can reinforce first while experiment mixed
The order of contact in liquid between each component is closed, there is certain mixing effect, and reduces fangchinoline and oxidation reaction generation occurs
Other substances generate new impurity, to improve the extraction effect of tetrandrine.
3, the present invention purifies the second solids using diethyl ether solution to extract tetrandrine, and the second solids is first dissolved in second
In ethereal solution, then Ether solution, during heating, ether is gradually reduced to no ether, Fourstamen Stephania Root in diethyl ether solution
Alkali is gradually decreased in the solubility of diethyl ether solution, and tetrandrine crystallization is precipitated, and obtains the tetrandrine of high-purity.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the water maze laboratory rats'swimming trajectory diagram of the present invention in one embodiment；
Fig. 2 is the bar shaped that the tetrandrine of the present invention in another embodiment influences rat average escape latency
Figure；
Fig. 3 is the bar chart that the tetrandrine of the present invention in another embodiment crosses over platform times influence to rat；
Fig. 4 is each group rat of the present invention in another embodiment typical path figure in spacious field experiment；
Fig. 5 is the bar chart of influence of the tetrandrine to rat total activity distance of the present invention in another embodiment；
Fig. 6 is the bar chart that the tetrandrine of the present invention in another embodiment influences rat average speed；
Fig. 7 is that the tetrandrine of the present invention in another embodiment accounts for total distance ratio to rat middle section activity distance
The bar chart of the influence of example；
Fig. 8 is the influence that the tetrandrine of the present invention in another embodiment damages Rat hippocampus morphosis
Electron microscope；
Fig. 9 is that the tetrandrine of the present invention in another embodiment is super to microwave radiation brain injury rat hippocampus neuron
The electron microscope that micro-structure influences；
Figure 10 is the tetrandrine of the present invention in another embodiment to microwave radiation brain injury in rats neuronal synapse
The electron microscope of ultrastructural influence；
Figure 11 is that the tetrandrine of the present invention in another embodiment is super to microwave radiation brain injury rat hippocampus blood vessel
The electron microscope that micro-structure influences.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention is described in further detail, to enable those skilled in the art's reference
Specification word can be implemented accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, it is described
Reagent and material commercially obtain unless otherwise specified.
Embodiment 1:
The present invention provides a kind of for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, comprising:
S1, it takes the Fourstamen Stephania Root of 100 parts by weight to clean, dry, crush, Fourstamen Stephania Root powder is obtained, then into Fourstamen Stephania Root powder
The water of 300 parts by weight and the solid sodium chloride of 30 parts by weight is added, then is placed in the ultrasonic wave of 30kHz and handles 20min, obtains first
Then first mixed liquor is placed in the electric field that electric current is 4mA, is passed through CO by mixed liquor2Gas, CO2The flow velocity that is passed through of gas is
20mL/min, being passed through the time is 15min, obtains the second mixed liquor；
S2, the second mixed liquor is placed in a centrifuge, 20min is centrifuged with the revolving speed of 5000r/min, sediment is removed, takes
Supernatant, then supernatant is dissolved in the first diethyl ether solution of 30 parts by weight, third mixed liquor is obtained, then sets third mixed liquor
The water-bath heating and refluxing extraction 40min at 70 DEG C, filtering, obtains the first solids, wherein ether and water in the first diethyl ether solution
Volume ratio is 2:1；
S3, the ether that 30 parts by weight are added into the first solids and 50 parts by weight water and be uniformly mixed, add 20
The quick lime of parts by weight filters after stirring 20min, obtains the second solids, then the second solids is dissolved in 4 DEG C of 30 parts by weight
In second diethyl ether solution, then diethyl ether solution to be heated to no ether smell evolution with the speed of 5 DEG C/min until, filtering is done
It is dry, obtain tetrandrine, wherein the volume ratio of ether and water is 1:3 in the second diethyl ether solution.
Embodiment 2:
The present invention provides a kind of for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, comprising:
S1, it takes the Fourstamen Stephania Root of 120 parts by weight to clean, dry, crush, Fourstamen Stephania Root powder is obtained, then into Fourstamen Stephania Root powder
The water of 500 parts by weight and the solid sodium chloride of 50 parts by weight is added, then is placed in the ultrasonic wave of 50kHz and handles 30min, obtains first
Then first mixed liquor is placed in the electric field that electric current is 8mA, is passed through CO by mixed liquor2Gas, CO2The flow velocity that is passed through of gas is
20mL/min, being passed through the time is 25min, obtains the second mixed liquor；
S2, the second mixed liquor is placed in a centrifuge, 30min is centrifuged with the revolving speed of 8000r/min, sediment is removed, takes
Supernatant, then supernatant is dissolved in the first diethyl ether solution of 50 parts by weight, third mixed liquor is obtained, then sets third mixed liquor
The water-bath heating and refluxing extraction 50min at 80 DEG C, filtering, obtains the first solids, wherein ether and water in the first diethyl ether solution
Volume ratio is 2:1；
S3, the ether that 50 parts by weight are added into the first solids and 80 parts by weight water and be uniformly mixed, add 30
The quick lime of parts by weight filters after stirring 30min, obtains the second solids, then the second solids is dissolved in 4 DEG C of 50 parts by weight
In second diethyl ether solution, then diethyl ether solution to be heated to no ether smell evolution with the speed of 5 DEG C/min until, filtering is done
It is dry, obtain tetrandrine, wherein the volume ratio of ether and water is 1:3 in the second diethyl ether solution.
Embodiment 3:
The present invention provides a kind of for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, comprising:
S1, it takes the Fourstamen Stephania Root of 110 parts by weight to clean, dry, crush, Fourstamen Stephania Root powder is obtained, then into Fourstamen Stephania Root powder
The water of 400 parts by weight and the solid sodium chloride of 40 parts by weight is added, then is placed in the ultrasonic wave of 40kHz and handles 25min, obtains first
Then first mixed liquor is placed in the electric field that electric current is 6mA, is passed through CO by mixed liquor2Gas, CO2The flow velocity that is passed through of gas is
20mL/min, being passed through the time is 20min, obtains the second mixed liquor；
S2, the second mixed liquor is placed in a centrifuge, 25min is centrifuged with the revolving speed of 6500r/min, sediment is removed, takes
Supernatant, then supernatant is dissolved in the first diethyl ether solution of 40 parts by weight, third mixed liquor is obtained, then sets third mixed liquor
The water-bath heating and refluxing extraction 45min at 75 DEG C, filtering, obtains the first solids, wherein ether and water in the first diethyl ether solution
Volume ratio is 2:1；
S3, the ether that 40 parts by weight are added into the first solids and 65 parts by weight water and be uniformly mixed, add 25
The quick lime of parts by weight filters after stirring 25min, obtains the second solids, then the second solids is dissolved in 4 DEG C of 45 parts by weight
In second diethyl ether solution, then diethyl ether solution to be heated to no ether smell evolution with the speed of 5 DEG C/min until, filtering is done
It is dry, obtain tetrandrine, wherein the volume ratio of ether and water is 1:3 in the second diethyl ether solution.
Embodiment 4:
On the basis of embodiment 3, the partial size of Fourstamen Stephania Root powder is 90nm in step S1.
Embodiment 5:
On the basis of embodiment 3, in step S3 when quick lime is added, N is passed through with the flow velocity of 10mL/min2Gas, N2
The time that is passed through of gas is 30min.
Embodiment 6:
On the basis of embodiment 3, the first solids needs to be purified before ether and water is added in step S3, purifies
Process are as follows: the first solids is dissolved in the ether of 12 parts by weight, then hands over the cation of the ether dissolved with the first solids
Change resins exchange absorption, then with diethylether-ethanol solution elute adsorption saturation cation exchange resin column, flow velocity 5mL/min,
Until outflow eluent inanimate object alkali reaction, the volume ratio of ether and ethyl alcohol is 1:2 in diethylether-ethanol solution.
Embodiment 7:
On the basis of embodiment 3, dry in step S3 is that cryogenic vacuum is spray-dried, and cryogenic vacuum is spray-dried true
Reciprocal of duty cycle is 7Pa, and temperature is 60 DEG C.
Comparative example 1:
Compared with Example 3, the mass fraction that 200 parts by weight are directly added in step S1 into the first mixed liquor is 20%
Carbon acid solution, obtain the second mixed liquor, other are same as Example 3.
Comparative example 2:
Compared with Example 3, the second solids is dissolved in 4 DEG C of the water of 45 parts by weight again in step S3, filtering is done
It is dry, tetrandrine is obtained, other are same as Example 3.
Interpretation of result
1, it using the yield and purity of HPLC chromatogram measurement embodiment 3~7 and the resulting tetrandrine of comparative example 1~2, produces
Rate=(weight/Fourstamen Stephania Root weight of tetrandrine) × 100%, purity=(weight/product weight of tetrandrine) ×
100%, it the results are shown in Table 1.
The yield and purity of 1 tetrandrine of table
As known from Table 1, the yield and purity of embodiment 3~7 and the resulting tetrandrine of comparative example 1~2 have reached one
Fixed height, wherein compared to embodiment 3~4, embodiment 7 and comparative example 1~2, the resulting tetrandrine of embodiment 5~6 is produced
Rate and purity are higher, and the yield of embodiment 5 is up to 0.83%, and the purity of embodiment 6 is up to 98.4%, illustrate in step
N is passed through when quick lime is added in S32Gas, it is possible to prevente effectively from the other compositions in Fourstamen Stephania Root are oxidized generation other impurities
And be blended in product and be difficult to separate, so that the yield of tetrandrine is improved, and cation exchange resin exchange can have
Effect improves the yield of tetrandrine in product, improves the purity of product；It is directly added into carbon acid solution in comparative example 1, does not there is stirring
Effect, to reduce the extraction effect of tetrandrine；The yield more a height of 0.80% of the resulting tetrandrine of comparative example 2, but
Purity is 83.7%, this is because the second solids to be dissolved in 4 DEG C of 45 parts by weight again in step S3 compared to embodiment 5~6
Water in, it is filtering, dry, obtain tetrandrine, rather than in the second diethyl ether solution of 45 parts by weight, be a mistake directly filtered
The process of journey rather than recrystallization, also mixed with impurity not soluble in water in the product of acquisition.
2, therapeutic effect of the tetrandrine to microwave radiation brain injury in rats
2.1, experimental material:
Tetrandrine thermo-sensitive gel (weigh 12g poloxamer188,2g PLURONICS F87,1g HP- β-CD in beaker,
Appropriate amount of deionized water is added, magnetic agitation makes it be uniformly dispersed, and places in 4 DEG C for 24 hours, is sufficiently swollen, it is former to form transparent blank
Position gel solution.It weighs the resulting tetrandrine of appropriate embodiment 6 to be dissolved in a certain amount of 0.1mol/L hydrochloric acid, keeps it sufficiently molten
Then solution adjusts pH=5 or so with 0.1mol/L sodium hydroxide, the above-mentioned blank being swollen is added in tetrandrine suspension
In situ-gel solution, magnetic agitation makes it be uniformly dispersed, and 4 DEG C are placed for 24 hours, and supplement deionized water to 50mL is final to be made
50mL tetrandrine thermo-sensitive gel)；
Formalin；
Wistar rat, male, weight 180-200g, SPF grade.
2.2, experimental method:
After rat adaptive feeding 5d, 6 groups are randomly divided into, i.e., normal group, model group, Blank gel group, tetrandrine temperature
Quick gel 10mgkg-1、20mg·kg-1、30mg·kg-1Group, every group 10.
(water maze laboratory is the Classic Experiments for evaluating animal learning memory capability for water maze laboratory training.Water maze analysis
System is made of full-automatic track acquisition system and stainless steel round pool, the high 40cm in pond, diameter 180cm, depth of water 30cm, water
Temperature is kept for 22 DEG C or so, is divided into tetra- quadrants of I, II, III, IV.There is a moveable platform in pond, platform diameter is
8cm is lower than liquid level 2cm, keeps position of platform to fix in experiment.Video camera is hung right above pond, records activities in rats track,
Peripheral pool is blocked with the door curtain made of cloth, and interior is kept quite.), after 3d, model is begun setting up, using the self-built high power of Academy of Military Sciences
Microwave radiation simulation source, centre frequency 2.856GHz, average power density 30mW/cm2；Pulsewidth fixes 500ns；Repetition rate
1000Hz；Peak power density is respectively 200W/cm2, irradiation time 15min.Specific method of radiating are as follows: fix each group rat
In organic glass radiation seed, radiation seed is placed in rotatable radiation table in microwave dark room, microwave source uniform spoke from top to bottom
It penetrates, the irradiation of normal rats row equal conditions puppet.The same day after microwave radiation, administration group rat are respectively adopted self-control delivery device and carry out
Nasal-cavity administration, continuous 8d, is administered twice a day, and being administered daily volume is 0.5mLkg-1；Normal group and model group are not processed.
Experiment, which is divided into, finds platform training, orientation navigation experiment and space exploration experiment three parts.
1. finding platform training: rat being close on the inside of I quadrant cell wall, gently into the water towards cell wall, is started simultaneously at
Record track of swimming in rat 60s.If platform is found and rested on to rat in 60s, trajectory track stops at once, if rat 60s
Inside fail to find platform, it is guided to appear on the stage, and it is kept to stop 20s in platform with consolidating memory, is then sequentially completed II~IV
Quadrant, taking-up put back to mouse cage after drying.Training 1 time daily, continuous 3d.
2. orientation navigation experiment: the 3rd, 4,5d orientation navigation experiment is carried out after administration, by rat successively from four quadrants
Into the water, the time that platform is found in 60s, as average escape latency AEL (the Average Escape of the quadrant are recorded
Latency), if failing to find platform in rat 60s, AEL will be recorded as 60s, and rat is not required to stop in platform, into it is next as
Limit test.The average value of four quadrant AEL is the AEL that this tests the animal.During test, water is replaced daily, keeps water
The clean free from extraneous odour in pond.
3. space exploration is tested: 7d progress space exploration is tested to detect the spatial memory capacity of rat after administration,
Platform is removed, using the opposite side quadrant for being placed with platform quadrant as water-drop point, into the water by rat, observes rat in 60s
Across the number of platform position.
4. spacious field is tested: for evaluating autonomic activities and exploratory behavior of the rat in free environments, experimental provision is by spacious field
Reaction chamber and automatic data collection, processing system composition.Square of the spacious field reaction chamber bottom surface for long 100cm, the high 40cm of surrounding,
Bottom edge and surrounding are grey black, are equipped with digital camera head at 150cm right above reaction chamber to monitor tracking rat behavior.Experiment
In, bottom surface square area (40cm × 40cm) is defined as middle section, rat is placed in spacious field center, while opening automatic view
Frequency tracking system, total activity distance, central movable distance, average speed and activity trajectory in record rat five minutes.Experiment
Room is kept quite, and human interference is excluded.After every rat experiment, 75% ethanol solution wipes bottom and tank wall, keeps clear
It is clean, the interference of rat smell before exclusion.
After 8d is administered, after the anesthesia of 1% yellow Jackets is injected intraperitoneally according to 30mg/kg in each group rat, sacrificed by decapitation, i.e.,
It carves and takes out rat whole brain, every group 3.After 10% formalin solution fixes 2 weeks, along hippocampus maximum coronal-plane, thickness about 3mm is cut
Thin slice is placed in embedded box, and flowing water rinses fixture and precipitating, through conventional dehydration with it is transparent, waxdip with embed after be cut into thickness
For 3 μm of thin slice, conventional H E dyeing is carried out after dry, neutral gum mounting is observed with optical microscopy and acquires picture.
2.3 interpretation of result
2.3.1, influence of the tetrandrine to microwave radiation brain injury in rats cognitive function
As shown in Figures 1 to 3, orientation navigation experiment is the results show that the 4th, 5d, model group rats average escape after microwave radiation
More normal group of incubation period dramatically increases (P < 0.05), shows 30mW/cm2Microwave radiation can lead under learning and memory in rats ability
Drop, tetrandrine 10,20mgkg-1Administration group, average escape latency significantly reduce (P < 0.05) compared with model group, Blank gel
Group, tetrandrine 30mgkg-1Administration group no difference of science of statistics.The space exploration experimental result of 7d is shown after microwave radiation,
The number of model group rats across position of platform more normally organizes significant decrease (P < 0.01), tetrandrine 10,20mgkg-1Administration
The number of group, rat across position of platform dramatically increases (P < 0.05) compared with model group, Blank gel group, tetrandrine 30mg
kg-1Administration group no significant difference (P > 0.05) compared with model group.Tetrandrine 10,20mgkg-1Administration group can significantly change
The cognitive function of kind microwave radiation brain injury in rats, improves Spatial memory ability.
2.3.2, tetrandrine probes into microwave radiation brain injury in rats and the influence of independent behaviour
In spacious field experiment, the motion profile record of rat such as Fig. 4~7, through statistical analysis, model group rats total activity road
Journey, average speed and middle section distance account for the ratio of total distance, and substantially less than normal group (P < 0.01), tetrandrine 10,
20mg·kg-1Administration group rat total activity distance (P < 0.01), average speed (P < 0.01) and middle section distance account for total distance
Ratio (P < 0.05) dramatically increased compared with model group.Blank gel group, tetrandrine 30mgkg-1Administration group and model group
Compared to no significant difference (P > 0.05).Tetrandrine 10,20mgkg-1It is big that administration group can dramatically increase microwave radiation cerebral injury
The independent behaviour and spirit of exploration of mouse.
2.3.3, the influence that tetrandrine damages Rat hippocampus morphosis
As shown in figure 8, the hippocampal tissue of normal rats is in normal morphology, neuron sharpness of border, nucleus caryogram rule
Whole, the light dye of cytoplasm, model group rats nucleus is triangular in shape or shuttle shape, karyopycnosis contaminate deeply, to scheme sea in C and figure A in such as Fig. 8
Ma Qu is more obvious, tetrandrine 10,20mgkg-1Administration group can significantly improve this phenomenon.
2.3.4, influence of the tetrandrine to Rat hippocampus ultrastructure damage
Neuron is as shown in Figure 9: normal rats, and hippocampal neuron chromatin is uniform, and caryogram is regular, and endochylema is abundant, carefully
Born of the same parents' device is clear in structure；Side collection is concentrated in model group rats nuclei dyeing chromaticness, and caryogram is not whole, nuclear membrane obscure boundary, mitochondrial swelling,
Reticulum dilatation；Tetrandrine is low, middle dosage can significantly improve this phenomenon, and tetrandrine high dose slightly improves.
Cynapse is as shown in Figure 10: normal rats hippocampal synapse double membrane structure is clear, it is seen that presynaptic vesica and cynapse
Fine and close object afterwards, model group synaptic cleft is broadening, and postsynaptic density increases, and cynapse double membrane structure is smudgy, tetrandrine
Low, middle dosage is obviously improved, and high dose improves unobvious.
Blood vessel is as shown in figure 11: normal rats perivascular canal is normal, and after microwave radiation, perivascular canal is significantly broadening, powder
Menispermine is low, middle dosage is obviously improved, and high dose improves unobvious.
In conclusion the continuous nasal-cavity administration of tetrandrine nasal thermosensitive gel 8 days, it is investigated by behaviouristics and pathology is seen
It examines, show that tetrandrine nasal thermosensitive gel dosage is 10~20mgkg-1Rat average escape latency can be significantly reduced,
Spatial memory ability is improved, rat spirit of exploration is enhanced and increases autonomic activities behavior.Pass through pathologyofbraintissue, optics
Microscope observes institutional framework, and transmission electron microscope observing ultra micro institutional framework can find microwave radiation to rat cerebral tissue and mind
It causes obviously to damage through member, and tetrandrine has therapeutic effect to microwave radiation cerebral injury.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and legend shown and described herein.
Claims (6)
1. the application of tetrandrine, which is characterized in that application of the tetrandrine in treatment rat microwave radiation cerebral injury.
2. the extracting method for the tetrandrine for treating rat microwave radiation cerebral injury characterized by comprising
S1, it takes the Fourstamen Stephania Root of 100~120 parts by weight to clean, dry, crush, Fourstamen Stephania Root powder is obtained, then into Fourstamen Stephania Root powder
The water of 300~500 parts by weight and the solid sodium chloride of 30~50 parts by weight is added, then is placed in the ultrasonic wave of 30~50kHz
20~30min is managed, the first mixed liquor is obtained, then the first mixed liquor is placed in the electric field that electric current is 4~8mA, is passed through CO2Gas
Body, CO2The flow velocity that is passed through of gas is 20mL/min, and being passed through the time is 15~25min, obtains the second mixed liquor；
S2, the second mixed liquor is placed in a centrifuge, 20~30min, removal precipitating is centrifuged with the revolving speed of 5000~8000r/min
Object takes supernatant, then supernatant is dissolved in the first diethyl ether solution of 30~50 parts by weight, obtains third mixed liquor, then by
Three mixed liquors are placed in 40~50min of water-bath heating and refluxing extraction at 70~80 DEG C, and filtering obtains the first solids, wherein the first second
The volume ratio of ether and water is 2:1 in ethereal solution；
S3, the ether of 30~50 parts by weight and the water of 50~80 parts by weight are added into the first solids and is uniformly mixed, then plus
Enter the quick lime of 20~30 parts by weight, is filtered after stirring 20~30min, obtain the second solids, then the second solids is dissolved in 4 DEG C
30~50 parts by weight the second diethyl ether solution in, diethyl ether solution is then heated to by no ether smell with the speed of 5 DEG C/min
Until evolution, filtering, drying obtain tetrandrine, wherein the volume ratio of ether and water is 1:3 in the second diethyl ether solution.
3. as claimed in claim 2 for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, feature
It is, the partial size of Fourstamen Stephania Root powder is 80~100nm in step S1.
4. as claimed in claim 2 for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, feature
It is, in step S3 when quick lime is added, N is passed through with the flow velocity of 10mL/min2Gas, N2Gas be passed through the time be 25~
35min。
5. as claimed in claim 2 for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, feature
It is, the first solids needs to be purified before ether and water is added in step S3, purification process are as follows: the first solids is molten
In the ether of 10~15 parts by weight, it then will be dissolved with the ether cation exchange resin exchange adsorption of the first solids, then
The cation exchange resin column of adsorption saturation, flow velocity 5mL/min are eluted with diethylether-ethanol solution, until outflow eluent is without life
Until alkaloids are reacted, the volume ratio of ether and ethyl alcohol is 1:2 in diethylether-ethanol solution.
6. as claimed in claim 2 for treating the extracting method of the tetrandrine of rat microwave radiation cerebral injury, feature
It is, dry in step S3 is that cryogenic vacuum is spray-dried, and the vacuum degree of cryogenic vacuum spray drying is 6~8Pa, temperature 60
℃。
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