CN109620852A - A kind of application for grafting storesin in the extension clotting time - Google Patents
A kind of application for grafting storesin in the extension clotting time Download PDFInfo
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- CN109620852A CN109620852A CN201910049453.6A CN201910049453A CN109620852A CN 109620852 A CN109620852 A CN 109620852A CN 201910049453 A CN201910049453 A CN 201910049453A CN 109620852 A CN109620852 A CN 109620852A
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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Abstract
The present invention provides a kind of grafting storesins in the application extended in the clotting time, is related to the applied technical field of storesin.The difference on volatile ingredient and non-volatile component is larger with commodity storesin for grafting storesin of the present invention, and has the extremely significant extension clotting time to act on fibrinogen (FIB), thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) in 5~50mg/mL dosage range.
Description
Technical field
The invention belongs to the applied technical fields of storesin, and in particular to a kind of grafting storesin is solidifying in extension
Application in the blood time.
Background technique
Storax (Storax) is Hamamelidaceae sweetgum platymiscium storax tree Liquidambar orientalis
Mill. resin secreted by trunk is that the common import Chinese medicine in China first recorded in " Mingyi Bielu " is classified as top grade." China
Pharmacopeia " it formally records since the nineteen ninety.Although clinically big to the demand of storax, storax all needs import,
The high price of medicinal material also greatly limits the application of storax.
China's tropical, subtropical zone and temperate regions belong to tropical monsson climate, subtropical monsoon climate, temperate zone
Monsoon climate, temperate continental climate and plateau mountain band weather, and storax original plant only warms up (minimum temperature 4~10 to the winter
DEG C), Xia Liang (22 DEG C of maximum temperature or so), annual range of temperature be small, uniform (the annual precipitation 300~1000 of annual precipitation season distribution
Millimeter, rainfall in winter accounts for about 60~70%, Summer Precipitation 30~40%) Etesian climate compare adaptation, therefore it is at me
The growth of state is restricted.It is chatted in small sub- West Asia southern areas, the Turkey of three continent of Europe, Asia and Africa boundary storax main producing region
Various countries near Leah, Egypt, Somalia and the Persian Gulf, though there is the storax introduced a fine variety point in China in Yunnan, Guangdong, Guangxi province
Cloth, but not formed industry size.Jiangsu Province once introduced a fine variety storax in the last century 80's, but survival rate is very low, only
5%.Sweetgum (Liquidambar formosana Hance.) is Hamamelidaceae sweetgum platymiscium, likes warm and moist weather, compared with
Drought-resistant hungry soil, germination rate is high, and growth is fast, adaptable, and distribution is wide, in China Shaanxi, Henan, Hebei, Anhui, river
The provinces such as Soviet Union, Zhejiang, Fujian, Taiwan, Guangxi, Guangdong, Jiangxi, Hunan, Sichuan, Yunnan, Guizhou, Qinghai, Tibet are distributed, main
Originate in the ground such as Jiangsu, Zhejiang, Jiangxi, Fujian, Guangdong, Guangxi.
For the growth question for solving storax, expanding propagation is carried out with its branch, the trial of Ye Jinhang tissue cultures, but this is not
It can solve storax and be not suitable with the low problem of China's weather, survival rate.In the eighties in last century, the branch of storax is transferred in trial
Be connected on the closer equal congener sweetgum of affinity, this measure achieve it is ideal as a result, grafting after storax at
Motility rate is up to 80%.Although the storax tree survival rate after grafting improves, the effective component and effect of storesin are grafted
Fruit is simultaneously indefinite.
Summary of the invention
In view of this, the application it is an object of the invention to a kind of grafting storesin in the extension clotting time, mentions
Height influences fibrinogen, thrombin time, prothrombin time, activated partial thromboplastin time etc..
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
It is described to transfer the present invention provides a kind of application of grafting storesin in the extension fibrinogen clotting time
It connects storesin and picks up from the storax after grafting, the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in thrombin time, and the grafting Soviet Union closes
Botany bar gum picks up from the storax after grafting, and the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in prothrombin time, the grafting Soviet Union
Blending resin picks up from the storax after grafting, and the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in activated partial thromboplastin time, institute
It states grafting storesin and picks up from the storax after grafting, the grafting is using storax as scion, using sweetgum as stock.
Preferably, the application concentration of the grafting storesin is 5~50mg/mL.
The present invention provides a kind of grafting storesins in the application extended in the clotting time, specifically includes extension fiber
Proteinogen clotting time, thrombin time, prothrombin time and activated partial thromboplastin time.In embodiments of the present invention
Storesin is grafted under the additional amount of 5mg/mL, thrombin time, prothrombin time and activated partial thromboplastin
Time extends to 10.62 ± 0.60s, 58.22 ± 0.75s and 17.77 ± 0.74s respectively, shows extremely significant extension blood coagulation
Chronergy.
Detailed description of the invention
Fig. 1 is grafting storesin GC-MS ion flow graph；
Fig. 2 is the total ion current figure for grafting storesin；
Fig. 3 is the HLPC figure for grafting storesin；
Fig. 4 is the GC-MS ion flow graph of commodity storax.
Specific embodiment
It is described to transfer the present invention provides a kind of application of grafting storesin in the extension fibrinogen clotting time
It connects storesin and picks up from the storax after grafting, the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in thrombin time, and the grafting Soviet Union closes
Botany bar gum picks up from the storax after grafting, and the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in prothrombin time, the grafting Soviet Union
Blending resin picks up from the storax after grafting, and the grafting is using storax as scion, using sweetgum as stock.
The present invention also provides a kind of grafting storesins in the application extended in activated partial thromboplastin time, institute
It states grafting storesin and picks up from the storax after grafting, the grafting is using storax as scion, using sweetgum as stock.
The application concentration of grafting storesin of the present invention is preferably 5~50mg/mL.In embodiments of the present invention,
When the diameter for grafting storax tree reaches 10cm or more, can carry out adopting glue, in 7~October, the gum yield of each tree for 10g with
On.
Grafting storesin provided by the invention is carried out in the application extended in the clotting time below with reference to embodiment
Detailed description, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Select the Chinese sweet gum of the diameter of a cross-section of a tree trunk 1.3 meters above the ground about 2~8cm as stock, by the grafting of storax branch on Chinese sweet gum, 5 years to be grown
More than, and when diameter reaches 10cm or more, 7 annual~October carries out adopting glue, and gum yield is about each tree 10g or more.
1, to the collected grafting storesin pharmacology activity research of embodiment 1
(1) laboratory apparatus
LG-PABER-I type Standard for semi-Automated Blood Coagulation Analyzer (Beijing Shi Di scientific instrument Co., Ltd)；Centrifuge (Town in Shanghai
Pavilion scientific instrument factory)；Blood rheological instrument (Beijing Zhong Qinshidi scientific instrument Co., Ltd)；The analysis of platelet aggregation coagulation factor
Instrument (Beijing Steellex Scientific Instrument Company).
(2) experiment reagent
Fibrinogen (FIB, lot number STG20401-34), thrombin time (TT, lot number STG20301-35A), blood coagulation
Zymogen time (PT, lot number STG20101-49), activated partial thromboplastin time (APTT, lot number ST20201-56), diphosphonic acid
Adenosine (1 bottle of ADP pulvis, 10mL/ bottles of buffer) is purchased from Beijing Zhong Qinshidi scientific instrument Co., Ltd.Rabbit is purchased from south
Capital Qinglongshan experimental animal breeding base.
(3) experimental method
After rabbit is fixed, the hair of neck is shaved off, until seeing neck arteries blood vessel, after disinfection, is surgically inserted into blood sampling
Pipe.When being tested, while comparative example is set, the storesin produced with Turkey formed levant storax oil after ethyl alcohol refines
Carry out identical experiment.
Prothrombin time (PT) assay kit (freezing method) STY20101-786:
Testing principle:
Excessive tissue thromboplastin and Ca are added in blood plasma to be detected2+, compound is formed with FVIIa, triggers external source
Property blood coagulation system, make prothombin at fibrin ferment, fibrin ferment make fibrinogen change fibroblast cells.Surveyed blood plasma is solidifying
Solid time, i.e. prothrombin time.The extrinsic coagulation factor content in time and blood plasma that grumeleuse is formed is negatively correlated.
Sample requirement: after venous blood collection, 109mmol/L sodium citrate and whole blood should be uniformly mixed in 1:9 ratio immediately.
The whole blood for being mixed with anti-coagulants is centrifuged 10min with 3000*g, and takes out blood plasma with plastic suction pipet immediately.Plasma sample room temperature
Under need to be tested in 2h, to guarantee its stability.If test sample is delayed over 2h or more, need to protect plasma sample
In the presence of being stored under the conditions of -20 DEG C (2 weeks) or -70 DEG C (one months).When testing again, the plasma sample of freezing should be in 37 DEG C of items
Quick-thawing under part, and test immediately.It should be avoided when blood sampling and haemolysis and hyperlipidemic occur, the use of bilious sample.
Detection method: blood coagulation analyzer, centrifuge, test cup, distilled water, Quality Control blood plasma.PT reagent is placed in instrument
In 37 DEG C of reagent pre-temperature holes, reagent pre-heating time has to be larger than 10min, must shake up before.Test pearl is added, it then will be to
It surveys 50 μ L of blood plasma to be added in test cup, 10 μ L of sample is added, sample is in 37 DEG C of pre- accurate pre-temperature 180s of warm area, by the PT after pre-temperature
Above-mentioned 100 μ L of test cup is added in reagent, starts instrument immediately and is tested automatically.
Test result is as shown in table 1:
Table 1 sample prothrombin time (PT) measurement result
Note: * indicates there is significant difference, and * * indicates there is extremely significant difference
Sample prothrombin time measurement result shows that influence of the solvent (dimethyl sulfoxide) to blank plasma is little, sun
Property control aspirin have a significant difference when concentration is 5mg/mL, commodity storax and grafting storesin it is high, in,
It is compared under low concentration with blank plasma, there is extremely significant difference.
Thrombin time (TT) assay kit (freezing method) STY20301-49:
Desired use: gauged blood coagulation is added for detecting test plasma in thrombin time test kit (freezing method)
Time needed for fibrinogen is changed into fibrin after enzyme.
Testing principle: under the action of the thrombin solution that calibration is added in test plasma does not have fibrin ferment again, in blood plasma
Fibrinogen is changed into fibrin, and the time of volume needed for the clotting of plasma is the clotting time.When anticoagulant in test plasma
Substance increases or when fibrinogen content exception, and thrombin time can occur to change accordingly.
Sample requirement: sample collection:, should be immediately by 109mmol/L sodium citrate and whole blood in 1:9 ratio after venous blood collection
It is uniformly mixed.Sample preparation: the whole blood for being mixed with anti-coagulants is centrifuged 10min with 3000*g, and is taken immediately with plastic suction pipet
Blood plasma out.Plasma sample need to be tested in 2h at room temperature, to guarantee its stability.If test sample be delayed over 2h with
On, then it needs for plasma sample to be stored in and stores under the conditions of -20 DEG C (2 weeks) or -70 DEG C (one months).When testing again, freezing
Plasma sample should under the conditions of 37 DEG C quick-thawing, and test immediately.It should be avoided when blood sampling and haemolysis and hyperlipidemic occur,
The use of bilious sample.
Detection method: blood coagulation analyzer, centrifuge, test cup, distilled water, Quality Control blood plasma.TT reagent is gently shaken up, room
Temperature stands 10min, spare after equalized temperature.Test pearl is added, then 100 μ L of test plasma is added in test cup, sample is added
Above-mentioned 100 μ L of test cup is added in 37 DEG C of pre- accurate pre-temperature 180s of warm area, by the TT reagent after pre-temperature in 10 μ L of product, sample, immediately
Starting instrument is tested automatically.
Test result is as shown in table 2:
Table 2 sample thrombin time (TT) measurement result
Note: * indicates there is significant difference, and * * indicates there is extremely significant difference
Sample thrombin time test the result shows that, influence of the solvent (dimethyl sulfoxide) to blank plasma is little, positive
Control aspirin has significant difference when concentration is 5mg/mL, and commodity storax and grafting storesin are high, medium and low
It is compared under concentration with blank plasma, there is extremely significant difference, and the more high and low concentration clotting time in clotting time of middle concentration
It is long.
Activated partial thromboplastin time (APTT) assay kit (freezing method) STY20201-57:
Desired use: the product is for Quantitative in vitro measurement activated partial thromboplastin time (APTT)
Inspection principle: being added activator in 37 DEG C of downloading platelet poor plasmas (PPP), F Ⅻ and F Ⅺ is activated, with brain phosphorus
Rouge substitutes blood platelet and provides the catalytic surface of blood coagulation, in Ca2+Under participation, the time required for the clotting of plasma is observed, it is as to be measured
Blood plasma Activated partial thromboplastin time (APTT).
Detection method: blood coagulation analyzer, centrifuge, test cup, distilled water, 25mmol/LCaCl2, Quality Control blood plasma.It will
APTT reagent takes out from refrigerator, gently shakes up, spare after equalized temperature.CaCl2Reagent is using preceding must be placed into 37 DEG C of pre-temperatures
Area, pre-heating time have to be larger than 20min.Test pearl is added, then 50 μ L of test plasma is added in test cup, APTT examination is added
50 μ L of agent is eventually adding 10 μ L of sample, sample is in 37 DEG C of pre- accurate pre-temperature 180s of warm area, after pre-temperature in test cup
CaCl2Above-mentioned 50 μ L of test cup is added in reagent, starts instrument immediately and is tested automatically.
Test result is as shown in table 3:
Table 3 sample activated partial thromboplastin time (APTT) measurement result
Note: * indicates there is significant difference, and * * indicates there is extremely significant difference
Sample activated partial thromboplastin time assay the result shows that, solvent (dimethyl sulfoxide) has one to blank plasma
Fixing is rung, and positive control aspirin has significant difference when concentration is 5mg/mL.Commodity storax and grafting storax tree
Rouge is compared under high, medium and low concentration with blank plasma, has extremely significant difference, and concentration is higher, the clotting time is longer.
2, grafting storesin volatile component identification:
(1) laboratory apparatus
Water-bath (Jiangsu high honour instrument manufacturing Co., Ltd), heating mantle (Nanjing Cole's experimental instruments and equipment limited), water
Steam distillation device (Nanjing Cole's experimental instruments and equipment limited), gas-chromatography ion trap combination mass spectrograph (the silent winged generation of match you,
ITQ1100)。
(2) experiment reagent
Ether (Chinese medicines group), anhydrous sodium sulfate (Chinese medicines group), standard control (J&K lark prestige).
(3) experimental method
The extraction of volatile oil: grafting storesin 50g is taken respectively, is put into flask, volatile oil extractor and cold is installed
Solidifying device, electricity consumption heating mantle heats extract volatile oil.Volatile oil is dry with anhydrous sodium sulfate.Volatile oil has fragranced.Use ether
It is stand-by after dissolution.
GC-MS analysis condition: quartz capillary column SE-54 (25m × 0.25mm, 0.25 μm, the Chinese Academy of Sciences
Dalian Physical and Chemical Inst.), chromatography column and programmed temperature, after keeping 5min, is warming up to 250 with 5 DEG C/min by 50 DEG C of initial temperature
DEG C and keep 50min, carrier gas N2, pressure is 50kPa before column, and split ratio 1:30, vaporizer and detector temperature are 270 DEG C, into
0.3 μ L of sample amount.Mass spectrum carrier gas He, pressure is 50kPa, split ratio 1:30 before column, 270 DEG C of interface temperature, ion source temperature 200
DEG C, ionization voltage 70eV, mass scan range, 30~400amu.Graft storesin GC-MS ion flow graph as shown in Figure 1,
Chemical component qualification result is shown in Table 4.
Table 4 grafts storesin chemical analysis GC-MS qualification result
3, storesin involatile constituent is grafted:
Grafting storax UPLC/QTOF-MS qualification result is shown in Table 5.
The total ion current figure of storesin is grafted as shown in Fig. 2, the HLPC of grafting storesin is as shown in Figure 3.
Table 5 grafts storesin LC-MS qualification result
4, to the storesin of Turkey's production, formed levant storax oil is identified after ethyl alcohol refines, commodity Soviet Union
Blending GC-MS ion flow graph as shown in figure 4, the GC-MS of volatile component the results are shown in Table 6:
6 commodity storax chemical analysis GC-MS qualification result of table
Analysis through table 4 and table 6 finds that the resin volatile component for grafting storax is mainly longifolene, secondly
For caryophyllene, the two is its main ingredient.α-cubebene, copaene, α-pinene and D-Limonene contain
It measures also relatively high.The main component of commodity levant storax oil is caryophyllene, is secondly Isosorbide-5-Nitrae-Methanoazulene,
Decahydro-4,8,8-trimethyl-9-methylene-, [1S- (1 α, 3a á, 4 α, 8a á)]-, the two be its mainly at
Part.Secondly, 1-Naphthalenol, 1,2,3,4,4a, 7,8,8a-octahydro-1,6-dimethyl-4- (1-
methylethyl)-,[1R-(1α,4á,4aá,8aá)]-、Azetidine,3-methyl-3-phenyl-、α-Cubebene、
The content of Bicyclo [3.1.1] hept-2-ene, 3,6,6-trimethy is also relatively high.
5, involatile constituent detection is carried out to identical commodity levant storax oil, testing result is as shown in table 7:
7 commodity storax LC-MS qualification result of table
The present invention provides a kind of grafting storesins in the application extended in the clotting time, the grafting storax tree
The difference on volatile ingredient and non-volatile component is larger with commodity storesin for rouge, and in 5~50mg/mL dosage range to fibre
Fibrillarin original (FIB), thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) have
Extremely significant extension clotting time effect.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (5)
1. a kind of grafting storesin is in the application extended in the fibrinogen clotting time, the grafting storesin is adopted
From the storax after grafting, the grafting is using storax as scion, using sweetgum as stock.
2. a kind of grafting storesin is in the application extended in thrombin time, after the grafting storesin picks up from grafting
Storax, it is described grafting using storax as scion, using sweetgum as stock.
3. a kind of grafting storesin is in the application extended in prothrombin time, the grafting storesin picks up from grafting
Storax afterwards, the grafting is using storax as scion, using sweetgum as stock.
4. a kind of grafting storesin is in the application extended in activated partial thromboplastin time, the grafting storesin
Storax after picking up from grafting, the grafting is using storax as scion, using sweetgum as stock.
5. the application of any one of Claims 1 to 4, which is characterized in that the application concentration of the grafting storesin for 5~
50mg/mL。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115778990A (en) * | 2022-11-15 | 2023-03-14 | 浙江中医药大学 | Application of storax in preparation of antithrombotic drugs |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105325182A (en) * | 2015-11-23 | 2016-02-17 | 江苏省中国科学院植物研究所 | Propagation method for introducing plant storax into Mediterranean region |
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