CN102579515B - Folium turpiniae ethanol percolating extract, preparation method and application thereof - Google Patents
Folium turpiniae ethanol percolating extract, preparation method and application thereof Download PDFInfo
- Publication number
- CN102579515B CN102579515B CN 201110290522 CN201110290522A CN102579515B CN 102579515 B CN102579515 B CN 102579515B CN 201110290522 CN201110290522 CN 201110290522 CN 201110290522 A CN201110290522 A CN 201110290522A CN 102579515 B CN102579515 B CN 102579515B
- Authority
- CN
- China
- Prior art keywords
- ethanol
- percolation
- extract
- leaf
- percolate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to an application of a folium turpiniae ethanol percolating extract which is used for preparation of drugs capable of preventing and treating flu complication diseases. The complications refer to splenic injury and/or lung injury. The extract is obtained through the extraction of the ethanol percolation of the folium turpiniae. The extracting method includes selecting coarse power of the folium turpiniae, adding 25%-90% of ethanol as an extractant, impregnating the coarse powder for 12-72 hours, percolating at a speed of 1-8ml/kg/minute, collecting percolate with an amount 4-9 times of the crude drug, recycling the ethanol, reducing pressure and concentrating the percolate to thick paste, or drying the percolate to obtain the folium turpiniae ethanol percolating extract.
Description
Technical field
The present invention relates to a kind of acute turpinia leaf ethanol percolation extract and its production and use.
Background technology
Influenza virus is day by day serious at present, and influenza virus and respiratory tract disease and the systemic disease that causes are closely related, China is one of influenza country occurred frequently, not only populous, and living habit also is conducive to the relay of influenza virus, the number of times of falling ill for each person every year reaches 0.3～0.7 time not to be waited, and key population reaches 2～4 times.There is serious threat in influenza to the mankind, especially new kind influenza virus, and it not only causes other infected by microbes of Secondary cases, and can directly cause organ to destroy and the allergy causing death.But effectively do not treat at present method and the treatment means of influenza virus and disease (influenza) thereof, as carry out the inflammatory reaction treatment for clinical symptoms, cure the symptoms, not the disease.The method of present modal prevention and treatment influenza virus is to use influenza virus vaccine inoculation, non-specific anti virus herb or Western medicine oseltamivir phosphate (oseltamivir phosphate capsule)-neuraminidase inhibitor.Yet influenza virus vaccine prophylactic immunization drawback is that the inoculation crowd is selective, is not that everybody can inoculate.In addition, the influenza virus vaccine protective rate is not high, and guard time is shorter (3～6 months) also.Although the non-specific anti virus herb of most lays claim to antivirus action, is not for influenza virus, and antivirus action mechanism is unclear.At many kind treatment affection of exogenous wind-cold of our former studies, the Chinese prescription of the flu that doctor trained in Western medicine is thought does not have the effect of direct resisiting influenza virus.Briefly, the Chinese prescription of not every treatment flu has the effect of clear and definite inhibition influenza virus.Western medicine " oseltamivir phosphate (oseltamivir phosphate capsule; (3R; 4R; amino-3 (1-the third 2-ethoxyethyl acetate)-1-cyclohexene-1 carboxylic acid, ethyl ester phosphate of 5S)-4-acetamide-5-) " be neuraminidase inhibitor, infected by influenza enters cell specific inhibitory effect, but not only expensive, and have that some patients take rear appearance vomiting, feel sick, the side reactions such as insomnia, headache, stomachache, diarrhoea, dizziness, fatigue, nasal obstruction, pharyngalgia and cough.Oseltamivir phosphate capsule is that the esterase that is positioned in vivo liver and intestinal is converted into active metabolite and plays the neuraminidase effect that suppresses, if undesired its drug effect that also will affect of patient's liver and intestinal organ dysfunction, the renal insufficiency patient also will be cautious use of.
Therefore, specific aim suppresses influenza virus and copies, suppresses influenza neuraminidase hydrolysis cell surface sialic acid, thereby cause influenza virus not to be combined with cell surface receptor and to enter in cell, and the minimizing influenza virus will be the cause of disease that treatment influenza virus sexuality is emitted in intracellular generation.
Summary of the invention
The purpose of this invention is to provide a kind of acute turpinia leaf ethanol percolation extract, suppress influenza neuraminidase hydrolysis cell surface sialic acid, cause influenza virus not to be combined with cell surface receptor and to enter in cell, copy thereby can effectively suppress influenza virus, more importantly overcome the side reaction drawback of medicine.
Another object of the present invention has been to provide a kind of preparation method of acute turpinia leaf ethanol percolation extract.
Another object of the present invention has been to provide the purposes of the medicine of acute turpinia leaf ethanol percolation extract preparation prevention and treatment influenza and complication disease thereof.
Another object of the present invention has been to provide the pharmaceutical composition that contains acute turpinia leaf ethanol percolation extract.
Acute turpinia leaf ethanol percolation extract of the present invention, by being extracted, the Turpinia pomifera(Roxb) D O. alcohol percolation obtains, the alcohol percolation extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 25～90% ethanol as extractant, flood after 12～72 hours, carry out percolation with the speed of 1～8ml/kg/ minute, collect the percolate of 4～14 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferably, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 30～85% ethanol as extractant, flood after 24～72 hours, carry out percolation with the speed of 1～7ml/kg/ minute, collect the percolate of 5～13 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferred, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 35～80% ethanol as extractant, flood after 24～48 hours, carry out percolation with the speed of 1～6m/kg/ minute, collect the percolate of 5～11 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferred, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 45～75% ethanol as extractant, flood after 48 hours, carry out percolation with the speed of 1～5ml/kg/ minute, collect the percolate of 5～10 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferably, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 55～70% ethanol as extractant, flood after 36 hours, carry out percolation with the speed of 2～5ml/kg/ minute, collect the percolate of 5～8 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferred, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 65% ethanol as extractant, flood after 36 hours, carry out percolation with the speed of 3～5ml/kg/ minute, collect the percolate of 5 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferably, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 60～70% ethanol as extractant, flood after 12 hours, carry out percolation with the speed of 3～5ml/kg/ minute, collect the percolate of 4～6 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Preferably, concrete extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 70% ethanol as extractant, flood after 72 hours, carry out percolation with the speed of 4～5ml/kg/ minute, collect the percolate of 4 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
The present invention also provides the purposes of acute turpinia leaf ethanol percolation extract for the preparation of the medicine of prevention and treatment influenza and complication disease thereof.
Preferably, be used for suppressing influenza virus.
Preferably, be used for suppressing influenza virus FM1.
Preferably, be used for suppressing neuraminidase.
Preferably, its complication refers to renal failure.
Preferably, its complication refers to splenic trauma or/and injury of lung.
The present invention also provides a kind of acute turpinia leaf ethanol percolation extract preparation, and said preparation is take described acute turpinia leaf ethanol percolation extract as main active.
acute turpinia leaf ethanol percolation extract of the present invention can extract effective inhibition neuraminic acid enzyme component, acute turpinia leaf ethanol percolation extract is along with the using dosage size variation, it suppresses the ability of neuraminic acid enzymatic activity, be also corresponding the changing of height of neuraminic acid enzyme inhibition rate, and one-tenth positive correlation, acute turpinia leaf ethanol percolation extract can be by suppressing the surperficial neuraminidase of influenza virus, and then the inhibition influenza virus enters inside cell, the influenza virus that inhibition has entered inside cell is copied, propagation, thereby reduced the infection of influenza virus to cell, growth, and prevention and treatment influenza and complication thereof.
The specific embodiment
Acute turpinia leaf ethanol percolation extract of the present invention, it is to obtain by the Turpinia pomifera(Roxb) D O. alcohol percolation is extracted, the alcohol percolation extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 25～90% ethanol as extractant, flood after 12～72 hours, carry out percolation with the speed of 1～8ml/kg/ minute, collect the percolate of 4～14 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 1
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 24 hours with 50% ethanol, carry out percolation with the speed of 7ml/kg/ minute, collect the percolate of 11 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 2
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 48 hours with 60% ethanol, carry out percolation with the speed of 2ml/kg/ minute, collect the percolate of 9 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 3
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 48 hours with 65% ethanol, carry out percolation with the speed of 5ml/kg/ minute, collect the percolate of 8 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 4
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 36 hours with 70% ethanol, carry out percolation with the speed of 2ml/kg/ minute, collect the percolate of 6 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 5
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 36 hours with 65% ethanol, carry out percolation with the speed of 4ml/kg/ minute, collect the percolate of 5 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 6
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 12 hours with 70% ethanol, carry out percolation with the speed of 3ml/kg/ minute, collect the percolate of 4 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 7
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 24 hours with 80% ethanol, carry out percolation with the speed of 6ml/kg/ minute, collect the percolate of 5 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Embodiment 8
Preparation method: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, as extractant, flood after 72 hours with 70% ethanol, carry out percolation with the speed of 4ml/kg/ minute, collect the percolate of 4 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Experimental data 1, extraction process optimization research
Leaf of Turpinia pomifera (Roxb) D O. mainly contains the compounds such as flavonoid, organic acid, flavone compound is better with the ethanol extraction effect, therefore, we with concentration of alcohol (A), collect four of amount of alcohol (B), percolation speed (C), dip time (D) as the investigation factor, each factor is established three levels, presses L9 (3
4) orthogonal table carries out orthogonal (table 1, table 2) extraction process is optimized research, investigating index is the total flavones extraction ratio.
Table 1 factor level table
Get 27 parts of leaf of Turpinia pomifera (Roxb) D O. medical materials, every group average three parts, totally nine groups, by table 1L9 (3
4) the orthogonal test table design experiment.The results are shown in Table 2.
Table 2L9 (3
4) orthogonal test table
Variance analysis
Orthogonal experiments is carried out variance analysis, the results are shown in Table 3.
The variance analysis of table 3 take the total flavones extraction ratio as index
＊F
1-0.05(2，2)＝19.0
＊＊F
1-0.01(2，2)＝99.0
Conclusion: intuitive analysis is collected amount of alcohol (B) larger on the extraction effect impact as can be known, and dip time (A), percolation speed (C) and concentration of alcohol (D) are less on the extraction effect impact.Table 3 the results of analysis of variance has further been verified the intuitive analysis conclusion, and namely factor B has considerable influence to extraction effect.Analyze in conjunction with the K value, determine that extraction conditions is: get leaf of Turpinia pomifera (Roxb) D O., after 72 hours, with 3～6ml/kg/ minute percolation, collect 10 times of amount percolates of medical material with 70% alcohol dipping, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Experimental data 2: the inhibitory action of acute turpinia leaf ethanol percolation extract to the neuraminic acid enzymatic activity
Get the acute turpinia leaf ethanol percolation extract that embodiment 4 preparation methoies obtain, add suitable quantity of water and make dissolving, use tiring of neuraminidase inhibitor identification kit mensuration acute turpinia leaf ethanol percolation extract inhibition neuraminidase (N1) and see Table 1.
(1). standard curve is prepared: a. every hole in 96 hole luciferase targets adds 70 μ l neuraminidases to detect buffer; B. every hole adds respectively 0,1,2,5,7.5,10 μ l H5N1 neuraminidases again; C. every hole adds 0～20 μ l Milli-Q water again.
(2). the preparation of sample detection: a. every hole in 96 hole luciferase targets adds 70 μ l neuraminidases to detect buffer; B. every hole adds 10 μ l H5N1 neuraminidases again; C. every hole adds 0～10 μ l acute turpinia leaf ethanol percolation extract sample again; D. every hole adds 0～10 μ l Milli-Q water again.
(3). detecting step:
A. vibrate approximately 1min of mixing;
B.37 ℃ hatch 2min inhibitor and H5N1 neuraminidase are fully interacted, the sample of doing standard curve is also hatched together;
C. every hole adds 10 μ l neuraminidase fluorogenic substrates;
D. vibrate again approximately 1min of mixing;
E.37 carry out fluoremetry after ℃ hatching 20～30min.Excitation wavelength is 360nm, and emission wavelength is 440nm.
(4). calculate sample for H5N1 neuraminic acid enzymeinhibition percentage ratio according to standard curve, and calculate acute turpinia leaf ethanol percolation extract for the IC50 of H5N1 neuraminidase after doing concentration curve.The suppression ratio IC50 that acute turpinia leaf ethanol percolation extract reaches neuraminidase is 13.35mg/ml.See Table 4.
Table 4. acute turpinia leaf ethanol percolation extract suppresses the activity of neuraminidase
Can be clear that according to above-mentioned experimental result:
(1). acute turpinia leaf ethanol percolation extract can extract effective inhibition neuraminic acid enzyme component;
(2). acute turpinia leaf ethanol percolation extract is along with the using dosage size variation, and it suppresses the ability of neuraminic acid enzymatic activity, i.e. also corresponding changing of the height of neuraminic acid enzyme inhibition rate, and become positive correlation;
(3). by above-mentioned experiment as seen, acute turpinia leaf ethanol percolation extract can be by suppressing the surperficial neuraminidase of influenza virus, and then suppress that influenza virus enters the cell the inside, the influenza virus that suppresses to have entered the cell the inside copies, breeds, thereby reduced infection, the growth of influenza virus to cell, and prevention and treatment influenza and complication thereof.
Medical science and study of pharmacy personnel can't not do the inhibition influenza infection, copy in advance, or under the prerequisite of the experiment of inhibition neuraminidase, learn that in advance acute turpinia leaf ethanol percolation extract has prevention and treats the good result that the influenza virus sexuality is emitted.
Experimental data 3: the acute turpinia leaf ethanol percolation extract infected by influenza infects the inhibitory action of Embryo Gallus domesticus
Get acute turpinia leaf ethanol percolation extract, add suitable quantity of water and make dissolving, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that acute turpinia leaf ethanol percolation extract suppresses the ability that the FM1 influenza virus is copied and suppresses in Embryo Gallus domesticus.(1). FM1 influenza virus liquid is inoculated in no-special pathogen chick embryo allantois intracavity in 10 day age, and every embryo 0.2ml was hatched 72 hours for 37 ℃, observed and calculate half Embryo Gallus domesticus infective dose (EID50).(2). acute turpinia leaf ethanol percolation extract adopts the toxic action of Embryo Gallus domesticus, physiological saline solution is done to be inoculated in no-special pathogen chick embryo allantois intracavity in 10 day age after serial dilution to acute turpinia leaf ethanol percolation extract, every embryo 0.2ml, each concentration inoculation 6 embryo, hatch for 37 ℃, observe Embryo Gallus domesticus growth promoter situation, the Cmax that can survive 96 hours with Embryo Gallus domesticus is as the TD of medicine.(3). the inhibitory action of acute turpinia leaf ethanol percolation extract infected by influenza in Embryo Gallus domesticus adopts, 0.1ml influenza virus liquid and different dilution acute turpinia leaf ethanol percolation extract mix, 37 ℃ act on 2 hours, be inoculated in no-special pathogen chick embryo allantoic cavity in 10 day age, every winding kind 6 embryos were hatched 72 hours for 37 ℃.The virus attack amount is 50EID50, establishes simultaneously virus control, physiological saline solution normal control, calculates acute turpinia leaf ethanol percolation extract to the median effective dose (ED50) of viral inhibition.
(1) the .FM1 influenza virus is calculated through the Reed-Muench method the virulence of Embryo Gallus domesticus, and its EID50 is 10
-5.07(2). after acute turpinia leaf ethanol percolation extract was inoculated in Embryo Gallus domesticus, its growth promoter and Normal group were basically identical.Embryo Gallus domesticus was all survived in 96 hours.Embryo Gallus domesticus gives acute turpinia leaf ethanol percolation extract stock solution and has no chicken embryo death, so can think that TD0 is 9.2g/100ml.(3). acute turpinia leaf ethanol percolation extract inhibitory action of infected by influenza in Embryo Gallus domesticus sees Table 5.
Table 5. acute turpinia leaf ethanol percolation extract infected by influenza infects the inhibitory action of Embryo Gallus domesticus
Compare with the virus control group:
*P＜0.05
As shown in Table 5, acute turpinia leaf ethanol percolation extract has significant inhibitory action (P＜0.05) at 22.5～90.0mg/ml infected by influenza, and ED50 is 31.5mg ± 2.76mg/ml, and TI is 13.94 ± 1.42.
Experimental data 4: acute turpinia leaf ethanol percolation extract affects the FM1 influenza virus
Get acute turpinia leaf ethanol percolation extract, add suitable quantity of water and make dissolving, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that acute turpinia leaf ethanol percolation extract suppresses the ability of FM1 influenza virus virulence.(1) .FM1 adopts cell median infective dose (TCID50) micromethod to the toxicity test of Testis et Pentis Canis passage cell (MDCK).(2). acute turpinia leaf ethanol percolation extract adopts serum-free to the toxicity test of mdck cell DMEM does to be inoculated in after serial dilution in the mdck cell hole that forms monolayer to acute turpinia leaf ethanol percolation extract, every hole 100 μ l, each dilution factor repeats 4 holes, establishes simultaneously the normal cell contrast.Culture plate is put 37 ℃, 5%CO
2Cultivate in incubator, observation of cell pathological changes every day (CPE), Continuous Observation 3 days, with " +～++ ++ " record result, press the Reed-Muench method and calculate medicine median toxic concentration (TD50) and maximal non-toxic concentration (TD0).(3). acute turpinia leaf ethanol percolation extract suppresses the effect of FM1 influenza virus and measures: mdck cell 5 * 10
5/ ml, every hole 100 μ l, in 96 orifice plates, 37 ℃, 5%CO
2Cultivate in incubator, suck culture fluid in the hole next day, add 100 TCID50 influenza virus liquid, every hole 100 μ l suck supernatant after 37 ℃ of absorption 1h.Wash 2 times with phosphate buffer (PBS), take the TD0 of medicine as the 1st hole, then with the DMEM liquid of serum-free, acute turpinia leaf ethanol percolation extract is made serial dilution, add respectively in the above-mentioned cell that has infected virus, establish simultaneously virus control and Normal group, 37 ℃, 5%CO
2Cultivate in incubator, observe the mdck cell characteristics of lesion that influenza virus produces every day, i.e. cell monolayer degeneration becomes circle etc., for three days on end, calculates 50% of medicine and suppresses pathological changes concentration (IC50) and therapeutic index (TI).The calculating of TI: TI=TD50/IC50, the TI value is larger, shows that the safety range of medicine is larger.With Kruskal-Walis and Mann-Whitney method of inspection comparative test group and the cytopathic difference of virus control group, drug dose and suppression ratio that virus infected cell is avoided cytopathy (CPE) occurs are carried out correlation analysis, judge whether amount validity response relation.
(1) the .FM1 influenza virus is calculated through the Reed-Muench method the virulence of mdck cell, and its TCID50 is 10
-4.81(2). the TD0 of acute turpinia leaf ethanol percolation extract mdck cell is respectively 1105.8mg ± 23.55mg/100ml.(3). after acute turpinia leaf ethanol percolation extract is made serial dilution, the 100TCID50 influenza virus is carried out inhibition test, calculate median effective dose IC50 and the TI value size of medicine, the results are shown in Table 6.
Table 6. acute turpinia leaf ethanol percolation extract is to the IC50 (mg/100ml) of FM1 influenza virus and TI (x ± s)
As shown in Table 6, the cytopathogenic effect of acute turpinia leaf ethanol percolation extract inhibition FM1 influenza virus all strengthens along with the increase of drug dose.Drug dose and medicine are shown the correlation analysis that the suppression ratio of CPE carries out, and the dosage of acute turpinia leaf ethanol percolation extract and medicine are to being obvious positive correlation between the CPE suppression ratio.
Experimental data 5: the death protection result of acute turpinia leaf ethanol percolation extract to influenza virus infection FM1 strain in Mice Body
Get acute turpinia leaf ethanol percolation extract, use influenza virus A-prime Mus lung adapted strain (FM1) and (H1N1) identify that acute turpinia leaf ethanol percolation extract is to the dead protective effect of influenza virus infection FM1 strain in Mice Body.(1). influenza virus FM1 strain virus is inoculated respectively every group of 10 BALB/C mice, male and female half and half after doing 10 times of doubling dilutions.After the slight anesthesia of ether, give and different dilution viruses respectively every mice collunarium inoculation 20 μ l for every group.Observe the dead mouse situation of 10 days, calculating LD50 by the Reed-Muench method is 10
-1.36Therefore determine that experiment modeling concentration used is 10LD50.(2). the dead protective effect of acute turpinia leaf ethanol percolation extract to influenza virus infection FM1 strain in Mice Body: Normal group, influenza virus FM1 strain virus matched group, acute turpinia leaf ethanol percolation extract 11.25mg/ml, 22.5mg/ml, 45.0mg/ml, 90.0mg/ml dosage group philosophy gavage, gavage capacity only are 0.4ml/.After 3 days, each group is only used 10LD50 influenza virus FM1 strain collunarium infecting mouse 20 μ l/ under the slight anesthesia of ether except Normal group.Normal group gives the normal saline with volume simultaneously.4 groups of administration are continued administration, Normal group and influenza virus FM1 strain virus control group administered physiological saline 8 days, and dosage is the same.Day by day observe the animal morbidity and record death toll, observing altogether 14 days, calculating mortality rate (mortality rate=every group of death toll/every group of total mice * 100%), the results are shown in Table 7.(3). the impact of acute turpinia leaf ethanol percolation extract on influenza virus infection FM1 strain lung index in Mice Body: Normal group, influenza virus FM1 strain virus matched group, acute turpinia leaf ethanol percolation extract 11.25mg/ml, 22.5mg/ml, 45.0mg/ml, 90.0mg/ml dosage group philosophy gavage, gavage capacity only are 0.4ml/.After 3 days, each group is only used 1.0LD50 influenza virus FM1 strain collunarium infecting mouse 20 μ l/ under the slight anesthesia of ether except Normal group.Normal group gives the normal saline with volume simultaneously.4 groups of administration are continued administration, Normal group and influenza virus FM1 strain virus control group administered physiological saline 8 days, and dosage is the same.Put to death mice on the 8th day after viral infection, weigh, get lung and claim lung heavy, calculate lung index (lung index=lung quality/weight * 100%); In addition, get spleen and claim spleen heavy, calculate spleen index (spleen index=spleen quality/weight * 100%), the results are shown in Table 7.
The death protection result of table 7. acute turpinia leaf ethanol percolation extract to influenza virus infection FM1 strain in Mice Body
Annotate: ※ ※ P＜0.01VS influenza virus model group
The impact of table 8. acute turpinia leaf ethanol percolation extract on spleen index and the lung index of influenza virus infection FM1 strain in Mice Body
Annotate: ##P＜0.01VS Normal group ※ p＜0.05VS influenza virus model group
(1). as shown in Table 7, acute turpinia leaf ethanol percolation extract has significant protective effect (p＜0.01) at 45.0～90.0mg/ml influenza virus infected.
(2). as shown in Table 8, acute turpinia leaf ethanol percolation extract has significant effect (p＜0.05) at the lung index of 90.0mg/ml influenza virus infected.
Experimental data 6: the impact of acute turpinia leaf ethanol percolation extract on chronic renal failure
1) SD rat, 200～240g, Shanghai west pul-Bi Kai laboratory animal responsibility company limited, the animal quality certification number: SCXK (Shanghai) 2003-0002
2) reagent and medicine adenine, adenine (content＞98% is the import packing, Chinese Shanghai, lot number 20010520), the acute turpinia leaf ethanol percolation extract that embodiment 4 preparation methoies obtain;
3) test method: male SD rat, about body weight 220g after first feeding 10 days normal growths with normal diet, is divided into Normal group, administration experimental group and modeling matched group by body weight, 12 every group at random.Administration experimental group and modeling matched group are made chronic renal failure (CRF) model with the adenine gavage, with adenine 320mg/ (kgd) gavage, make approximately 2ml/ of suspension, totally 20 days with distilled water; Administration after 20 days, the administration experimental group is with acute turpinia leaf ethanol percolation extract 1.2g/ (kgd) gavage, acute turpinia leaf ethanol percolation extract is made suspension solution (0.2g/ml) with distilled water, each approximately 2ml/ only, the gradation gavage, Normal group and modeling matched group be with equal-volume water gavage, administration after 35 days with the rat etherization. indices is observed in the blood sampling of Mus arteria caudalis.
4) result:
The impact of table 9. acute turpinia leaf ethanol percolation extract on the CRF Renal Function in Rats
Annotate: through the T check, administration experimental group and the comparison of modeling matched group "
*" expression P＜0.05, "
*" expression P＜0.01
This experiment is looked sidelong at after the purine gavage sets up rat CRF model with gland, modeling control rats lethargy, body weight obviously reduces, serum Bun, Ser obviously raise, Hb obviously descends, Ret obviously raises, show that Renal Function in Rats is impaired, after giving the acute turpinia leaf ethanol percolation extract treatment. rat spirit is obviously bestirred oneself, and body weight is recovered gradually, improves creatinine clearance rate, promote the discharge of metabolite, reduce serum BUN, SCr concentration, renal function and anemia are obviously improved, and show that acute turpinia leaf ethanol percolation extract has preventive and therapeutic effect to kidney of rats damage due to adenine.
Can be clear that according to above-mentioned experimental result:
(1). acute turpinia leaf ethanol percolation extract can extract effectively preventing chronic renal failure composition;
(2). acute turpinia leaf ethanol percolation extract has the effect while of suppressing the neuraminic acid enzymatic activity, the effect that also has the chronic renal failure controlled, overcome oseltamivir phosphate capsule sternly to the clear ability of creatinine and renal failure crowd's side effect, otherwise also had prevention effect.
Claims (8)
1. acute turpinia leaf ethanol percolation extract is for the preparation of the purposes of the medicine of prevention and treatment influenza complication disease, described complication refers to splenic trauma or/and injury of lung, wherein said extract obtains by the Turpinia pomifera(Roxb) D O. alcohol percolation is extracted, the alcohol percolation extracting method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 25～90% ethanol as extractant, flood after 12～72 hours, carry out percolation with the speed of 1～8ml/kg/ minute, collect the percolate of 4～9 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
2. purposes as claimed in claim 1, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 30～85% ethanol as extractant, flood after 24～72 hours, carry out percolation with the speed of 1～7ml/kg/ minute, collect the percolate of 5～9 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
3. purposes as claimed in claim 1 or 2, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 35～80% ethanol as extractant, flood after 24～48 hours, carry out percolation with the speed of 1～6ml/kg/ minute, collect the percolate of 5～9 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
4. purposes as claimed in claim 3, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 45～75% ethanol as extractant, flood after 48 hours, carry out percolation with the speed of 1～5ml/kg/ minute, collect the percolate of 5～9 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
5. purposes as claimed in claim 3, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 55～70% ethanol as extractant, flood after 36 hours, carry out percolation with the speed of 2～5ml/kg/ minute, collect the percolate of 5～8 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
6. purposes as claimed in claim 5, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 65% ethanol as extractant, flood after 36 hours, carry out percolation with the speed of 3～5ml/kg/ minute, collect the percolate of 5 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
7. purposes as claimed in claim 1, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 60～70% ethanol as extractant, flood after 12 hours, carry out percolation with the speed of 3～5ml/kg/ minute, collect the percolate of 4～6 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
8. purposes as claimed in claim 1, concrete percolation extraction method is as follows: get the leaf of Turpinia pomifera (Roxb) D O. coarse powder, with 70% ethanol as extractant, flood after 72 hours, carry out percolation with the speed of 4～5ml/kg/ minute, collect the percolate of 4 times of amounts of medical material, reclaim ethanol, be evaporated to thick paste or the dry acute turpinia leaf ethanol percolation extract that gets.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110290522 CN102579515B (en) | 2010-04-13 | 2010-04-13 | Folium turpiniae ethanol percolating extract, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110290522 CN102579515B (en) | 2010-04-13 | 2010-04-13 | Folium turpiniae ethanol percolating extract, preparation method and application thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101448854A Division CN101862358B (en) | 2010-04-13 | 2010-04-13 | Acute turpinia leaf ethanol percolation extract, and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102579515A CN102579515A (en) | 2012-07-18 |
| CN102579515B true CN102579515B (en) | 2013-06-26 |
Family
ID=46469120
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110290522 Active CN102579515B (en) | 2010-04-13 | 2010-04-13 | Folium turpiniae ethanol percolating extract, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102579515B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857343A (en) * | 2006-04-11 | 2006-11-08 | 徐蕾 | Chinese medicine composition for preventing and treating viral cold and throat disease and its preparing method |
| CN1872106A (en) * | 2006-04-18 | 2006-12-06 | 徐蕾 | Application of wild basil circle leaves in treating disease of virulence cold |
| CN101249118A (en) * | 2007-02-25 | 2008-08-27 | 胡军 | Chinese medicine extract and medicine use thereof |
-
2010
- 2010-04-13 CN CN 201110290522 patent/CN102579515B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857343A (en) * | 2006-04-11 | 2006-11-08 | 徐蕾 | Chinese medicine composition for preventing and treating viral cold and throat disease and its preparing method |
| CN1872106A (en) * | 2006-04-18 | 2006-12-06 | 徐蕾 | Application of wild basil circle leaves in treating disease of virulence cold |
| CN101249118A (en) * | 2007-02-25 | 2008-08-27 | 胡军 | Chinese medicine extract and medicine use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102579515A (en) | 2012-07-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bailly et al. | Glycyrrhizin: An alternative drug for the treatment of COVID-19 infection and the associated respiratory syndrome? | |
| ES2882588T3 (en) | Antiviral activity of medicinal mushrooms and their active constituents | |
| Wang et al. | Antiviral activity of puerarin as potent inhibitor of influenza virus neuraminidase | |
| CN101890038B (en) | Nuezhenoside-rhoifolin-hyperin composite and application in medicinal preparation thereof | |
| CN101862357B (en) | Acute turpinia leaf general flavone ethanol reflux extract, and preparation method and application thereof | |
| CN101906122B (en) | Method for extracting rhoifolin and prepared medicament application thereof | |
| CN101891785B (en) | Extraction method of hyperin and application thereof in medicament preparation | |
| CN101884651B (en) | Water extract of turpinia formosana leaves and preparation method and application thereof | |
| CN102633850A (en) | Rhoifolin extraction method and usage of drug prepared by rhoifolin | |
| CN101879197B (en) | Turpinia Montana leaf ethanol reflux extractive, preparation method and application thereof | |
| CN102579515B (en) | Folium turpiniae ethanol percolating extract, preparation method and application thereof | |
| CN101897715B (en) | Nuezhenoside and rhoifolin composition and use thereof in preparation of drugs | |
| CN101899078B (en) | Extraction method of glossy privet fruit glycoside and application of glossy privet fruit glycoside in preparation of medicament | |
| CN101862358B (en) | Acute turpinia leaf ethanol percolation extract, and preparation method and application thereof | |
| CN102935098A (en) | Acute turpinia leaf general flavone ethanol reflux extract, and preparation method and application thereof | |
| CN102920751A (en) | Turpinia arguta leaf total flavonoid ethanol reflux extract, preparation method and uses thereof | |
| CN102526086A (en) | Method for extracting ligustroflavone and application of ligustroflavone to preparation of medicines | |
| CN101890037B (en) | Nuezhenoside and hyperin composition and application thereof for preparing medicine | |
| CN101904859B (en) | Rhoifolin and hyperin composition and application of medicines prepared therefrom | |
| CN102631383A (en) | Turpinia arguta seen leaf ethanol diacolation extract, preparation method thereof and application | |
| CN102631382A (en) | Turpinia arguta seen leaf ethanol diacolation extract, preparation method thereof and application | |
| CN102920752A (en) | Turpinia arguta leaf ethanol reflux extract, preparation method and uses thereof | |
| CN101862359A (en) | Acute turpinia leaf general flavone ethanol reflux extract, and preparation method and application thereof | |
| CN102784177A (en) | Turpinia montana leaf ethanol reflux extractive, preparation method and application thereof | |
| CN102784178A (en) | Turpinia montana leaf general flavone ethanol reflux extractive, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |