CN103393752A

CN103393752A - New application of palchouli oil

Info

Publication number: CN103393752A
Application number: CN2013103228941A
Authority: CN
Inventors: 彭成; 魏晓露; 万峰; 林大胜
Original assignee: Chengdu Huanshen Group Co Ltd; Chengdu University of Traditional Chinese Medicine
Current assignee: Chengdu Huanshen Group Co Ltd; Chengdu University of Traditional Chinese Medicine
Priority date: 2012-07-30
Filing date: 2013-07-29
Publication date: 2013-11-20
Also published as: WO2014019486A1; CN103191183A

Abstract

The invention discloses application of palchouli oil in preparation of antiviral healthcare foods, foods, cosmetics, disinfectants or everyday cosmetics. The invention also discloses the antiviral healthcare food, food, the cosmetic, the disinfectant or the everyday cosmetic which comprises palchouli oil (an active ingredient), as well as acceptable materials or auxiliary ingredients of the healthcare foods, the foods, the cosmetics, the disinfectants or the everyday cosmetics. The palchouli oil disclosed by the invention has remarkable antiviral effect, can treat the diseases caused by viruses, has definite treatment effect on on viral myocarditis and pulmonitis, and has strong practical application value.

Description

The new purposes of oil of Herba Pogostemonis

Technical field

The present invention relates to the new purposes of oil of Herba Pogostemonis, particularly the purposes in the purposes of oil of Herba Pogostemonis in preparing antiviral health food, food, cosmetics, disinfectant or cosmetics of everyday use.

Background technology

By the infectious disease that virus causes, serious threat always the mankind's life with healthy.According to statistics, the infectious disease more than 80% is to be caused by virus.Viral disease has related to clinical medical every field, and viral and disease that cause becomes the focus that many subjects are paid close attention at present, and modal in virus infection is exactly by catching that Respirovirus and enterovirus are caused.

Respirovirus is that a large class can be invaded respiratory tract and causes the respiratory tract local patholoic change, or only take respiratory tract as portal of entry, causes the virus of respiratory tissues organ lesion, is the main pathogens that causes the acute respiration road transmission.At present, knownly can cause that the virus of acute respiratory infection has kind more than 10, comprises influenza virus, parainfluenza virus, adenovirus, herpes simplex virus, respiratory syncytial virus etc.According to statistics, 90% above acute respiratory infection causes by virus, and take upper respiratory tract infection as clinically commonly encountered diseases and frequently-occurring disease, in the infected, severe patient is bronchiolitis and pneumonia, can cause acute respiratory distress and heart failure, even causes death especially.Respirovirus easily morphs, and it is pathogenic thereby increase, or causes some new diseases.Wherein influenza virus (influenza) is the main Causative virus that causes acute infections of respiratory tract disease influenza, influenza have infectiousness strong, propagate fast, the crowd characteristics such as susceptible repeatedly.According to the up-to-date epidemic situation circular of World Health Organization (WHO) issue, only this year so far first type HINI influenza oneself causes in the whole world that to surpass 10,000 people dead, its in the Central Asia, the activity of South Asia region continues to strengthen especially.China is the multiple district of influenza, and popular or local the breaking out of influenza all can occur every year basically, makes people more than 100,000,000 suffer the puzzlement of influenza, and because influenza surpasses 500,000 people to the hospitalier of hospital, the society that causes and financial burden are difficult to statistics.The harm of adenovirus also can not be ignored, adenovirus AdV(Adneovirus) be the common virus of childrens respiratory tract infection, the disease that the most often causes is pharyngoamygdalitis, secondly be pneumonia, gastroenteritis, bronchitis and otitis media, also can cause pharynx-conjunctivitis, encephalitis, keratitis, enteritis etc., case fatality rate is very high.

Coxsackie virus (Coxsaekieviurs) belongs to Picornaviridae (Picomaviridea), and enterovirus genus (Enterroviurs) can be divided into A group and B group according to its difference to the neonatal rat pathogenecity.Coxsackie B virus (Coxsackievirusgroup B, CVB) can cause epidemic pleurodynia, aseptic encephalitis, meningoencephalitis and pericarditis, especially CVB3 is the main pathogens of viral myocarditis, can cause that focal necrosis occurs in cardiac muscular tissue, and with inflammatory cell infiltration, the pathological changes such as myocardial cell cracking, persistent viral infection can cause DCM (dilated cardiomyopathy), to the mankind, has brought great threat.

Since the nineties in 20th century, research worker is found many new antiviral drugs, the antiviral drugs of application has surpassed more than 20 kinds clinically at present, but its definite curative effect still has very large dispute, and after antiviral chemosynthesis class long term combined use, virus easily produces drug resistance to it, has now become the thorny problem for the treatment of clinically viral disease.The conventional medicine virazole (ribavirin) that is used for the control viral infection, by suppressing phosphoric acid time fast core dehydrogenase, thereby suppress virus replication now, and early stage medication is effective.But there is medullary cell toxicity in the virazole vein while giving, therefore preferably only by a small amount of aerosol, through respiratory tract, give, but leukopenia often appears in clinical practice, can cause the side effect such as headache, irreversible anemia, serum bilirubin rising when dosage is excessive, zoopery has the report of teratogenesis.Therefore, the research of antiviral drugs has important practical significance, and the research of antiviral drugs of finding a kind of efficient, safety, few side effects is imperative.

The natural resources of Chinese medicinal materials of China is very abundant, and applicating history is long, has accumulated a lot of data and experience in long-term practice.Certainly, toxic and side effects is little for Chinese Herbs, and the medicine source is abundant and cheap.Propose to have heat clearing away in theory of Chinese medical science, Detoxication, as single medicines such as Flos Lonicerae, Folium Isatidis, Rhizoma Atractylodis, the Radixs Astragali, and the compound medicine such as Radix Isatidis electuary can be made the most of the advantage at anti-virus aspect.Experiment in vitro and clinical practice prove, most viruses are responsive to Chinese herbal medicine, as use treatments by Chinese herbs such as Respirovirus (influenza virus, Measles virus, mumps virus), enterovirus (the scorching virus of ridge ash, rotavirus, Coxsackie virus), sooty mould poison (epidemic encephalitis B virus, encephalitis), can obtain good therapeutic effect.The general pharmacological action principle is by suppressing virus replication, stop virus to cause the pathological changes of cell, regulate immunization, improve pulmonary circulation, the comprehensive functions such as minimizing and the pathogenic position of elimination inflammatory exudate, make disease of viral infection sx↓ or recovery from illness, so Traditional Chinese Medicine Anti virus there are potential advantages and wide prospect.

Herba Pogostemonis is the perennial thorn stamen of Labiatae (Labiatae) herbaceous plant Herba Pogostemonis (Pogostemon cablin (Blanco) Benth.), have another name called branch fragrant, originate in the Tropical Asia such as Philippine, existing China, India, Japan, Indonesia, Malaysia, Madagascar, Brazil, Paraguay and the Russia etc. of extensively being distributed in.At present wide flower bud perfume (or spice) in Guangzhou, Zhaoqing, Zhanjiang and the Hainan in Guangdong, all there are cultivation in the provinces (district) such as Guangxi, Sichuan, be one of " ten Da Nan medicines ", its medical material commodity are divided into board perfume (or spice) (Guangzhou product), branch fragrant (Zhaoqing product), fragrant (Hainan product) 4 kinds of profound fragrant (Zhanjiang product) and south by place of production difference.Its acrid in the mouth, slightly warm in nature, return spleen, stomach, lung meridian,, with all herbal medicine, has eliminating turbid pathogen with aromatics, the preventing or arresting vomiting that whets the appetite, delivers the effect of expelling summer-heat.Be usually used in the vomiting of turbid damp obstructing in middle-JIAO, gastral cavity painful abdominal mass, the heat-damp in summer asthenia, uncomfortable in chestly do not relax, cold-damp is closed summer-heat, stomachache vomiting and diarrhoea, nasosinusitis headache, affection of exogenous wind-cold.The volatile oil of Herba Pogostemonis has antiinflammatory, analgesia and antibacterial activity.

Herba Pogostemonis is the dry aerial parts of labiate Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., contains volatile oil, the same Herba Pogostemonis of effect.Modern pharmacology shows, Herba Pogostemonis has the effects such as resisting pathogenic microbes, antiinflammatory, analgesic, analgesia, be the main component of the treatment such as HUOXIANG ZHENGQI SHUI, antivirus oral liquid influenzas, flu Chinese patent medicine commonly used, but its effective substance is still not clear and there are no resisiting influenza virus effect report in its body.The volatile oil of Herba Pogostemonis has antibacterial action.

Summary of the invention

The object of the present invention is to provide the new purposes of oil of Herba Pogostemonis, and the health food take oil of Herba Pogostemonis as active component, food, cosmetics, disinfectant or cosmetics of everyday use.

Purposes in the purposes of oil of Herba Pogostemonis of the present invention in preparing antiviral health food, food, cosmetics, disinfectant or cosmetics of everyday use.

Wherein, described virus is influenza virus, CVB-3 and/or adenovirus.

Wherein, described health food, food, cosmetics, disinfectant or cosmetics of everyday use are health food, food, cosmetics, disinfectant or the cosmeticses of everyday use of prevention, treatment or auxiliary treatment viral infection.

Described health food, food, cosmetics, disinfectant or cosmetics of everyday use are that prevention, treatment or auxiliary treatment respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis are or/and the health food of enteritis, food, cosmetics, disinfectant or cosmetics of everyday use.

Described respiratory tract infectious disease is that rhinitis, pharyngitis, tonsillitis, laryngitis, tracheitis are or/and bronchitis.

Described oil of Herba Pogostemonis derives from the volatile oil of Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. extraction.

The described oil of Herba Pogostemonis that comes from Herba Pogostemonis Pogostemon cablin (Blanco) Benth., patchouli alcohol content is not less than 40%(w/w), Pogostone content is not less than 20%(w/w).

Described oil of Herba Pogostemonis is to be prepared as follows: the herb of getting Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., be ground into coarse powder, add water, soak, adopt extraction by steam distillation, obtain.

The antiviral health food of the present invention, food, cosmetics, disinfectant or cosmetics of everyday use, it is take oil of Herba Pogostemonis as active component, adds the preparation that acceptable adjuvant on health food, food, cosmetics, disinfectant or cosmetics of everyday use or complementary composition are prepared from.

Described preparation is liquid preparation, gas preparation, solid preparation, semi-solid preparation.

In described preparation, the content of oil of Herba Pogostemonis is 0.1%～100%(w/w).

Oil of Herba Pogostemonis of the present invention has antiviral effect, especially the inhibitory action of infected by influenza, CVB-3 and/or adenovirus is remarkable, acute respiratory infection and other inflammation that virus is caused have therapeutical effect, wherein,, to the determined curative effect of viral myocarditis, pneumonia, has good potential applicability in clinical practice.

Obviously,, according to foregoing of the present invention,, according to ordinary skill knowledge and the customary means of this area,, not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite, can also make modification, replacement or the change of other various ways.

The specific embodiment of form, be described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.

The specific embodiment

The preparation of embodiment 1 oil of Herba Pogostemonis of the present invention

Get Herba Pogostemonis Pogostemon cablin (Blanco) Benth. herb, pulverized the 20-40 mesh sieve, the distilled water that adds 10-14 times of weight, soak after 1-5 hour, adopt extraction by steam distillation 2-6 hour, obtain oil of Herba Pogostemonis of the present invention, claim again patchouli oil, wherein, contain patchouli alcohol (C ₁₅H ₂₆O) must not be lower than 40%w/w, Pogostone must not be lower than 20%w/w.

The preparation of embodiment 2 oil of Herba Pogostemonis of the present invention

Get Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. herb, pulverized the 20-40 mesh sieve, add the distilled water of 10-14 times of weight, soak after 1-5 hour, adopt extraction by steam distillation 2-6 hour, obtain oil of Herba Pogostemonis of the present invention.

Oil of Herba Pogostemonis of the present invention also can extract according to pharmacopeia (version in 2005) appendix XD determination of volatile oil method, also can adopt organic solvent extraction, supercritical CO ₂The prior aries such as extraction are extracted, and perhaps by buying commercially available prod, obtain.

The preparation of embodiment 3 collutorys of the present invention

Get the oil of Herba Pogostemonis of embodiment 1 or embodiment 2 preparations,, with propylene glycol, Tween-60, distilled water, glycerol, surfactant, isopropyl alcohol, after being uniformly dispersed, filter, get filtrate, packing namely obtains collutory.The preparation of embodiment 4 cosmetics of the present invention

Get the oil of Herba Pogostemonis of embodiment 1 or embodiment 2 preparations, with appropriate lanoline, glyceryl monostearate, vaseline, Cera Chinensis, stearic acid, liquid paraffin, benzoic acid, triethanolamine, propylene glycol, distilled water, Tween-60, sodium metabisulfite, essence, antiseptic, cerebrin peptide, fruit acid sodium, hexadecanol, after evenly mixing, cooling storage, obtain cosmetics.

The preparation of embodiment 5 disinfectants of the present invention

The oil of Herba Pogostemonis of getting embodiment 1 or embodiment 2 preparations is appropriate, with Tween-60, sodium carboxymethyl cellulose, Macrogol 200, distilled water, mixes, and obtains disinfectant.

Embodiment 6 cosmetics of everyday use of the present invention---the preparation of abluent

Get the oil of Herba Pogostemonis of embodiment 1 or embodiment 2 preparations, add appropriate carboxylic acid polyalcohol, non-ionic surface active agent, anion surfactant, enzyme, enzyme stabilizers, foam controller, tween 80, water to prepare.

Embodiment 7 cosmetics of everyday use of the present invention---the preparation of handwashing liquid

Get the oil of Herba Pogostemonis of embodiment 1 or embodiment 2 preparations, with glycerol, sodium benzoate, sodium chloride, deionized water, essence, triethanolamine, glyceryl monostearate, hydrogenated lanolin, stearic acid, ethyl hydroxybenzoate, michelia essence, after being uniformly dispersed, fill, obtain handwashing liquid.

Embodiment 8 food of the present invention

Get oil of Herba Pogostemonis and the Fructus Myricae rubrae of embodiment 1 or embodiment 2 preparations, add appropriate Pericarpium Citri tangerinae powder, Cortex cinnamomi japonici powder, Saccharum Sinensis Roxb., Flos Caryophylli XIANGFEN, Radix Glycyrrhizae powder, Fructus Foeniculi XIANGFEN, Sal and Alumen, prepare glazed waxberry.

Get Fructus Myricae rubrae and be ready for and first put the approximately thick Fructus Myricae rubrae of 20cm of one deck afterwards, then put Sal, Alumen, patchouli alcohol that one deck mixes, compress at once.Later same alternate Fructus Myricae rubrae and the Sal Alumen patchouli alcohol mixture put, after compression is pickled the Fructus Myricae rubrae base.Getting the Fructus Myricae rubrae base need be with after Saccharum Sinensis Roxb., spice powder, Pericarpium Citri tangerinae powder, Cortex cinnamomi japonici powder, Flos Caryophylli XIANGFEN, Radix Glycyrrhizae powder, Fructus Foeniculi XIANGFEN, then, through immersion, dry in the sun, spice, packing, obtains glazed waxberry.

Embodiment 9 health food of the present invention

Get the oil of Herba Pogostemonis of embodiment 1 or embodiment 2 preparations, add appropriate sodium carboxymethyl cellulose, with in right amount through withering trough wither, two pots of green-keeping machines complete, Herba Artemisiae Scopariae, Fructus Amomi, Fructus Lycii, Fructus Citri Sarcodactylis, Folium Isatidis fresh tea, Flos Chrysanthemi, Semen Cassiae, Flos Lonicerae, the Rhizoma Atractylodis Macrocephalae, Fructus Corni, the Radix Paeoniae Alba, Folium Mori, Fructus Schisandrae Chinensis, Fructus Foeniculi, the Rhizoma Dioscoreae of kneading machine after kneading mixed homogeneously, and prepare liver-nourishing tea.

Below prove beneficial effect of the present invention by the test of pesticide effectiveness:

Experimental example 1 oil of Herba Pogostemonis antiviral study in vitro of the present invention

1, material and instrument

1.1 cell strain and Strain

1.1.1 cell strain:

Mdck cell (Madin-Darby canine kidney(cell line)), Hep-2 cell (people's laryngeal carcinoma epithelial cell), HeLa cell (cervical cancer cell), available from Chinese medicine and biological products assay institute.

1.1.2 Strain:

Influenza A virus is selected the reassortant virus strain, and serotype is that H1N1(is hereinafter to be referred as influenza virus), be derived from national influenza center; Coxsackie virus is selected B group 3 types (hereinafter to be referred as CVB-3), is derived from visiting center, Sichuan Province; Adenovirus is selected common 3 types (Ad-3), by Gansu Province Disease Control and Prevention Center, is presented.

1.2 medicine and control drug

Trial drug: the patchouli oil of invention medicine: embodiment 1 preparation.

Positive drug: ribavirin injection: specification is 100mg/ml, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H19992467 of traditional Chinese medicines, product batch number: 10110521; The injection acyclovir: specification is the 0.25g/ bottle, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H20034034 of traditional Chinese medicines, product batch number: 10122403.

1.3 culture medium, reagent and consumptive material

RPMI-1640 culture medium (GibcoTM company, lot number 1313945); DMEM culture medium (Gibco ^TMCompany, lot number 1345538); Hyclone (HyClone biochemistry goods (Beijing) company limited, lot number NVM0347); Trypsin Amresco company); Penicillin G sodium, the aseptic Tissue Culture Flask of streptomycin and 96 porocyte culture plates (U.S. Corning company).1,5, the 10ml Dispette, the disposable rifle head of 200 μ l, 1ml (the biological consumptive material company in Haimen, Jiangsu)

1.4 key instrument

Water isolation type constant temperature CO ₂Incubator (model MCO-15AC, SANYO GS company);

OLYMPUS inverted microscope (model C KX41, Japanese Olympus company);

Micropipettor (French GILSON company produce);

Water isolation type electro-heating standing-temperature cultivator (model GSV-DA-1, Huangshi, Hubei Province medical apparatus and instruments factory);

Horizontal centrifuge (model LXJ-II, Shanghai medical analytical instrument factory);

Electronic balance (model JA-2603, Shanghai balance equipment factory).

2, test method

2.1 the pre-treatment of medicine and positive control

With 0.5% tween 80 hydrotropy, be first the white liquid preparation with invention medicine patchouli oil, its initial concentration is that 20%(patchouli oil density is 1.012g/ml, concentration 202.4mg/ml).With 0.5% tween 80, this white liquid is done 1/10 dilution before using, standby after filtration sterilization.

To and for the positive control medicine injection acyclovir of adenovirus, with sterilized water, do 1/10 dilution (being 10mg/ml) with standby for the positive control drug 'Libaweilin ' injection of influenza virus and Coxsackie virus.

The mensuration of 2.250% histiocyte infective dose

Influenza virus and adenovirus are diluted to the virus liquid of variable concentrations with 3% hyclone DMEM, Coxsackie B virus 3 use 3% hyclone RPMI-1640.

Influenza virus inoculation mdck cell, Coxsackie B virus 3 strain inoculation HeLa cells, adenovirus inoculation Hep-2 cell, in 37 ℃, 5%CO ₂After cultivating 7d in incubator, the observation infection cell is justified cytopathy (cytopathiceffect such as contracting, come off, CPE) situation, cause the infective dose (50%tissue culture infective dose, TCID50) of 50% pathogenic histiocyte to measure each virus.

2.3 antiviral experiment

Add 100 * TCID50 influenza virus liquid 0.1ml in the MDCK of 96 well culture plates cell monolayer, add 100 * TCID50CVB-3 virus liquid 0.1ml and add 100 * TCID50Ad-3 virus liquid 0.1ml in the HeLa cell monolayer in the Hep-2 cell monolayer, in 37 ℃, 5%CO ₂Adsorbed 2 hours, and inhaled and abandon virus liquid, add respectively the invention medicine and the positive control drug that have diluted, each concentration is done 3 multiple holes.Experiment arranges the infection contrast of not administration of infection equivalent virus and the normal cell contrast of virus-free infection simultaneously.Each culture plate is in 37 ℃, 5%CO ₂Continue to cultivate 5d, every day, microscopically was observed CPE.

3. experimental result and pharmacodynamic index calculate

3.1 the TCID that each is viral ₅₀Value

Approximately occur CPE in 3 days after influenza virus inoculation mdck cell, approximately occurred CPE in 5 days after Coxsackie B virus 3 inoculation HeLa cells and adenovirus Ad-3 inoculation Hep-2 cell.It is 10 that the TCID50 of influenza virus tires ^-4, the TCID50 of Coxsackie B virus 3 and adenovirus Ad-3 tires and is 10 ^-12

3.3 invention Antiviral Effect experiment effect and pharmacodynamic index calculate

3.3.1 experimental result

Experimental result is as shown in table 1:

Table 1 invention Antiviral Effect experimental result (n=3)

As can be seen from Table 1, the inhibition of oil of Herba Pogostemonis infected by influenza of the present invention, Coxsackie B virus 3 strains and adenovirus is good.Reach identical viral suppression ratio (75%), the consumption of oil of Herba Pogostemonis of the present invention is 0.092mg/ml, and the consumption of positive drug is 0.156mg/ml, is 1.7 times of consumption of oil of Herba Pogostemonis of the present invention.

Description of test, the effect that oil of Herba Pogostemonis of the present invention suppresses influenza A virus, CVB-3 and adenovirus is good.

Experimental example 2 oil of Herba Pogostemonis interior resisting virus experiment of the present invention

1, material and instrument

1.1 Strain

1.2 medicine and control drug

Trial drug: the patchouli oil of invention medicine: embodiment 1 preparation.

1.3 animal: mice Bab/c level is male, body weight 10 scholar 2g, male and female half and half

Mice Bab/c level is male, body weight 18 scholar 2g, male and female half and half

1.4 culture medium, reagent and consumptive material

RPMI-1640 culture medium (Gibco ^TMCompany, lot number 1313945); DMEM culture medium (Gibco ^TMCompany, lot number 1345538); Hyclone (HyClone biochemistry goods (Beijing) company limited, lot number NVM0347); Trypsin Amresco company); Penicillin G sodium, the aseptic Tissue Culture Flask of streptomycin and 96 porocyte culture plates (U.S. Corning company).1,5, the 10ml Dispette, the disposable rifle head of 200 μ l, 1ml (the biological consumptive material company in Haimen, Jiangsu)

Ether (analytical pure, the Long Huagongshijichang of Chengdu section, lot number: 20111215); 0.9% normal saline (Cologne, Chengdu pharmaceutical Co. Ltd, lot number: H120115)

Lactic acid dehydrogenase (LDH) is measured test kit, and bio-engineering research institute, lot number: 20121013 are built up in Nanjing; Superoxide dismutase (SOD) is measured test kit, and bio-engineering research institute, lot number: 20121012 are built up in Nanjing; Malonaldehyde (MDA) is measured test kit, and bio-engineering research institute, lot number: 20121010 are built up in Nanjing; The CK-MB(creatine kinase isozyme), TNF-α (tumor-activated factor-alpha) measures test kit, U.S. RD company, lot number: 201210

1.4 key instrument

OLYMPUS inverted microscope (model C KX41, Japanese Olympus company);

Micropipettor (French GILSON company produce);

Electronic balance (model JA-2603, Shanghai balance equipment factory);

Microplate reader, U.S. Thermo Scientific company, model: Varioskan Flash.

2, test method

2.1 the pre-treatment of medicine and positive control

With 0.5% tween 80 hydrotropy, be first the white liquid preparation with invention medicine oil of Herba Pogostemonis, its initial concentration is that 20%(oil of Herba Pogostemonis density is 1.012g/ml, concentration 202.4mg/ml).With 0.5% tween 80, this white liquid is done 1/10 dilution before using, standby after filtration sterilization.

2.2 invention medicine interior resisting virus experiment

The foundation of model 2.2.1 the viral infection animal is caused a disease

Get 40 of Bab/c mices, the SPF level, body weight 10 scholar 2g, be divided into 4 groups at random, 10 every group.After the injection site routine disinfection, by group respectively former times of lumbar injection, 1/2 and 1/4 CVB3 virus liquid 0.3ml, Normal group adopts 0.3ml normal saline lumbar injection.

Separately get the Bab/c mice, SPF level, body weight 18 scholar 2g, random packet, 10 every group.First with ether, slightly anaesthetize by group, then respectively the collunarium infector doubly, 1/2 and 1/4 influenza virus liquid or adenopathy venom 1ml, Normal group adopts 1ml normal saline collunarium.

Animal housing keeps air fresh, relative humidity 60%, 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, observe morbidity and the death condition of mice, and record it, to determine whether successfully to set up the pathogenic model of CVB3, influenza virus and adenovirus infection animal.

The model test 2.2.2 invention medicine interior resisting virus causes a disease

2.2.2.1 invention medicine interior resisting virus Myocarditis Model test

Get 140 of Bab/c mices, body weight 10 scholar 2g, be divided into 7 groups at random, 20 every group.Make marks.

With patchouli oil clinical injection consumption and LD ₅₀For foundation, set up respectively the medication therapy groups of high, medium and low, ultralow 4 kinds of concentration in the safe dose scope, i.e. the 1.25%(0.127g/kg of medicine original concentration), 0.95%(0.096g/kg), 0.625%(0.063g/kg) and 0.48%(0.049g/kg).Normal group, CVB3 virus model matched group and ribavirin positive drug matched group (positive drug concentration is 0.15g/kg) are set simultaneously.

With injected in mice position routine disinfection, except Normal group, other respectively organize the required CVB3 virus liquid of lumbar injection virus model, and Normal group adopts the normal saline lumbar injection, and injection volume is identical with other papova liquid injection volumes.Start intraperitoneal injection after 24h, injection volume is 0.1ml/10g, every day 1 time.Animal housing keeps air fresh, relative humidity 60%, and 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, after treating continuously 7 days, stop administration and observe the 15th day.Observe morbidity sign and the death condition of mice every day, take body weight and record.

Started at the same day by the treatment administration, and put to death 4 from each treated animal is random respectively when the 5th day, the 10th day and the 15th day, eye socket is got blood and is collected, and takes out heart, liver, lung, kidney, and after removing surperficial blood with filter paper, weighing, calculate conscience lung kidney body rate.Then each internal organs are got 100mg, with the abundant homogenate of RPMI-1640 cell maintenance medium 2ml.After tissue homogenate is used the filtering with microporous membrane degerming, multigelation three times, the centrifugal 30min of 3000rPm, get supernatant 0.1ml and be inoculated in 96 orifice plates that grow into monolayer Hela cell, and each sample is done 3 multiple holes, 37 ℃, 5%CO2 discards liquid in hole after hatching 2h, and PBS adds 1640 cell maintenance mediums after washing 3 times, 37 ℃, 5%C02 cultivates 72h-120h.Every day, observation of cell growing state and corresponding CPE degree, do not occur that CPE person still is the virus-free infection person without CPE 3 times through blind passage.Eye socket is got the blood that blood collects in 4 ℃ of placements, spend the night, the centrifugal 20min of 3000rpm, get serum and carry out Biochemical Indices In Serum and detect.

2.2.2.2 anti-pulmonary inflammation model test in invention medicine body

Get 140 of Bab/c mices, body weight 18 scholar 2g, random packet, 20 every group.Make marks.

Take the clinical gastric infusion consumption of patchouli oil as foundation, set up respectively the medication therapy groups of high, medium and low, ultralow 4 kinds of concentration in the safe dose scope, i.e. the 10%(1.27g/kg of medicine original concentration), 5%(0.508g/kg), 2.5%(0.254g/kg) and 1.25%(0.127g/kg).Normal group, influenza virus model control group, adenovirus model control group, ribavirin positive drug matched group and acyclovir positive drug matched group are set simultaneously.

Except Normal group, model group infects for respectively the mice collunarium by modeling success required influenza virus and adenovirus virus quantity, and Normal group adopts the normal saline collunarium, and the collunarium amount is identical with other papova liquid infective doses.Start gastric infusion after 24h, the gavage amount is 0.3ml/10g, every day 1 time.

Animal housing keeps air fresh, relative humidity 60%, and 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, after treating continuously 7 days, stop administration and observe the 15th day.Observe morbidity sign and the death condition of mice every day, take body weight and record.

Started at the same day by the treatment administration, put to death 4 dissections from each treated animal is random respectively when the 5th day, the 10th day and the 15th day, perusal phase pulmonary lesion situation, after winning full lung and removing surperficial blood with filter paper, weighing, calculate its lung body rate.Then grind the corresponding sensitive cells of Mus lung inoculation, to determine that whether the pneumonopathy change is by due to this test viral infection.

3, result of the test

3.1 Mice Body inner virus model is set up

With former times, 1/2 and 1/4 CVB3 virus liquid intraperitoneal injection of mice, 0.3ml/ only, observes and finds that mice hair before injecting virus is glossy, and activity, diet, defecation are normal.After injectable drug, compare with the Normal group mice, the minimizing that came into play at the 2nd～3 day of former times of virus liquid group mice, dietary amount descends, and loses weight, hair tarnishes, and frizzle appears, and died off in the 6th～7 day, all dead during by the 8th day, dissect and find that heart is obviously little than normal group mice, simultaneously visible milky point-like, streak class fat-like pathological changes under the heart adventitia.1/2 former times of concentration group and 1/4 former times of concentration group mice also appearance activity minimizing in the time of the 2nd～3 day, dietary amount descends, and loses weight, but started the increase activity in the time of the 5th day, and dietary amount increases, and state returns to normal group mice state gradually.Illustrate set up viral myocarditis model need to be with former times of CVB3 virus liquid intraperitoneal injection of mice, 0.3ml/ is only.

With former times, 1/2 and 1/4 influenza virus liquid, adenopathy venom collunarium infecting mouse respectively, 1ml/ only, observe to find that model mice compares with the Normal group mice, former times of virus liquid group mice started to occur successively death at collunarium the same day, and be all dead in about the 4th day; 1/2 former times of concentration group reduces appearance activity in the 2nd～3 day, and dietary amount descends, and hair tarnishes, and frizzle occurs, death in succession occurs on the 5th～6 day, and is all dead in the time of the 7th day, dissects and finds that pulmonary lesion is obvious.Malaise symptoms appearred in 1/4 former times of concentration group at the 2nd day, but in the 3rd～4 day state, in succession took a turn for the better.Illustrate that influenza virus and the adenovirus of setting up the pulmonary inflammation model needs are 1/2 former times of concentration group, 1ml/ only.

3.2 invention medicine interior resisting virus effect

3.2.1 the effect of invention medicine to myocarditis mice model

3.2.1.1 ordinary circumstance impact

Experimental result is as shown in table 2:

Table 2 each experimental mice body weight and death condition

Group	0d	5d	10d	14d	Total death toll
						Normal group	12.0±0.5	13.3±0.5	15.0±0.5	18.9±2.1	0/20
Model group	12.5±0.4	10.3±0.5	12.0±1.5	17.5±0.5	18/20
						Positive group (0.15g/kg)	12.0±0.5	11.8±0.7	12.6±1.2	15.3±1.0	8/20
Invention medicine 1.25% (0.127g/kg)	11.5±0.6	9.8±0.7	15.0±1.5	16.8±1.5	14/20
						Invention medicine 0.95% (0.096g/kg)	12.0±0.5	9.9±0.2	10.8±0.7	17.2±1.6	6/20
Invention medicine 0.63% (0.063g/kg)	11.8±0.5	10.5±0.5	12.1±1.2	14.5±1.2	8/20
						Invention medicine 0.48%(0.049g/kg)	12.0±0.5	10.5±0.5	11.5±1.0	13.8±1.2	13/20

The action of normal group mice is normal, and while to observation, finishing, mortality rate is 0%;

Model group, positive group and invention medicine group mice when modeling finishes, all occur losing weight in various degree, and companion's fur is not damp, movable, the minimizing of ingesting, the increase of urinating;

The model group mice started to die off in the 6th～7 day, observed while finishing, and mortality rate is up to 90%;

The symptom of positive drug group and invention medicine group decimal all alleviates, and activity, food ration increase, and body weight starts to rise:

Wherein, the mortality rate of positive group is 40%; The mortality rate of invention medicine 1.25% is 70%; The mortality rate of invention medicine 1.25% is 30%; The mortality rate of invention medicine 0.63% is 40%; The mortality rate of invention medicine 0.48% is 65%.

Oil of Herba Pogostemonis of the present invention, to the increase along with consumption of the therapeutic effect of viral myocarditis, first raises, rear reduction, and the effect take consumption as 0.096g/kg is optimum.

When patchouli oil using dosage of the present invention was 0.063～0.096g/kg, therapeutic effect was better than positive drug (0.15kg/g), illustrated that medicine of the present invention is good to the therapeutic effect of viral myocarditis.

3.2.1.2 the impact of invention medicine on mice cardiopulmonary kidney liver body rate

Experimental result is as shown in table 3:

Body weight and the cardiopulmonary kidney liver body rate result (n=4) of each experimental mice of table 3 different sample times

Time

Project

Normal group

Model group

Positive group

1.250%

0.950%

0.625%

0.480%

5d

Body weight

12.3±0.5

10.3±0.5

11.8±0.7

9.8±0.7

9.9±0.2

10.5±0.5

?

Heart body rate

0.6315±0.0235

0.6597±0.0663

0.7184±0.0042

0.5996±0.0046

0.6821±0.0112

0.6076±0.0848

0.6419±0.0109

?

Lung body rate

0.8166±0.0450

0.8887±0.0783

0.8332±0.0399

0.9207±0.0077

0.7389±0.0453

0.9248±0.0257

0.8686±0.0932

?

Kidney body rate

1.3447±0.0286

1.6533±0.0867

1.5537±0.102

1.4497±0.0234

1.1958±0.0611

1.5233±0.1719

1.5800±0.0665

?

Liver body rate

4.4300±0.3908

3.7796±0.3976

4.1572±0.4694

4.4444±0.5096

5.0274±0.9181

3.8457±0.2019

5.2962±1.0982

10d

Body weight

15.0±0.5

12.0±1.5

12.6±1.2

15.0±1.5

10.8±0.7

12.1±1.2

11.5±1.0

?

Heart body rate

0.5877±0.0601

0.5232±0.0546

0.5922±0.0922

0.5253±0.0192

0.5843±0.0277

0.5388±0.0812

0.5606±0.0564

?

Lung body rate

0.7314±0.0653

0.7073±0.1145

0.7642±0.0773

0.7402±0.0998

0.7428±0.0332

0.8637±0.1201

0.7274±0.0124

?

Kidney body rate

1.3330±0.0909

1.2767±0.2100

1.4587±0.1702

1.2835±0.1712

1.3658±0.0299

1.4435±0.9087

1.2995±0.1021

?

Liver body rate

5.5674±0.9655

4.7726±1.3837

4.4343±0.9207

4.8972±0.2366

5.1760±0.8779

3.8525±2.0364

4.3800±0.34

14d

Body weight

18.9±2.1

17.5±2.0

15.3±1.0

16.8±1.5

17.2±1.6

14.5±1.2

12.8±1.2

?

Heart body rate

0.4601±0.0312

0.4960±0.0213

0.4881±0.0505

0.4555±0.0368

0.4807±0.0093

0.5966±0.0557

0.4641±0.0634

?

Lung body rate

0.6754±0.0846

0.6954±0.264

0.7439±0.0892

0.8072±0.0385

0.6291±0.0150

0.7214±0.0854

0.7187±0.1611

?

Kidney body rate

1.0997±0.0570

1.1669±0.0690

1.3179±0.1507

1.2525±0.0993

1.3243±0.0551

1.2476±0.0229

1.1804±0.1223

?

Liver body rate

4.3529±0.2331

4.4303±0.8590

4.5229±1.2381

4.1260±0.8362

5.5778±0.0487

5.0745±0.5338

4.0730±1.0212

Normal group mice cardiopulmonary kidney liver device is red and gloss all;

Model group heart color is dim and dwindle and simultaneously with visceral pericardium, milky point-like, streak class fat-like occur, the lungs enlargement, and Liver and kidney is aobvious the variation;

The heart of positive group and each concentration group of invention medicine occurs that the dim color that narrows down to of color recovers gradually, the trend that visceral pericardium milky point-like, streak class fat-like reduce, and lungs enlargement degree alleviates.

Patchouli oil of the present invention can be alleviated every organ disease, and the color that can effectively dwindle heart is dim to recovering gradually normal, further illustrates patchouli oil of the present invention and can alleviate viral myocarditis.

3.2.1.3 the impact of invention medicine on the mice serum biochemical indicator

The mice serum of virus inoculation 5d, 10d and 14d is done Biochemical Indices In Serum detect, normal group is compared with model group, and LDH, CK-MB and TNF-alpha levels and MDA content are significantly low, SOD active obviously high (P＜0.01); Positive group is compared with model group with each concentration group of invention medicine, LDH, CK-MB and TNF-alpha levels and MDA content have reduction in various degree, the SOD activity increases (P＜0.01) in various degree, and positive group and each concentration group of invention medicine are compared LDH, CK-MB and TNF-alpha levels and MDA content in various degree increase are arranged with normal group, and the SOD activity reduces (P＜0.05) in various degree.The LDH of normal group and CK-MB level are along with the increase of time changes not quite, and the SOD activity has less increase, and MDA content and TNF-alpha levels obviously reduce; The LDH of model group, CK-MB level are along with the increase of time obviously reduces, and SOD activity, MDA content and TNF-alpha levels significantly increase; The LDH of positive group, 0.95% and 0.625% concentration group and CK-MB level are along with the less reduction of the increase of time, and SOD is active to be increased, and MDA content and TNF-alpha levels obviously reduce, and the index variation all is tending towards the normal group level; The LDH of 1.25% and 0.48% concentration group, CK-MB level, MDA content and TNF-alpha levels are along with the increase of time reduces, and SOD is active to be increased, and the index variation is tending towards the model group level.Each experimental mice Biochemical Indices In Serum the results are shown in Table 4.

The Biochemical Indices In Serum (n=4) of each experimental mice of table 4 different sample times

(* and model group compare, P＜0.01, # and normal group compare, P＜0.05)

As can be seen from Table 4, the invention medicine can significantly reduce LDH, CK-MB and TNF-alpha levels and MDA content, improves SOD active, with model group, compares, significant difference (P＜0.01), further verified the therapeutical effect of patchouli oil of the present invention to viral myocarditis.

3.2.2 the effect of invention medicine to the Pneumonia Mice model

3.2.2.1 ordinary circumstance impact

After viral collunarium infected 2～3 days, losing weight all appearred in virus group mice in various degree, and companion's fur is not damp, movable, the minimizing of ingesting, the increase of urinating; The model group mice started to die off in the 6th～7 day, only remained 2 after 12 days until observe end; 4 concentration group Mouse Weights of invention medicine alleviate obvious than model group, Mortality appearred in ribavirin, the positive group of acyclovir and each concentration group mice of invention medicine in 5～6 days, positive group mice takes a turn for the better gradually in 7～8 days (after being drug withdrawal 1 day) initial states, activity, food ration increase, and body weight starts to rise.Although and each concentration group mice partial status of invention medicine takes a turn for the better to some extent.

3.2.2.2 the impact of invention medicine on the mouse lung index

Invention Drug therapy pneumonia the results are shown in Table 5,6.

In table 5 medicine body, resisiting influenza virus effect body weight and lung index compare (n=4)

As can be seen from Table 5, the invention drug level greater than 5% the time, can effectively improve the lung index, illustrates that the invention medicine can effectively treat the pneumonia that influenza virus causes.

In table 6 medicine body, Antiadenovirus body weight and lung index compare (n=4)

As can be seen from Table 6, the invention medicine can effectively improve the lung index, illustrates that the invention medicine can effectively treat the pneumonia that adenovirus causes.

To sum up, oil of Herba Pogostemonis has antiviral effect, the resistance of infected by influenza, CVB-3 and adenovirus is clear and definite, can treat respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis and enteritis, wherein, to the determined curative effect of viral myocarditis, pneumonia, pharmacological action is strong, and market application foreground is good.

Claims

1. the purposes in the purposes of oil of Herba Pogostemonis in preparing antiviral health food, food, cosmetics, disinfectant or cosmetics of everyday use.

2. purposes according to claim 1, it is characterized in that: described virus is influenza virus, CVB-3 and/or adenovirus.

3. purposes according to claim 1, it is characterized in that: described health food, food, cosmetics, disinfectant or cosmetics of everyday use are health food, food, cosmetics, disinfectant or the cosmeticses of everyday use of prevention, treatment or auxiliary treatment viral infection.

4. purposes according to claim 3, it is characterized in that: described health food, food, cosmetics, disinfectant or cosmetics of everyday use are that prevention, treatment or auxiliary treatment respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis are or/and the health food of enteritis, food, cosmetics, disinfectant or cosmetics of everyday use.

According to claim 4 for, it is characterized in that: described respiratory tract infectious disease is that rhinitis, pharyngitis, tonsillitis, laryngitis, tracheitis are or/and bronchitis.

6. according to claim 1～5 described purposes of any one is characterized in that: described oil of Herba Pogostemonis derives from the volatile oil that Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. extract.

7. purposes according to claim 6 is characterized in that: the described oil of Herba Pogostemonis that comes from Herba Pogostemonis Pogostemon cablin (Blanco) Benth., patchouli alcohol content is not less than 40%(w/w), Pogostone content is not less than 20%(w/w).

8. according to claim 1～7 described purposes of any one, it is characterized in that: described oil of Herba Pogostemonis is to be prepared as follows: the herb of getting Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., be ground into coarse powder, add water, soak, adopt extraction by steam distillation, obtain.

9. an antiviral health food, food, cosmetics, disinfectant or cosmetics of everyday use, it is characterized in that: it is take oil of Herba Pogostemonis as active component, adds the preparation that acceptable adjuvant on health food, food, cosmetics, disinfectant or cosmetics of everyday use or complementary composition are prepared from.

10. health food according to claim 9, food, cosmetics, disinfectant or cosmetics of everyday use, it is characterized in that: described preparation is liquid preparation, gas preparation, solid preparation, semi-solid preparation.

11. health food according to claim 10, food, cosmetics, disinfectant or cosmetics of everyday use is characterized in that: in described preparation, the content of oil of Herba Pogostemonis is 0.1%～100%(w/w).