CN102743432A - Application of patchouli oil in preparation of medicines used for treating colpitis - Google Patents
Application of patchouli oil in preparation of medicines used for treating colpitis Download PDFInfo
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- CN102743432A CN102743432A CN2012102315651A CN201210231565A CN102743432A CN 102743432 A CN102743432 A CN 102743432A CN 2012102315651 A CN2012102315651 A CN 2012102315651A CN 201210231565 A CN201210231565 A CN 201210231565A CN 102743432 A CN102743432 A CN 102743432A
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Abstract
The invention discloses application of patchouli oil in preparation of medicines used for treating colpitis. The invention also discloses a pharmaceutical composition which is a medicament prepared from the active component of patchouli oil and pharmaceutically acceptable adjuvants or adjuvant components. Patchouli oil has a substantial curative effect on colpitis and has a strong practical application value.
Description
Technical field
The present invention relates to the new purposes of oil of Herba Pogostemonis, belong to pharmaceutical sanitary field.
Background technology
Vaginitis is the inflammation of connective tissue under vaginal mucosa and the mucosa, is the common disease of gynecological's outpatient service.Normal health women, because anatomy and biochemistry characteristics, vagina has the nature defense function to the intrusion of pathogen, when the natural defense function of vagina is destroyed, then pathogen is easy to invade, and causes colpitis.At present, be main to colpitic treatment with infection, antiinflammatory, be aided with naturopathy simultaneously, Therapeutic Method commonly used has medicinal vaginadouche, part to put medicine, oral medication etc.Common vaginitis has bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, senile vaginitis.
Bacterial vaginitis is a kind of mixed infection due to the intravaginal normal flora imbalance, is one group of syndrome that intravaginal has a large amount of antibacterials, changes with the vaginal secretion properties; Cardinal symptom is that vaginal secretions increases, and is lean homogeneous charge shape or scattered paste shape, is canescence or lark; The stench flavor of fish is arranged; The sexual intercourse postemphasis can be with slight pruritus vulvae or burn feeling, and lower abdominal pain, dyspareunia or dysurea can take place the somebody.The common medicine of treatment bacterial vaginitis is wide spectrum antibacterium chemical medicine such as antibiotic such as tetracycline and fasigyne.
Monilial vaginitis is the vaginitis that is caused by monilial infection, and main pathogenic bacterium are Candida albicans.Whether candidiasis is a yeast, is the condition pathogenic fungus, and it is pathogenic to be relative, the fall ill height that depends on body immunity and quantity, the virulence of infectious bacteria.In normal body, its quantity is few, is oval, is in commensalism with body, does not cause disease.When body in the influence of some physiology of body, pathological factor down, when Abwehrkraft des Koepers or immunity reduced, poised state was destroyed, candidiasis transfers the mycelia phase to mutually by yeast, breeds at local raised growth, causes vaginitis.Show as leucorrhoea grow in quantity, pudendum, pruritus of vagina, burning sensation, painful urination, Chang Fahong, edema around the pudendum, epidermis changes varied; Very shallow vesicle pimple can take place, and occurs in groups; It is rotten to the corn also can to form the eczema shape, and be confined to pudendum or extend to around perineum, the anus towards periphery and strand reproduction pleat, until femoribus internus, appearance, the acute or subacute eczema of all fours; Mucosa thickens near labia and the clitoris, and the skin surface flushing that contacts with each other is rotten to the corn; Can cause small white pustule individually, ulcer, vulvodynia and regional glandular enlargement take place when serious.The common medicine of treatment monilial vaginitis is antibiotic and broad-spectrum antifungal chemical medicine such as fluconazol, itraconazole such as nystatin.
Trichomonal vaginitis (trichomonas vaginitis) is common vaginitis, and per vaginam trichomonacide causes.With leucorrhoea grow in quantity, the rare foam of matter, dirty smelly, pruritus of vagina is main performance clinically.Visible vaginal mucosa is congested during inspection, and severe patient has the hemorrhage speckle that is dispersed in, and posterior fornix has the volume leucorrhea, is lark, yellow-white wash or is the yellow green purulent secretion, normal show bubble.The common treatment medicine is deinsectization chemical medicine such as metronidazole.
Senile vaginitis is common in postclimacteric elderly woman, and because of the ovarian function decline, estrogen level reduces; The vaginal wall atrophy, the mucosa attenuation, glycogen content reduces in the epithelial cell; The intravaginal pH value rises, and local resistance reduces, and pathogenic bacterium are invaded breeding easily and cause inflammation.Cardinal symptom is that vaginal secretions increases and pruritus vulvae, burning sensation.Inspection sees that vagina is senile change, last atrophoderma, and pleat disappears, epithelium smoothed, poor.Vaginal mucosa is congested, and little petechia is arranged, and sees shallow table ulcer sometimes; If ulcer surface and offside adhesion; Adhesion can be separated and causes bleeding during examination per vagina, can cause stricture of vagina even locking when adhesion is serious, even inflammation secretions inadequate drainage can form the pyrocolpos pyometra.Replenishing a small amount of estrogen is the Therapeutic Principle of senile vaginitis, and vaginal mucosa is thickened, and builds up resistance.
At present, the colpitic medicine of treatment commonly used is antibiotic and extensive pedigree antibiotic mostly, and side effect is big, and the meeting of being widely used produces drug resistance.The Chinese medicine wide material sources, toxicity is little, and drug effect is long, and therefore, the colpitic new drug that from Chinese herbal medicine, seeks treatment is very necessary.
Herba Pogostemonis Pogostemon cablin (Blanco) Benth. is the Labiatae herbaceos perennial; It is fragrant to have another name called branch; Originate in Philippine; Existing China, India, Japan, Indonesia, Malaysia, Madagascar, Brazil, Paraguay and the Russia etc. of extensively being distributed in, patchouli oil is that leaf, branch bar and the root of Labiatae herbaceos perennial Herba Pogostemonis Pogostemon cablin (Blanco) Benth. perhaps carried oil through the cured leaf of fermentation process through the vapor distillation gained.Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. is a labiate, contains volatile oil, the same Herba Pogostemonis of effect.
Patchouli oil mainly contains patchouli alcohol and Pogostone (total amount accounts for more than 40%); Have effects such as antiinflammatory, antiallergic, raising are immune, antibiotic, analgesia, spasmolytic, antioxidation, emesis; Oil of Herba Pogostemonis mainly contains estragole (accounting for more than 80%); Effect (Liu Liangfeng etc., " research overview of patchouli oil and oil of Herba Pogostemonis ", Chinese Chinese medicine information magazine the 16th the 2nd phase of volume of February in 2009) with antibiotic, spasmolytic, calmness and leukocyte increasing.Wherein, Bibliographical information about patchouli oil and oil of Herba Pogostemonis in-vitro antibacterial is more, as, path etc. not; " Herba Pogostemonis essential oil is to the bacteriostatic activity research of plant pathogenic fungi "; Chinese crude drug volume o. 11th November the 27th in 2004 has reported that patchouli oil all has inhibitory action in various degree to 13 kind of plant pathogenic fungi, and is wherein, stronger to the inhibitory action of Lignum Santali Albi stey, tomato early blight bacterium, sclerotinite; Path etc. not; " the antibacterial action researchs of two kinds of patchouli oils "; Contemporary Chinese Chinese medicine the 13rd the 9th phase of volume of JIUYUE in 2011 discloses patchouli oil and can suppress escherichia coli, staphylococcus epidermidis, staphylococcus aureus, proteus vulgaris, bacillus subtilis, bacillus pyocyaneus, 8 kinds of antibacterials of enterococcus faecalis, and Penicllium chrysogenum, aspergillus niger and 3 kinds of funguses of Candida albicans; Yang Depo etc.; " the anti-skin bacterium activity and the The Chemical Constituents of Herba Pogostemonis and Herba Pogostemonis Volatile oil "; The 18th the 4th phase of volume of JOURNAL OF MICROBIOLOGY December in 1998 discloses the vitro inhibition effect to 16 kinds of skin bacteriums of Herba Pogostemonis and Herba Pogostemonis Volatile oil, like drying rod bacillus, set micrococcus luteus etc.; " Herba Pogostemonis and Herba Pogostemonis Volatile oil to dermatophytosis and condition cure the disease the inhibitory action of fungus ", Chinese Pharmaceutical Journal the 35th the 1st phase of volume of January in 2000 has reported that oil of Herba Pogostemonis and patchouli oil have inhibitory action to mycetes such as dermatophytosis such as red mentagrophytes and Aspergillus fumigatus; Zhang Guangwen etc., " Herba Pogostemonis essential oil chemical composition analysis and antibacterial activity thereof (II) ", Chinese herbal medicine 2002 the 33rd the 3rd phase of volume discloses the antibacterial action of patchouli oil to antibacterials such as pathomycete such as aspergillus niger, Aspergillus flavus and escherichia coli.Existing research about patchouli oil and oil of Herba Pogostemonis antibacterial bacteriostatic focuses mostly in plant pathogen and dermatophytosis, and main purpose is to provide antibiotic external used new drug.
Summary of the invention
The object of the present invention is to provide the new purposes of oil of Herba Pogostemonis, and be the pharmaceutical composition of active component with the oil of Herba Pogostemonis.
At first, the invention provides the purposes of oil of Herba Pogostemonis in the colpitic medicine of preparation treatment.
Wherein, said medicine is the medicine of treatment monilial vaginitis or trichomonal vaginitis.Preferably, said medicine is the monilial vaginitis that the treatment Candida albicans causes.
Wherein, 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.
Wherein, described oil of Herba Pogostemonis derives from the volatile oil that Herba Pogostemonis Pogostemon cablin (Blanco) Benth. extracts, and contains patchouli alcohol in this volatile oil and must not be lower than 26.0%w/w.
Wherein, Described oil of Herba Pogostemonis is prepared by following method: 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; The employing steam distillation extracts, and promptly gets oil of Herba Pogostemonis.
The present invention also provides a kind of treatment colpitic pharmaceutical composition, and it is to be active component with the oil of Herba Pogostemonis, adds the medicament that acceptable accessories or complementary composition are prepared from.
Wherein, described medicament is external preparation, oral formulations or ejection preparation.Preferably, said external preparation is lotion, suppository, liniment or liniment.
Wherein, the content of oil of Herba Pogostemonis is 0.1%～100%w/w in the said preparation.
Oil of Herba Pogostemonis of the present invention has the colpitic effect of treatment, especially monilial vaginitis and trichomonal vaginitis is had definite curative effect, has good potential applicability in clinical practice.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.
Below, foregoing of the present invention is remake further detailed description through the specific embodiment of embodiment form.But should this be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following instance.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
Description of drawings
Fig. 1 medicine of the present invention is to the trichomonal vaginitis ultrastructure effects.Figure A is the normal ultrastructure without any pharmaceutically-active trichomonal vaginitis, and wherein 1 is nucleus, and 2 is rough endoplasmic reticulum, and 3 is over hydrogenation enzyme body, and 4 is the flagellum transverse section; Figure B is to scheming the Change of Ultrastructure that H is a trichomonal vaginitis behind the drug effect, a: cavity, b: autophagic vacuole, c: perinuclear space broadening; D: dilatation of rough endoplasmic reticula, e: chromatin gathers, f: ribosome disappears, g: nuclear membrane disappears; Nucleus is disintegrate gradually, h: karyopycnosis, i: organelle disintegrate; J: after birth damages, and endochylema spills, and polypide is dead.
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, add the distilled water of 10-14 times of weight, soak after 1-5 hour, adopt steam distillation to extract 2-6 hour, promptly get oil of Herba Pogostemonis of the present invention, wherein contain patchouli alcohol (C
15H
26O) must not be lower than 26.0%.
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 steam distillation to extract 2-6 hour, promptly get 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
2Prior aries such as extraction are extracted, and perhaps obtain through buying the commercially available prod.
Below prove beneficial effect of the present invention through the test of pesticide effectiveness:
The activity of experimental example 1 oil of Herba Pogostemonis treatment monilial vaginitis of the present invention
1, material and instrument
1.1 experimental strain
1. type strain: Candida albicans type strain (ATCC14053), from DSMZ of U.S. clinical laboratory; Candida albicans type strain (CICC32819) and Candida albicans type strain (CICC31284) are available from Chinese industrial microorganism fungus kind preservation administrative center.
2. clinical separation strain: 14 strain Candida albicans are located away from the out-patient of Sichuan Province gynecological of healthcare hospital for women & children vaginal secretions, and identify that through the KC-16 of BIO-KONT company lath and drug sensitive test paper are accredited as Candida albicans during in JIUYUE, 2011 to the December in.
1.2 medicine
Trial drug: oil of Herba Pogostemonis of the present invention (according to the method preparation of embodiment 1).
Positive drug: the Mycoporin vaginal sheet (the favorable to the people pharmaceutical Co. Ltd in Jinan, lot number: 1108148), JIEERYIN XIYE (Sichuan En Wei pharmaceutical Co. Ltd, lot number: 1107102).
1.3 laboratory animal
Mice (SPF level) [Chengdu University of Traditional Chinese Medicine Animal Experimental Study center, production licence number: SCXK (river) 2008-11 use the quality certification number: SCXK (river) 2008-049].
1.4 culture medium, reagent and consumptive material
Sabouraud's agar (every liter contains chloromycetin 0.1g, Huankai Microbes Tech Co., Ltd., Guangdong).
Estradiol benzoate injection (Tianjin gold credit aminoacid company limited, lot number: 1007061), disposable sterilized culture dish (the healthy biological company limited in Jiangsu) etc.
1.5 key instrument
Multiple spot inoculation appearance (Japan's assistant is made institute, SAKUMA MIT-P type for a long time), laboratory is with autoclave (SANYO; The MLS-3780 type); CO2INCUBATOR (SANYO, MOC-15A), energy-saving and purifying workbench (the non-blue ultra-clean chemical industry journey company limited of the new light in Chengdu).
2, experimental technique
2.1 the activity in vivo of Drug therapy monilial vaginitis of the present invention
2.1.1 the foundation of mice monilial vaginitis model
Tried the bacterium activation: CICC32819 is inoculated in sabouraud's agar with the Candida albicans type strain, cultivates 48h for 37 ℃, and is subsequent use after the activation.
Model preparation: through preliminary experiment each item factor that influences modelling is optimized, confirms the method for building up of monilial vaginitis model.In formal experiment; Choose 70 of SPF kunming mices, body weight (20 ± 2) g, wherein 60 before infection 6d begin; Back subcutaneous injection 1mg/ml estradiol benzoate oil preparation 0.1ml next day of every mice, and in last 1 injection back inoculation Candida albicans 1.5 * 10
7CFU/, repeated infection is 1 time behind the 24h.And 48h and 72h get vaginal secretions respectively and carry out smear for microscopic examination behind last 1 subinfection, observe mycelia and blastopore and generate situation, are inoculated in simultaneously on the sabouraud's agar flat board, observe the growing state of bacterial strain; And observe variation such as vaginal secretions.In the end get mouse vagina secretions with inoculating loop behind the 1 subinfection 48h and be inoculated on the sabouraud's agar flat board and cultivate 24h ~ 48h, observe the dull and stereotyped colony growth situation that goes up Candida albicans, microscopic examination mycelial growth situation.If growth has typical Candida albicans in flat board, microscopically is observed mycelia and blastopore, explains that the monilial vaginitis model mice prepares successfully.
2.1.2 experiment is divided into groups
After treating mice modeling success; Every mice carries out vaginadouche with the 0.1ml physiological saline solution, and flushing liquor is carried out colony counting, according to mouse vagina colony counting result mice is divided into 6 groups at random; Comprise high, medium and low three dose groups of medicine of the present invention; Positive drug clotrimazole group and JIEERYIN group, model group, 10 every group.In addition with 10 mices that do not infect Candida albicans as blank.Wherein respectively organize the situation analysis of infecting Candida albicans in the mouse vagina and see table 1.
Table 1: each organizes the situation analysis that mouse vagina infects Candida albicans
Annotate:
AExpression relatively has extremely significantly statistical significance (P＜0.01) with blank control group.
2.1.2 the treatment of monilial vaginitis model mice
Adopt medicine intravaginal administration method that the monilial vaginitis model mice is treated, promptly adopt every day the invention medicine of high, medium and low three dosage that the infection model mice is treated respectively, every day 1 time, 20d continuously.Positive drug is used clotrimazole and JIEERYIN respectively, and model group is used normal saline.Before administration (0d) respectively; And behind 3d, 5d and the 7d administration 6h with 0.1ml physiological saline solution washing vagina; Candida albicans in the flushing liquor is carried out colony counting; The situation of change of clump count in the mouse vagina is observed medicine killing or inhibitory action infecting mouse intravaginal Candida albicans before the statistical analysis administration behind (0d), the administration 3d, behind the administration 5d and behind the administration 7d.Every day, mycelia and the blastopore growing state and the growing state in the Sha Shi flat board of vaginal secretions were observed in inoculation behind administration 7d, and the situation of change of observing every day such as mouse vagina hyperemia, redness and secretions etc.Add up healing time and the cure rate of administration 14d and administration 20d respectively.On the sabouraud's agar flat board, do not see albicans growth with continuous 3d vaginal content, and vagina do not have secretions, do not have clinical symptoms such as obviously hyperemia, swelling and be judged to be healing, the no longer medication of healing mice.
2.2 the external activity of medicine anti-candida albicans of the present invention
The preparation that pastille is dull and stereotyped: adopt coubling dilution that medicine oil of Herba Pogostemonis of the present invention is diluted; Be diluted to 1:2 ~ 1:2048 totally 11 gradients respectively; In disposable sterilized culture dish, add the medicinal liquid 1ml of variable concentrations gradient and the sabouraud's agar of 14ml sterilization respectively, promptly the dilution gradient of each medicine in flat board is respectively 35.00mg/ml ~ 34.18 μ g/ml, fully mixing; The oven dry back is subsequent use, and is dull and stereotyped to add equivalent physiologic saline for substitute medication preparation positive control in dull and stereotyped.
Bacterium liquid configuration: Candida albicans type strain and clinical separation strain are transferred to bacterial concentration 1.5 * 10 with physiological saline solution
6CFU/ml.
The mensuration of minimum inhibitory concentration (MIC): adopt the pastille dull and stereotyped and positive control flat board of multiple spot inoculation appearance, and make negative control with physiologic saline for substitute bacterium liquid with the bacterium liquid adding variable concentrations gradient of experimental strain.37 ℃ of constant temperature culture 24h ~ 48h observe the growing state of each bacterial strain in containing variable concentrations gradient medicine flat board.
The result judges: with the medicine Cmin of no albicans growth in dull and stereotyped for this reason medicine to the minimum inhibitory concentration of this bacterial strain.
3, experimental result
3.1 medicine activity in vivo of the present invention
Medicine of the present invention is seen table 2 to the result that influences of monilial vaginitis model mice intravaginal albicans growth, healing time and cure rate analysis in table 3.
Table 2: medicine of the present invention is to the influence of monilial vaginitis model mice intravaginal albicans growth
Annotate: before 0d, 3d, 5d, 7d represent administration respectively, after the administration the 3rd day, the 5th day and the 7th day.
Compare with blank control group,
*P＜0.01; With comparison before the administration,
▲P＜0.05,
▲ ▲P＜0.01; Compare with 3d after the administration,
★P＜0.05.
Can know by table 2; Before the administration; Compare with blank control group, model group, positive drug group and the high, medium and low dose groups intravaginal of the present invention clump count obviously increase, and have obvious statistical significance (P＜0.01); And the clump count between each group of model group and experimental group relatively, all not statistically significant (P＞0.05).
After the administration, compare with model group, high, medium and low dose groups of medicine of the present invention and clotrimazole group intravaginal clump count obviously descend; Compare with positive group JIEERYIN, high, medium and low dose groups of medicine of the present invention and clotrimazole group intravaginal clump count obviously descend; Compare with the positive drug clotrimazole, high, medium and low three dose groups of medicine of the present invention have anti-candida albicans activity in the similar body, and along with the prolongation of administration time, clump count is obvious downward trend, and the state of an illness takes a turn for the better gradually, and curative effect is superior to JIEERYIN.
Comparison each dose groups administration front and back and clump count situation of change of different dosing time, with comparison before the administration, 3d, 5d, 7d bacterium colony number average significance reduce (P＜0.05) after the high dose group administration; 3d, 5d, 7d bacterium colony number average significance reduce (P＜0.05) after the middle dose groups administration; The 3d clump count significantly reduces (P＜0.05) after the low dose group administration, and 5d and 7d clump count utmost point significance reduce (P＜0.01) after the administration.Compare with administration 3d, the bacterium colony number average of 5d and 7d significantly reduces (P＜0.05) after the middle dose groups administration.
Experimental result explanation medicine oil of Herba Pogostemonis of the present invention can significantly be killed the intravaginal Candida albicans of monilial vaginitis.
Table 3: medicine of the present invention is to the healing situation analysis of monilial vaginitis
Annotate: in medication 14d statistical result, the healing time of not curing mice in the 14d is designated as 15d.
In medication 20d statistical result, the healing time of not curing mice in the 20d is designated as 21d.
Compare with model group,
*P＜0.05,
*P＜0.01.
Can know by table 3:
During medication 14d, the high, medium and low dose groups of medicine of the present invention average cure time to the vaginitis model mice in 14d is respectively 11.50d, 11.70d and 13.10d, and cure rate is respectively 70.00%, 60.00% and 40.00%; Relatively, medicine high dose group healing time of the present invention significantly reduces (P＜0.05) with model group (the average spontaneous recovery time is 14.30d, and the spontaneous recovery rate is 10%), and the high, medium and low dose groups cure rate of the present invention significantly improves; Compare there was no significant difference with the positive drug JIEERYIN; Relatively, present dose-effect relationship to a certain degree between in each dose groups.
During medication 20d, the high, medium and low dose groups of medicine of the present invention average cure time to the vaginitis model mice in 20d is respectively 12.90d, 13.20d and 15.40d, and cure rate is 80%; Relatively, medicine of the present invention is high, all utmost point significance minimizings (P＜0.01) of middle dose groups healing time with model group (the average spontaneous recovery time is 18.90d, and the spontaneous recovery rate is 30%), and the high, medium and low dose groups cure rate of the present invention significantly improves; Compare there was no significant difference with the positive drug JIEERYIN; Relatively, all there is not obvious statistical significance between in each dose groups.
Experimental result explanation oil of Herba Pogostemonis of the present invention has the effect of treatment monilial vaginitis.
3.2 medicine external activity of the present invention
Adopt the agar plate doubling dilution to measure invention medicine, positive drug clotrimazole and JIEERYIN 3 strain Candida albicans type strains and 14 strains are come from the minimum inhibitory concentration of the clinical separation strain of vaginitis infected patient, the result sees table 4.
Table 4: medicine of the present invention is to the oidiomycetic antibacterial activity in vitro of white
Annotate: "-" representes that the Cmax of this medicine in flat board do not have obvious antibacterial activity to the Candida albicans of correspondence.Wherein inventing the Cmax of medicine in flat board is 35.00mg/ml; The Cmax of clotrimazole in flat board is 1.67mg/ml; JIEERYIN is done 15 times of dilutions with former medicine in flat board.
Can know by last table; Medicine of the present invention all has stronger antifungal activity to 3 strain Candida albicans type strains and clinical separation strain; Wherein the minimum inhibitory concentration to 3 strain Candida albicans type strains is 0.07mg/ml, is that (meansigma methods of minimum inhibitory concentration is 0.10 ± 0.06mg/ml) to 0.07 ~ 0.27mg/ml to the minimum inhibitory concentration that comes from the scorching infected patient separated strain of clinical vaginal.The positive drug clotrimazole does not have obvious antibacterial activity to 1 strain Candida albicans type strain ATCC14053 and 2 strain clinical separation strains, explains that strain has produced drug resistance to a certain degree to clotrimazole to the part Candida albicans.And JIEERYIN does not have obvious antibacterial activity external to Candida albicans.Description of test medicine oil of Herba Pogostemonis of the present invention has obvious external bacteriostasis to Candida albicans.
Experiment and experiment in vitro presentation of results oil of Herba Pogostemonis of the present invention can significantly suppress Candida albicans in the above-mentioned body, have the effect of treatment monilial vaginitis.
The activity of experimental example 2 oil of Herba Pogostemonis treatment trichomonal vaginitis of the present invention
1 material and instrument
1.1 experiment worm strain
Trichomonal vaginitis: 2 strains (numbering is respectively Tv1 and Tv2).This experiment worm strain through with No. 2 culture medium of OXOID trichomonacide in April, 2012 out-patient of gynecological of healthcare hospital for women & children of Sichuan Province vaginal secretions separate, purification is cultivated and microscopical identification obtains.
1.2 medicine
Trial drug: medicine of the present invention: oil of Herba Pogostemonis (according to the method preparation of embodiment 1).
Positive drug: metronidazole (Kelun Pharm Ind Co., Ltd., Sichuan, lot number: M11120602), JIEERYIN XIYE (Sichuan En Wei pharmaceutical Co. Ltd, lot number: 1107102).
1.3 culture medium, reagent and consumptive material
TRICHOMONAS MEDIUM No.2 (OXOID Ltd. lot number: 1180675), PBS buffer, 96 porocyte culture plates, cell counting count board etc.
1.4 key instrument
Hitachi's transmission electron microscope (H-600IV type), ultramicrotome (ULTRACUT-E type), laboratory is with autoclave (SANYO, MLS-3780 type), CO
2INCUBATOR (SANYO, MOC-15A), energy-saving and purifying workbench (the non-blue ultra-clean chemical industry journey company limited of the new light in Chengdu); High speed centrifuge (HC-3518; Keda Innovation Co., Ltd), low speed centrifuge (model KDC-40, Keda Innovation Co., Ltd).
2 experimental techniques
2.1 medicine trichomonas vaginalis resisting activity research of the present invention
The preparation of worm suspension: get trichomonal vaginitis 37 ℃ of cultivation 48h in No. 2 trichomonacides are cultivated of evaluation, as primary culture, count the worm density and the worm rate of living behind the jolting mixing with cell counting count board.Primary culture worm Shuai alive>95%, cellular morphology is intact, and motion is active, worm density 2.0 * 10
6Worm/ml.Get the 1ml primary culture, the centrifugal 10min of 500rpm, with the warm in advance resuspended polypide of new culture fluid of 10ml, making worm density is 2.0 * 10
5Worm/ml.
Medicine preparation: at first medicine oil of Herba Pogostemonis of the present invention is carried out emulsifying, prepare 10% emulsion.With No. 2 trichomonacides cultivations the invention medicine for preparing is carried out doubling dilution, be diluted to 1:2 ~ 1:1024 totally 10 gradients respectively.
The mensuration of minimum parasite killing concentration (MLC): every hole adds the medicine of the present invention of worm suspension 100 μ l and 100 μ l variable concentrations gradient dilutions in 96 orifice plates, and 37 ℃ of wet boxes are cultivated 48h, pursues sampling smear for microscopic examination in hole with capillary tube from each medicine, adds up dead worm rate.If the positive medicine of metronidazole, drug does not only contain the negative contrast in hole of culture medium and trichomonal vaginitis.With dead worm rate>95% minimum drug level is the minimum lethal concentration (MLC) of medicine to this trichomonal vaginitis.The experiment triplicate.
2.2 medicine of the present invention is to the trichomonal vaginitis ultrastructure effects
The preparation of worm strain and fixing: medicine adding of the present invention is contained in the infusorian culture medium of Tv2 worm strain, and wherein worm density is 2.0 * 10
6Worm/ml, drug concentrations of the present invention is 1/2MLC.At CO
237 ℃ act on 1h, 3h and 5h respectively in the incubator, and the centrifugal 10min of 2000rpm abandons supernatant; Slowly add cold 0.5% glutaraldehyde 1ml along tube wall, 4 ℃ leave standstill 30min, and the centrifugal 10min of 12000rpm abandons supernatant, slowly add cold 3% glutaraldehyde 1ml along tube wall and pre-fix, and 1% Osmic acid. is fixed.The Tv2 worm strain that can replace medicine of the present invention with normal saline is as the normal control group.
Ultrathin section and dyeing: the sample that fixes is dewatered step by step Epon812 embedding, semithin section optical alignment, ultrathin section, acetic acid uranium and lead citrate double staining with acetone.
Analysis of Ultrastructure: adopt the Change of Ultrastructure of the H-600IV of Hitachi type transmission electron microscope observing infusorian strain after 1/2 MLC medicine of the present invention acts on 1h, 3h and 5h respectively, inquire into its mechanism of action.
3 experimental results
3.1 medicine of the present invention is to the insecticidal activity of trichomonal vaginitis
Adopt 96-orifice plate micro-dilution method to measure medicine of the present invention to being located away from the scorching infected patient trichomonacide of institute's per vaginam insecticidal activity, the result sees table 5.
Table 5: medicine of the present invention is to the minimum parasite killing concentration of trichomonal vaginitis
Can know by last table; Medicine of the present invention has tangible insecticidal activity to trichomonal vaginitis; Minimum parasite killing concentration (MLC) to coming from 2 strain trichomonal vaginitis in the scorching patient's secretions of clinical vaginal is 0.41mg/ml, explains that medicine of the present invention has trichomonal effect of killing.
3.2 medicine of the present invention is to the mechanism of action of trichomonal vaginitis
Adopt transmission electron microscope to observe and analyzed Tv2 worm strain Ultrastructural variation of worm strain behind 1/2MLC drug effect 1h of the present invention, 3h and 5h respectively.The result finds the worm strain under medicine different effects time point of the present invention, and identical Change of Ultrastructure appears in the worm strain, explains that medicine length action time of the present invention is to polypide ultrastructure and mechanism of action no significant difference.
Wherein, the Change of Ultrastructure behind the drug effect 1h of the present invention is as shown in Figure 1, and the trichomonal vaginitis ultrastructure more normally contrasts and significant change occurs.Variation from the inside to the outside mainly contains: multiple variation appears in nucleus, like perinuclear space broadening (figure D), and karyopycnosis (figure G), chromatin gathers (figure E), and last nuclear membrane disappears (figure G), and nucleus dissolves gradually.A large amount of cavitys (figure B and figure C) appear in the kytoplasm, dilatation of rough endoplasmic reticula (figure D), and ribosome is reduced to disappearance (figure E and figure F), autophagic vacuole (figure C, figure E and figure F) occurs, organelle disintegrate (figure G).The local cells film is damaged (figure H), and endochylema spills (figure H), last cell disruption downright bad (figure H).Explain that medicine of the present invention passes through pair cell nuclear and organelle structural damage, finally kills polypide.
Can know that to the experimental result of trichomonal vaginitis insecticidal activity and ultrastructural influence medicine of the present invention has the effect of the trichomonal vaginitis killed by the invention described above medicine, can be used for treating trichomonal vaginitis.
To sum up, oil of Herba Pogostemonis has the colpitic effect of treatment, evident in efficacy to monilial vaginitis and trichomonal vaginitis especially, and pharmacological action is strong, and market application foreground is good.
Claims (10)
1. oil of Herba Pogostemonis is treated the purposes in the colpitic medicine in preparation.
2. purposes according to claim 1 is characterized in that: said medicine is the medicine of treatment monilial vaginitis or trichomonal vaginitis.
3. purposes according to claim 2 is characterized in that: said medicine is the monilial vaginitis that the treatment Candida albicans causes.
4. according to any described purposes of claim 1 ~ 3, it is characterized in that: 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.
5. according to any described purposes of claim 1 ~ 3, it is characterized in that: described oil of Herba Pogostemonis derives from the volatile oil that Herba Pogostemonis Pogostemon cablin (Blanco) Benth. extracts, and contains patchouli alcohol in this volatile oil and must not be lower than 26.0%w/w.
6. according to any described purposes of claim 1 ~ 3; It is characterized in that: described oil of Herba Pogostemonis is prepared by following method: 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; The employing steam distillation extracts, and promptly gets oil of Herba Pogostemonis.
7. colpitic pharmaceutical composition of treatment, it is characterized in that: it is to be active component with the oil of Herba Pogostemonis, adds the medicament that acceptable accessories or complementary composition are prepared from.
8. pharmaceutical composition according to claim 7 is characterized in that: described medicament is external preparation, oral formulations or ejection preparation.
9. pharmaceutical composition according to claim 8 is characterized in that: said external preparation is lotion, suppository, liniment or liniment.
10. pharmaceutical composition according to claim 7 is characterized in that: the content of oil of Herba Pogostemonis is 0.1%～100%w/w in the said preparation.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103223009A (en) * | 2013-05-10 | 2013-07-31 | 成都华神集团股份有限公司 | New uses of Pogostemon cablin oil |
| CN114259459A (en) * | 2021-11-11 | 2022-04-01 | 成都中医药大学 | Patchouli suppository, preparation method and application |
| CN114903878A (en) * | 2022-04-22 | 2022-08-16 | 广州医科大学 | Application of sesquiterpenoids in inhibiting activity of TRPA1 channel |
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| CN102058659A (en) * | 2009-12-28 | 2011-05-18 | 成都中医药大学 | Application of palchouli oil in preparing drug for treating bovine mastitis |
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| CN102058659A (en) * | 2009-12-28 | 2011-05-18 | 成都中医药大学 | Application of palchouli oil in preparing drug for treating bovine mastitis |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103223009A (en) * | 2013-05-10 | 2013-07-31 | 成都华神集团股份有限公司 | New uses of Pogostemon cablin oil |
| CN114259459A (en) * | 2021-11-11 | 2022-04-01 | 成都中医药大学 | Patchouli suppository, preparation method and application |
| CN114903878A (en) * | 2022-04-22 | 2022-08-16 | 广州医科大学 | Application of sesquiterpenoids in inhibiting activity of TRPA1 channel |
| CN114903878B (en) * | 2022-04-22 | 2023-09-22 | 广州医科大学 | Application of sesquiterpenoids in inhibiting activity of TRPA1 channel |
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