CN105687342A - Powder capable of clearing lung, resisting inflammation and relieving cough and preparation method thereof - Google Patents

Abstract

The invention discloses powder capable of clearing the lung, resisting inflammation and relieving a cough. The powder is composed of four traditional Chinese medicines of, by weight, 26.3% of herba centellae, 15.8% of radix platycodonis, 31.6% of roots and leaves of rumex nepalensis Spreng. and 26.3% of rhizoma fagopyri dibotryis. The powder can clear heat, promote diuresis, remove toxin, disperse swelling, facilitate the lung, relieve sore throat, reduce phlegm, expel pus, eliminate phlegm, relieve the cough, cool the blood, clear the lung and promote blood circulation to remove blood stasis, the good curative effect is achieved in clinically curing a lung heat cough, a significant curative effect on symptoms such as pain, fever and cough caused by lung diseases is achieved, the raw materials are wide in source and low in price, and the powder is beneficial for popularization.

Description

A kind of lung heat clearing antiinflammatory ZHIKE SAN and preparation method thereof

Technical field

The present invention relates to medical treatment and pharmacy technical field, be specifically related to a kind of lung heat clearing antiinflammatory ZHIKE SAN and preparation method thereof。

Background technology

In recent years, due to factors such as increasing the weight of of various pollutions, the abuse of feed additive and the changes of weather, cause that the animal lung disease caused is more and more many。The traditional Chinese medical science is thought, due to lung governing qi, department breathes, the road of the lifting that comes in and goes out for mechanism of qi, lung being a delicate viscus again, and weak to cold and heat, exopathogen is attacked, then sluggishness of lung-QI is not declared, clear respectful not normal, abnormal rising of lung-QI, then produce cough, pant or make original cough, panting and increase the weight of^[1]。Body, when whole body or respiratory tract local defense function reduce, is occurred by pathogenic infections such as virus, antibacterials generating heat, coughs, spits, the disease such as pharyngalgia, if clinical symptoms continues not heal, it is possible to can cause the serious diseases such as pneumonia, asthma and respiratory failure。The cause of disease of cough is many answers determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs caused by diseases caused by exogenous pathogenic factor or internal injury, according to lungs by evil difference with different Therapeutic Principle, can be summarized as eliminating evil profit lung, lung qi dispersing cough-relieving, invigorating the spleen for dissipating phlegm。

Modern medicine thinks that cough is a kind of pathological state, when the vagus nerve being distributed in respiratory mucosa and pleura is subject to the stimulation of inflammation, warm, mechanically or chemically factor, causes cough by medulla oblongata respiratory center and brainstem network reflexive。Cough is a kind of important defense mechanism of animal body, it is possible to remove the foreign body in respiratory tract and secretions, stops the diffusion of respiratory tract infection。Meanwhile, cough is also a kind of strong physiological reflex, and it is the main cause causing cough to occur that different paathogenic factors makes reflex arc related link impaired。If cough is violent or overfrequency, all it is likely to result in the infringement of animal respiratory tissue and function, even causes whole body multiple complications。The antitussive applied clinically can suppress coughre flex, is divided into central antitussive, peripheral antitussive and facultative antitussive by its site of action。Such as codeine, narcotine, pentoxyverine etc., they have gastrointestinal reaction, toleration and additive more。And Chinese medicine thinks that cough is caused by diseases caused by exogenous pathogenic factor and internal injury, i.e. exopathogen invasion and attack, lung is defended and is felt, impairment of dispersing and descending function of the lung, and lungs virtual loss or his disease of ZANG-organs become involves in caused by lung, takes different method for the treatment of according to characteristic of disease and disease position。

The local organization that inflammation is the live body with vascular system is main to the defence caused by local lesion is reacted, inflammation early stage, some inflammatory mediator stimulates blood vessel, distend the blood vessels, blood vessel endothelium gap enlargement, vascular permeability increases, and liquid in blood plasma, protein and leukocyte etc. are seeped into interstice, along with the increase of transudate, cause swollen tissue。Although inflammation can resist the defensive reaction of external irritant as body, but excessive inflammatory reaction also can cause tissue injury, promotes advancing of disease。The reason caused inflammation has a lot, and endotoxin is a kind of critically important triggering agent exciting inflammatory reaction。Current Western medicine anti-inflammatory drug is broadly divided into two big classes, i.e. steroidal anti-inflammatory drugs and nonsteroidal antiinflammatory drug on the whole。Steroidal anti-inflammatory drugs mainly has glucocorticoid medicine, has physiology and pharmacological action widely；Non-steroidal anti-inflammatory drug is with aspirin for representative。Side effect produced by these medicines can not look down upon, as anti-inflammatory agent abuse causes that the situation of cross infection can occur often；The steroidal of some traditional source or NSAID (non-steroidal anti-inflammatory drug) can lead to complications while representing good efficacy, therefore in the urgent need to the appearance of new anti-inflammatory agent。Relative to current Western medicine simply for the action character of target spot, Chinese Traditional Medicine can while for target spot comprehensive adjustment target spot correlation factor, thus regulate the links that inflammation occurs comprehensively, can better avoid therapeutic process is attended to one thing and lose sight of another the appearance of phenomenon, reduce rate of side effects, self immune system can be stimulated simultaneously, reach the purpose that righting is got rid of evils。

At present, the antipyretic-antalgic agent being representative with nonsteroidal antiinflammatory drug be clinically for antipyretic-antalgic being applied at most, a class medicine the most widely。But, there is a lot of untoward reaction in life-time service, and the natural drug being representative with Chinese medicine is increasingly subject to acceptance and the attention of the world of medicine because its side reaction is few。Therefore, it is necessary for selecting Chinese medicine to carry out researching and developing。

Summary of the invention

It is an object of the present invention to provide a kind of lung heat clearing antiinflammatory ZHIKE SAN and preparation method thereof, it can efficiently solve problem existing in background technology。

A kind of lung heat clearing antiinflammatory ZHIKE SAN, it is made up of Herba Centellae, Radix Platycodonis, Radix Rumicis Crispi and Rhizoma Fagopyri Dibotryis four taste Chinese medicine, and wherein, the percentage by weight of each composition is:

The summation of the percentage by weight of each composition is 100% (weight)。

The preparation method of a kind of lung heat clearing antiinflammatory ZHIKE SAN, specifically comprises the steps of

(1) taking Herba Centellae 25g, Rhizoma Fagopyri Dibotryis 25g, Radix Rumicis Crispi 30g, Radix Platycodonis 15g, respectively, be placed in large beaker, the cold distilled water adding 600mL soaks 1.5 hours；

(2), soaked Chinese medicine is decocted together with the distilled water soaking Chinese medicine, boil 30min, carry out first time filtration, prepare first time medicinal liquid；

(3) cold distilled water adding 400ml in medicinal residues, after filtration carries out continuing to decoct, and boils 20min, and then decompression carries out second time filtration, prepares second time medicinal liquid；

(4), first time medicinal liquid and second time medicinal liquid are merged, prepare third time medicinal liquid；

(5), by three medicine liquid heatings be condensed into respectively and be equivalent to the high, normal, basic three kinds of concentration liquids of crude drug 2.0g/mL, 1.0g/mL, 0.5g/mL, and with 4 DEG C of refrigerators in save backup。

Owing to have employed above technical scheme, the method have the advantages that energy clearing away heat-damp and promoting diuresis, removing toxic substances and promoting subsidence of swelling, lung qi dispersing sore-throat relieving, dispersing lung-QI and dissipating phlegm, evacuation of pus, eliminating phlegm and relieving cough, clearing away heat and cooling blood, preventing phlegm from forming and stopping coughing, energy heat-clearing and toxic substances removing, lung heat clearing evacuation of pus, blood circulation promoting and blood stasis dispelling, demonstrate,prove clinical treatment cough due to lung-heat, there is good curative effect, and the symptom such as the pain that pulmonary disease is caused, heating, cough has significant curative effect, and crude drug source is wide, low price, is beneficial to popularization。

Accompanying drawing explanation

In order to be illustrated more clearly that the present invention, below in conjunction with accompanying drawing, embodiment is briefly described。

Fig. 1 is each group antipyretic trendgram of 4h in detailed description of the invention embodiment 2。

Detailed description of the invention

For the technological means making the present invention realize, creation characteristic, reach purpose and effect and be easy to understand, below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described。

Embodiment 1

Laboratory animal

Healthy mice 50, ♀ ♂ dual-purpose, body weight (18-22g), Chengdu reach rich fruit and Company of Animals Ltd.'s offer, credit number: SCXK (river) 2013-24 are provided。

Medicine and reagent

Mice normal feedstuff；Radix Rumicis Crispi (is plucked in Sichuan Agricultural University boss mountain, identify through teacher Fan Qiaojia), Herba Centellae (is plucked in Sichuan Agricultural University boss mountain, identify through teacher Fan Qiaojia), Rhizoma Fagopyri Dibotryis (is purchased from Huimin hall pharmacy, Ya'an), and Radix Platycodonis (is purchased from Huimin hall pharmacy, Ya'an)；Dimethylbenzene, Chengdu Ke Long chemical reagent factory, lot number: 20120208；26% ammonia, Chengdu Ke Long chemical reagent factory, lot number: 20120321；Normal saline, Sichuan Guorui Pharmaceutical Co., Ltd., lot number: 11092232；Qingfeiyihuo Tablets, Xiuzheng Pharmaceutical Group Co., Ltd, lot number: 101043；The blue Pianomide in Pu ground, Yunnan Paiyao Group Corp., Ltd, lot number: ZL1104。

Experimental apparatus

ESJ200-4 electronic balance, Shenyang Longteng Electronic Co., Ltd.；Water fountain, little mouse cage, tweezers, 25mL graduated cylinder, 50mL volumetric flask 7, cotton balls, marking pen, 400mL beaker, 50mL beaker, 2000mL beaker, 3 Mus casees, gastric perfusion needle, 6mm card punch, eye scissors。

Experimental period and place

Experiment is completed in Sichuan Agricultural University's national level animal class experiment teaching center in the 6-7 month in 2012。

Experimental technique

The preparation of " lung heat clearing antiinflammatory ZHIKE SAN " decocting liquid

Taking Herba Centellae 25g, Rhizoma Fagopyri Dibotryis 25g, Radix Rumicis Crispi 30g, Radix Platycodonis 15g respectively, be placed in large beaker, the cold distilled water adding 600mL soaks 1.5 hours, boils 30 minutes, filters。The distilled water adding 400mL in medicinal residues continues to decoct, and boils 20 minutes, filtration under diminished pressure, merges twice filtrate, and heating is condensed into respectively and is equivalent to the high, normal, basic three kinds of concentration liquids of crude drug 2.0g/mL, 1.0g/mL, 0.5g/mL, saves backup in 4 DEG C of refrigerators。

The feeding and management of mice

Mice being placed on the quiet place of ventilation, dark, allows mice freely drink water, take food, the food ration of every day is 4-6g, and amount of drinking water is about 6mL。Bedding and padding are changed once for every 2-3 days, and drinking bottle to change sterilization water every day once, raise box and clean once for every 3 days。

Statistical data is analyzed

Data SPSS19.0 analyzes, and carries out one factor analysis of variance, compares and check by t value between group。If P < 0.01, then it represents that have obvious curative effect；If P < 0.05, then it represents that effective in cure；P > 0.05, then it represents that curative effect is inconspicuous or is of no curative effect。

" lung heat clearing antiinflammatory ZHIKE SAN " impact on mice ammonia stimulus method

Taking 25 mices, be randomly divided into 5 groups, often group 5, is " lung heat clearing antiinflammatory ZHIKE SAN " high dose group, middle dosage group, low dose group, normal saline group and Qingfeiyihuo Tablets positive controls respectively。Doing a gavage, continuous gastric infusion 4 days noon every day, each given low is 0.2mL/10g。Water 12h is can't help in 25 mice fasting, is respectively placed in 500mL beaker, puts a cotton balls and inject 0.2mL ammonia in cotton balls in diameter is less than the surface plate of beaker, rapidly beaker is inverted, making ammonia stimulate mice to cause cough, with mice abdominal muscle strong contraction, expiration of simultaneously opening one's mouth is as the criterion。The success of cough model Preliminary screening it is judged to more than or equal to 3 times with number of times of coughing in 2min。Observe and record cough latent period (stopping the modeling time to cough) and the cough number of times (with contraction of abdominal muscle, magnify mouth and hear that cough is as the criterion) of mice in two minutes of mice。

" lung heat clearing antiinflammatory ZHIKE SAN " xylol causes the impact of mouse ear edema

Taking 25 mices, be randomly divided into 5 groups, often group 5, is " lung heat clearing antiinflammatory ZHIKE SAN " high dose group, middle dosage group, low dose group respectively, the blue Pianomide positive controls of normal saline group and Pu ground。Doing a gavage, continuous gastric infusion 3 days noon every day, each given low is 0.2mL/10g。After being administered half an hour for the last time, mouse right ear portion for often organizing is coated with and only spreads dimethylbenzene 50 μ L/, cause inflammation, left ear is not painted with normal ear, with the card punch of 6mm, the same area of the left and right ear of mice is cut after 20 minutes, and the sub-balance of electricity consumption immediately weighs respectively, record and calculate ear swelling degree (mg) and inhibitory rate of intumesce (%)。

Swelling (mg)=auris dextra auricle weight (mg)-left ear auricle weight (mg)

Suppression ratio (%)=(the average swelling of normal saline group-average swelling of administration group)/average swelling × 100% of normal saline group

Result of the test

" lung heat clearing antiinflammatory ZHIKE SAN " is to mice ammonia stimulus method experimental result

Mice ammonia is drawn and coughs stimulus method experimental result in Table 1 by " lung heat clearing antiinflammatory ZHIKE SAN "。Result shows: compared with normal saline group, " lung heat clearing antiinflammatory ZHIKE SAN " high dose group, middle dosage group, low dose group, Qingfeiyihuo Tablets positive controls, it is obviously prolonged mice to hide Cough length (P < 0.01, P < 0.05), substantially reduce the cough number of times (P < 0.01) in mice 2min, and along with dosage increases, mice Cough length of hiding is more long, and cough number of times is more few。

Ammonia is caused the impact of mouse cough by table 1 " lung heat clearing antiinflammatory ZHIKE SAN "

Note: with column data shoulder mark^*Represent significant difference (P < 0.05) compared with normal saline group, with column data shoulder mark^*Represent difference extremely notable (P < 0.01) compared with normal saline group。

Mice ear is affected mice ear experimental result by " lung heat clearing antiinflammatory ZHIKE SAN "

Mice ear experimental result is in Table 2。Result shows: normal saline group mice ear is obvious, compared with normal saline group, mice ear is had significant inhibitory action (P < 0.01) by the blue Pianomide positive controls in Pu ground, " lung heat clearing antiinflammatory ZHIKE SAN " high (0.4g crude drug/10g), in (0.2g crude drug/10g), low (0.1g crude drug/10g) three dosage groups mice ear is had obvious inhibitory action (P < 0.01 or P < 0.05), and along with dosage to increase it more high to the suppression ratio of mice ear。

Table 2 " lung heat clearing antiinflammatory ZHIKE SAN " xylol causes the impact of mouse ear edema

Note: with column data shoulder mark^*Represent significant difference (P < 0.05) compared with normal saline group,^*Represent difference extremely notable (P < 0.01) compared with normal saline group。

Analyze and discuss

By mice ammonia being drawn the data analysis coughing stimulation test, find all have significant difference (P < 0.05) or pole significant difference (P < 0.01) between decocting liquid group and the normal saline group of " lung heat clearing antiinflammatory ZHIKE SAN ", proving that " lung heat clearing antiinflammatory ZHIKE SAN " has good antitussive action, its antitussive action is likely to relevant with Radix Platycodonis and Radix Rumicis Crispi。The main active of Radix Platycodonis is triterpene saponin, and platycodin is the characteristic chemical constituent of Radix Platycodonis, and its pharmacological action is mainly eliminated the phlegm, antitussive, antiinflammatory。Radix Rumicis Crispi nature and flavor are bitter, cold, enter the heart, liver, large intestine channel, have clearing away heat and cooling blood, effect of preventing phlegm from forming and stopping coughing。Grandson is slipped by, and the cloud et al. that is filled finds that Radix Platycodi total saponins can significantly reduce animal lung tissue's BALF total white blood cells and neutrophilic granulocyte, and acts on smooth muscle cough-relieving。

By the data analysis that mice ear is tested, this experiment finds that the decocting liquid group of " lung heat clearing antiinflammatory ZHIKE SAN " is compared with normal saline group, significant difference (P < 0.05 or P < 0.01), showing that acute inflammation is had good antiinflammatory action by " lung heat clearing antiinflammatory ZHIKE SAN ", its effect is likely to relevant with Multiple components therein。The monarch etc. that shows one's high ideals have studied Herba Centellae total glycosides xylol and causes the impact of mice ear, and result high dose group can be obviously improved mice ear, it was shown that Herba Centellae total glycosides has obvious antiinflammatory action。Wherein asiaticoside can reduce high-caliber COX-2 expression and PGE in Collagen-induced Arthritis mice (CIA) ankle joint soft tissue₂Content, reduces the level of inflammatory factor TNF-and IL-6 in blood plasma, increases the level of IL-1O simultaneously, illustrate that asiaticoside can effectively suppress CIA mice inflammatory reaction。It is taken as that asiaticoside be Herba Centellae treatment inflammation main active。The research of Cheng Youbin shows^[27], Rhizoma Fagopyri Dibotryis chloroform extract and water liquid position can suppress the increase of capillary permeability significantly, it is suppressed that the directed of inflammation part leukocyte assembles and suppress inflammatory factor PEG₂Release。Being present in the former cyanidin aglycon of double focusing of chloroform extract, flavonol (catechin) has the effect of antiinflammatory, and a class condensation tannin of the flavonol derivant and polymer composition thereof being concurrently present in water liquid position has antiinflammatory action。Radix Rumicis Crispi root and rhizome contains rheum emodin。Research finds, the inflammatory cytokines such as rheum emodin energy activated mononuclear phagocyte secretion interleukin-11 (IL-1), interleukin 6 (IL-6), interleukin 8 (IL-8), moreover it is possible to suppress by the secretion of the above-mentioned inflammatory factor of endotaxin induction。Rheum emodin can also play antiinflammatory action by the secretion of other cellular inflammation factors of suppression, such as TNF-。Jin Jiu finds in the antiinflammatory action of research Radix Platycodonis: 2 "-0-acetyl group polygalic acid D, platycodin A, Platycodin D and polygalic acid D all can suppress iNOS and COX-2。The inhibitory action of iNOS and COX-2 is the activity by stoping NF-к B (nuclear factor к B) by platycodin, reduces iNOS and COX-2 gene expression and plays antiinflammatory action。Various inflammatory models are all had stronger antiinflammatory action by Radix Platycodonis, main and suppression prostaglandin E₂(PGE₂) path, nitric oxide (NO) secrete relevant。

Cough due to lung-heat's card treated by experience proved recipe " lung heat clearing antiinflammatory ZHIKE SAN " on veterinary clinic, and card is shown in cough, and expectorant is thick or yellow thick, coughs up phlegm not well, thirsty, pharyngalgia, nose stream yellow thick nasal discharge, heating, sweating, headache, aversion to wind, tongue fur BOHUANG, floating and rapid pulse or slick and frivolous。Radix Rumicis Crispi clearing away heat and cooling blood in side, preventing phlegm from forming and stopping coughing, for monarch drug。Rhizoma Fagopyri Dibotryis heat-clearing and toxic substances removing, lung heat clearing evacuation of pus, blood circulation promoting and blood stasis dispelling, Herba Centellae clearing away heat-damp and promoting diuresis, removing toxic substances and promoting subsidence of swelling, is ministerial drug altogether。Radix Platycodonis lung qi dispersing sore-throat relieving, dispersing lung-QI and dissipating phlegm, medicine carrying is up, and is adjuvant。

This experiment stimulates by ammonia to draw coughs method experiment and the antiinflammatory of mice caused by dimethylbenzene xylene ear swelling test, have studied " lung heat clearing antiinflammatory ZHIKE SAN " to the cough of manually pathogenic white mice and antiinflammatory action。Test result indicate that " lung heat clearing antiinflammatory ZHIKE SAN " has good antitussive effect and acute inflammation has good antiinflammatory action。But other effect of " lung heat clearing antiinflammatory ZHIKE SAN " and the antiinflammatory action to chronic inflammatory disease need to be studied further。

Embodiment 2

Antipyretic experiment

Healthy white big ear rabbit, male and female dual-purpose, body weight 1.7-2.0 kilogram, by pharmacopeia heat-original determinating method^[9]Screening body temperature eligible (normal body temperature should in the scope of 38-39.6 DEG C, and between each rabbit, regular using warming therapy difference must not exceed 1 DEG C), be randomly divided into 5 groups, often group 5, blank group is normal saline；Positive controls is aspirin (100mg/kg)；The basic, normal, high dosage component of trial drug is not: crude drug 0.5g/ml, 1.0g/ml, 2.0g/ml。Experiment starts to first pass through anus and measures rabbit basal body temperature, and the time that electronic clinical thermometer inserts when measuring body temperature rabbit anus is respectively organized all identical with the degree of depth, and the degree of depth is typically about 6cm, and the time is no less than 1 point of half。Then respectively group rabbit is measured body temperature rising 0.5 DEG C and is heating after injecting skim milk 1ml/kg, 30min from auricular vein respectively, then prove that fever model is successfully established。It is pressed in rabbit back side with gag, takes No. 8 stomach catheter and be about 15cm by aperture insertion esophagus in the middle part of gag。For avoiding being strayed into trachea, being immersed in the water by stomach catheter collar extension, lose bubble and show that conduit inserts in stomach, be then injected into medicinal liquid, every rabbit dosage is 5ml/kg。Measure and record after first time gastric infusion 1 respectively, 2,3, the rabbit anus temperature of 4h。Result of the test represents with body temperature range of decrease average。

Antibacterial experiment in vitro

The preparation of test reagent and article

Nutrient broth medium: NaCl10g, Carnis Bovis seu Bubali cream 3g, peptone 10g, pure water 1000ml。In pure water, add Carnis Bovis seu Bubali cream, peptone and NaCl successively, be sufficiently stirred for glass rod and make it be completely dissolved。PH value 7.2-7.4 is regulated with 4%NaOH。121 DEG C of autoclaving 30min are standby。

Nutrient agar: NaCl10g, Carnis Bovis seu Bubali cream 3g, peptone 10g, agar 15g, pure water 1000ml。Being placed on by beaker on the electric furnace added with asbestos gauge on the basis prepare nutrient broth and heat, add agar powder while stirring, heating is boiled to micro-, liquid-transparent。PH value 7.2-7.4 is regulated with 4%NaOH。121 DEG C of autoclaving 30min。In clean bench, the agar culture medium of heat is poured in the test tube after sterilizing, less than 1/3rd of test tube length, so as to tilt condensation, become slant medium；To pour into before the culture medium solidifying of heat in the clean flat board after sterilizing, every plate 10-15ml, stand so as to condense, become agar plate standby。

Sterilized water: be loaded in triangular flask by tertiary effluent, volume is less than 2/3rds of triangular flask volume, and sealing, 121 DEG C of autoclaving 30min are standby。

Physiological saline solution: be loaded in triangular flask by the normal saline configured, volume is less than 2/3rds of triangular flask volume, and sealing, 121 DEG C of autoclaving 30min are standby。

The preparation of medicine to be measured

Hereinafter work and all carry out in clean bench。

By clearing away lung-heat to relieve cough aproll decocting liquid filtration sterilization, it is 9 concentration with sterilized water doubling dilution, crude drug content respectively 1280mg/ml, 640mg/ml, 320mg/ml, 160mg/ml, 80mg/ml, 40mg/ml, 20mg/ml, 10mg/ml, 5mg/ml。

Collecting cells

Strains tested is inoculated in respectively slant medium activation, after 37 DEG C are cultivated 24h, separates typical single bacterium colony, turn and be inoculated in broth bouillon, be placed in 37 DEG C of calorstats and cultivate 24h。Take 1ml culture fluid normal saline and be sequentially carried out dilution with 10 times of dilution methods, choose on the 0.1ml bacterium solution horizontalization ware agar culture medium of debita spissitudo, gently bacterium solution is smoothened with aseptic spreader, 37 DEG C of constant temperature culture 24h, calculate clump count。Calculate the viable count of every milliliter of bacterium solution according to bacterium solution diluted concentration, make bacteria suspension concentration reach 10⁶-10⁷Cfu/ml, as test bacterium solution。

Drug susceptibility is tested

Employing cup-plate method carries out。Dipping concentration with aseptic cotton carrier is 10⁶-10⁷The dilution of bacteria of cfu/mL, coats on General nutrition agar plate, is uniformly put on flat board by steel pipe with aseptic nipper, stands 10min～15min, drips " lung heat clearing antiinflammatory ZHIKE SAN " low middle high three kinds of concentration liquids, adds to liquid level parallel with steel tube surface。After flat board 37 DEG C is cultivated 18h～24h, by vernier caliper measurement inhibition zone size。Antibacterial circle diameter d < 9.0mm is less sensitive, and 9.1 < d < 11mm are medium sensitivity, and d >=12mm is extremely sensitive。

Minimal inhibitory concentration (MIC) measures

Adopt test tube doubling dilution。To there is the medicinal liquid normal saline doubling dilution of fungistatic effect, be made into the broth bouillon test tube that concentration is 9 concentration in 2.2 successively, and in each pipe, add bacteria suspension 50 μ l, each pipe is placed in constant temperature culture under 37 DEG C of conditions。After 18～24h, the whether long bacterium of each test tube of observed and recorded and growing state。Each Concentraton gradient does three groups of parallel tests, the minimum inhibitory concentration that least concentration is this medicine of the not long bacterium of perusal。

Minimal bactericidal concentration (MBC) measures

First measure the MIC of test organisms, according to its result, the more each pipe culture fluid of bacterial growth will be had no successively draw 50 μ l respectively, and carefully coat in aseptic fresh agar plate, put 37 DEG C of incubators and cultivate observed result after 24h。On plate, the clump count minimum dilution drug level less than 5 is MBC。

Result and analysis

Antipyretic experimental result

Experimental result is in Table 1。After the display administration of table 1 result, body temperature all declines to some extent, compared with blank group, and positive controls refrigeration function extremely notable (P < 0.01)；During clearing away lung-heat to relieve cough dissipates, low dose group refrigeration function notable (P < 0.05), high dose group refrigeration function notable (P > 0.05)。And compare with aspirin group, trial drug refrigeration function difference not notable (P > 0.05), each group antipyretic trend of 4h is shown in Fig. 1。

" clearing away lung-heat to relieve cough dissipates the " refrigeration function to skim milk pyrogenicity rabbit to table 1

Note: * positive group compares P < 0.01 with blank group；In * trial drug, low dose group compare P < 0.05 with blank group；* * trial drug high dose group compares P > 0.05 with blank group；Test group compares P > 0.05 with positive group。

Antibacterial experiment in vitro result

Determination of drug sensitivity result

" lung heat clearing antiinflammatory ZHIKE SAN " three kinds of concentration liquids all have bacteriostatic activity, and staphylococcus aureus and escherichia coli are had certain fungistatic effect, and result is in Table 2。

Table 2 " lung heat clearing antiinflammatory ZHIKE SAN " is to the average diameter of inhibition zone (unit: mm) for examination bacterium

Minimal inhibitory concentration (MIC) measurement result

After cultivation terminates, naked eyes visible high concentration medicine test tube, positive control test tube and blank test tube all keep original color, have no change。There is metachromatism in low concentration medicine test tube, white opacity thing occurs。Liquid turns turbid in negative control test tube, produces white precipitate。Illustrative experiment suitable operation credible result。Visual results, in Table 3, can determine whether that " lung heat clearing antiinflammatory ZHIKE SAN " is to staphylococcus aureus, colibacillary MIC respectively 80mg/ml and 80mg/ml。

Table 3 " lung heat clearing antiinflammatory ZHIKE SAN " minimal inhibitory concentration (MIC)

Note: "-" asepsis growth, "+" thalline is less, " ++ " thalline is more, and " +++ " thalline is many。

Minimal bactericidal concentration (MBC) measurement result

The two groups of 1-5 pipe culture fluid having no bacterial growth are drawn respectively and coats in aseptic fresh agar plate, put the result observed after 37 DEG C of incubators cultivate 24h in Table 4。As shown in Table 4, staphylococcus aureus and colibacillary MBC are 160mg/ml and 640mg/ml by " lung heat clearing antiinflammatory ZHIKE SAN " respectively。Bactericidal activity is: staphylococcus aureus > escherichia coli。

Table 4 " lung heat clearing antiinflammatory ZHIKE SAN " minimal bactericidal concentration (MBC)

Note: "-" asepsis growth, "+" thalline is less, the more discussion of " ++ " thalline

The refrigeration function of " lung heat clearing antiinflammatory ZHIKE SAN "

Theory of Chinese medical science thinks between lung and large intestine the communication by meridians, and Pathophysiology activity influences each other, and forms the relation of " the lung and the large intestine being interior-exteriorly related "。Lung covering other viscera, occupies the part of the body cavity above the diaphragm housing the heart and lungs, for sucking the upper orifices of clearing heat in QI system, exhalation foul smell；Large intestine occupies the part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels, and for transmitting waste matter, discharging the lower orifices of foul smell, both the exterior and the interior are correlated with, close fit, the common effect playing ascending the clear and descending the turbid。In the four Chinese medicine of " lung heat clearing antiinflammatory ZHIKE SAN ", Radix Et Rhizoma Rhei bitter cold, enter spleen, stomach, large intestine, pericardium, Liver Channel, main attack is stagnant, has clearing away damp-heat, pathogenic fire purging, removing heat from blood, blood stasis dispelling, function of detoxification, is Chinese medicine simply, is monarch drug because improving respiratory function by purging heat and dredging bowels。Rhizoma Fagopyri Dibotryis has effect of heat-clearing and toxic substances removing, invigorating spleen to remove dampness, therapeutic method to keep the adverse QI flowing downwards removing food stagnancy；Herba Centellae has clearing away heat-damp and promoting diuresis, effect of removing toxic substances and promoting subsidence of swelling。Both energy heat-clearing and toxic substances removing, help Radix Et Rhizoma Rhei to remove lung-heat, for ministerial drug。And lunar lung meridian returned by Radix Platycodonis, can eliminate the phlegm by lung qi dispersing, moreover it is possible to dispersing and elevation lung qi, dredge gastrointestinal, for the principal agent that medicine carrying is up, make each drug effect can arrive pulmonary, for adjuvant。All medicines share, and based on eliminating evil, take into account healthy energy, to reach effect of clearing away lung-heat to relieve cough。

Modern pharmacological research shows, Radix Et Rhizoma Rhei can substantially suppress secreted by alveolar macrophages tumor necrosis factor and interleukin-1, the interleukin II that endotoxin induces, thus alleviating the infringement to lungs, has obvious refrigeration function。Modern pharmacological research shows that it also has the multiple efficacies such as antitumor, blood sugar lowering, blood fat reducing, antioxidation, Antipyretic。Research at present thinks that its active component playing drug effect is mainly flavones ingredient, but more ingredient medical mechanism need further research。Fitochemical studies finds that the herb main chemical compositions of Herba Centellae is cupreol, daucosterol, asiatic acid, vanillic acid and asiaticoside。Asiaticoside is possibly through suppressing COX-2/PGE₂System, reduces pyrogenicity medium PGE₂Content, it is suppressed that COX-2 protein expression and play prevention heating functioin。Platycodin also has significant refrigeration function。

The In Vitro Bacteriostasis of " lung heat clearing antiinflammatory ZHIKE SAN "

This test selects common pathogen as object of study。Staphylococcus aureus is a kind of important pathogen of animals and humans, and can cause many severe infections, traditional prevention and controls is use antibiotics。But along with the development of medical and health cause, antibiotic, synthesising bacteria anti-reflecting medicine use in a large number and abuse causes, drug residue, Drug resistance and the animal food safety problem thus brought are becoming increasingly acute。Herbal control pandemic, solution antibiotic remains and resistance problems is utilized to become one of pharmacology of Chinese materia medica study hotspot。Antibacterials are to certain sensitive bacteria or drug resistance, it is decided by the mechanism of action of antibacterials and the genotype of antibacterial, drug resistance can be directly predicted in the detection of drug resistant gene, but owing to monitoring the molecular biology method of drug resistance relative to drug sensitivity testing in vitro with greater need for complicated equipment and instrument。Bacterial resistance relates to many factors, and its mechanism not yet illustrates completely, and therefore, current Clinical microorganism laboratory is main or predicts bacterial resistance by drug sensitivity testing in vitro。Extracorporeal bacteria inhibitor test method has cup-plate method, paper disk method, punch method etc.。But punch method is big because of herb liquid stickiness, it is difficult to spread so that dropping in the medicinal liquid in hole。Paper disk method is simple, but the dose that the scraps of paper adsorb is less, and the medicinal liquid being adsorbed onto on the scraps of paper not easily spreads, and can affect the fungistatic effect of medicine。And cup-plate method taken amount is accurate, result of the test is comparatively accurate, therefore this test adopts the method to carry out bacteriostatic test。

Plant is the natural treasure-house of Substance, and Grange and Ahmed report there are about 2400 kind of plant and has the activity of preventing and treating harmful organism。Histological research shows, Radix Et Rhizoma Rhei can protect the epithelial integrity of small intestinal mucosa, promotes a large amount of hypertrophy of goblet cell in intestinal villi, and the alkaline mucus of the latter's secretion can reinforce gut barrier, it is suppressed that bacterial translocation from intestine。Staphylococcus aureus, escherichia coli, bacillus subtilis, Su Yun bacillus cereus, OK a karaoke club diplococcus are had obvious bacteriostasis by Rhizoma Fagopyri Dibotryis。Asiaticoside is external has antibacterial action, especially various drug-resistant bacterias is had stronger antibacterial activity, and platycodin also has certain bacteriostasis。This test primarily determines that both the above antibacterial is all had inhibitory action by " lung heat clearing antiinflammatory ZHIKE SAN ", but owing to " lung heat clearing antiinflammatory ZHIKE SAN " decocting liquid has certain yellowish-brown, therefore test tube doubling dilution measures its minimal inhibitory concentration and there is certain error。

Last it is noted that above example is only in order to illustrate technical scheme, it is not intended to limit；Although the present invention being described in detail with reference to previous embodiment, it will be understood by those within the art that, the technical scheme described in foregoing embodiments still can be modified by it, or wherein portion of techniques feature carries out equivalent replacement；And these amendments or replacement, do not make the essence of appropriate technical solution depart from the spirit and scope of various embodiments of the present invention technical scheme。

Claims (2)

1. a lung heat clearing antiinflammatory ZHIKE SAN, it is characterised in that it is made up of Herba Centellae, Radix Platycodonis, Radix Rumicis Crispi and Rhizoma Fagopyri Dibotryis four taste Chinese medicine, wherein, the percentage by weight of each composition is:

Herba Centellae 26.3% (weight)

Radix Platycodonis 15.8% (weight)

Radix Rumicis Crispi 31.6% (weight)

Rhizoma Fagopyri Dibotryis 26.3% (weight)

The summation of the percentage by weight of each composition is 100% (weight)。

2. the preparation method of a lung heat clearing antiinflammatory ZHIKE SAN as claimed in claim 1, it is characterised in that specifically comprise and specifically comprise the steps of

(1) taking Herba Centellae 25g, Rhizoma Fagopyri Dibotryis 25g, Radix Rumicis Crispi 30g, Radix Platycodonis 15g, respectively, be placed in large beaker, the cold distilled water adding 600mL soaks 1.5 hours；

(2), soaked Chinese medicine is decocted together with the distilled water soaking Chinese medicine, boil 30min, carry out first time filtration, prepare first time medicinal liquid；

(3) cold distilled water adding 400ml in medicinal residues, after filtration carries out continuing to decoct, and boils 20min, and then decompression carries out second time filtration, prepares second time medicinal liquid；

(4), first time medicinal liquid and second time medicinal liquid are merged, prepare third time medicinal liquid；

(5), by three medicine liquid heatings be condensed into respectively and be equivalent to the high, normal, basic three kinds of concentration liquids of crude drug 2.0g/mL, 1.0g/mL, 0.5g/mL, and with 4 DEG C of refrigerators in save backup。