CN113384647B - Application of traditional Chinese medicine composition in preparation of medicine for treating acute lung injury - Google Patents

Abstract

The invention discloses an application of a traditional Chinese medicine composition in preparing a medicine for treating acute lung injury. The traditional Chinese medicine composition mainly comprises ephedra, gypsum, fructus forsythiae, radix scutellariae, cortex mori, bitter apricot kernel, peucedanum root and the like, plays the integral regulation advantage of the compound traditional Chinese medicine, and has the effects of eliminating pathogenic factors, relieving symptoms and regulating immunity through organic combination, and clinical experiments prove that the traditional Chinese medicine composition has remarkable curative effect on acute lung injury.

Description

Application of traditional Chinese medicine composition in preparation of medicine for treating acute lung injury

Technical Field

The invention relates to an application of a traditional Chinese medicine composition in preparing a medicine for treating acute lung injury, and belongs to the field of Chinese herbal medicines.

Background

Acute lung injury (acute lung injury, ALI) is a clinically common critical condition. The main factors causing acute lung injury are systemic infection, sepsis, trauma, shock, poisoning, etc. The main pathological manifestations are damaged endothelial cells of pulmonary blood vessels, damaged endothelial integrity, dysfunction of alveolar-capillary barrier, increased vascular permeability, extensive inflammatory cell infiltration of the lung, pulmonary edema and the like, and if the acute respiratory distress syndrome is not treated in time, the mortality rate is up to 40%. At present, ALI diagnosis still maintains the standards proposed by European and American conference in 1994 (1) acute onset, existence of causative factors, (2) oxygenation index (arterial partial pressure/concentration of inhaled oxygen, paO2/Fi 02) <300mmHg (1 mmHg=0.133 kPa) without reference to positive end expiratory pressure (PEEP, positive end expiratory pressure) level, (3) positive X-ray chest film shows that both lungs are patchy shaded, (4) pulmonary artery incarceration pressure <18mmHg, or no clinical evidence of left atrial pressure increase; (5) acute paroxysmal respiratory failure.

At present, the treatment of acute lung injury is considered to be comprehensive treatment, not only is to correct refractory hypoxia, improve alveolar oxygenation, but also is comprehensive treatment centered on a proper respiratory support technology, including multiple auxiliary treatments such as infection control, early nutrition support treatment, stress ulcer prevention, deep vein thrombosis prevention and the like, and also has fine care, and no specific drug treatment exists at present. The western medicine treatment is frequently administered for symptomatic and supportive treatment, and although clinical symptoms can be alleviated, the disease progress cannot be obviously controlled, and certain limitations exist. The Chinese medicine therapy has the characteristics of dialectical treatment, multiple ways, multiple target pharmacological actions and the like, and plays a unique advantage in the treatment of diseases. The acute lung injury caused by sepsis is classified in the category of "sudden asthma" according to the theory of traditional Chinese medicine, and the failure of viscera qi to circulate lung qi is considered as the main pathogenesis of the acute lung injury.

The traditional Chinese medicine composition has the effects of clearing lung heat, resolving phlegm and relieving cough. The present invention is an improved invention based on the application number 2008100894475, which is incorporated by reference in its entirety. The above patent does not disclose the use of the traditional Chinese medicine composition in the preparation of a medicament for treating acute lung injury.

Disclosure of Invention

The invention provides application of a traditional Chinese medicine composition in preparing a medicine for treating acute lung injury, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:

52-86 of ephedra; gypsum 194-324; fructus forsythiae 194-324; 78-130 parts of radix scutellariae; white mulberry root bark 194-324; 78-130 parts of bitter apricot kernel; 78-130 parts of radix peucedani; 78-130 parts of pinellia ternate; 78-130 parts of dried orange peel; 78-130 of fritillary bulb; 78-130 parts of burdock; 78-130 parts of honeysuckle; 39-65 parts of rheum officinale; radix Platycodi 46-76; 39-65 parts of liquorice.

The traditional Chinese medicine composition comprises the following raw materials in parts by weight:

herba Ephedrae 52; gypsum 324; fructus forsythiae 194; radix Scutellariae 78; cortex Mori 194; bitter apricot seed 130; radix Peucedani 78; pinellia ternate 130; dried orange peel 78; fritillary bulb 78; fructus Arctii 130; honeysuckle 130; rhubarb 39; radix Platycodi 76; licorice 65.

The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:

herba Ephedrae 86; gypsum 194; fructus forsythiae 324; radix Scutellariae 130; cortex Mori 324; bitter apricot kernel 78; radix Peucedani 130; pinellia tuber 78; dried orange peel 130; fritillary 130; burdock 78; honeysuckle 78; rhubarb 65; radix Platycodi 46; licorice 39.

The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:

ephedra 69; gypsum 259; fructus forsythiae 259; radix Scutellariae 104; cortex Mori 259; bitter almonds 104; radix Peucedani 104; pinellia ternate 104; dried orange peel 104; fritillary 104; fructus Arctii 104; honeysuckle 104; rhubarb 52; radix Platycodi 61; licorice 52.

The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:

herba Ephedrae 55; gypsum 254; fructus forsythiae 318; radix Scutellariae 107; white mulberry root bark 203; bitter apricot kernel 107; radix Peucedani 82; pinellia ternate 105; dried orange peel 84; fritillary bulb 125; burdock 122; honeysuckle 113; rhubarb 42; radix Platycodi 60; licorice root 50.

In the traditional Chinese medicine composition, the bitter apricot kernel is fried bitter apricot kernel, the fritillaria is thunberg fritillary bulb, the honeysuckle is lonicera japonica and the pinellia tuber is purified pinellia tuber.

The traditional Chinese medicine composition mainly comprises ephedra, gypsum, fructus forsythiae, radix scutellariae, cortex mori radicis and the like, plays the integral regulation advantage of the compound traditional Chinese medicine, and has the effects of eliminating pathogenic factors, relieving symptoms and regulating immunity through organic combination, and clinical experiments prove that the traditional Chinese medicine composition has remarkable curative effects on acute lung injury.

The traditional Chinese medicine can be replaced by traditional Chinese medicines with the same or similar effects, and the traditional Chinese medicines can be processed according to national traditional Chinese medicine processing standards or Chinese medicine dictionary.

The active ingredients of the traditional Chinese medicine composition are prepared by the following steps:

A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 40-70% ethanol for 2 times, each for 1-4 hr, adding 8-10 times of the extractive solution for the first time and adding 6-9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14-1.16 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, honeysuckle, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water for decoction twice, each time for 1-4 hours, adding 9-11 times of the weight of the first time and 7-9 times of the weight of the second time, merging decoction, filtering, concentrating to obtain clear paste with the relative density of 1.14-1.16 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;

the fine powder obtained in the step A and the combined fluid extract obtained in the step C jointly form the active ingredient of the pharmaceutical composition.

The medicament of the invention is in the dosage forms of capsules, tablets, powder, granules, oral liquid, soft capsules, pills, tinctures, syrups, suppositories, gels, sprays or injections.

To enable the above dosage forms, pharmaceutically acceptable excipients are added in the preparation of these dosage forms, for example: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, matrices, and the like. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, and the like; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, crosslinked sodium carboxymethyl cellulose, and the like; the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like; the binder includes starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, etc.; the flavoring agent comprises: sweetener and various flavors; the preservative comprises: nipagin, benzoic acid, sodium benzoate, sorbic acid and salts thereof, benzalkonium bromide, chlorhexidine acetate, eucalyptus oil and the like; the matrix comprises: PEG6000, PEG4000, insect wax, and the like.

Wherein the tablet is prepared by the following steps:

A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 40-70% ethanol for 2 times, each for 1-4 hr, adding 8-10 times of the extractive solution for the first time and adding 6-9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14-1.16 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, honeysuckle, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water for decoction twice, each time for 1-4 hours, adding 9-11 times of the weight of the first time and 7-9 times of the weight of the second time, merging decoction, filtering, concentrating to obtain clear paste with the relative density of 1.14-1.16 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. the spray powder obtained in the step D and the fine powder obtained in the step A are used for preparing soft materials by taking ethanol as an adhesive, and sieving and granulating are carried out; tabletting according to conventional pharmaceutical method.

The preparation method of the preferred tablet comprises the following steps:

A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 50% ethanol for 2 times, each for 3 hr, adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, honeysuckle, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water for decoction twice, each time for 2 hours, adding 10 times of the weight for the first time, adding 7 times of the weight for the second time, merging the decoction, filtering, concentrating to obtain clear paste with the relative density of 1.15 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. and D, preparing soft materials by taking ethanol as an adhesive from the spray powder obtained in the step A, sieving, granulating, drying, finishing, adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, uniformly mixing, and tabletting.

The preparation method of other dosage forms of the medicine comprises the following steps: the raw materials are weighed according to the proportion and prepared by adopting a conventional preparation method, for example, a preparation process recorded in Fan Biting traditional Chinese medicine pharmacy (1 st edition of 1997 of Shanghai science publishing Co., ltd.) is adopted to prepare a pharmaceutically acceptable conventional dosage form.

The traditional Chinese medicine composition can also reduce the rise of cell inflammatory factors caused by acute lung injury.

The traditional Chinese medicine composition can reduce cell inflammatory factors caused by acute lung injury, such as IL-6, TNF-alpha, NE and MCP-1.

The Chinese medicinal composition can also inhibit signal transduction of protein expression caused by acute lung injury.

The protein which can inhibit acute lung injury is NF-kappa B P65, ikB-alpha, p-IkB-alpha and IKKK beta.

Description of the drawings:

fig. 1: effect of groups on NF- κ B P65, IκB- α, p-IκB- α, IKKKβ protein expression in acute Lung injury lung

Fig. 2: mice model acute lung injury were observed by HE staining of their lungs (x 100).

Detailed Description

The following examples are presented to illustrate the preparation of the medicaments of the present invention, but are not intended to limit the scope of the invention in any way.

Example 1

Prescription:

52 g of ephedra; 324 grams of gypsum; 194 g of weeping forsythia; 78 g of baikal skullcap root; 194 g of white mulberry root-bark; 130 g of bitter almond; radix peucedani 78 g; 130 g of pinellia ternate; 78 g of dried orange peel; 78 g of fritillary bulb; 130 g of burdock; 130 g of lonicera japonica; 39 g of rheum officinale; 76 g of platycodon grandiflorum; 65 g of liquorice.

The preparation method comprises the following steps:

A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 50% ethanol for 2 times, each for 3 hr, adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for 2 hours each time, adding 10 times of the weight for the first time and 7 times of the weight for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.15 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.

Example 2

Prescription:

86 g of ephedra; 194 g of gypsum; 324 g of fructus forsythiae; 130 g of radix scutellariae; 324 g of white mulberry root-bark; 78 g of fried bitter apricot kernel; 130 g of radix peucedani; 78 g of pinellia ternate; 130 g of dried orange peel; 130 g of fritillary bulb; 78 g of burdock; 78 g of lonicera japonica; 65 g of rheum officinale; 46 g of platycodon grandiflorum; licorice root, radix Glycyrrhizae Praeparata 39 g.

The preparation method comprises the following steps:

A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 40% ethanol for 2 times and 4 hr each time, adding 8 times of the extractive solution for the first time and 9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for two times, wherein the amount of the water is 9 times for each time, the amount of the water is 7 times for the first time, the decoction is combined, filtered and concentrated to the clear paste with the relative density of 1.16 measured by heat at 60 ℃ for standby;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.

Example 3

Prescription:

69 g of ephedra; 259 grams of gypsum; 259 g of weeping forsythia; 104 g of radix scutellariae; 259 g of white mulberry root-bark; 104 g of fried bitter apricot kernel; 104 g of radix peucedani; 104 g of purified pinellia tuber; 104 g of dried orange peel; 104 g of fritillary bulb; 104 g of burdock; 104 g of lonicera japonica; 52 g of rheum officinale; 61 g of platycodon grandiflorum; licorice root, radix Glycyrrhizae Praeparata 52 g.

The preparation method comprises the following steps:

A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, purified pinellia Tuber, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 70% ethanol for 2 times (1 hr each time), adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.16 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for two times, 1 hour each time, adding 11 times of the weight for the first time, adding 7 times of the weight for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.14 measured by heat at 60 ℃, and merging with the clear paste obtained in the step B for standby;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.

Example 4:

the formula of the raw materials is as follows:

ephedra herb 55 g; 254 g of gypsum; 318 g of fructus forsythiae; baical skullcap root 107 g; 203 g of white mulberry root-bark; stir-frying 107 g of bitter apricot seeds; radix peucedani 82 g; 105 g of pinellia ternate; 84 g of dried orange peel; 125 g of fritillary bulb; 122 g of burdock; 113 g of lonicera japonica; 42 g of rheum officinale; 60 g of platycodon grandiflorum; 50 g of liquorice.

The preparation method comprises the following steps:

A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;

B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 60% ethanol for 2 times and 2 hr each time, adding 9 times of the extractive solution for the first time and 7 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;

C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the formula, adding water and decocting for 2.5 hours each time, adding 10 times of the weight of the mixture for the first time, adding 7 times of the weight of the mixture for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.14 measured by heat at 60 ℃, merging the clear paste obtained in the step B for later use;

D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;

E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.

Example 5:

62 g of ephedra, 220 g of gypsum, 256 g of weeping forsythia, 90 g of baikal skullcap root, 300 g of white mulberry root-bark

90 g of bitter apricot kernel, 90 g of peucedanum root, 90 g of pinellia tuber, 100 g of dried orange peel, 100 g of fritillary bulb, 100 g of burdock, 100 g of honeysuckle, 50 g of rhubarb, 66 g of platycodon root and 50 g of liquorice

The above materials are made into capsule by conventional method.

Example 6:

ephedra 68 g gypsum 215 g weeping forsythia 215 g baikal skullcap root 100 g mulberry bark 220 g

90 g of bitter apricot kernel, 90 g of peucedanum root, 90 g of pinellia tuber, 90 g of dried orange peel, 90 g of fritillary bulb and 90 g of

90 g of burdock, 90 g of honeysuckle, 50 g of rhubarb, 50 g of platycodon root, 50 g of liquorice

The above materials are made into granule by conventional method.

Example 7:

50 g of ephedra, 200 g of gypsum, 300 g of weeping forsythia, 100 g of baikal skullcap root, 250 g of mulberry bark

100 g of bitter apricot kernel, 100 g of peucedanum root, 100 g of pinellia tuber, 100 g of dried orange peel, 100 g of fritillary bulb and 100 g of

100 g of burdock, 100 g of honeysuckle, 50 g of rheum officinale, 50 g of platycodon grandiflorum, 50 g of liquorice

The above materials are made into injection by conventional method.

Example 8:

ephedra herb 60 g gypsum 200 g weeping forsythia fruit 200 g baikal skullcap root 95 g mulberry bark 230 g

Semen armeniacae amarae 95 g radix peucedani 95 g pinellia ternate 95 g dried orange peel 95 g fritillary bulb 95 g

95 g of burdock, 95 g of honeysuckle, 50 g of rhubarb, 50 g of platycodon root, 50 g of liquorice

The above materials are made into pill by conventional method.

Experimental example:

in order to confirm the efficacy of the present invention in treating acute lung injury, the following pharmacological test studies were conducted using the tablet (hereinafter referred to as the present drug) prepared in the method of example 3:

data and method

1. Experimental materials

1.1 Medicine

1.1.1 Test article

The invention has the following specification: 1.843g crude drug/tablet, shijia is available from Kadsura Co.

1.1.2 Solvent(s)

Distilled water, self-made by the center, stainless steel water-break automatic control electric heating distilled water device (center number: 241), preservation condition: room temperature.

1.2 laboratory animals

BALB/c mice 60, SPF grade, female halves, body weight 17.6-23.4 g, received on month 09 of 2018, offered by Hunan Srilickida Biotechnology Co., ltd., quality certificate of laboratory animals: no.43004700050920, experimental animal production license number: SCXK (xiang) 2016-0002; the Hunan drug safety evaluation research center barrier A is raised, and the experimental animals use license number: SYXK (Hunan) 2015-0016.

1.3 Main reagent

Lipopolysaccharide (LPS, cat# L2880, sigma Co., ltd.), mouse IL-6 ELISA kit (lot # 201811), mouse TNF-alpha ELISA kit (lot # 201811), mouse Neutrophil Elastase (NE) ELISA kit (lot # 201811), mouse SOD ELISA kit (lot # 201811), mouse MDA ELISA kit (lot # 201811), mouse NO ELISA kit (lot # 201811), mouse monocyte chemotactic protein-1 (MCP-1) ELISA kit (lot # 201811), all of which are Bio-Swamp products; human nuclear factor kappa B inhibitor protein alpha (IKB-alpha) primary antibody (product number: ab 32518), phosphorylated human nuclear factor kappa B inhibitor protein alpha (p-IKB-alpha) primary antibody (product number: ab 133462), human nuclear factor kappa B inhibitor kinase beta (IKK-beta) primary antibody (product number: ab 124957), all of which are products of abcam corporation; nuclear factor kappa B P (NF-kappa B P) primary antibody (cat# 10745-1-AP) and actin (beta-actin) primary antibody (cat# 60008-1-Ig), all of which are products of Proteintech company.

1.4 Main instrument

Spectra Max i3x type multifunctional enzyme labeling instrument (present center number: 525, manufactured by Austrian Mey Valley photon Co.); DFC 420C pathology imaging System (this center number: 017, manufactured by Leica, germany); JY04S-3C gel imaging analysis system (present center number: 048, manufactured by Beijing junyi Oriental Co.).

2. Experimental method

2.1 Modeling, grouping and administration

BALB/c mice 60, SPF grade, male and female half, body weight 17.6-23.4 g, according to the body weight random divided into normal control group, model control group, the low, middle and high drug of the invention (1.83, 3.67, 7.33g crude drug/kg) dose group, each group of 10 animals. The medicine (ground into powder) is prepared into liquid medicine with corresponding concentration by distilled water before daily administration. Each group of mice was given the corresponding liquid medicine by gavage of 20mL/kg for 1 time/day for 7 consecutive days, and the normal control group and the model control group were given the same volume of distilled water by gavage. 1h after the last administration, injecting 300mg/kg of 10% chloral hydrate into the abdominal cavity of each group of mice for anesthesia, fixing the mice on an operation plate, disinfecting the neck, longitudinally cutting the skin of the neck, stripping subcutaneous tissues and exposing the trachea; the mice of each group except the normal group control group were injected with lipopolysaccharide at 0.5mg/kg into the trachea to induce an acute lung injury model. Immediately after injection, the mice were stood upright and gently rocked several times to allow the liquid to be evenly distributed in the lungs, and then the wounds were sutured, and the normal control group was injected with the same dose of 0.9% sodium chloride injection, and the mice were left to wake naturally.

2.2 Detection index

Mice of each group were sacrificed 5h after the end of molding, lungs were dissected and removed, and left lung was weighed (right lung was left for later use), placed in a homogenizer, and lung mass (g): physiological saline (mL) =1:9, adding 0.9% sodium chloride injection, grinding to homogenate, preparing into a rat lung suspension, centrifuging at 2500rpm in 4 ℃ environment for 20min, collecting supernatant, and detecting MDA, SOD, NO, MCP-1 content by ELISA kit; orbital blood collection, ELISA kit detects in serum: IL-6, TNF- α, NE content; dissecting, taking lung tissue, and detecting NF-kappa B P, IκB-alpha, p-IκB-alpha and IKKKbeta protein expression in the lung tissue by WB; dissecting right lung tissue, formalin fixation, paraffin embedding, slicing, HE staining, observing pathological changes of the lung by using a light microscope, and scoring the injury condition in the lung and inflammatory cell infiltration condition, wherein the specific scoring criteria are as follows: 0 point: normal; 1, the method comprises the following steps: mild (less than 30% lesions); 2, the method comprises the following steps: moderate (lesion 30% -50%); 3, the method comprises the following steps: severe (lesions greater than 50%).

2.3 Dose design

The specific groupings and dose designs are detailed in Table 1.

2.4 Statistical method

The significant numerical modification of this test data was performed by rounding and statistical analysis according to the SOP specifications. The software used for statistics was SPSS 16.0. The metrology data is expressed as mean ± standard deviation (), and the normalization and variance alignment are checked using the level's test method. If there is no statistical significance (P > 0.05), statistical analysis is performed using one-way analysis of variance (ANOVA). If ANOVA is statistically significant (P.ltoreq.0.05), a comparison analysis is performed using LSD test (parametric method). If the variance is not uniform (P.ltoreq.0.05), then the Kruskal-Wallis test is used. If the Kruskal-Wallis Test is statistically significant (P.ltoreq.0.05), a comparative analysis is performed using Dunnett's Test (nonparametric method). The statistical result takes alpha=0.05 as a checking limit, wherein p.ltoreq.0.05 represents a statistical significance and p.ltoreq.0.01 represents a very significant significance of the checked differences.

3. Results

3.1 Influence on antioxidant capacity of lung of acute lung injury model

As shown in Table 2, compared with the normal control group, the MDA content in the alveolar lavage fluid of the mice in the model control group is obviously increased (P is less than or equal to 0.01), and the SOD and NO are obviously reduced (P is less than or equal to 0.01); compared with a model control group, the MDA content in the low, medium and high dose group mouse alveolar lavage fluid is obviously reduced (P is less than or equal to 0.01), the SOD content is obviously increased (P is less than or equal to 0.01), and the NO content in the high dose group mouse alveolar lavage fluid is obviously increased (P is less than or equal to 0.01), so that the drug can reduce the deposition of lipid peroxide in the lung of an acute lung injury model mouse to relieve oxidative stress, and can improve the content of antioxidant enzyme to enhance the antioxidant capacity.

Effect on acute lung injury inflammatory cytokine release

As shown in Table 3, compared with the normal control group, the serum of the mice in the model control group has significantly increased IL-6, TNF-alpha, NE and MCP-1 contentsP Not more than 0.05 orP Less than or equal to 0.01); compared with the model control group, the invention has the advantages that the IL-6 and TNF-alpha content in the serum of the mice in the low, medium and high dose groups is obviously reducedP Not more than 0.05 orP Less than or equal to 0.01), the content of NE and MCP-1 in the high dose group is obviously reducedP Less than or equal to 0.05), the medicine provided by the invention can obviously relieve inflammatory reaction in lung.

Effect on NF- κ B P65, IκB- α, p-IκB- α, IKKKβ protein expression in lung in acute lung injury model

As shown in Table 4 and FIG. 1, the expression levels of IkB-alpha, p-IkB-alpha, IKKK beta and NF- κ B P65 in the lung tissue of the model control group mice were compared with those of the normal control groupAll have obvious riseP Less than or equal to 0.01); compared with the model control group, the invention has obviously reduced expression level of IkB-alpha and p-IkB-alpha in the lungs of mice in low, medium and high dose groupsP Not more than 0.05 orP Less than or equal to 0.01), the expression of IKKK beta and NF-kappa B P65 in the medium-dose group is obviously reducedP Less than or equal to 0.01), the medicine of the invention is suggested to inhibit the transduction of IKK/IκB/NF- κB.

Impact on acute Lung injury mice Lung

As shown in table 5, fig. 2, the normal control mice had intact alveolar structure, no apparent congestion, no inflammatory cell infiltration, and normal alveolar wall structure. Compared with the normal control group, the pathological score of the mice in the model control group is obviously increasedP Less than or equal to 0.01), the pathological changes are mainly characterized by massive inflammatory cell infiltration in alveolar cavities, red blood cell exudation, obvious thickening of alveolar walls, obvious hyperplasia and edema of alveolar interstitium. Compared with the model control group, the pathology score of the medium-high dosage group of the medicine is obviously reducedP Not more than 0.05 orP Less than or equal to 0.01), the alveolar structure of the mice in the dosage group is complete, a small amount of inflammatory cells infiltrate, proliferation and edema phenomena are obviously reduced, and the alveolar results of the mice in the high dosage group are complete, and various indexes are close to those of a normal control group.

4. Conclusion(s)

The test results show that the medicine can obviously reduce the release of inflammatory factors in the lung of acute lung injury to relieve inflammatory reaction in the lung, improve the content of antioxidant enzyme to enhance antioxidant capacity, inhibit signal transduction of protein expression, and has obvious clinical curative effect without any clinical adverse reaction and toxic or side effect in the whole treatment process. The medicine has good treatment effect on acute lung injury.

Claims (9)

1. The application of the traditional Chinese medicine composition in preparing the medicine for treating the acute lung injury is characterized in that the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:

52-86 of ephedra; gypsum 194-324; fructus forsythiae 194-324; 78-130 parts of radix scutellariae; white mulberry root bark 194-324; 78-130 parts of bitter apricot kernel; 78-130 parts of radix peucedani; 78-130 parts of pinellia ternate; 78-130 parts of dried orange peel; 78-130 of fritillary bulb; 78-130 parts of burdock; 78-130 parts of honeysuckle; 39-65 parts of rheum officinale; radix Platycodi 46-76; 39-65 parts of liquorice.

2. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:

herba Ephedrae 52; gypsum 324; fructus forsythiae 194; radix Scutellariae 78; cortex Mori 194; bitter apricot seed 130; radix Peucedani 78; pinellia ternate 130; dried orange peel 78; fritillary bulb 78; fructus Arctii 130; honeysuckle 130; rhubarb 39; radix Platycodi 76; licorice 65.

3. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:

herba Ephedrae 86; gypsum 194; fructus forsythiae 324; radix Scutellariae 130; cortex Mori 324; bitter apricot kernel 78; radix Peucedani 130; pinellia tuber 78; dried orange peel 130; fritillary 130; burdock 78; honeysuckle 78; rhubarb 65; radix Platycodi 46; licorice 39.

4. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:

ephedra 69; gypsum 259; fructus forsythiae 259; radix Scutellariae 104; cortex Mori 259; bitter almonds 104; radix Peucedani 104; pinellia ternate 104; dried orange peel 104; fritillary 104; fructus Arctii 104; honeysuckle 104; rhubarb 52; radix Platycodi 61; licorice 52.

5. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:

herba Ephedrae 55; gypsum 254; fructus forsythiae 318; radix Scutellariae 107; white mulberry root bark 203; bitter apricot kernel 107; radix Peucedani 82; pinellia ternate 105; dried orange peel 84; fritillary bulb 125; burdock 122; honeysuckle 113; rhubarb 42; radix Platycodi 60; licorice root 50.

6. The use according to any one of claims 1-5, characterized in that the bitter almonds are fried bitter almonds, fritillary bulbs, honeysuckle flowers, lonicera confusa and pinellia ternate.

7. The use according to any one of claims 1-5, characterized in that the pharmaceutical dosage form is a tablet, capsule, powder, granule, oral liquid, pill, tincture, syrup, suppository, gel, spray.

8. Use according to any one of claims 1 to 5, for the preparation of a medicament for the treatment of elevated protein expression caused by acute lung injury.

9. The use according to claim 8, characterized in that the protein is NF- κ B P65, ikb- α, p-ikb- α, ikkβ.