CN105079227B - Application of traditional Chinese medicine composition in preparation of medicine for treating radioactive lung injury - Google Patents
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Abstract
The invention discloses application of a traditional Chinese medicine composition in preparation of a medicine for treating radioactive lung injury. The traditional Chinese medicine composition mainly comprises fructus forsythiae, honeysuckle, isatis root, rhubarb, patchouli, male fern rhizome, rhodiola rosea and the like, plays the integral regulation advantage of the compound traditional Chinese medicine, organically combines the functions of removing disease evil, relieving symptoms and regulating immunity, and has obvious curative effect on treating radioactive lung injury proved by clinical experiments.
Description
Technical Field
The invention relates to application of a traditional Chinese medicine composition in preparation of a medicine for treating radiation-induced lung injury, and belongs to the field of Chinese herbal medicines.
Background
Radiation-induced lung injury (radio-pulmonary injury) is a complication caused by radiotherapy of malignant tumors of the chest, most commonly seen in lung cancer and breast cancer, and secondly in radiotherapy of esophageal cancer and mediastinal malignant tumors. The common clinical manifestations are: the symptoms can generally appear 4-16 weeks after the beginning of radiotherapy, most of the symptoms occur within 2-4 weeks after the end of radiotherapy, and a few of the symptoms can also occur within half a year after the end of radiotherapy, and are mainly dry cough, little sputum, dysphagia, chest distress and chest pain, dyspnea of severe patients, low fever, normal leucocyte, ineffective antibacterial treatment, lung smelling and wet hiccup when receiving radiation in the acute period, and the respiratory sound is coarse; the chronic period is manifested as voiced sound due to local percussion and low respiration or retch sound. The inspection method mainly comprises the following steps: chest X-ray examination, (1) early stage radiation pneumonitis: the early stage of radiotherapy causes pulmonary congestion and exudation change, and the corresponding parts of the radiotherapy have increased texture and small flaky images, the edges are blurred, and the images are gradually fused with each other when the edges are serious. (2) Pulmonary fibrosis: the radiation pneumonitis appears in the radiation area, part of the radiation pneumonitis is absorbed through treatment, but obvious fibrosis appears in the later period of part of patients, the pulmonary veins in the radiation area are rough and dense, the fibers shrink, and adjacent tissue displacement is pulled, such as mediastinum and trachea moving towards the patient side, diaphragm pulling up, intercostal space narrowing, lung volume reduction and the like. Compensatory emphysema in healthy areas. (3) Other changes: radiation induced mediastinitis in the mediastinal region, manifesting as widened mediastinal space and unclear margin, sometimes fused with the radiation zone in the lung; large-area radiotherapy can cause pachynsis and adhesion of pleura, or small amount of pleural effusion, leaf fissure broadening and the like; extensive fibrosis leads to symptoms of arterial hypertension, lung portal enlargement and right heart involvement.
The mechanism of the development of radiation-induced lung injury has not been fully elucidated at present, it is generally believed that radiation-induced lung injury is the result of a multi-factor involved multi-cell-factor cascade effect, which is related to target cell injury and active oxygen free radicals, and to cytokine production, expression and signal transduction [ Lifengyu, Liu Xiu, Zhang Jianyu ] research progress of the mechanism of the development of radiation-induced lung injury [ J ]. modern oncology, 2009, 17(3):576-578.], early stages of radiation-induced lung injury are manifested as radiation pneumonitis, which occur more than 1 to 3 months after radiotherapy, and are related to Tumor Necrosis Factor (TNF), interleukin 1 (interleukin-1, I L-1) and monocyte chemotactic protein (monocyte protein)
MCP), in particular TNF and I L-1 [ CaoJingxu, Zhangzhi. cell factor and radiation lung injury [ J ]. Wayao Med. Radiology Nuclear medicine, 2001, 25, (4):181-
The main current therapeutic means of western medicine is to use large dose of glucocorticoid plus antibiotic, and although a certain effect is achieved, the curative effect of part of patients is still not good [ Ghafoori P, Marks L B, Vujaskovic Z, et al, radiation-induced lung in therapy, Assessment, and preservation. Oncology (Willison Park) 2008, 22(1):37-47; study 52-53. ].
Western medicine for treating radioactive lung injury has no effective treatment means except for the use of adrenocortical hormone and antibiotics. According to clinical symptoms, radioactive lung injury can be classified into cough, asthma and consumptive lung disease in traditional Chinese medicine. Radiation belongs to the pathogenic factor of heat toxin in traditional Chinese medicine, easily burns body fluid of human body and damages healthy qi and yin blood of human body. The pathogenic factors of radiation are different from the pathogenic factors of general heat-toxicity, and they do not follow the transmission and transformation rule of wei-defensive qi, ying-nutrient blood, but directly enter the blood collaterals of zang-fu organs. The lung is the delicate organ, and the pathogenic toxin of radiation invades the lung, which is easily damaged. Firstly, the pathogenic heat-toxicity most easily fumigates lung yin, the deficient heat is internally exuberant, consuming healthy qi of human body and damaging both qi and yin, and secondly, the qi deficiency fails to circulate blood vessels, yin deficiency and fluid do not nourish blood, the blood is sticky, unsmooth blood circulation is caused, blood stasis is generated internally, and the lung collaterals are blocked. Qi and yin deficiency and toxic stasis obstructing the lung can affect the function of dispersing, descending and directing all the meridians of the lung, which can cause difficulty in entering clear qi and difficulty in discharging turbid qi, so that patients with radiation lung injury can have symptoms of dyspnea and chest distress, and serious patients can die due to respiratory failure. In conclusion, toxic heat is the main cause of radiation-induced lung injury, and qi-yin injury and toxic stasis are the main pathogenesis of the disease. Traditional Chinese medicine has certain advantages in this field. Traditional Chinese medicine focuses on integral adjustment, and can increase radiotherapy effect, improve immunity, protect hematopoietic system and improve life quality of patients while preventing and treating radiation lung injury. The treatment of the disease in traditional Chinese medicine follows the principle of treating the symptoms in case of emergency and treating the root cause in case of slow, and the treatment should be carried out according to different types of symptoms and signs no matter the symptoms and root causes are treated or the symptoms and root causes are treated in case of temporary symptoms.
The traditional Chinese medicine composition has the effects of clearing pestilence, detoxifying, ventilating lung and clearing heat, and is prepared from fructus forsythiae, honeysuckle, mix-fried ephedra herb, mint and cablin potchouli herb, and has the effects of relieving exterior syndrome and expelling pathogenic factors, and diffusing lung and defensive qi externally, gypsum, isatis root, male fern rhizome, houttuynia cordata and liquorice, and clearing heat, detoxifying, clearing internal heat, nourishing yin, clearing lung-heat and removing blood stasis, and clearing internal heat, so that the traditional Chinese medicine composition is a good medicine for treating defensive qi and qi, relieving both exterior and interior, and clearing heat and detoxifying, and experiments show that the medicine has a remarkable effect of treating radiation lung injury.
The invention is an improved invention based on the Z L03143211 patent, the content of which is described in the patent document is cited in the whole.
Disclosure of Invention
The invention provides application of a traditional Chinese medicine composition in preparing a medicine for treating radiation-induced lung injury, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight:
200 pieces of forsythia, 200 pieces of honeysuckle, 200 pieces of 300 pieces of radix isatidis and 40-60 pieces of 300-rhubarb
Patchouli 60-100 aspidium crassirhizoma 200-300 rhodiola rosea 60-100 menthol 5-9
60-100 fried Chinese ephedra, 60-100 fried bitter apricot seeds, 60-100 heartleaf houttuynia herb, 200-
Licorice 60-100, gypsum 200-300.
The traditional Chinese medicine composition disclosed by the invention preferably comprises the following raw material medicines in parts by weight:
forsythia fruit 200 honeysuckle 300 isatis root 200 rhubarb 60 patchouli 60
Rhizoma dryopteris crassirhizomae 300 rhodiola rosea 60 menthol crystal 9 mix-fried ephedra 60
Parched semen Armeniacae amarum 100, herba Houttuyniae 200, Glycyrrhrizae radix 100, and Gypsum Fibrosum 200.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
forsythia fruit 300 honeysuckle 200 isatis root 300 rhubarb 60 patchouli 100
Rhizoma Dryopteris Crassirhizomatis 200 radix Rhodiolae 60 Mentholum 5 herba Ephedrae 100
Parched semen Armeniacae amarum 60 herba Houttuyniae 300 Glycyrrhrizae radix 60 Gypsum Fibrosum 300.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
weeping forsythia 278 honeysuckle 294, isatis root 285, rhubarb 55 and patchouli 95
Rhizoma dryopteris crassirhizomae 290 rhodiola rosea 87 menthol crystal 8.5 mix-fried ephedra 88
Parched semen Armeniacae amarum 80, herba Houttuyniae 284, Glycyrrhrizae radix 95 Gypsum Fibrosum 277.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
255 portions of forsythia, 255 portions of honeysuckle, 255 portions of isatis root, 255 portions of rhubarb, 51 portions of patchouli, 85
Rhizoma dryopteris crassirhizomae 255 rhodiola rosea 85 menthol crystal 7.5 mix-fried ephedra 85
Parched semen Armeniacae amarum 85 herba Houttuyniae 255 Glycyrrhrizae radix 85 Gypsum Fibrosum 255.
The traditional Chinese medicine composition mainly comprises fructus forsythiae, honeysuckle, isatis root, rhubarb, patchouli, male fern rhizome, rhodiola rosea and the like, plays the integral regulation advantage of the compound traditional Chinese medicine, organically combines the functions of removing disease evil, relieving symptoms and regulating immunity, and has obvious curative effect on radioactive lung injury proved by clinical experiments.
The traditional Chinese medicine of the invention 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 standard or traditional Chinese medicine dictionary.
The active ingredients of the traditional Chinese medicine composition are prepared by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
the dry paste powder obtained in the step (5), the volatile oil obtained in the step (2) and the menthol jointly form the active ingredients of the traditional Chinese medicine composition.
The medicament of the invention is in the form of capsules, tablets, powders, granules, oral liquid, soft capsules, pills, tinctures, syrups, suppositories, gels, sprays or injections.
In order to make the above dosage forms possible, pharmaceutically acceptable excipients, such as: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, bases, and the like. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crospolyvinylpyrrolidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, etc.; 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 adhesive comprises starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, and the like; the flavoring agent comprises: sweeteners and various essences; the preservative comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chloroacetidine acetate, eucalyptus oil, etc.; the matrix comprises: PEG6000, PEG4000, insect wax, etc.
The capsule is prepared by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the particles obtained in the step (6), sealing, uniformly mixing, and encapsulating to obtain the capsule.
The preparation method of the granules comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot kernel, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to be 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the granules obtained in the step (6), sealing, uniformly mixing and bagging to obtain the menthol-containing capsule.
The preparation method of the preferred granules comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 6 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 8 times of 70% ethanol for 2 times, 2 hr for the first time, and 1.5 hr for the second time, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 9 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, combining extracting solutions, filtering, combining obtained filtrate with water filtrate obtained after oil extraction of pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding 95% ethanol, adjusting the alcohol concentration to be 70%, refrigerating, standing, filtering, and recovering ethanol until no alcohol smell exists, so as to obtain clear paste filtrate;
(5) mixing the fluid extract filtrate obtained in step (4) with the ethanol extract obtained in step (3), and concentrating to obtain soft extract with relative density of 1.25-1.35 at 60 deg.C;
(6) adding appropriate pharmaceutically acceptable adjuvants into the soft extract obtained in step (5), and granulating;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the granules obtained in the step (6), sealing, uniformly mixing and bagging to obtain the menthol-containing capsule.
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 a conventional preparation method, for example, a preparation process recorded in Vanbitsin traditional Chinese medicine pharmacy (1 st 12 months in 1997 of Shanghai scientific Press) to prepare a conventional dosage form acceptable in pharmacy.
The Chinese medicinal composition can also reduce the expression level and the protein level of interleukin-6 andor tumor necrosis factor- α caused by radioactive lung injury.
Description of the drawings:
FIG. 1 shows the effect of pathological changes of the lung tissue of rats with radioactive lung injury at different times (HE staining × 200), C. is blank control group, R is simple irradiation group, L HQWH is irradiation and high dose group of the traditional Chinese medicine composition, and L HQWM is irradiation and high dose group of the traditional Chinese medicine composition.
FIG. 2 shows the DNA bands I L-6 and TNF- α of each experimental group at each time point after irradiation, C. is blank control group, R is simple irradiation group, L HQWH is irradiation and high dose group of the traditional Chinese medicine composition, and L HQWM is irradiation and high dose group of the traditional Chinese medicine composition.
Detailed Description
The following examples are intended to illustrate the preparation of the medicaments according to the invention, but they are not intended to limit the scope of the invention in any way.
Example 1
Prescription:
255 g of forsythia, 255 g of honeysuckle, 255 g of isatis root, 255 g of rhubarb and 51 g of rhubarb
Pogostemon cablin 85 g male fern rhizome 255 g rhodiola rosea 85 g menthol crystal 7.5 g
85 g of mix-fried ephedra herb, 85 g of fried bitter apricot seed, 85 g of heartleaf houttuynia herb and 255 g of heartleaf houttuynia herb
Licorice root 85 g gypsum 255 g.
The preparation method comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the prescription, and cleaning;
(2) crushing herba Agastaches, extracting volatile oil with 6 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 8 times of 70% ethanol for 2 times, 2 hr for the first time, and 1.5 hr for the second time, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 9 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating and standing for 24 hours, filtering, and recovering the ethanol until no alcohol smell exists, thereby obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and spray drying to obtain dry extract powder;
(6) adding 142 g of starch into the dry paste powder obtained in the step (5), and granulating by using 85% ethanol;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the particles obtained in the step (6), sealing, uniformly mixing, and filling into 1000 capsules to obtain the capsule.
Example 2
Prescription:
200g of forsythia, 300 g of honeysuckle, 200g of isatis root, 200g of rhubarb, 60 g of rhubarb
Patchouli 60 g male fern rhizome 300 g rhodiola 60 g menthol 9 g
Mix-fried ephedra 60 g, bitter apricot kernel 100 g, cordate houttuynia 200g
Licorice root 100 g plaster 200 g.
The preparation method comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the prescription, and cleaning;
(2) crushing herba Agastaches, extracting volatile oil with 5 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6 times of 50% ethanol for 2 times, 1 hr for the first time, and 2.5 hr for the second time, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating and standing for 24 hours, filtering, and recovering the ethanol until no alcohol smell exists, thereby obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and spray drying to obtain dry extract powder;
(6) adding 151 g of starch into the dry paste powder obtained in the step (5), and granulating by using 85% ethanol;
(7) and (3) adding menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the granules obtained in the step (6), sealing, uniformly mixing, and pressing into tablets to obtain 935 tablets.
Example 3
Prescription:
300 g of forsythia, 200g of honeysuckle, 300 g of isatis root, 300 g of rhubarb and 40 g of rhubarb
Patchouli 100 g male fern rhizome 200g rhodiola root 100 g menthol 5g
Roasted ephedra 100 g, fried bitter apricot kernel 60 g, cordate houttuynia 300 g
Licorice root 60 g plaster 300 g.
The preparation method comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the prescription, and cleaning;
(2) crushing herba Agastaches, extracting volatile oil with 8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 10 times of 90% ethanol for 2 times, 3 hr for the first time, and 2.5 hr for the second time, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating and standing for 24 hours, filtering, and recovering the ethanol until no alcohol smell exists, thereby obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and spray drying to obtain dry extract powder;
(6) and (3) preparing the dry paste powder obtained in the step (5), the volatile oil obtained in the step (2) and the menthol into pills according to a conventional method to obtain pills 905.
Example 4:
the formula of the raw material medicine is as follows:
weeping forsythia 170 g honeysuckle flower 170 g mix-fried Chinese ephedra 57 g stir-fried bitter apricot kernel 57 g
170 g of gypsum, 170 g of isatis root, 170 g of male fern rhizome, 170 g of cordate houttuynia
Patchouli 57 g rhubarb 34 g rhodiola 57 g menthol 5.0 g
Licorice root, radix Glycyrrhizae 57 g
The preparation method comprises the following steps:
the extraction process comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the prescription, cleaning, and cutting according to the requirement;
(2) extracting herba Agastaches with 6 times of water for 4 hr, collecting volatile oil with oil yield of 0.33 + -0.05%, filtering the extractive solution, and removing residue;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 8 times of 70% ethanol for 2 times, 2 hr for the first time, and 1.5 hr for the second time, filtering the extractive solutions, mixing the filtrates, and recovering ethanol until no ethanol smell exists;
(4) adding 9 times of water into honeysuckle, fried bitter almond, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding the fried bitter almond, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, filtering an extracting solution, combining filtrates, simultaneously adding an aqueous solution obtained after oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 determined at 60 ℃, adding 95% ethanol, stirring while adding until the alcohol concentration is 70%, refrigerating and standing for 24 hours, filtering, recovering ethanol from the filtrate until no alcohol taste exists, combining with an alcohol extracting solution, concentrating into thick paste with the relative density of 1.25-1.35 determined at 60 ℃ for later use;
(II) preparation process:
(5) the formula of the preparation is as follows: 335.5g of thick paste obtained in step (4) and 5g of menthol
0.2ml of patchouli oil obtained in the step (2) and 342.5g of powdered sugar 514.0g of dextrin
(6) And (3) granulating: mixing sugar powder and dextrin, making soft mass with soft extract as binder, granulating with 14 mesh screen, oven drying at 60-65 deg.C, and grading with 10 mesh screen;
(7) subpackaging: sieving to obtain fine powder, adding appropriate amount of ethanol into Mentholum and herba Agastaches volatile oil, dissolving, spraying into the fine powder, mixing with the granule, sealing for half an hour, and packaging to obtain 1000g granule.
Experimental example:
to confirm that the Chinese medicinal composition of the present invention has the therapeutic effect on radiation-induced lung injury, the following pharmacological test studies were conducted on the capsules (hereinafter referred to as the present invention) prepared according to the method of example 1:
data and method
Materials and methods
1. The experimental animals and groups comprise 48 healthy, adult, SPF-grade, female and Wistar rats with the weight of 180-200 g purchased from the research center of experimental animals in Hubei province (certificate number: SCXK (Hubei) 2008. 0005), the rats are raised in a clean animal room for 7 days before the experiment, and the rats are observed to enter the Group, wherein the rats are randomly divided into a irradiated high dose Group (irradiated by high-dose L HQW with radiation, L HQ Group), a irradiated medium dose Group (irradiated by kymiddose-dose L HQW with radiation, L HQWM Group), a simply irradiated Group (radiation on Group, R Group) and a Blank Control Group (Blank taste Group, C Group), and each Group is 12 rats and are marked by picric acid.
2. The main reagents and drugs of the present invention are provided by Chijiazhuang Ling pharmaceutical products Ltd (production lot: 130232), and commercially available reagents Trizol (Invitrogen, USA), MM L V reverse transcriptase and dNTPs (TOYOBO, Japan), RnaseA enzyme inhibitor (Takara, Dalian), DNA polymerase (TransGen, Beijing), I L-6 and TNF- α E L ISA detection kit (PeproTech, USA), and oligodT, β -actin, I L-6 and TNF- α primers were synthesized by Shanghai Biotechnology Ltd.
3. The irradiation method comprises the steps of carrying out intraperitoneal injection anesthesia on rats by using 10% chloral hydrate physiological saline (350 mg. kg < -1 >), lying on the back, unfolding and fixing four limbs on a self-made fixing plate, fully exposing the chest, observing the irradiated rats by a monitor, wherein the size of an irradiation field is 2.8cm × 4.2.2 cm, the total dose DT = 20Gy and SSD =100.0 cm., and removing groups of the rats when wriggling occurs in the irradiation period.
4. Animal treatment, wherein the gavage starts from 3 days before the irradiation and ends at 28 days after the irradiation, 31 days are carried out, L HQWH group comprises 0.84 g.kg-1The weight is 1 time per day, L HQWM group comprises 0.42 g/kg of the Chinese medicinal composition powder for intragastric administration-1Body weight, 1 time daily; group C and group R: the normal saline is infused into the stomach 1 time a day.
5. Section and HE staining after lung tissue is fixed in 10% formaldehyde fixing solution for 24h, conventional paraffin embedding, section, sticking, dewaxing, HE staining, dehydration, transparency, mounting, and section observation and image acquisition are performed with an optical microscope (O L YMPUS IX81, Japan).
6. Cell counting of alveolar lavage fluid the rat dissects the chest after sacrifice, ligates the left main bronchus, lavages the right lung with PBS 3 times 2m L each time after tracheal intubation, the recovery rate is about 90%
Centrifuge at 1200 r/min for 5min, discard the supernatant, repeat washing 2 times, finally resuspend the cells with 4ml PBS and count.
7. Measurement of Interleukin-6 (I L-6) and tumor necrosis factor- α (TNF- α) in serum includes taking blood from rat heart at 4m L/min and 3000r/min after anesthesia, centrifuging for 10min, taking serum and storing at-80 deg.c, and detecting I L-6 and TNF- α in serum sample according to E L ISA kit.
8. An RT-PCR method for detecting the expression of lung tissue I L-6 and TNF- α mRNA includes extracting 50mg of lung tissue total RNA by 1ml of Trizol, measuring the purity (A260/A280 = 1.6-1.8) and concentration of the RNA by an ultraviolet spectrophotometer, carrying out reverse transcription reaction on 0.5 mu g of total RNA according to instructions, carrying out reaction at 30 ℃ for 10min, carrying out reaction at 42 ℃ for 60min, carrying out reaction at 70 ℃ for 15min, carrying out PCR on 1 mu l of RT product, carrying out pre-denaturation on PCR reaction at 94 ℃ for 3.5min, carrying out pre-denaturation at 94 ℃ for 30s, carrying out pre-denaturation at 53 ℃ for 15s, carrying out pre-denaturation at 72 ℃ for 20s, carrying out pre-denaturation for 33 cycles, carrying out photo-taking on PCR amplification products at the temperature of 50% agarose gel, and.
9. Statistical analysis
The analysis is carried out by using SPSS 17.0 statistical software, and the experimental data are all calculated by mean +/-standard deviation (xS), and the comparison between groups was performed by t-test and analysis of variance between groups, as P<0.05 indicates that the difference is statistically significant, P<0.01 indicates that the difference is statistically significant.
Second, experimental results
1. General conditions in rats
The rats in the single irradiation group exhibited listlessness, decreased food intake and activity, yellow secretion in eyes and dry stool on day 3 after irradiation, and partially had hair loss after 14 days after irradiation, and some of the rats exhibited cough symptoms, which began to gradually decrease on day 26 after irradiation. The rats with high and medium dose of the medicine of the invention have better mental states than the rats with simple irradiation, low incidence rate, light degree, late appearance time, short duration and the like in the aspects of eye secretion, dry and hard stool and the like, and the above manifestations begin to disappear gradually and no obvious cough symptom is caused by the 18 th day after irradiation.
2. Pathological tissue observation
The high and medium dose group after irradiation only shows a small amount of lung interstitium inflammatory cell infiltration on the 1 st day after irradiation, acute inflammatory changes such as light to moderate edema and inflammatory cell infiltration of the lung interstitium and the like on the 14 th day, the inflammation is obviously relieved at 28 days, the lung interstitium is thickened, and the alveolar structure is slightly damaged. In the simple irradiation group, the pulmonary interstitium is thickened and inflammatory cell infiltration is seen on the 1 st day after irradiation, the inflammatory reaction is serious on the 14 th day, and the pulmonary interstitium is hyperemia, hemorrhage and edema, a large amount of inflammatory cell infiltration is caused, the alveolar structure is damaged, the inflammation is slightly relieved on the 28 th day, the pulmonary interstitium is thickened, and the alveolar space is reduced (as shown in figure 1).
3. Alveolar lavage fluid cell count: after irradiation, the total number of alveolar lavage fluid cells of rats was significantly increased, and the total number of alveolar lavage fluid cells was decreased at each time point after irradiation in the high dose group of the present drug compared to the single irradiation group (see table 2), wherein the difference between day 14 and day 28 was statistically significant (t =3.237, 3.431, P < 0.05). The total number of alveolar lavage fluid cells in the dose group of the drug of the present invention was reduced compared with the single-irradiation group, but the difference was not statistically significant at day 14 after irradiation (t =1.710, P > 0.05), while the difference was significant at day 28 after irradiation (t =2.658, P < 0.05).
TABLE 2 alveolar lavage fluid cell count (x + -S, × 104/ml) of rats with radioactive lung injury under the action of the Chinese medicinal composition of the invention
Note that L HQWH group compared with the R group, t =3.237, # P <0.05, t =3.431, # P <0.05.
L HQWM group was compared with R group, t =1.710, # # P >0.05, t =2.658, # # # # P <0.05.
4. Determination of Interleukin-6 (I L-6) and tumor necrosis factor- α (TNF- α) levels in serum
Acute radiation lung injury rat model serum levels of both I L-6 and TNF-a were significantly elevated, but after the use of the inventive drugs, levels of both I L-6 and TNF-a were reduced (see table 3) wherein at the level of I L-6, the inhibitory effect of the inventive drugs at high doses was more significant, and the differences at days 1, 14 and 28 after irradiation compared to the single irradiation group were statistically significant (t =7.748, 15.119, 6.902, P < 0.01), whereas at the level of TNF-a, the inhibitory effect of the inventive drugs at medium doses was more significant, and the differences at day 1 after irradiation compared to the single irradiation group were statistically significant (t =3.940, P < 0.01), and the differences at days 14 and 28 after irradiation were also statistically significant (t =3.333, 2.259, P < 0.05).
TABLE 3 serum TNF-a and I L-6 levels (x + -S, ng/ml) in rats with radiation lung injury caused by the Chinese medicinal composition of the invention
Note that L HQWH group compared with R group t =3.940, IV P <0.01, t =3.333, t =2.259, VP <0.05, L HQWM group compared with R group t =7.748, t =15.119, t =6.902, III P <0.01.
5. Expression of lung tissue interleukin-6 (I L-6) and tumor necrosis factor- α (TNF- α) mRNA
At each time point after irradiation, the relative gray values of the I L-6 and TNF- α DNA bands and the relative β -actin internal reference band of each experimental group (see figure 2 and table 4) compared with the single irradiation group, the relative gray values of the I L-6 and TNF- α bands of the irradiation and honeysuckle antipyretic high and medium dose groups are all reduced, and the difference has statistical significance (P < 0.01).
TABLE 4 Effect of the Chinese medicinal composition of the present invention on the expression of TNF- α and I L-6 mRNA in lung tissue of rats with radiation-induced lung injury
C. A blank control group, a L HQWH irradiation group and a high dosage group of the traditional Chinese medicine composition, a L HQWM irradiation group and a R simple irradiation group
Third, conclusion
The experiments show that the traditional Chinese medicine composition can improve the general condition of rats, relieve the inflammatory reaction of lung tissues, inhibit the increase of the total number of cells in alveolar lavage fluid caused by acute radioactive lung injury, and play a role in preventing and treating the radioactive lung injury, and the traditional Chinese medicine composition can inhibit the protein levels of rat serum I L-6 and TNF- α and can down regulate the expression of I L-6 and TNF- α mRNA in the injured lung tissues.
Experimental results fully prove that the medicament can effectively treat the radioactive lung injury, is particularly suitable for increasing interleukin-6 (I L-6) and tumor necrosis factor- α (TNF- α) caused by the radioactive lung injury, does not have any clinical adverse reaction and toxic or side effect in the whole treatment process, and has remarkable clinical curative effect.
Claims (6)
1. The application of a traditional Chinese medicine composition in preparing a medicine for treating radiation-induced lung injury is characterized in that the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight:
255 portions of forsythia, 255 portions of honeysuckle, 255 portions of isatis root, 255 portions of rhubarb, 51 portions of patchouli, 85
Rhizoma dryopteris crassirhizomae 255 rhodiola rosea 85 menthol crystal 7.5 mix-fried ephedra 85
Parched semen Armeniacae amarum 85 herba Houttuyniae 255 Glycyrrhrizae radix 85 Gypsum Fibrosum 255.
2. Use according to claim 1, characterized in that the active ingredient of the medicament is made by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot kernel, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) the dry paste powder obtained in the step (5), the volatile oil obtained in the step (2) and the menthol jointly form the active ingredients of the traditional Chinese medicine composition.
3. The use according to claim 1, characterized in that the pharmaceutical dosage form is a capsule, tablet, powder, granule, oral liquid, pill, tincture, syrup, suppository, gel, spray or injection.
4. Use according to claim 3, characterized in that the capsules are made by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot kernel, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the particles obtained in the step (6), sealing, uniformly mixing, and encapsulating to obtain the capsule.
5. Use according to claim 3, characterized in that said granules are made by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot kernel, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to be 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the granules obtained in the step (6), sealing, uniformly mixing and bagging to obtain the menthol-containing capsule.
6. The use of claim 1, wherein the use of the composition is in the preparation of a medicament for treating an increase in interleukin-6 and/or TNF- α caused by radiation lung injury.
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