CN108126134B - Traditional Chinese medicine compound composition for treating liver injury, preparation method and application thereof - Google Patents
Traditional Chinese medicine compound composition for treating liver injury, preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/076—Poria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/484—Glycyrrhiza (licorice)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/535—Perilla (beefsteak plant)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
- A61K36/8998—Hordeum (barley)
Abstract
The invention discloses a traditional Chinese medicine compound composition for treating liver injury, which is prepared from the following raw materials in parts by weight: 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos. In addition, the invention also provides a preparation method and application of the traditional Chinese medicine compound composition for treating liver injury. The traditional Chinese medicine composition disclosed by the invention is simple in composition and reasonable in proportion, and can obviously relieve symptoms such as inappetence, tiredness, hypodynamia, abdominal distension, eructation, nausea, vomiting, weight loss, fullness and dull pain in the liver area or right upper abdomen and the like during liver injury, and the accumulation of liver fat is obviously reduced. The traditional Chinese medicine composition has good curative effect, no side effect and convenient taking after being taken for a long time, and provides a good choice for the patients with liver injury.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine compound composition for treating liver injury, a preparation method and application thereof.
Background
Liver damage is one of the most important common diseases affecting human health, and various harmful factors such as virus, biology, medicine, physicochemical, alcohol and the like can cause liver damage. The etiology of liver injury diseases is complex, and the mechanism thereof is not clear so far, but at present, it is considered that organelle damage, change in cell structure, lipid peroxidation, lipodystrophy, change in enzyme activity, disruption of ion balance, and the like are mainly involved. Among them, the disorder of lipid metabolism directly causes hepatic steatosis, causing damage, and at least the links involved in the assembly of triglyceride apolipoprotein complex, the transport function of cell membrane, the increase of lipid storage, and the oxidation function of mitochondria are considered to be involved.
The prevention and treatment of liver injury diseases by drugs has become an important issue of social and medical attention. Although the development of drugs for treating liver injury diseases is still going deep, the application of the existing chemical drugs such as interferon, arabinoside and the like is limited due to the fact that the price is high, the popularization is difficult, the adverse reactions are many and the like.
The traditional Chinese medicine has a long history of treating liver injury diseases, and belongs to the categories of symptoms such as hypochondriac pain, jaundice, accumulation and tympanites in traditional Chinese medicine according to the clinical manifestations of liver injury, and the etiology of the diseases is usually the external infection of damp-heat and epidemic disease; improper diet and overeating of fat and sweet food; excessive drinking; the drug toxicity and liver poison in industry and environment act on human body. Therefore, the pathogenic factors of damp-toxin are the main causes of this disease, and liver depression and spleen deficiency, and lingering pathogen due to deficiency of vital qi are the basis of the pathological changes of this disease. According to the common pathogenesis of liver injury, the treatment rules of clearing heat, promoting diuresis, removing toxicity, soothing liver, relieving depression, promoting blood circulation, removing blood stasis, strengthening spleen, eliminating phlegm and the like are mostly adopted, but the curative effect needs to be further improved. Therefore, under the guidance of the traditional Chinese medicine theory, a traditional Chinese medicine compound composition of natural medicines with good liver protection effect is preferably selected as a treatment means for patients with liver injury.
Disclosure of Invention
The invention provides a traditional Chinese medicine compound composition for treating liver injury, which solves the problems that the chemical medicine for treating liver injury in the prior art is expensive, difficult to popularize and has more adverse reactions, and also solves the problem that the traditional Chinese medicine for treating liver injury has an unsatisfactory curative effect.
The invention aims to provide a traditional Chinese medicine compound composition for treating liver injury, which is prepared from the following raw materials in parts by weight: 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos.
Preferably, the preparation raw materials of the traditional Chinese medicine compound composition for treating liver injury comprise the following components in parts by weight: 30 parts of purslane, 30 parts of cortex albiziae, 20 parts of raw malt, 10 parts of honey-fried licorice root, 20 parts of perilla stem and 20 parts of poria cocos.
The second purpose of the invention is to provide a preparation method of a traditional Chinese medicine compound composition for treating liver injury, which is implemented according to the following steps:
step 1, weighing 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos for later use;
step 2, crushing the purslane weighed in the step 1, sieving the crushed purslane with a 30-mesh sieve to obtain purslane coarse powder, adding petroleum ether with the weight 10 times that of the purslane coarse powder into the purslane coarse powder, performing reflux extraction for 1 hour to perform degreasing, and performing reduced pressure concentration after the reflux is finished to obtain degreased purslane coarse powder;
adding an ethanol solution which is 10 times of the weight of the defatted purslane coarse powder and has a mass concentration of 80% into the defatted purslane coarse powder, soaking for 0.5h, performing reflux extraction for 2 times after soaking, extracting for 1h each time, filtering after extraction, combining the two filtrates, and performing reduced pressure concentration until no ethanol smell exists to obtain a concentrated solution;
adding distilled water into the concentrated solution, and dispersing the concentrated solution into a liquid medicine containing crude drugs of 0.3 g/ml; loading the liquid medicine on AB-8 type macroporous adsorbent resin, eluting with 2 times column volume of distilled water, and removing water eluate; then eluting with ethanol solution with 4 times column volume and mass concentration of 10%, and discarding the first ethanol eluate; eluting with 80% ethanol solution with 6 times column volume, collecting second alcohol eluate, concentrating under reduced pressure, drying, and pulverizing to obtain herba Portulacae extract;
step 3, mixing the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos weighed in the step 1, adding water which is 5 times of the total weight of the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos into the mixture, soaking the mixture for 30min, decocting the mixture for 1h after soaking, and filtering the decoction to obtain first filtrate and filter residue; adding water with weight 3 times of the residue into the residue, decocting for 0.5 hr, filtering to obtain second filtrate, mixing the first filtrate and the second filtrate, mixing, and concentrating to relative density of 1.2-1.4 to obtain mixed extract;
and 4, uniformly mixing the purslane extract obtained in the step 2 and the mixed extract obtained in the step 3 to obtain the traditional Chinese medicine compound composition for treating liver injury.
The third purpose of the invention is to provide the application of the traditional Chinese medicine compound composition for treating liver injury in preparing a medicinal preparation for treating liver injury.
Preferably, the traditional Chinese medicine compound composition for treating liver injury is prepared into a pharmaceutically acceptable pharmaceutical preparation according to a conventional method.
Preferably, the pharmaceutical preparation is oral liquid, capsules, granules, pills, powder, microcapsules or dropping pills.
The pharmacological action and the drug effect of the traditional Chinese medicine of the invention are as follows:
1. purslane: cold in nature, sweet and sour in taste; enter heart, liver, spleen and large intestine meridians. Clearing away heat and toxic material, inducing diuresis, removing dampness, dispelling blood stasis, relieving swelling, diminishing inflammation, relieving pain, stopping bleeding, and cooling blood. Can be used for treating dysentery, enteritis, nephritis, puerperal metrorrhagia, hematochezia, mastitis, etc.
2. Silktree albizzia bark: sweet and neutral. It enters heart, liver and lung meridians. Relieve depression, soothe the nerves, activate blood and relieve swelling. Can be used for treating uneasiness, melancholy insomnia, lung carbuncle, sore, swelling, traumatic injury and pain.
3. Raw malt: sweet and neutral. It enters spleen and stomach meridians, has the actions of soothing liver, moving qi and relieving qi stagnation, and can be used for hypochondriac pain and abdominal pain due to liver qi stagnation or liver-stomach disharmony, and it is combined with other liver-soothing and qi-regulating herbs. Zhang xi is used in modern famous physicians to soothe the liver and promote liver qi. Zhang xi is indicated that although it is a spleen and stomach herb, it excels in soothing liver qi (unprocessed for soothing liver and not effective when stir-baked). The liver is spring when it covers, wood in five elements is originally the bud of qi transformation of human body, and malt and liver are in the same qi phase, so they are good at relieving. Fu gan governing smoothing flow of qi is the function of kidney moving qi, which can soothe liver and help liver wood smoothing flow of qi to move kidney qi.
4. Honey-fried licorice root: and (4) tonifying qi. Sweet in flavor and neutral in nature. Has the functions of invigorating spleen and replenishing qi, relieving cough and eliminating phlegm, clearing away heat and toxic material, relieving spasm and pain and blending various medicines. Can be used for treating weakness of spleen and stomach, deficiency of middle-warmer energy, cough, asthma, carbuncle, sore, abdominal spasm, and pain, relieving drug toxicity, and removing drug toxicity.
5. Perilla stem: pungent and warm. It enters lung and spleen meridians. The purple perilla has the effects of pungent and warm nature and moving powder, has light leaves like the lung, can disperse wind-cold, ventilate the lung and relieve cough; the stalks, like the spleen and stomach, are good at moving qi and regulating the middle warmer, regulating qi and preventing abortion.
6. Tuckahoe, poria cocos: sweet, bland and flat, has the effects of promoting diuresis, eliminating dampness, strengthening spleen and stomach, calming heart and tranquilizing mind, and is used for treating edema oliguria, phlegm and fluid retention and dizziness and palpitation, spleen deficiency and poor appetite, loose stool and diarrhea, uneasiness and the like. Compendium of materia Medica: poria cocos, Poria; dongyuan means that it is the yin in yang and descending, so it is also called the action.
The purslane is selected as a monarch drug, and has the effects of clearing away heat and toxic materials, nourishing the liver, dredging collaterals, and eliminating dampness and harmonizing the stomach. Sour liver enters liver to nourish liver yin and nourish liver, and purslane has the action of resolving dampness and harmonizing stomach to help food transportation and transformation to generate qi and blood. Cortex albiziae, raw malt and honey-fried licorice root are taken as the ministerial drugs, the cortex albiziae is sweet and mild in nature, has the effects of tonifying spleen and excreting dampness, and expels toxin by virtue of the property of excreting dampness of the cortex albiziae when the spleen and the stomach are protected; the raw malt is sweet and neutral in nature and has the effects of promoting qi circulation to promote digestion, assisting the transportation and transformation of the spleen and stomach, soothing the liver and relieving depression and regulating the liver qi; the prepared licorice root, radix glycyrrhizae preparata, sweet and neutral, enters spleen and stomach channels, has the effects of protecting the spleen and stomach, clearing heat and promoting fluid production, and has double effects of tonifying and eliminating evil for patients with stomach deficiency and body fluid deficiency. The perilla stem, the tuckahoe and the perilla stem which is pungent and warm and disperses, promote the circulation of qi and regulate the middle warmer, regulate the flow of qi and prevent miscarriage are assisted; poria cocos wolf can not only sooth liver and raise yang, but also clear head and eyes, clear up and descend. The medicines are used together to nourish the liver, invigorate the spleen, detoxify and excrete dampness.
Compared with the prior art, the invention has the beneficial effects that:
the traditional Chinese medicine compound composition is used for treating liver injury and regulating and reducing the accumulation of fat in the liver. Based on the theory of traditional Chinese medicine, syndrome differentiation and disease differentiation are combined, and the medicine application is relatively focused on the effects of soothing liver, regulating qi, strengthening spleen, reducing phlegm, promoting diuresis, reducing turbidity, promoting blood circulation, removing blood stasis, removing food retention, removing food stagnation and the like, and the effects of soothing liver, activating stagnancy, reducing dampness, reducing turbidity, strengthening spleen, tonifying kidney, promoting blood circulation and removing blood stasis.
The traditional Chinese medicine composition has simple composition and reasonable proportion, and experiments prove that the traditional Chinese medicine composition obviously relieves symptoms of inappetence, tiredness, hypodynamia, abdominal distension, eructation, nausea, vomiting, weight loss, fullness and dull pain in the liver area or the right upper abdomen and the like during liver injury, and obviously reduces the accumulation of liver fat. The traditional Chinese medicine composition has good curative effect, no side effect and convenient taking after being taken for a long time, and provides a good choice for the patients with liver injury.
Detailed Description
In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.
Purslane, cortex albiziae, raw malt, honey-fried licorice root, perilla stem and poria cocos used in the embodiments of the invention are all commercially available, and the test methods are all conventional methods unless otherwise specified.
Example 1
A traditional Chinese medicine compound composition for treating liver injury is prepared from the following raw materials in parts by weight: 30 parts of purslane, 30 parts of cortex albiziae, 20 parts of raw malt, 10 parts of honey-fried licorice root, 20 parts of perilla stem and 20 parts of poria cocos.
The method is implemented according to the following steps:
step 1, weighing 30 parts of purslane, 30 parts of cortex albiziae, 20 parts of raw malt, 10 parts of honey-fried licorice root, 20 parts of perilla stem and 20 parts of poria cocos for later use;
step 2, crushing the purslane weighed in the step 1, sieving the crushed purslane with a 30-mesh sieve to obtain purslane coarse powder, adding petroleum ether with the weight 10 times that of the purslane coarse powder into the purslane coarse powder, performing reflux extraction for 1 hour to perform degreasing, and performing reduced pressure concentration after the reflux is finished to obtain degreased purslane coarse powder;
adding an ethanol solution which is 10 times of the weight of the defatted purslane coarse powder and has a mass concentration of 80% into the defatted purslane coarse powder, soaking for 0.5h, performing reflux extraction for 2 times after soaking, extracting for 1h each time, filtering after extraction, combining the two filtrates, and performing reduced pressure concentration until no ethanol smell exists to obtain a concentrated solution;
adding distilled water into the concentrated solution, and dispersing the concentrated solution into a liquid medicine containing crude drugs of 0.3 g/ml; loading the liquid medicine on AB-8 type macroporous adsorbent resin, eluting with 2 times column volume of distilled water, and removing water eluate; then eluting with ethanol solution with 4 times column volume and mass concentration of 10%, and discarding the first ethanol eluate; eluting with 80% ethanol solution with 6 times column volume, collecting second alcohol eluate, concentrating under reduced pressure, drying, and pulverizing to obtain herba Portulacae extract;
step 3, mixing the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos weighed in the step 1, adding water which is 5 times of the total weight of the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos into the mixture, soaking the mixture for 30min, decocting the mixture for 1h after soaking, and filtering the decoction to obtain first filtrate and filter residue; adding water with weight 3 times of the residue into the residue, decocting for 0.5 hr, filtering to obtain second filtrate, mixing the first filtrate and the second filtrate, mixing, and concentrating to relative density of 1.2-1.4 to obtain mixed extract;
step 4, uniformly mixing the purslane extract obtained in the step 2 and the mixed extract obtained in the step 3 to obtain a traditional Chinese medicine compound composition for treating liver injury;
pulverizing the Chinese medicinal compound composition, adding appropriate amount of adjuvants, mixing, granulating, sieving, and drying to obtain granular Chinese medicinal compound composition for treating liver injury.
Example 2
A traditional Chinese medicine compound composition for treating liver injury is prepared from the following raw materials in parts by weight: 20 parts of purslane, 15 parts of cortex albiziae, 15 parts of raw malt, 5 parts of honey-fried licorice root, 10 parts of perilla stem and 15 parts of poria cocos.
Example 2 was prepared as in example 1 except that the formulation in example 1 was changed to that of example 2.
The traditional Chinese medicine compound composition prepared in the embodiment 2 is crushed, added with a proper amount of auxiliary materials, mixed uniformly, sieved, granulated and dried to prepare the traditional Chinese medicine compound composition for treating liver injury in the form of pills.
Example 3
A traditional Chinese medicine compound composition for treating liver injury is prepared from the following raw materials in parts by weight: 25 parts of purslane, 20 parts of cortex albiziae, 18 parts of raw malt, 8 parts of honey-fried licorice root, 15 parts of perilla stem and 15 parts of poria cocos.
Example 3 was prepared as in example 1 except that the formulation in example 1 was changed to that of example 3.
The compound traditional Chinese medicine composition prepared in the embodiment 3 is crushed and added with a proper amount of auxiliary materials, the mixture is uniformly mixed, granulated, sieved and dried, the prepared particles are filled into a capsule shell, and capsules are prepared, so that the compound traditional Chinese medicine composition for treating liver injury in a capsule dosage form is prepared.
The traditional Chinese medicine compound composition for treating liver injury prepared in the embodiments 1 to 3 has good effects, and the effects of the invention are explained only by using the traditional Chinese medicine composition prepared in the embodiment 1 because the effects of the traditional Chinese medicine compound composition for treating liver injury prepared in the embodiments 1 to 3 are basically parallel.
The effects of the present invention will be further described below using an animal test.
Influence on mouse acute alcoholic liver injury model
1. Experimental Material
1.1 Experimental drugs
The traditional Chinese medicine compound composition for treating liver injury prepared in the embodiment 1;
bifendate dripping pills, batch number: a020141206, Vanbond pharmaceutical group, Inc.
1.2 Experimental reagents
Ethanol, Tianjin Yuansheng chemical Co., Ltd., Lot 2015122012; alanine Aminotransferase (ALT) kit, lot No.: 1507053, respectively; aspartate Aminotransferase (AST) kit, lot No.: 1507073, respectively; the medicines are all provided by Shanghai Fuxing Changcheng medical science Co., Ltd.
Coomassie brilliant blue protein kit, lot No.: 20160118, respectively; superoxide dismutase (SOD) kit, lot No.: 20160121, respectively; malondialdehyde (MDA) kit, lot No.: 20160119, respectively; the medicines are provided by Nanjing institute of bioengineering.
1.3 Experimental instruments
AU400 full-automatic biochemical analyzer, olympus, japan; cgtation3 CYT3 MFUDG microplate reader, beth instruments ltd, usa; UV-2000 UV-visible spectrophotometer, Youchi (Shanghai) instruments, Inc.; TGL-16G bench centrifuge, shanghai anting scientific instruments factory; electronic balance, mettler-toledo instruments (shanghai) ltd; DMI4000B intelligent inverted fluorescence microscope, laika (Leica) instruments ltd, germany.
1.4 Experimental animals
Kunming mouse, SPF grade, half male and female, weight 18-22g, provided by Jinan Pengyue laboratory animal Breeding Limited company, production license number: SCXK (Lu) 20140007, license number SYXK (Yu) 2015-.
2. Experimental methods
2.1 preparation and administration of acute alcoholic liver injury model
60 KM mice were randomly and uniformly divided into 6 groups, namely a normal control group, a model group, a low, medium and high dosage group (12.58, 25.16 and 50.32g/kg) and a bifendate group (5.625mg/kg) of the traditional Chinese medicine compound composition. Each group comprises 10 male and female halves. The administration group of the traditional Chinese medicine compound composition and the bifendate group are respectively intragastrically administrated with different dosages of the traditional Chinese medicine compound composition and the bifendate according to the volume of 20ml/kg, and the normal control group and the model group are respectively administered with distilled water with the same volume every day. The administration was 1 time per day for 7 days. 1h after the last administration, the rest groups except the normal control group are intragastrically administered with 14ml/kg of 50% ethanol to establish an acute alcoholic liver injury model.
2.2 measurement of index
After the stomach is perfused with ethanol, fasting is performed without water prohibition, eyeballs are picked up after 12h, blood is taken, serum is separated, and serum liver function indexes ALT and AST levels are measured by adopting a full-automatic biochemical analyzer. Adding normal saline into the right liver leaf to prepare 10% liver homogenate, centrifuging to obtain supernatant, detecting SOD activity in the liver homogenate by microplate method, and detecting MDA content by colorimetric method. After the serum was collected, the liver was removed, and the left lobe of the liver was fixed in a 10% formaldehyde solution for histopathological examination. Liver tissues are taken and fixed in 10% formaldehyde solution, and after the liver tissues are sliced by conventional paraffin embedding, the slices are stained by HE, and the pathological histological change is observed under a light microscope.
Wherein, the classification condition of the alcoholic liver injury in the histopathological examination of the liver is as follows:
"-" the hepatocyte structure is complete, the boundary is clear, the liver cord is arranged orderly, and the hepatocyte has no edema, fatty degeneration and necrosis;
the "+" hepatocytes showed mild edema, slightly increased cell volume, and small amount of inflammatory cells infiltrated into the hepatocytes;
the liver cells of "+" showed moderate edema, disorder of the liver cell cord arrangement, and significant increase of the lipid drop cell volume in part of cytoplasm, which was seen in infiltration of multiple inflammatory cells;
the +++ "liver cells show severe edema, the volume of lipid drop cells in partial cytoplasm is obviously increased, the structure of liver lobules is fuzzy, inflammatory cell infiltration is obvious, and liver cell necrosis is caused.
2.3 statistical methods
Using SPSS17.0 statistical software, the data measured is the average plus or minus standard deviation
"shows that the comparison between two groups adopts t test, the comparison between multiple groups adopts a one-factor variance method, the comparison between two groups adopts an LSD method when the variances are equal, and the comparison between two groups adopts a Dunnett's method when the variances are not equal. The semi-quantitative pathological analysis adopts a radius method.
3. Results of the experiment
The influence of the traditional Chinese medicine compound composition on the liver function of the mice with acute alcoholic liver injury is shown in tables 1-3.
TABLE 1 Effect on liver function in mice with acute alcoholic liver injury
(n＝10)
Note: p <0.05, P <0.01 compared to model groups.
As can be seen from Table 1, compared with the normal control group, the ALT and AST values in the serum of the model group mice are obviously increased (P is less than 0.01), which indicates that the alcoholic liver injury of the mice is successfully modeled. Compared with the model group, the ALT and AST values of the traditional Chinese medicine compound composition in the low, medium and high dose groups and the bifendate group are obviously reduced (P is less than 0.01 or P is less than 0.05).
TABLE 2 Effect on liver tissue SOD and MDA in mice with acute liver injury
(n＝10)
Note: p <0.05, P <0.01 compared to model groups.
As can be seen from Table 2, compared with the normal control group, the liver tissues of the model mice have significantly reduced levels of SOD and GSH (P <0.01) and significantly increased MDA content (P < 0.01). Compared with a model group, the traditional Chinese medicine compound composition has the advantages that the levels of SOD and GSH in liver tissues of mice in each dosage group are obviously increased, the content of MDA is obviously reduced (P is less than 0.01), and the surface traditional Chinese medicine compound composition has a certain protection effect on oxidative damage of the liver tissues.
Table 3 histopathological examination of mouse liver (n ═ 10)
As can be seen from Table 3, the lobular structure of the liver of the normal control mice was normal, the liver cell cords were radially arranged, there was no edema, degeneration and necrosis of the liver cells, and there was no infiltration of inflammatory cells in the area of the sink. In the model group, mice have disorder liver cord arrangement, loose cytoplasm, enlarged liver cells with edema, obvious steatosis and fat drops with different sizes in the cells, and the phenomena of liver cell necrosis and inflammatory cell infiltration can be seen. Compared with the model group, the traditional Chinese medicine compound composition has the advantages that the mouse liver cell lesion degree of each dose group is obviously reduced, the intracytoplasmic lipid drops are reduced, the steatosis is obviously reduced, and the liver cell edema and the necrosis are not obvious. The results show that each dosage group of the traditional Chinese medicine compound composition can inhibit the formation of hepatocyte necrosis and steatosis to different degrees, wherein the dosage group with high dosage is more obvious.
Second, the influence on the mouse drug-induced liver injury model caused by paracetamol
1 materials of the experiment
1.1 Experimental drugs
The traditional Chinese medicine compound composition for treating liver injury prepared in the embodiment 1;
bifendate dripping pills, batch number: a020141206, Vanbond pharmaceutical group, Inc.;
paracetamol sustained release tablets, batch number: h20010394, shanghai qiangsheng pharmaceuticals, inc.
1.2 Experimental reagents
Alanine Aminotransferase (ALT) kit, lot No.: d1507053; aspartate Aminotransferase (AST) kit, lot No.: d1507073; the above are provided by Shanghai Fuxing Long-character medical science, Inc. Coomassie brilliant blue protein kit, lot No.: 20160323, respectively; superoxide dismutase (SOD) kit, lot No.: 20160322, respectively; malondialdehyde (MDA) kit, lot No.: 20160311, respectively; are provided by Nanjing institute of biological engineering.
1.3 Experimental instruments
AU400 full-automatic biochemical analyzer, olympus, japan; rotation 3 CYT3 MFUDG microplate reader, Berton instruments Inc. USA; UV-2000 UV-visible spectrophotometer, Youchi (Shanghai) instruments, Inc.; TGL-16G bench centrifuge, shanghai anting scientific instruments factory; AX3242H electronic balance, mettler-toledo instruments (shanghai) ltd; DMI4000B intelligent inverted fluorescence microscope, laika (Leica) instruments ltd, germany.
1.4 Experimental animals
Kunming mouse, SPF grade, half male and female, weight 18-22g, provided by Jinan Pengyue laboratory animal Breeding Limited company, production license number: SCXK (Lu) 20140007, license number SYXK (Yu) 2015-.
2 method of experiment
2.1 Molding and treatment
60 mice are taken and randomly divided into 6 groups, each group comprises 10 mice, each half of the mice are male and female, and the mice are respectively a normal control group, a paracetamol model group, a bifendate group (5.625mg/kg), and a low, medium and high dosage group (12.58, 25.16, 50.32g/kg) of the traditional Chinese medicine compound composition. The administration groups of the traditional Chinese medicine compound composition and the bifendate group are respectively intragastrically administered with different dosages of the traditional Chinese medicine compound composition and the bifendate according to the volume of 20ml/kg, and the normal control group and the model group are intragastrically administered with distilled water with the same volume every day. The administration was 1 time per day for 7 days. The last administration is 6h, except for the normal control group, the administration of paracetamol is 0.2ml/10g, the intragastric administration is carried out once, so as to cause acute liver injury, and the intragastric administration of the normal control group is carried out with distilled water with the same volume. Animals were treated by weighing 12 hours after the last administration without water deprivation. Blood was collected from the eyeball, serum was separated, and liver was rapidly collected. Taking the liver right leaf to prepare 10% liver homogenate, centrifuging and taking supernatant for later use. The left lobe of the liver was fixed in 10% formaldehyde solution and examined histopathologically.
Wherein, the classification condition of the alcoholic liver injury in the histopathological examination of the liver is as follows:
"-" hepatocyte structure is normal, hepatocyte chord and hepatocyte are all normal;
the structure of the + hepatic cells is normal, the arrangement and the hierarchy of the hepatic cells are regular, the hepatic cells are slightly degenerated, and a small amount of inflammatory cell infiltration can be seen;
the structure of the liver cells is deformed, the arrangement of the liver cells is disordered, the liver cells are moderately degenerated and necrotized, and a plurality of inflammatory cells are obviously infiltrated;
the structure of the liver cells is obviously changed, the arrangement of the liver cells is disordered, large-area degeneration and necrosis are seen, and inflammatory cell infiltration is obvious.
2.2 measurement of index
Alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) in serum were measured using a fully automatic biochemical analyzer. The activity of superoxide dismutase (SOD) in the liver homogenate is detected by a microplate method, and the content of Malondialdehyde (MDA) is detected by a colorimetric method. HE staining, and observing histopathological changes under light microscope.
2.3 statistical methods
Using SPSS17.0 statistical software, the data measured is the average plus or minus standard deviation
"shows that the comparison between two groups adopts t test, the comparison between multiple groups adopts a one-factor variance method, the comparison between two groups adopts an LSD method when the variances are equal, and the comparison between two groups adopts a Dunnett's method when the variances are not equal. The semi-quantitative pathological analysis adopts a radius method.
3. Results of the experiment
The effect of the compound traditional Chinese medicine composition on the liver function of the mouse with the liver injury caused by the paracetamol is shown in tables 1-3.
TABLE 4 Effect on serum ALT, AST of liver injury mice induced by Paracetamol
(n＝10)
Note: p <0.05, P <0.01 compared to model groups.
As shown in Table 4, the serum levels of ALT and AST in the model mice are significantly increased (P <0.01) compared with the normal control group, indicating that the modeling is successful. Compared with a model group, the bifendate and the traditional Chinese medicine compound composition can reduce the levels of ALT and AST (P is less than 0.01) in each dosage group, and the effect is most obvious in a high-dosage group of the traditional Chinese medicine compound composition.
TABLE 5 Effect on liver tissue SOD and MDA of mice with liver injury caused by paracetamol
(n＝10)
Note: p <0.05, P <0.01 compared to model groups.
As can be seen from Table 5, the liver tissue of the model mice had a significantly increased MDA content (P <0.01) and the SOD activity was significantly decreased (P <0.01) as compared to the normal control group. Compared with the model group, the bifendate and the traditional Chinese medicine compound composition can reduce the content of MDA and increase the activity of SOD (P <0.01 or P <0.05) in each dose group.
Table 6 grading of liver tissue lesions (n ═ 10) for each group of mice
As shown in Table 6, compared with the model group, the dosage groups and bifendate group of the compound Chinese medicinal composition have significant statistical differences (P <0.01 or P < 0.05). The pathological morphology examination of the liver tissue shows that the liver lobules of the normal control group have normal structure, regular arrangement of liver cells, uniform size of liver cells, obvious cell nucleus, rich cytoplasm and radial and regular arrangement of liver cell cables around the central vein. The paracetamol model group has the defects that the arrangement of liver cells is disordered, liver cell cords disappear, the liver cells have edema and enlarged volume, the liver cells have degeneration and necrosis with different degrees, and obvious inflammatory cell infiltration can be seen. The success of the mouse drug-induced liver injury model replication caused by paracetamol is demonstrated. The liver cell degeneration and necrosis degree of the traditional Chinese medicine compound composition administration group is light, and the liver rope has recovery of different degrees. Wherein the effect of the high-dose group of the traditional Chinese medicine compound composition is most obvious.
The adoption of the model of acute alcoholic liver injury of mice caused by ethanol and the model of liver injury of mice caused by paracetamol proves that the Chinese medicinal compound composition has an exact liver protection effect, so that the Chinese medicinal compound composition can be used for preparing medicaments for treating liver injury.
On the basis of ensuring the safety of animal experiments, the Chinese medicinal compound composition for treating liver injury prepared in the embodiment 1 of the invention is clinically applied, and good technical effects are obtained through clinical application of 46 cases of patients with liver injury. The traditional Chinese medicine compound composition is taken by patients for 2 times a day, and is continuously taken for 30 days, so that clinical symptoms disappear or basically disappear or symptoms are improved, liver functions are normal or obviously recovered, or the numerical values of glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase are reduced by more than 50%, the number of patients reaches 44, and the effective rate reaches 95.7%. Clinical tests show that the traditional Chinese medicine compound composition has the effect of treating liver injury, is effectively used for preparing the medicine for treating the liver injury, realizes the application of the traditional Chinese medicine compound composition in treating the liver injury, and has remarkable economic and social benefits
It should be noted that when the following claims refer to numerical ranges, it should be understood that both ends of each numerical range and any value between the two ends can be selected, and since the steps and methods used are the same as those in embodiments 1-3, the preferred embodiments of the present invention have been described for the purpose of preventing redundancy, but once the basic inventive concept is known, those skilled in the art can make other changes and modifications to these embodiments. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (6)
1. The traditional Chinese medicine compound composition for treating liver injury is characterized by comprising the following raw materials in parts by weight: 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos.
2. The traditional Chinese medicine compound composition for treating liver injury according to claim 1, which is prepared from the following raw materials in parts by weight: 30 parts of purslane, 30 parts of cortex albiziae, 20 parts of raw malt, 10 parts of honey-fried licorice root, 20 parts of perilla stem and 20 parts of poria cocos.
3. The preparation method of the traditional Chinese medicine compound composition for treating liver injury according to claim 1, which is implemented by the following steps:
step 1, weighing 20-30 parts of purslane, 15-30 parts of cortex albiziae, 15-20 parts of raw malt, 5-10 parts of honey-fried licorice root, 10-20 parts of perilla stem and 15-20 parts of poria cocos for later use;
step 2, crushing the purslane weighed in the step 1, sieving the crushed purslane with a 30-mesh sieve to obtain purslane coarse powder, adding petroleum ether with the weight 10 times that of the purslane coarse powder into the purslane coarse powder, performing reflux extraction for 1 hour to perform degreasing, and performing reduced pressure concentration after the reflux is finished to obtain degreased purslane coarse powder;
adding an ethanol solution which is 10 times of the weight of the defatted purslane coarse powder and has a mass concentration of 80% into the defatted purslane coarse powder, soaking for 0.5h, performing reflux extraction for 2 times after soaking, extracting for 1h each time, filtering after extraction, combining the two filtrates, and performing reduced pressure concentration until no ethanol smell exists to obtain a concentrated solution;
adding distilled water into the concentrated solution, and dispersing the concentrated solution into a liquid medicine containing crude drugs of 0.3 g/ml; loading the liquid medicine on AB-8 type macroporous adsorbent resin, eluting with 2 times column volume of distilled water, and removing water eluate; then eluting with ethanol solution with 4 times column volume and mass concentration of 10%, and discarding the first ethanol eluate; eluting with 80% ethanol solution with 6 times column volume, collecting second alcohol eluate, concentrating under reduced pressure, drying, and pulverizing to obtain herba Portulacae extract;
step 3, mixing the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos weighed in the step 1, adding water which is 5 times of the total weight of the cortex albiziae, the raw malt, the honey-fried licorice root, the perilla stem and the poria cocos into the mixture, soaking the mixture for 30min, decocting the mixture for 1h after soaking, and filtering the decoction to obtain first filtrate and filter residue; adding water with weight 3 times of the residue into the residue, decocting for 0.5 hr, filtering to obtain second filtrate, mixing the first filtrate and the second filtrate, mixing, and concentrating to relative density of 1.2-1.4 to obtain mixed extract;
and 4, uniformly mixing the purslane extract obtained in the step 2 and the mixed extract obtained in the step 3 to obtain the traditional Chinese medicine compound composition for treating liver injury.
4. The use of the compound Chinese medicinal composition for treating liver injury according to any one of claims 1-2 in the preparation of a pharmaceutical preparation for treating liver injury.
5. The application of the traditional Chinese medicine compound composition for treating liver injury in preparing a pharmaceutical preparation for treating liver injury according to claim 4, wherein the traditional Chinese medicine compound composition for treating liver injury is prepared into a pharmaceutically acceptable pharmaceutical preparation according to a conventional method.
6. The application of the traditional Chinese medicine compound composition for treating liver injury in preparing a medicinal preparation for treating liver injury according to claim 5, wherein the medicinal dosage form of the medicinal preparation is oral liquid, capsules, granules, pills, powder, microcapsules or dripping pills.
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| WO2004112483A1 (en) * | 2003-06-13 | 2004-12-29 | Bui Can V | Nutraceutical for the prevention and treatment of cancers and diseases affecting the liver |
| CN102139006A (en) * | 2011-03-30 | 2011-08-03 | 匡海学 | Compound picrorhiza rhizome composition for treating hepatic injury and preparation method and application thereof |
| CN102727787A (en) * | 2012-06-27 | 2012-10-17 | 王忠民 | Drug with characteristics of rapid transaminase lowering and live protection |
| CN106902174A (en) * | 2017-03-15 | 2017-06-30 | 河南科技大学 | A kind of Chinese medicine composition for treating liver injury and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004112483A1 (en) * | 2003-06-13 | 2004-12-29 | Bui Can V | Nutraceutical for the prevention and treatment of cancers and diseases affecting the liver |
| CN102139006A (en) * | 2011-03-30 | 2011-08-03 | 匡海学 | Compound picrorhiza rhizome composition for treating hepatic injury and preparation method and application thereof |
| CN102727787A (en) * | 2012-06-27 | 2012-10-17 | 王忠民 | Drug with characteristics of rapid transaminase lowering and live protection |
| CN106902174A (en) * | 2017-03-15 | 2017-06-30 | 河南科技大学 | A kind of Chinese medicine composition for treating liver injury and preparation method thereof |
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