CN102145025B - Pine pollen extract for treating fatty liver as well as preparation method and application thereof - Google Patents
Pine pollen extract for treating fatty liver as well as preparation method and application thereof Download PDFInfo
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- CN102145025B CN102145025B CN 201010607364 CN201010607364A CN102145025B CN 102145025 B CN102145025 B CN 102145025B CN 201010607364 CN201010607364 CN 201010607364 CN 201010607364 A CN201010607364 A CN 201010607364A CN 102145025 B CN102145025 B CN 102145025B
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Abstract
The invention provides a pine pollen extract for treating fatty liver as well as a preparation method and an application thereof, relating to the technical field of extracting effective substances from gymnosperm pine pollen. The preparation method comprises the following steps: the pine pollen is subjected to wall breaking at a low temperature, and impurities in the pine pollen are removed; the pine pollen is extracted with ultrasonic after the pine pollen is immersed in normal-pressure normal-temperature water; solid impurities are centrifugally removed to obtain an aqueous solution; and a gymnosperm pine pollen aqueous extract with the molecular weight lower than 3,000D is separated at the environmental temperature of between 1 and 5 DEG C to obtain the gymnosperm pine pollen aqueous extract for treating the fatty liver. The pine pollen aqueous extract can be used for general health-care for children, and can prevent occurrence of fatty liver. The pine pollen aqueous extract can be used for preparing an oral preparation for treating the fatty liver, has a good absorption effect and low price, and also can be used for preparing an injection preparation for treating the fatty liver, which can be absorbed quickly, has strong treatment pertinence.
Description
Technical field
The present invention relates to from plant, particularly from the gymnosperm Pollen Pini, extract the Technology field of active substance.
Background technology
Fatty liver is meant the pathological changes of fatty overheap in the hepatocyte that causes owing to a variety of causes, and the positive serious threat compatriots' of fatty liver disease health becomes the second largest hepatopathy that is only second to viral hepatitis, has been acknowledged as the common cause of disguised liver cirrhosis.Fatty liver is a kind of common clinical picture, but not a kind of independently disease.The lighter is asymptomatic in its clinical manifestation, and weight person's state of an illness is violent.Generally speaking, fatty liver belongs to the reversibility disease, and early diagnosis and treatment in time often can recover normal.The formation of fatty liver generally is because the decomposition and the anabolism disequilibrium of fat in the liver or transport the generation obstacle, fat will be in hepatocyte excess accumulation, when fat content increases to more than 30%, then be called fatty liver.Fatty liver is different by the etiopathogenesis that self voxel matter is different, dietary habit does not cause fatty liver on an equal basis; Generally can be divided into obesity, cross feeding habits fatty liver, fatty liver after the hepatitis, alcoholic fatty liver; The deficiency disease fatty liver; Medicine property fatty liver, diabetic fatty liver, the latent source property fatty liver of gestational fatty liver and unknown cause etc.Fatty liver is divided into slight fatty liver, moderate fatty liver and severe fatty liver again by the light and heavy degree difference.Slight fatty liver has become modern's common problem, and slight fatty liver is the best period of treatment fatty liver.The average attack rate of (white collar personage, taxi driver, professional manager, individual proprietor, government official, high level intellectual etc.) fatty liver is 25% in some professional population; The sickness rate of fatty liver is 50% among obese people and the type patient; The sickness rate of being addicted to drink with the alcoholic fatty liver is 58%; The sickness rate of fatty liver is about 60% in often insomnia, tired, the sub-health population of not thinking ordinary food and drink, gastrointestinal dysfunction.Fatty liver crowd's age also constantly descends in recent years, and the age has only 40 years old, and the patient about 30 years old is also more and more, and the fatty liver of male below 45 years old is obviously more than the women.Hepatic fibrosis takes place in wherein about 25% patient, and 1.5%～8.0% patient develops into liver cirrhosis, and the developed area sickness rate is higher, and morbidity has the trend of younger ones in an age groupization, has become the common hidden danger of a serious harm human health.Therefore, the control fatty liver has caused the great attention of medical circle.Because fatty liver is not to be independent sexually transmitted disease (STD) kind,, cause the variation of blood parameters and enzyme value through causing the damage of liver function.In the test item of reaction hepatocyte injury, the most common with the sero-enzyme value, like glutamate pyruvate transaminase (ALT), glutamic oxaloacetic transaminase, GOT (AST), alkali phosphatase (ALP), gamma glutamyl transpeptidase (GGT), lactic acid dehydrogenase (LDH) etc.When fatty the change takes place in liver, owing to hepatocyte suffers damage, cytopathy, necrosis, the permeability of cell membrane fragmentation or cell membrane increases, and ALT and AST contained in the hepatocyte will be released in the blood, make ALT in the blood, the active increase of AST.Because ALT, AST mainly are present in the hepatocyte, when its prompting often that obviously raises has hepatic injury.When liver fat takes place became, common ALT, AST raise, but were main with the ALT rising how.Glutamate pyruvate transaminase (ALT) is the main project of diagnosis hepatocyte material injury, and its height often parallels with state of an illness weight.Gamma glutamyl transpeptidase is the important indicator of alcoholic liver injury, and very responsive.
The treatment of current fatty liver is still removing the cause of disease, Cure the Primary Disease with adhere to that the reasonable diet system is main, medicine only plays the auxiliary treatment effect.So, the research of this disease has been become the common focus and the difficult point of domestic and international research.Up to the present, Western medicine does not still have lipotropic active drug, and for well, wherein Radix Polygoni Multiflori and Fructus Crataegi are best with Chinese medicine long-time conditioning property, can blood fat reducing, prevent that cholesterol is at intrahepatic deposition.Western medicine is often selected protection hepatocyte, degrease medicine and antioxidant etc. for use; Like vitamin B, C, E, lecithin, inosine, coenzyme A, reduced glutathion, taurine, carnitine Orotate, glucuronolatone, and some fat-reducing medicament (for example: the liver purport is clear) or the like.Though said medicine is a lot, still need further its curative effect of checking and safety mostly, therefore, should can't abuse in right selection under the physician guidance.
Someone likens into two locusts on rope of hyperlipemia to fatty liver and coronary heart disease because obesity, hyperlipemia, diabetes and excessive drinking be fatty liver commonly encountered diseases because of, and these factors are in close relations with atherosclerosis and coronary heart disease too.There are obvious myocardial damage and hemodynamics disorderly, the T cellular immune function decreased, liver biopsy research shows that 91.4% has hepatic cell fattydegeneration in these cases.Generally speaking, the ethanol fatty liver belongs to the reversibility disease, and early diagnosis and treatment in time often can recover normal.Western developed country infers that the prevalence of child's non-alcoholic fatty liver disease is at least 1% to 2%, and the Obese children prevalence is higher, reaches 22.5% to 52.8% sickness rate.Liver is to be a main organ with the substance metabolism functions in the health, and is playing the part of detoxifcation, metabolism, deoxidation in the health the inside, stores glycogen, synthetic or the like the function of secreted protein.Along with living standard improves, many people take in a large amount of ethanol and high lipid food every day, form fatty liver unconsciously.Modern life rhythm is fast, and everybody is busy with the work of oneself, and society generally believes that fatty liver is not a disease and ignore treatment.Yet fatty liver can cause a lot of diseases, particularly through causing the change of multinomial blood parameters, influences organism metabolism.Long-term edible high lipid food or heavy drinking, ethanol can directly be poisoned hepatocyte, influences its structure and function; Many people are exactly because insobriety is just suffered from alcoholic hepatitis, fatty liver causes liver damage.And hepatic injury can cause triglyceride (TG) to descend, and triglyceride is the fat molecule that LCFA and glycerol form.Be the maximum lipid of people's in-vivo content, most tissues all can be utilized triglyceride catabolite energize, is the main source of energy i (in vivo).The significant quantities of fat that fatty liver can cause eating absorbs minimizing, and bile secretion is not enough, can not decompose the fat of eating into, increases the weight of the state of an illness thereby fat forms fat drop at liver.
The gymnosperm Pollen Pini is a male sex-cell gymnospermous, is undertaking the important task of plant procreation, is comprising and is breeding the necessary complete nutrients matter of new life, and it is nutritious, balanced, collocation is reasonable, is the nutrition treasure-house of pure natural.Pollen Pini is pinaceae plant Pinus massoniana Lamb, Pinus tabuliformis or the pollen that belongs to seeds together, and resulting from the colored core of stamen is the place of pinaster body elite.Pollen Pini is the dietotherapeutic pollen kind in the Chinese medicine treasure-house; As the traditional medical material of China; The history of its dietotherapeutic has exceeded thousands of years; Shennong's Herbal before 2400 is all on the books to successive dynasties such as the Pharmacopoeia of the People's Republic of China of today medicine ancient books and records, and record in the Compendium of Material Medica of the Ming Dynasty " Flos pini, sweet, warm, nontoxic.The profit cardiopulmonary, QI invigorating removes the wind hemostasis." regarded as " elixir " with its magical effect especially among the people.The artificial Pollen Pini of gathering has pure, pollution-free, the no allosome pollen of quality or poisonous pollen mixes, the characteristics of no zoohormone, is the pollution-free food of pure natural.Contain multiple nutritional components and bioactive substances such as rich in protein, aminoacid, unsaturated fatty acid, polysaccharide, vitamin, dietary fiber, enzyme, flavone, nucleic acid, natural hormone in the Pollen Pini; Numerous researchs show that it has endocrine regulation; Improve the reproduction function, enhance metabolism; Effects such as human body immunity improving power are that the Nature is vouchsafed human natural nutrition storehouse.At present the sickness rate of China's fatty liver is very high, along with modern life level improves, people's overnutrition, obesity, excessive consumption of alcohol, high fat diet, lacks exercise, and causes fatty liver.In Chinese Obese children, the sickness rate of fatty liver is up to about eighty per cant, and fatty liver has become influences modern's major issue of quality of life.And the fatty liver treatment phase is long, and onset is slow, and patient's compliance is poor, and often treatment is prone to ignored by people.
Summary of the invention
The present invention's first purpose is to invent a kind of gymnosperm Pollen Pini water-soluble substances that can fundamentally treat fatty liver efficiently.
Water-soluble substances according to the invention is the water-soluble substances below the 3000D for the molecular weight that from the gymnosperm Pollen Pini, extracts.
Second method for preparing that purpose is to invent above-mentioned Pollen Pini water-soluble substances of the present invention.
May further comprise the steps:
1) with the gymnosperm Pollen Pini through comminution by gas stream broken wall treatment 10～20 minutes;
2) with behind the normal pressure and temperature water logging bubble 20～30 minutes, again with ultrasonic Treatment 20～40 minutes;
3) centrifuging is removed solid impurity, gets supernatant;
4) with said supernatant under 1～5 ℃ ambient temperature with the molecular sieve ultra-filtration and separation, obtain molecular weight and be the gymnosperm Pollen Pini water-soluble substances below the 3000D.
Through test, utilize gymnosperm Pollen Pini treatment fatty liver effect remarkable, and have no side effect, and cost, simple.The gymnosperm Pollen Pini is a gymnosperm stamen flower pesticide, is the male sex-cell of plant.The gymnosperm Pollen Pini includes various rich nutrient substances, has multiple BA.The gymnosperm Pollen Pini can be brought into play the pharmacological action of estrogen antagonist, androgen antagonist on the acceptor levels of hormone, but few in the gymnosperm Pollen Pini being the research at present of working of what material.The present invention is exactly that the method for utilizing water to carry is isolated the composition that fatty liver is had therapeutic effect from the gymnosperm Pollen Pini.
The comminution by gas stream breaking cellular wall of step 1) according to the invention is a kind of low temperature technology for broken wall, handles through 10～20 minutes, and purpose is to destroy the cell wall of gymnosperm Pollen Pini, is convenient to the extraction of active ingredient in the gymnosperm Pollen Pini and keeps active.
Step 2 of the present invention) first soak time is 20～30 minutes in, and ultrasonic extraction is 20～40 minutes then, and purpose is to be convenient in the gymnosperm Pollen Pini active ingredient fully dissolve, separate.
The method of removing solid impurity in the step 3) of the present invention is to be in 10000～16000 centrifuges that change centrifugal 10～15 minutes at rotating speed.Purpose is effective solid impurity in the place to go gymnosperm Pollen Pini, makes the extracting solution clarification, and is transparent, and extracting solution is further purified.
Still a further object of the present invention is to invent the application process of above-mentioned gymnosperm Pollen Pini water extract.
A kind of application process is: the gymnosperm Pollen Pini water extract that is used to treat fatty liver is used for preparation treatment fatty liver oral formulations.
Advantage as oral formulations is to use convenience, good absorbing effect, cheap, can be used as child's use that generally keeps healthy, the generation of prevention fatty liver.
Second kind of application process is: the gymnosperm Pollen Pini water extract that is used to treat fatty liver is used to prepare the ejection preparation of treating fatty liver.
Advantage as ejection preparation is that absorption is rapid, treatment is with strong points.
Description of drawings
Fig. 1 is natural health group liver tissue slices figure.
Fig. 2 is for irritating stomach Adeps Sus domestica Chinese liquor model control group liver tissue slices figure.
Fig. 3 is for irritating stomach Pollen Pini water extract group liver tissue slices figure.
Fig. 4 is injection Pollen Pini water extract group liver tissue slices figure.
The specific embodiment
One, preparation gymnosperm Pollen Pini treatment fatty liver water extract:
1, go-on-go gymnosperm Pollen Pini: remove pollen impurity at normal temperatures.
2, with the gymnosperm Pollen Pini comminution by gas stream broken wall treatment of remove impurity 10～20 minutes.
3, with the pollen of normal pressure and temperature water logging bubble after 20～30 minutes, ultrasonic grinding 20～40 minutes.
4, remove solid impurity, water intaking solution:
At rotating speed is in 10000～16000 centrifuges that change centrifugal 10～15 minutes, gets supernatant.
5, with supernatant under 1～5 ℃ ambient temperature, going out molecular weight with 3000D molecular sieve ultra-filtration and separation is the gymnosperm Pollen Pini water extract below the 3000D, is to be used to treat fatty liver Pollen Pini water extract.
Two, preparation oral formulations and ejection preparation:
1, preparation oral formulations A:
Gymnosperm Pollen Pini water extract and Mel or other food additive of treatment fatty liver are mixed well, and after ultrafiltration and pasteurization were sterilized, fill was subsequent use.
2, preparation ejection preparation B:
The gymnosperm Pollen Pini water extract pasteurization sterilization back of treatment fatty liver is mixed with the volume ratio of 1 ︰ 1～2 with injecting normal saline, and the evacuation sealing is for use after the uniform dissolution.
Three, use:
(1) subjects:
Rat (body weight 200～300 grams)
(2) using dosage:
1, oral formulations: oral 4-6 milliliter/sky connects and took 20～30 days.
2, injection: continuous 20～30 days of muscle or lumbar injection 2-4 milliliter/sky
(3) comparing result:
Two monthly age of test group SD rat liver tissue slice result compares:
1, with natural health group liver tissue slices, sees Fig. 1.
From the visible male white rat liver hepatocyte clear in structure of Fig. 1, hepatic sinusoid is obvious, liver rope marshalling.
2, will irritate stomach Adeps Sus domestica and Chinese liquor model group liver tissue slices, see Fig. 2.
From the visible hepatocyte obscure boundary of Fig. 2, becoming in the circle has cavity, and the hepatic sinusoid pressurized narrows down, liver rope arrangement disorder.
3, will irritate stomach Pollen Pini water extract group liver tissue slices, see Fig. 3.
A large amount of hepatic sinusoids occur from the visible liver of Fig. 3, the liver rope is more clear, and hepatocyte is arranged more neat, does not see fat vacuole in the cell.
4, will inject Pollen Pini water extract group liver tissue slices, see Fig. 4.
Be evenly distributed from the visible liver hepatic sinusoid of Fig. 4, the liver rope is high-visible, the hepatocyte marshalling, and nucleus is positioned at cell central authorities greatly.
After the intervention of gymnosperm Pollen Pini water extract to each rats in test groups blood enzyme value index comparative analysis, like following table:
After the intervention of Pollen Pini water extract to each rats in test groups blood enzyme value index comparative analysis table (unit: u/l)
| Group | ALT | ALP | AST | LDH | GGT |
| The nature matched group | 71.00±5.66 | 196.00±5.66 | 96.50±4.95 | 824.50±441.97 | 3.50±0.71 |
| Model control group | 70.00±7.07 | 229.50±29.00 | 99.50±2.12 | 888.50±385.37 | 5.50±2.12 |
| Irritate stomach pine group | 55.50±2.12 | 155.50±4.95 | 85.00±7.07 | 699.50±620.13 | 3.00±2.83 |
| Abdomen is annotated the pine group | 45.00±7.07 | 183.00±22.63 | 77.50±20.51 | 1060±626.50 | 4.00±2.83? |
Through the SPSS statistical analysis, it is extremely remarkable that natural matched group and model group ALT and abdomen are annotated pine group difference, with filling stomach pine group significant difference.Model group ALP is extremely remarkable with filling stomach pine group difference.
Long-term a large amount of ethanol is taken in several kinds of different lesions that can cause liver, and the lighter has only steatosis, and weight person forms alcoholic hepatitis and hepatic fibrosis, and development then causes irreversible liver cirrhosis again.The transaminase is the responsive sign of hepatocellular damage, also is the most responsive index of hepatic injury due to the ethanol.People's alcoholic fatty liver can have AST and ALT slightly to raise, and alcoholic hepatitis has distinctive zymetology and changes, and promptly AST raises and raises obviously than ALT.After using Pollen Pini water extract to intervene, ALT, ALP all have remarkable decline in the blood enzyme value index, and these two kinds of enzymes are key enzymes of weighing liver function; In addition, glutamic oxaloacetic transaminase, GOT (AST) has also obtained improvement to a certain degree.Can infer tentatively that gymnosperm Pollen Pini water extract becomes rat liver fat, tool improves significantly.
Above experimental result shows: gymnosperm Pollen Pini water extract becomes each experimental group rat liver fat has sizable therapeutical effect, and from liver tissue slices, gymnosperm Pollen Pini water extract has very strong effect to improving the liver organization structure; Pass through the comparison of enzyme value indexs such as ALT, ALP, AST, LDH in addition; Find that all experimental grouies and matched group have descended than all; And it is more much lower than the model group with the enzyme value of Pollen Pini water extract intervention group; Difference is significant, and this explains that better gymnosperm Pollen Pini water extract can not only improve the liver organization structure, and can improve liver function.
In a word, show through above test: product of the present invention can effectively be prevented and treated fatty liver, and the curative effect of ejection preparation is better than oral formulations.China is at the early-stage aspect the research of gymnosperm Pollen Pini control fatty liver at present, through experiment, finds that lumbar injection pollen water extract is best to the lipotropism effect, for the medicine of developing the treatment fatty liver proposes new approaches.The gymnosperm Pollen Pini is China's Traditional health care product, can improve hepatocyte activity, improves blood enzyme value index, and the prevention fatty liver is significant to the liver health care of safeguarding and improve China common people.
Claims (5)
1. be used to treat the Pollen Pini extract of fatty liver, be the molecular weight that from Pollen Pini, extracts water-soluble substances, may further comprise the steps less than 3000D:
1) with the gymnosperm Pollen Pini through comminution by gas stream broken wall treatment 10～20 minutes;
2) with behind the normal pressure and temperature water logging bubble 20～30 minutes, again with ultrasonic Treatment 20～40 minutes;
3) centrifuging is removed solid impurity, gets supernatant;
4) with said supernatant under 1～5 ℃ ambient temperature with the molecular sieve ultra-filtration and separation, obtain molecular weight and be the gymnosperm Pollen Pini water-soluble substances below the 3000D.
2. method for preparing that is used to treat the Pollen Pini water extract of fatty liver according to claim 1 is characterized in that may further comprise the steps:
1) with the gymnosperm Pollen Pini through comminution by gas stream broken wall treatment 10～20 minutes;
2) with behind the normal pressure and temperature water logging bubble 20～30 minutes, again with ultrasonic Treatment 20～40 minutes;
3) centrifuging is removed solid impurity, gets supernatant;
4) with said supernatant under 1～5 ℃ ambient temperature with the molecular sieve ultra-filtration and separation, obtain molecular weight and be the gymnosperm Pollen Pini water-soluble substances below the 3000D.
3. according to the said method for preparing of claim 2, it is characterized in that centrifuging described in the step 3) is is in 10000～16000 refrigerated centrifugers that change centrifugal 10～15 minutes at rotating speed.
4. the Pollen Pini extract that is used to treat fatty liver according to claim 1 is used to prepare the oral formulations of treating fatty liver.
5. the Pollen Pini extract that is used to treat fatty liver according to claim 1 is used to prepare the ejection preparation of treating fatty liver.
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| CN102940704B (en) * | 2012-11-30 | 2013-12-11 | 烟台新时代健康产业有限公司 | Pine pollen water extract buccal tablet for preventing and treating alcoholic fatty liver |
| CN102935105B (en) * | 2012-11-30 | 2014-08-13 | 烟台新时代健康产业有限公司 | Pollen pini water extract enteric-coated tablet for preventing alcoholic fatty liver (AFL) |
| CN106421550A (en) * | 2016-11-30 | 2017-02-22 | 上海市同仁医院 | Traditional Chinese medicine composition for treating fatty liver and preparation method of traditional Chinese medicine composition for treating fatty liver |
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Non-Patent Citations (1)
| Title |
|---|
| 蒋滢等.松花粉的研究.《氨基酸和生物资源》.1998,第20卷(第1期),8－11页. * |
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