CN102335186A - Application of icariin for preparing preparation for delaying aging and improving health state - Google Patents
Application of icariin for preparing preparation for delaying aging and improving health state Download PDFInfo
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- CN102335186A CN102335186A CN 201010238582 CN201010238582A CN102335186A CN 102335186 A CN102335186 A CN 102335186A CN 201010238582 CN201010238582 CN 201010238582 CN 201010238582 A CN201010238582 A CN 201010238582A CN 102335186 A CN102335186 A CN 102335186A
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Abstract
The invention belongs to the field of traditional Chinese medicines, relates to new application of icariin used as a traditional Chinese medicine active ingredient, and particularly relates to application of icariin used as an active monomer of epimedium for preparing a preparation for delaying aging and improving health state. In the invention, cell culture and passage technique proves that icariin has the action of increasing the passage frequency of human embryo lung diploid mechanocyte; the caenorhabditis elegans life test proves that icariin has the action of prolonging the life of lower animals; the mouse life test proves that icariin has the action of prolonging the life of mammals; and obvious pathological changes do not emerge in the vital organs such as the heart, liver, spleen, lung, kidneys and brain of the mouse intervened by icariin. The icariin provided by the invention can be used as a drug preparation raw material or a food additive.
Description
Technical field
The invention belongs to the field of Chinese medicines, the new purposes of pharmaceutically active ingredient icariin in relating to.The effective monomer icariin that is specifically related to the Chinese medicine Herba Epimedii is preparing slow down aging and is improving the purposes in the health status preparation.
Background technology
Aged tendency of population is the severe challenge that various countries, the whole world face jointly, has become the strategic problem of a national economy long term growth, realizes that the aged tendency of population of active and healthy also is international great public health problem.
By the end of the year 2008, China's aging population have increased to 1.69 hundred million, account for 12.79% of total population, and just with average annual 1000 ten thousand amplification " at the double, quick march ".To the year two thousand twenty, China's aging population will reach 2.48 hundred million, and the aging level will reach 17%.Expect China's entering severe aging stage the year two thousand fifty, China's aging population reach 4.37 hundred million when the time comes, account for total population more than 30%, and per three or four philtrums just have an old man.Do measuring and calculating according to national old-age group, at present the whole nation more than 80 years old the old man be about 2 times of aging population speedup just in superfast growth, expect that just to have 1 among the year two thousand fifty 5 old peoples be old man more than 80 years old.For this reason, Chinese society will face severe aged tendency of population challenge.
The aging of active and healthy will make the old people keep fit and vigor exactly, lets China old people in the long as far as possible time, have good quality of life.Keeping the elderly population health status, reducing disease is the burden on society that brings of minimizing aged tendency of population and the key link of financial burden.Therefore, explore simple and easy to doly, cost-effective to intervene old and feeble measure and method significant.
Slow down aging is two notions that closely link to each other with life-saving.Aging refers generally to physically-draining, and is handicapped, walks with difficulty; Amnesia, presentations such as thinking disorder; And the life-span refers to age, life time more.In general, the people that the life-span is long, its condition is healthy relatively; Healthy people, its life-span is long relatively.Therefore, the length in life-span has also been reacted the health status of health to a certain extent.
In recent years, a research expression on " nature " magazine online edition, a kind of medicine of from the argillous antibacterial of Easter Island (Easter Island), separating can fully prolong the life-span of mice.The medicine of this rapamycin by name (rapamycin) is first kind of found, as to have prolongation mammal life-span medicine, when getting into old age, takes this medicine and gets final product onset.Before this research, the unique method that prolongs the rodent life-span is through genetic engineering, and the perhaps absorption of caloric restriction is promptly improved but diet extremely low in calories through a kind of nutrition.Discover in mice 20 months (be equivalent to human longevity 60 years old) to give mice as a kind of food additive that can make the average life of male mice prolong 9%, the life-span of female mice prolongs 13% with rapamycin.Research worker is reminded simultaneously, it be not immediately clear whether rapamycin also has the identical effect of lengthening the life to the mankind, and rapamycin has well-known toxic and side effects, is prone to cause fungal infection and pneumonia, and the general population should arbitrarily not take this type of medicine.
Put down in writing the Chinese medicine and the method for many slow down aging in the Chinese medicine document, but most strong experimental evidence really that all lacks.Icariin (icariin, ICA, C
33H
40O
15) be the main effective ingredient of the total flavones that extracts in Berberidaceae (Berberidaceae) the Epimedium Epimedium plant Herba Epimedii stem and leaf.In recent years, the extraction process technology of icariin reaches its maturity.Herba Epimedii another name Herba Epimedii distributes extensively in China, mainly originates in ground such as Shaanxi, Jilin, Hunan; Be grown in the alpine belt, it is warm in nature, acrid in the mouth, sweet; Returning liver, kidney channel, is the traditional Chinese herbal medicine with kidney invigorating and YANG supporting, expelling wind and removing dampness effect, the effectiveness that Compendium of Material Medica is claimed it to be had " beneficial vital essence; hard muscles and bones is mended the waist knee joint, heart tonifying power "; Modern study confirms: effects such as icariin ability human body immunity improving function, adjusting bone metabolism, defying age, antitumor, can significantly suppress the inductive normal mouse of ConA external T lymphocyte early activation and propagation, and be dose dependent.The molecular weight of said icariin is 676.65, its molecular structure:
Summary of the invention
The new purposes of pharmaceutically active ingredient icariin in the purpose of this invention is to provide.The effective monomer icariin that relates in particular to the Chinese medicine Herba Epimedii is preparing slow down aging and is improving the purposes in the health status preparation.
Further purpose of the present invention provides icariin in biological integral slow down aging, the new purposes improving health status, prolong average life and maximum life.
The molecular weight of icariin according to the invention is 676.65, and it has the molecular structure of following formula (I):
The present invention is through cell in vitro go down to posterity experiment, nematicide life experiment and mouse life experiment; The result shows; The go down to posterity result of test, the beautiful nematicide life test of wild type, the life test of C57BL/6 male mice and the test of C57BL/6 male mice main organs pathology confirms human embryonic lung diploid fibroblast: icariin has the effect that prolongs the passage number of times, prolongs lower animal and mammal average life and maximum life, and toxic and side effects is minimum.
Experiment showed, that icariin can prolong the passage number of times, prolong lower animal and moving average life of suckling and maximum life, icariin has significantly slow down aging and improves health status so that life-saving effect.
Icariin of the present invention can be made into slow down aging, improves the raw material and the food additive of health status so that life-saving pharmaceutical preparation.
For the ease of understanding, below will describe in detail the new purposes of middle pharmaceutically active ingredient icariin of the present invention with embodiment through concrete accompanying drawing.What need particularly point out is; Instantiation and accompanying drawing only are in order to explain; Obviously those of ordinary skill in the art can explain according to this paper, within the scope of the invention the present invention is made various corrections and change, and these corrections and change are also included in the scope of the present invention.
Description of drawings
Fig. 1 Herba Epimedii total flavones, icariin be to the influence of MRC-5 passage,
Wherein: adopt SPSS15.0, one-way ANOVA analyzes; Compare * P＜0.001 with matched group.
Fig. 2 various dose Herba Epimedii total flavones is to the influence of wild type nematicide survival curve,
Wherein: adopt Kaplan-Meier survival analysi s, Log-rank test analyzes EF and can be significantly the survival curve of nematicide be moved to right and the Control comparison *
*P＜0.05 (Pi=0.06,
*P2=0.0014, P3=0.0435).
Fig. 3 various dose icariin is to the influence of the beautiful nematicide survival curve of wild type,
Wherein: adopt Kaplan-Meier survival analysis, Log-rank test analyzes
ICA can be significantly with the survival curve of nematicide move to right and Control relatively, *
*P＜0.05 (P1=0.1,
*P2=0.0083, P3=0.1224).
Fig. 4 EF, ICA end to the influence of the 760th day existence curve the male mice existing state,
Wherein: adopt Kaplan-Meier survival analysis, Log-rank Mantel-Cox test analyzes, and EF, ICA all can significantly move to right the survival curve of mice and Control compares * P=0.0319**P=0.0166.
Fig. 5 EF, ICA end to the influence of the 991st day existence curve the male mice existing state,
Wherein: adopt Kaplan-Meier survival analysis, Log-rank Mantel-Cox test analyzes, and EF, ICA all can significantly move to right the survival curve of mice and Control compares * P=0.0313**P=0.0288.
The specific embodiment:
The Herba Epimedii total flavones (EF) that the present invention adopts, icariin (ICA: purity 98.5%) but conventional method is extracted or commercial acquisition.
1, passage test
Herba Epimedii total flavones (EF), icariin (ICA) powder are dissolved into storage liquid with dimethyl sulfoxide (DMSO) respectively, place 4 ° of refrigerators to keep in Dark Place, and dilution is 100 times during use, make respectively to organize the whole content of DMSO and be below 1%.
Human embryonic lung diploid fibroblast MRC-5 (PD21 is available from cell institute of the Chinese Academy of Sciences) is incubated in the MEM culture fluid that contains 10% hyclone, grows in 37 ℃ of incubators, goes down to posterity in the time of at the bottom of cell covers with bottle, goes down to posterity once in average per 4～5 days.Digest with pancreatin during passage, process single cell suspension, count, transferring cell concentration is 3.5 * 10
5/ ml, each culture bottle inoculation 1ml.Cell carries out ICA and EF pharmaceutical intervention during since the 24th generation respectively.Wherein the administration concentration of ICA is 10
-6Mol/L, the administration concentration of EF is 3mg/L.If cell is at the bottom of can not covering with bottle then define those cells and be last cell in generation in 3 week.Adopt log2 (D/D0) formula to calculate CPD (cumulative population doubling), the cell density when wherein D and D0 represent collecting cell and inoculating cell respectively.SPSS15.0 is adopted in this experiment repetition 3 times, one-way ANOVA analytical control, and the result shows that Herba Epimedii total flavones, icariin all have the effect (like table 1, shown in Figure 1) that prolongs the maximum passage number of human embryonic lung diploid fibroblast MRC-5.
Table 1EF, ICA are to the influence of human embryonic lung diploid fibroblast MRC-5 (mean ± SD)
Annotate: compare with matched group
*P＜0.001
2, lower animal test
Herba Epimedii total flavones (EF), icariin (ICA) powder are dissolved into storage liquid with dimethyl sulfoxide (DMSO) respectively; Place 4 ° of refrigerators to keep in Dark Place; Dilution is 100 times during use; Make and respectively organize the whole content of DMSO and be 1%, the medicine final concentration is respectively EF 25,50,100ug/ml, ICA 10,30,50ug/ml.
Bacterial strain EColi.OP50, the beautiful nematicide C.elegans of wild type Bristol N2 is so kind as to give by CGC (U.S. nematicide heredity center).
Experimental technique: the L1 phase nematicide that is in same developmental stage adds 96 orifice plates (0h) at random; Add 5-fluorouracil (5-FU) when growing to the L4 phase (45-47h); (68h) adds EF, Ica solution respectively when growing to young adult, and life test this moment begins, and is designated as Day0.Microscope light source was opened to the prime time, and the nematicide that lives moves under the stimulation of high light, according to death/survival of this record nematicide, every other day observes once.Every hole total capacity 150ul in 96 orifice plates contains the about 8-15 bar of worm, E Coli.OP505mg/ml, the EF of 5-FU 0.12mM and various dose, ICA solution.
More than the experiment repetition is 3 times; Get wherein once the result as representative; Check through variance analysis; The result shows: EF50ug/ml, ICA 30ug/ml can prolong 33.4%, 30.0% respectively with nematicide average life, and the result shows: Herba Epimedii total flavones, icariin all have the effect (like table 2, shown in Figure 2) of significant prolongation nematicide average life.
Table 2 various dose EF, ICA are to the influence of wild type nematicide average life (mean ± SE)
Annotate: compare with matched group
*P＜0.01
The maximum life computational methods adopt this to organize the average life of last natural death 10% nematicide; Check through variance analysis; The result shows: EF 50ug/ml, ICA 30ug/ml can prolong 14%, 20.2% respectively with the nematicide maximum life, and the result shows: Herba Epimedii total flavones, icariin all have the effect (as shown in table 3) of significant prolongation nematicide maximum life.
Table 3 various dose EF, ICA are to the influence of wild type nematicide maximum life (mean ± SD)
Annotate: compare with matched group
*P＜0.01
3, mouse life test
Herba Epimedii total flavones (EF), icariin (ICA) powder respectively with normal control group mouse feed equivalent multiple proportions mixed diluting; Process and contain 0.06% Herba Epimedii total flavones mouse feed and 0.02% icariin mouse feed; 70～90 degree oven dry, sterilization, vacuum-packed for use.
The C57BL/6 male mice, SPF level, 8～9 ages (date of birth is 2007.2.26～2007.3.6, available from Beijing Vital River Experimental Animals Technology Co., Ltd., raises the animal center SPF level laboratory of the public health unit in Shanghai City) in week.Every single cage feedstuff of mice.150 mices are divided into 3 groups at random when whole 12 monthly ages: normal control group (matched group, 52), Herba Epimedii total flavones group (EF group, 49), icariin group (ICA group, 49).Control group mice gives competent feedstuff and drinking-water; EF group mice gives the competent 0.06% Herba Epimedii total flavones feedstuff that contains and feeds with drinking-water; ICA group mice gives the competent 0.02% icariin feedstuff that contains and feeds with drinking-water.Itemized record are respectively organized the date of each mice natural death, and calculate average life and the maximum life of respectively organizing mice.Experimental result adopts SPSS15.0, and one-way ANOVA analyzes.
The result shows: the average life of control group mice is 692.5 days; The average life of EF group mice is 724.5 days; The average life of ICA group mice is 768.1 days; EF, ICA make the average life of mice prolong 4.6%, 10.9% respectively, and the result shows: icariin has the effect (like table 4, shown in Figure 4) of obvious prolongation mice average life.
Table 4EF, ICA are to the influence of male mice average life (mean ± SD)
Annotate: compare with matched group
*P＜0.05
By maximum life computational methods commonly used at present; Promptly the average life of last natural death 10% mice of this group is maximum life; The result shows: the maximum life of control group mice is 922 days, and the maximum life of EF group mice is 954.5 days, and the maximum life of ICA group mice is 966.7 days; EF and ICA make the maximum life of mice prolong 3.5%, 4.8% respectively, and the result shows: icariin has the maximum life (like table 5, shown in Figure 5) of obvious prolongation mice.
Table 5EF, ICA are to the influence of male mice maximum life (mean ± SD)
Annotate: compare with matched group
*P＜0.05
4, Herba Epimedii total flavones, icariin are to the influence of mice main organs
The C57BL/6 male mice, the SPF level, 8～9 ages in week, (date of birth was 2007.2.26～2007.3.6), improved by Beijing Vital River Experimental Animals Technology Co., Ltd..Raise the animal center SPF level laboratory of public health unit in Shanghai City.Every single cage feedstuff of mice.36 12 monthly age C57BL/6 male mices are divided into 3 groups at random, normal control group (matched group, 11), Herba Epimedii total flavones group (EF group, 13), icariin group (ICA group, 12).Control group mice gives competent feedstuff and drinking-water; EF group mice gives the competent 0.06% Herba Epimedii total flavones feedstuff that contains and feeds with drinking-water; ICA group mice gives the competent 0.02% icariin feedstuff that contains and feeds with drinking-water.Each organizes tissues such as the execution of mice nursing to 24 monthly ages is cored, liver, spleen, lung, kidney, brain, utilizes tissue array technology, and above-mentioned tissue is produced in same the organization chip, adopts H&E dyeing, observes pathological change of respectively organizing.
Hepatic tissue pathology shows: the control group mice hepatic tissue has hydropic degeneration, Ke Schwann Cells hypertrophy, and part of hepatocytes occurs downright bad, and has a small amount of inflammatory cell to be pathological changes such as special mess property infiltration; EF group, ICA group murine liver tissue pathological change alleviate than control group mice.Nephridial tissue pathology shows: the few a small amount of cell infiltration of part mice found in renal tissue of matched group, and all the other mice nephridial tissues are normal basically; EF group, ICA group mice demonstrate basic normal kidney tissue pathological manifestations.Cardiac muscular tissue's pathology shows: a small amount of round cell soaks into sarolemma hyperplasia, accidental cardiac muscle fiber endochylema hydropic degeneration around the local visible special mess property blood vessel of matched group part mice; Cardiac muscular tissue's pathological change no significant difference between each group.Lung tissue pathology shows: matched group part mouse lung is organized a small amount of inflammatory cell infiltration, is dispersed in the distribution alveolar wall and thickens; Lung tissue pathology changes no evident difference between each group.The spleen histopathology shows: responding property of control group mice spleen tissue follicle hypertrophy, and surplus nothing is pathological change clearly; Spleen histo pathological change between each group does not have clearly difference.Cerebral tissue pathology shows: matched group part mice local neuron apoptosis kitchen range, accidental calcification kitchen range; Brain tissue pathology change's no significant difference between each group.Show: Herba Epimedii total flavones, icariin influence minimum to the toxic and side effects of the important organ of mice.
Experimental result shows; The go down to posterity result of study of test, the beautiful nematicide life test of wild type, the life test of C57BL/6 male mice and the test of C57BL/6 male mice main organs pathology of above-mentioned human embryonic lung diploid fibroblast confirms: icariin has the effect that prolongs the passage number of times, prolongs lower animal and mammal average life and maximum life, and toxic and side effects is minimum.
Claims (7)
1. the icariin of formula (I) is preparing slow down aging and is improving the purposes in the health status preparation,
2. by the described purposes of claim 1, it is characterized in that described icariin increases the passage number of times; Prolong lower animal or mammal average life or maximum life.
3. by the described purposes of claim 2, it is characterized in that described passage number of times is the passage number of human embryonic lung diploid fibroblast.
4. by the described purposes of claim 2, it is characterized in that average life and maximum life that the described lower animal life-span is the beautiful nematicide of wild type.
5. by the described purposes of claim 2, it is characterized in that average life or maximum life that the described mammal life-span is the C57BL/6 male mice.
6. by the described purposes of claim 1, it is characterized in that described preparation is a medicine material.
7. by the described purposes of claim 1, it is characterized in that described preparation is a food additive.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105920057A (en) * | 2016-06-03 | 2016-09-07 | 通化师范学院 | Application of icariin |
| CN108715829A (en) * | 2018-05-25 | 2018-10-30 | 大连医科大学 | A method of improving Human in vitro Fertilization embryo development rate |
| CN110251527A (en) * | 2019-06-06 | 2019-09-20 | 泓博元生命科技（深圳）有限公司 | The composition of niacinamide-containing mononucleotide is in the application of anti-aging drug/health care product |
-
2010
- 2010-07-26 CN CN 201010238582 patent/CN102335186A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 《中国中西医结合杂志》 20090331 沈自尹等 淫羊藿激活内源性干细胞及其机制研究 251-253 1-7 第29卷, 第3期 * |
| 《医学综述》 20090331 刘韶英 淫羊藿苷药理作用的研究进展 909-912 1-7 第15卷, 第6期 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105920057A (en) * | 2016-06-03 | 2016-09-07 | 通化师范学院 | Application of icariin |
| CN108715829A (en) * | 2018-05-25 | 2018-10-30 | 大连医科大学 | A method of improving Human in vitro Fertilization embryo development rate |
| CN110251527A (en) * | 2019-06-06 | 2019-09-20 | 泓博元生命科技（深圳）有限公司 | The composition of niacinamide-containing mononucleotide is in the application of anti-aging drug/health care product |
| WO2020244249A1 (en) * | 2019-06-06 | 2020-12-10 | 泓博元生命科技（深圳）有限公司 | Application of composition containing nicotinamide mononucleotide in anti-ageing drugs/healthcare products |
| CN110251527B (en) * | 2019-06-06 | 2021-06-25 | 泓博元生命科技（深圳）有限公司 | Application of composition containing nicotinamide mononucleotide in anti-aging drugs/health care products |
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Application publication date: 20120201 |