CN102978123A - Immune activity study on ergosterol of paecilomyces hepiali - Google Patents
Immune activity study on ergosterol of paecilomyces hepiali Download PDFInfo
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- CN102978123A CN102978123A CN2012105242690A CN201210524269A CN102978123A CN 102978123 A CN102978123 A CN 102978123A CN 2012105242690 A CN2012105242690 A CN 2012105242690A CN 201210524269 A CN201210524269 A CN 201210524269A CN 102978123 A CN102978123 A CN 102978123A
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- ergosterol
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- peacilomyce hepiahi
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- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 title claims abstract description 27
- RQOCXCFLRBRBCS-UHFFFAOYSA-N (22E)-cholesta-5,7,22-trien-3beta-ol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CCC(C)C)CCC33)C)C3=CC=C21 RQOCXCFLRBRBCS-UHFFFAOYSA-N 0.000 title claims abstract description 27
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 title claims abstract description 27
- DNVPQKQSNYMLRS-NXVQYWJNSA-N Ergosterol Natural products CC(C)[C@@H](C)C=C[C@H](C)[C@H]1CC[C@H]2C3=CC=C4C[C@@H](O)CC[C@]4(C)[C@@H]3CC[C@]12C DNVPQKQSNYMLRS-NXVQYWJNSA-N 0.000 title claims abstract description 27
- DNVPQKQSNYMLRS-SOWFXMKYSA-N ergosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H](CC[C@]3([C@H]([C@H](C)/C=C/[C@@H](C)C(C)C)CC[C@H]33)C)C3=CC=C21 DNVPQKQSNYMLRS-SOWFXMKYSA-N 0.000 title claims abstract description 27
- 230000005965 immune activity Effects 0.000 title abstract 2
- 241001416980 Paecilomyces hepiali Species 0.000 title 1
- 241000699666 Mus <mouse, genus> Species 0.000 claims abstract description 23
- 230000000242 pagocytic effect Effects 0.000 claims abstract description 8
- 241000699670 Mus sp. Species 0.000 claims abstract description 5
- 210000002864 mononuclear phagocyte Anatomy 0.000 claims abstract description 5
- 230000036039 immunity Effects 0.000 claims abstract description 4
- 230000000694 effects Effects 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 4
- 230000000452 restraining effect Effects 0.000 claims description 4
- 230000003637 steroidlike Effects 0.000 claims description 4
- 241000190633 Cordyceps Species 0.000 claims description 3
- 238000005728 strengthening Methods 0.000 claims description 3
- 229930182558 Sterol Natural products 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 150000001720 carbohydrates Chemical class 0.000 claims description 2
- 238000000855 fermentation Methods 0.000 claims description 2
- 230000004151 fermentation Effects 0.000 claims description 2
- 150000002632 lipids Chemical class 0.000 claims description 2
- 230000000527 lymphocytic effect Effects 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 150000003432 sterols Chemical class 0.000 claims description 2
- 235000003702 sterols Nutrition 0.000 claims description 2
- 150000003505 terpenes Chemical class 0.000 claims description 2
- 230000003053 immunization Effects 0.000 claims 1
- 230000009466 transformation Effects 0.000 abstract description 11
- 238000002474 experimental method Methods 0.000 abstract description 9
- 210000004698 lymphocyte Anatomy 0.000 abstract description 9
- 108010062580 Concanavalin A Proteins 0.000 abstract description 7
- 238000011160 research Methods 0.000 abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052799 carbon Inorganic materials 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 6
- 210000000952 spleen Anatomy 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract description 3
- 230000036737 immune function Effects 0.000 abstract description 3
- 241000233866 Fungi Species 0.000 abstract description 2
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 241001236817 Paecilomyces <Clavicipitaceae> Species 0.000 abstract 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000000976 ink Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 235000017550 sodium carbonate Nutrition 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 206010013183 Dislocation of vertebra Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000004988 splenocyte Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Steroid Compounds (AREA)
Abstract
The invention discloses an immune activity study on ergosterol of paecilomyces hepialid. According to the study, the immune functions of the ergosterol of the paecilomyces hepialid are tested by virtue of mouse spleen lymphocyte transformation experiments and mouse carbon clearance experiments, which are induced by concanavalin A (Con A). Study results indicate that the ergosterol of the paecilomyces hepialid has a function of inhibiting in-vitro T lymphocytes, and is capable of obviously improving the phagocytic function of mononuclear phagocyte in mice and boosting the whole immunity of the mice. The study belongs to application methods of fungus medicines in the fields of medicaments and health care products, lays a certain research foundation for the development of health care foods with the paecilomyces hepialid, and also lays a certain theoretical foundation for the development of new products containing the ergosterol in future.
Description
Technical field:
The invention discloses the immunocompetence research of ergosterol in the peacilomyce hepiahi, and the method that adopts mouse spleen lymphocyte transformation experiment that concanavalin A (ConA) induces and mouse carbon to clean up experiment is carried out the immunologic function check to it, result of study shows that ergosterol has the lymphocytic restraining effect of T that exsomatizes in the peacilomyce hepiahi, and can obviously strengthen the phagocytic function of mononuclear phagocyte in the Mice Body, mouse integral body is had the effect of strengthening immunity.This research belongs to the fungus crude drug at medicine and the application method of field of health care products, provide certain Research foundation for the exploitation of the health-care food of peacilomyce hepiahi, and the novel product that will develop following ergosterol provides certain theoretical basis.
Background technology:
Peacilomyce hepiahi is that the natural cordyceps separation obtains anamorphic strain, and through fermentation culture, the mycelial effect of gained is similar to natural cordyceps.Contain abundant carbohydrate, protein, steroidal compounds, lipid acid, terpenoid etc. in the peacilomyce hepiahi.Wherein, ergosterol is as a kind of steroidal compounds, and the feature sterol for Mycophyta but seldom receives publicity.The method that the mouse spleen lymphocyte transformation experiment that this literary grace employing concanavalin A (ConA) is induced and mouse carbon are cleaned up experiment is carried out the immunologic function check, and then verifies the enhancing immunocompetence of ergosterol.
Summary of the invention:
The invention discloses the immunocompetent research of ergosterol in the peacilomyce hepiahi,, method and result are as follows:
1, experimental technique (conventional methods)
(1) lymphocyte transformation test
Sample solution prepares precision and takes by weighing sample, 30% DMSO dissolving, and complete culture solution is diluted to 100 μ g/ml, and with 0.22 μ m membrane filtration degerming, complete culture solution is diluted to desired concn.
Get the BALB/C mouse, the cervical vertebra dislocation method is put to death, and the aseptic spleen of getting is made splenocyte suspension, and Hank ' s liquid is washed (centrifugal 1000rpm, 5min) 2 times, places RPMI 1640 complete culture solutions, counting, and it is 2 * 10 that cell dilution is become concentration
6Cell suspension, add in 96 well culture plates, every hole 100 μ l, 3 every group multiple holes are established negative control hole, positive control hole and blank well simultaneously.Test holes adds the sample solution of 100 μ l different concns, and negative control hole and blank well add nutrient solution, and the positive control hole adds 10 μ g/ml ConA, 100 μ l.37 ℃, cultivate 48h in the 5% CO2 incubator, add 5mg/ml MTT solution 20 μ l, continue to cultivate 4h, termination reaction.Take out culture plate, carefully suck supernatant liquor, add 150 μ l DMSO, fully dissolving crystallized, microplate reader 490nm measures each hole OD value.The result represents that with transformation efficiency calculation formula is as follows:
Transformation efficiency=(sample well OD value-blank well OD value)/(control wells OD value-blank well OD value) * 100%.
(2) mouse carbon is cleaned up experiment
Mouse is divided into control group, low dose group, high dose group gavages respectively physiological saline, 5mg/kg/d ergosterol, 10mg/kg/d ergosterol.Every day 1 time, continuous 15 days.Behind last administration 1 h, each organizes mouse by the india ink physiological salt liquid of 5 times of amount dilutions of the every 0.1ml/10g tail vein injection of body weight, 2min and 10min behind injection prepared Chinese ink, get blood 20 μ l with glass capillary from the mouse orbit rear vein beard, afterwards it is added immediately in the 0.1% Na2CO3 solution of 2ml, shake up, survey absorbancys (OD) value in spectrophotometer 600 nm wavelength places, with Na2CO3 solution as blank solution.Afterwards mouse is put to death, get spleen and liver, with the blood stains of filter paper absorption organ surface, weigh respectively and record.
Clean up index: K=(logOD 2-logOD10)/(t10-t2)
Phagocytic index: a=body weight/(liver weight+spleen is heavy) *
3√ K
Experimental data is used
Expression, and adopt SPSS18.0 for Windows statistical software to analyze.P＜0.05 explanation is remarkable difference statistically, and P＜0.01 explanation is utmost point significant difference statistically.
2. results and analysis
(1) ergosterol sees Table 1 to the impact of mouse lymphocyte transformation
Table 1 ergosterol on the impact of mouse lymphocyte transformation (
, n=6)
Annotate: compare * p＜0.05, * * p＜0.01, * * * p＜0.001 with control group
(2) mouse carbon is cleaned up experimental result and is seen Table 2
Table 2 ergosterol is on the impact of mouse phagocytic index
Result of study shows that the lymphocyte transformation experimental data shows that ergosterol is inhibited to the isolated mouse lymphocyte transformation, and inhibiting rate reached 85% when concentration was 50 μ g/ml, with control group utmost point significant difference (p＜0.001) is arranged relatively.When concentration was low, concentration and restraining effect had dose-effect relationship, reached certain limit but work as concentration, and restraining effect is strengthened (being the affect figure of ergosterol on the mouse lymphocyte transformation such as accompanying drawing 1) greatly.Carbon is cleaned up experimental result and is shown, the phagocytic index of mouse significantly increases behind the drug effect, be that Function of mononuclear phagocyte obviously strengthens, the phagocytic index of the high low dose group of ergosterol reaches respectively 6.429 and 7.300, all with control group utmost point significant difference (P＜0.01) is arranged, it is not clearly to the effect of macrophages in vitro function that document record ergosterol is arranged, but still produces effect significantly by integral experiment.This shows that ergosterol is inhibited to the T lymphocyte that exsomatizes in the peacilomyce hepiahi, also can obviously strengthen the phagocytic function of mononuclear phagocyte in the Mice Body, mouse integral body is had the effect of strengthening immunity.For the exploitation of the health-care food of peacilomyce hepiahi provides certain Research foundation, provide certain theoretical basis to the exploitation of the novel product that contain ergosterol future.
Claims (2)
1. the used peacilomyce hepiahi of the present invention is to separate in the natural cordyceps to obtain anamorphic strain, through fermentation culture, contain abundant carbohydrate, protein, steroidal compounds, lipid acid, terpenoid etc. in the gained mycelium, wherein, the content of ergosterol reaches 15.65%.
2. described according to claim 1, the ergosterol that separation obtains from peacilomyce hepiahi is a kind of steroidal compounds, is the feature sterol of Mycophyta, and its check of carrying out immunizing power is verified whether ergosterol can bring benefit to our human body.The present invention has verified that first ergosterol has the stripped lymphocytic restraining effect of T of mouse in the peacilomyce hepiahi, and can obviously strengthen the phagocytic function of mononuclear phagocyte in the Mice Body, mouse integral body is had the effect of strengthening immunity.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012105242690A CN102978123A (en) | 2012-12-08 | 2012-12-08 | Immune activity study on ergosterol of paecilomyces hepiali |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012105242690A CN102978123A (en) | 2012-12-08 | 2012-12-08 | Immune activity study on ergosterol of paecilomyces hepiali |
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| CN102978123A true CN102978123A (en) | 2013-03-20 |
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| CN2012105242690A Pending CN102978123A (en) | 2012-12-08 | 2012-12-08 | Immune activity study on ergosterol of paecilomyces hepiali |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104911109A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | High-yield paecilomyces hepiali mutant strain and cultivation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1095103A (en) * | 1993-05-07 | 1994-11-16 | 中国中医研究院 | Producing process of Chinese caterpillar fungus hypha fermentation |
| CN101214266A (en) * | 2007-12-28 | 2008-07-09 | 王辉 | Paecilomyces hepiali mycelium mixed powder and preparation |
-
2012
- 2012-12-08 CN CN2012105242690A patent/CN102978123A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1095103A (en) * | 1993-05-07 | 1994-11-16 | 中国中医研究院 | Producing process of Chinese caterpillar fungus hypha fermentation |
| CN101214266A (en) * | 2007-12-28 | 2008-07-09 | 王辉 | Paecilomyces hepiali mycelium mixed powder and preparation |
Non-Patent Citations (3)
| Title |
|---|
| 傅惠英等: "中国被毛孢和蝙蝠蛾拟青霉小鼠免疫功能调节作用的比较", 《中国比较医学杂志》, 30 September 2012 (2012-09-30), pages 17 * |
| 苏日古格等: "蒙古口蘑子实体的抗肿瘤活性", 《食品科学》 * |
| 陈林琤等: "高效液相色谱与液相色谱-质谱分析冬虫夏草相关产品之麦角固醇与其衍生物", 《分析化学》, 30 September 2011 (2011-09-30), pages 1380 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104911109A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | High-yield paecilomyces hepiali mutant strain and cultivation method |
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Owner name: JIANGXI JINMIN KEXIN JINSHUIBAO PHARMACEUTICAL CO. Free format text: FORMER OWNER: BAO HAIYING Effective date: 20130321 |
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Free format text: CORRECT: ADDRESS; FROM: 130118 CHANGCHUN, JILIN PROVINCE TO: 330096 NANCHANG, JIANGXI PROVINCE |
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Effective date of registration: 20130321 Address after: 330096 No. seven, No. 888, hi tech Development Zone, Jiangxi, Nanchang Applicant after: Jiangxi Jimin Kexin Jinshuibao Pharmaceutical Co., Ltd. Address before: 130118 Changchun, South Gate District, Jilin city street,, Jilin Agricultural University Applicant before: Bao Haiying |
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