CN104222527B - The application process of belulinic acid Betulinic acid - Google Patents
The application process of belulinic acid Betulinic acid Download PDFInfo
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- CN104222527B CN104222527B CN201410520978.0A CN201410520978A CN104222527B CN 104222527 B CN104222527 B CN 104222527B CN 201410520978 A CN201410520978 A CN 201410520978A CN 104222527 B CN104222527 B CN 104222527B
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Abstract
The invention belongs to the technical field of cattle breeding, specifically disclose belulinic acid Betulinic acid in preparation prevention and the purposes of the preparation for the treatment of domestic animal, especially diarrhea of weaned piglets.Belulinic acid Betulinic acid adds in ablactational baby pig feedstuff with the dosage of 0.1 0.5%, can significantly reduce diarrhea of weaned piglets rate, improves Gut Mucosa Morphology, protects intestinal mucosal barrier, is effectively improved the disease resistance ability that piglet is overall, improves daily gain.
Description
Technical field
The invention belongs to the technical field of cattle breeding, be specifically related to belulinic acid Betulinic acid and in preparation prevention and treat domestic animal, especially wean
The purposes of the preparation of piglet diarrhea.
Background technology
In present level intensive culture, the disease that harm that piglet diarrhea is common is the biggest.This disease directly results in survival of piglets
Rate decline, poor growth, arrest of development and become cad pig, even dead, the speed of growth and the feedstuff that affect whole feeding period turn
Rate, carrys out heavy economic losses finally to cultivation industrial belt.According to statistics, it is dead that the piglet death that China causes because of diarrhoea every year accounts for piglet
Die sum 39.8%.The average attack rate of piglet diarrhea is 46.5%, and mortality rate is 15%, and the direct economic loss caused is high
Reach 1,000,000,000 yuan.The reason of post-weaning diarrhea in piglet is a lot, mainly has dyspepsia, immunity degradation, electrolyte imbalance, intestinal to cause
Nutrition and environmental stress etc. after the infecting and wean of characteristic of disease escherichia coli and other pathogenic bacterium.For a long time, the preventing and treating master of this disease
Antibiotic to be depended on and chemicals solve, and the abuse of antibiotic causes drug resistance to produce, endogenous infection and superinfection,
Drug residue etc. in immunity of organisms decline and food brings huge threat to human health.Therefore, find effectively
Substitutes For Antibiotic is major issue urgently to be resolved hurrily.
Belulinic acid Betulinic acid belongs to lupinane type pentacyclic triterpene material, is distributed widely in as many in Rhamnaceae, Myrtaceae and Betulaceae etc.
Planting in plant, its amount is the lowest, higher with Japanese birch bark amount.Cortex Betulae Luminiferae described in Compendium of Material Medica can be used for yellow cellulitis, acute mastitis,
The treatment of the diseases such as scabies.As far back as 19th century, Japanese birch bark is just placed in tea and Other Drinks by American and drinks；Silver birch
Juice is referred to as " natural beer " and " forest beverage " by European.Nineteen ninety-five finds that belulinic acid Betulinic acid has extremely strong choosing to melanoma cell
Selecting sexual cell toxicity, the duplication of HIV in suppression H9 lymphocyte, its sphere of action almost affects each of viral lifecycle
Link.Owing to belulinic acid Betulinic acid has superior antitumor and HIV (human immunodeficiency virus)-resistant activity, cause people to the widely studied of belulinic acid Betulinic acid and concern.
Studying discovery subsequently, belulinic acid Betulinic acid has multiple stronger biological activitys such as immunomodulating, anti-oxidation stress, antiinflammatory, and belulinic acid Betulinic acid
Application in terms of piglet diarrhea is rarely reported.It is reported, belulinic acid Betulinic acid is approximately Normocellular 10 times to the sensitive intensity of cancerous cell,
When belulinic acid Betulinic acid dosage is up to 500mg/kg, animal does not has an obvious poisoning symptom, and belulinic acid Betulinic acid extensively to derive from nature many
Plant plant, illustrate that belulinic acid Betulinic acid has safely, is not likely to produce drug resistance, there is not the advantages such as drug residue, be preferable antibiotic
One of succedaneum.
Summary of the invention
It is an object of the invention to provide a kind of belulinic acid Betulinic acid and in preparation prevention and treat domestic animal, especially diarrhea of weaned piglets preparation
Purposes, the belulinic acid Betulinic acid of use safely, have no drug resistance, with strong points, it is possible to effectively control diarrhea of weaned piglets, have concurrently simultaneously and carry
The immunity of high body and production performance effect, alternative antibiotic carries out feeding and interpolation.
The application process of belulinic acid Betulinic acid, described belulinic acid Betulinic acid is for preparing the antidiarrhoea formulation of domestic animal.Described antidiarrhoea formulation can also
Improve the daily gain of domestic animal, reduce feedstuff-meat ratio.
Described domestic animal is pig, especially ablactational baby pig.
By described belulinic acid Betulinic acid direct feeding domestic animal or add domestic animal to basal diet in feeding.
Described domestic animal is pig, especially ablactational baby pig.
It is 0.1～0.5% that described belulinic acid Betulinic acid adds to the mass ratio in ablactational baby pig basal diet.Preferably 0.1-0.2%.
The preparation process of described belulinic acid Betulinic acid is: extracts betulinol from Japanese birch bark, prepares intermediate product through Jones reagent oxidation
After betulonic acid, synthesize belulinic acid Betulinic acid with sodium borohydride reduction.
The content using HPLC to measure belulinic acid Betulinic acid is 96.53%.
The belulinic acid Betulinic acid used in the present invention is to utilize existing technology path to prepare, and extracts betulinol, through Jones from Japanese birch bark
After reagent oxidation prepares intermediate product betulonic acid, synthesize belulinic acid Betulinic acid with sodium borohydride reduction.(list of references: Yi Jine, Wen Li
Newly, Yuan Liyun, Obminska-MrukowiczBozena, Yuan Hui. the extraction of betulinol and belulinic acid Betulinic acid study on the synthesis in silver birch. lake
South agriculture university journal, 2010,36 (5): 574-580.)
The present invention has the advantage that:
The belulinic acid Betulinic acid that the present invention uses, has various biological activity, and has disease-proof functions concurrently, not only preventable disease but also can improve animal
Production performance, belulinic acid Betulinic acid is difficult to remain in animal body, is not likely to produce drug resistance, it is ensured that the quality of livestock products.
1. can improve Growth Performance of Weaning Piglets: belulinic acid Betulinic acid additive group is compared with matched group (not having additive), and raising increases day by day
Weight, reduces feedstuff-meat ratio.
The most substantially suppress diarrhea of weaned piglets: belulinic acid Betulinic acid additive group is compared with matched group (not having additive), and diarrhea rate is notable
Reduce；
3. can promote intestinal regeneration: be obviously improved ablactational baby pig Mucosal Morphology in Small, alleviate intestinal villi atrophy, belulinic acid Betulinic acid adds
Agent group significantly improves duodenal height of naps, significantly reduces Crypt depth, improve height of naps and Crypt depth it
Ratio.
Accompanying drawing explanation
Fig. 1 is the belulinic acid Betulinic acid impact on Growth Performance of Weaning Piglets；
Compared with blank group, " * " significant difference (P < 0.05)；
Fig. 2 is the belulinic acid Betulinic acid impact on ablactational baby pig feed-weight ratio；
Compared with blank group, " * " significant difference (P < 0.05), " * * " difference is extremely notable (P < 0.01)；
Fig. 3 is the belulinic acid Betulinic acid impact on piglet diarrhea rate；
Compared with blank group, " * * " difference is extremely notable (P < 0.01)；
Fig. 4 is that belulinic acid Betulinic acid is on ablactational baby pig small intestinal mucosa height of naps and the impact of Crypt depth；
Compared with blank group, " * " significant difference (P < 0.05)；
Fig. 5 is that belulinic acid Betulinic acid is on ablactational baby pig small intestinal mucosa height of naps and the impact of Crypt depth ratio.
Compared with blank group, " * * " difference is extremely notable (P < 0.01).
Detailed description of the invention
The belulinic acid Betulinic acid that the present invention uses belongs to pentacyclic triterpenoid, derives from Japanese birch bark, extracts birch from Japanese birch bark
Alcohol, after intermediate product betulonic acid is prepared in Jones reagent oxidation, synthesizes belulinic acid Betulinic acid with sodium borohydride reduction.Concrete preparation method
As follows: to collect the bark of Betula platyphylla Suk. wood, through 60 DEG C of drying, it is housed in dark, is dried place.Semi-synthesis method is used to prepare belulinic acid Betulinic acid.
1) from Japanese birch bark, belulinic acid Betulinic acid is extracted: weigh Japanese birch bark powder, be heated to reflux 3h, drying under reduced pressure with methanol 70 DEG C, add
2moL/L NaOH stirs, and collects subnatant, evaporated under reduced pressure with ether dissolution, adds appropriate water and fully shake up, collection upper strata
Liquid, drying under reduced pressure, silica gel column chromatography, drying under reduced pressure obtains betulinol sterling.2) using betulonic acid is that intermediate synthesizes belulinic acid Betulinic acid
Method: quickly stir with acetone solution betulinol, after being cooled to 5-10 DEG C, be added dropwise over the Jones reagent (betulinol of brand-new
Jones reagent=1 6), 20 DEG C of reaction 3h, it is filtered to remove blackish green precipitation, evaporated under reduced pressure acetone, residual liquid ethyl acetate
Extraction, washing, 10%NaHCO3Washing, the anhydrous MgSO of organic facies4It is dried, obtains faint yellow solid (betulonic acid crude product)
With ether dissolution, flash column chromatography separates, and drying under reduced pressure obtains betulonic acid sterling.Again with sodium borohydride/oxolane (NaBH4/
THF) reduction, is dissolved in gained solid methanol and boils 15-20min, and cooling recrystallization obtains belulinic acid Betulinic acid sterling.Use HPLC
The content measuring belulinic acid Betulinic acid is 96.53%.
By following pharmacodynamic experiment, the present invention proves that belulinic acid Betulinic acid may be used for improving the diarrhoea of ablactational baby pig, and improve ablactational baby pig
Growth performance.
Example 1:
Healthy male KM mice 30,4-5w, SPF level, body weight (20 ± 2) g, credit number: (Hunan) 2009-0004, raise
Material is commonly bred as material for M02 mice, and mice and feedstuff are purchased from Hunan Si Laike Jing Da laboratory animal company.Mice is placed in (22-25)
DEG C, relative humidity is in the feeding room of 50%～70%, after raising 1w, is randomly divided into 3 groups, i.e. blank group (A group),
Cyclophosphamide group (B group, intestinal mucosal injury model), (C group, belulinic acid Betulinic acid presses 0.5mg/kg to belulinic acid Betulinic acid with cyclophosphamide combined group
B w give with).Belulinic acid Betulinic acid is suspended in the soluble starch of 1% by C group, uses the mode of gavage to be administered, and A group is with B group only
Gavage the soluble starch of 1%, all by 20ml/kg bw give with, 1/d, continuous 14d, observe clinical manifestation and body weight becomes
Change.Last give with 1h after modeling, B group and C group mouse peritoneal inject cyclophosphamide (50mg/kg bw), A group note
Penetrate isopyknic sterile saline.Raising terminates, and water 12h is can't help in mice fasting, and de-cervical vertebra is put to death, and draws materials and carries out every
Indexs measure.
1) belulinic acid Betulinic acid impact on Mouse Weight, thymus index and spleen index
Belulinic acid Betulinic acid is shown in Table 1 to the impact of Mouse Weight, thymus index and spleen index, compared with blank group, and cyclophosphamide group
Body weight and spleen index decrease, but difference is the most notable；But thymus index substantially reduces (P < 0.05), prompting cyclophosphamide makes
Obtain normal mouse immunologic function to be suppressed, cause body weight to decrease；Compared with cyclophosphamide group, the body of belulinic acid Betulinic acid group mice
Weight, thymus index and spleen index all obtain a certain degree of recovery, and thymus index significant difference (P < 0.05), show birch
Acid can improve the immunologic function of hypoimmunity mice.
The impact on Mouse Weight, thymus index and spleen index of table 1 belulinic acid Betulinic acid
Compared with blank group, " * " significant difference (P < 0.05)；Compared with cyclophosphamide group, " # " significant difference (P < 0.05).
2) belulinic acid Betulinic acid is on the impact of IL-2 and IL-10 content in mouse intestinal
It is stably immune response that Thl/Th2 system keeps, and the ratio of conventional IL-2/IL-10 represents.Belulinic acid Betulinic acid is to intestinal
Thl/Th2 impact is shown in Table 2, and compared with blank group, cyclophosphamide group mice intestinal mucosa IL-2/IL-10 ratio significantly reduces
(P < 0.05), shows that cellullar immunologic response is suppressed, and immunologic balance is broken；Compared with model group, belulinic acid Betulinic acid group intestinal
IL-2/IL-10 ratio significantly raises, and prompting belulinic acid Betulinic acid can reverse its drifting state by raising Th1/Th2 level, thus extensive
Multiple mucosa balance.
Table 2 belulinic acid Betulinic acid is on the impact of IL-2 and IL-10 content in mouse intestinal
Compared with blank group, " * " significant difference (P < 0.05), " * * " difference is extremely notable (P < 0.01)；Compared with cyclophosphamide group, " # " is poor
Different notable (P < 0.05), " ## " difference is extremely notable (P < 0.01).
3) belulinic acid Betulinic acid impact on mice intestinal mucosa morphology
Cyclophosphamide can cause small intestinal villous height to shorten, and height of naps/Crypt depth ratio declines, and is shown in Table 3, shows that small intestinal disappears
Change absorption and opposing foreign substance intrusion function is suppressed.Belulinic acid Betulinic acid can improve the intestinal mechanical barrier damage that cyclophosphamide causes,
Be conducive to maintaining the normal organizational structure of small intestinal and physiological function.
The impact on mice intestinal mucosa morphology of table 3 belulinic acid Betulinic acid
Compared with blank group, " * " significant difference (P < 0.05), " * * " difference is extremely notable (P < 0.01)；Compared with cyclophosphamide group, " # " is poor
Different notable (P < 0.05), " ## " difference is extremely notable (P < 0.01).
4) belulinic acid Betulinic acid impact on mice intestinal mucosa SlgA content
The change of the change display intestinal immunological barrier function that in small intestinal mucosa layer, SigA sIgA (SlgA) is measured.See
Table 4, compared with matched group, the content of cyclophosphamide group mouse intestinal SIgA significantly reduces (P < 0.01), points out cyclophosphamide
Cause mice intestinal mucosal immune barrier injury；Compared with cyclophosphamide group, belulinic acid Betulinic acid can significantly improve SIgA content (P < 0.01),
The SIgA of intestinal mucosal injury mice is secreted and has improvement result by prompting belulinic acid Betulinic acid, plays intestinal immunological barrier and necessarily protects work
With.
The impact on mice intestinal mucosa SlgA content of table 4 belulinic acid Betulinic acid
Compared with blank group, " * * " difference is extremely notable (P < 0.01)；Compared with cyclophosphamide group, " ## " difference is extremely notable (P < 0.01).
5) belulinic acid Betulinic acid impact on mouse intestinal anti-oxidation function
As shown in Table 5, compared with blank group, cyclophosphamide group small intestinal SOD activity is remarkably decreased, and MDA content shows
Writing and raise (P < 0.01), prompting cyclophosphamide reduces the oxidation resistance of mucous membrane of small intestine；Compared with cyclophosphamide group, belulinic acid Betulinic acid
Group mucous membrane of small intestine SOD activity is obviously enhanced, and MDA content significantly reduces (P < 0.01), illustrates that belulinic acid Betulinic acid has antioxidation
Ability, can substantially alleviate the oxidative stress caused by cyclophosphamide, play a protective role intestinal.
Table 5 belulinic acid Betulinic acid is on mouse intestinal SOD and the impact of MDA level
Compared with blank group, " * * " difference is extremely notable (P < 0.01)；Compared with cyclophosphamide group, " ## " difference is extremely notable (P < 0.01).
Example 2:
Feeding experiment selects 90 28 age in days wean of same nursery house, Du × big × long ternary of body weight (8.4 ± 0.72) kg
Hybridization ablactational baby pig, is randomly divided into 3 experimental grouies, each experimental group 3 repetition, and wherein A group is blank group (base
Plinth daily ration, without any medicine), B group is belulinic acid Betulinic acid group (basal diet+0.1% belulinic acid Betulinic acid)；C group is antibiotic group (base
Plinth daily ration+50ppm oxytetracycline+50ppm chlortetracycline), experimental period 21d.Test first 1 week pig house is comprehensively cleaned,
Sterilization, before test, the anthelmintic of each group is once, uses closed management model management colony house.Early, middle and late three concentrations are divided to throw something and feed,
Having a small amount of feedstuff residue to be advisable with hopper, every day is stepped up.Use automatic drinking bowl freely to drink water, clear up feces in time, protect
Hold colony house well-ventilated.Ground keeps being dried, and every day checks feces.
1) belulinic acid Betulinic acid impact on piglet growth performance
From Fig. 1 and Fig. 2 it can be seen that during off-test, compared with blank group, it is young that belulinic acid Betulinic acid can significantly improve 60 ages in days
The net gain of pig and Average weight increasing a day, improve significantly (P < 0.05) to feed-weight ratio；Illustrate that belulinic acid Betulinic acid can improve piglet
Production performance, improves food conversion ratio.
2) belulinic acid Betulinic acid impact on piglet diarrhea
From figure 3, it can be seen that compared with blank group, belulinic acid Betulinic acid can substantially reduce the diarrhea rate (P < 0.05) of 60 age in days piglets；
Illustrate that in the present invention, belulinic acid Betulinic acid has Antidiarrheal efficacy, and effect is preferable.
3) belulinic acid Betulinic acid impact on ablactational baby pig Mucosal Morphology in Small
As can be seen from Figure 4 and Figure 5, compared with blank group, belulinic acid Betulinic acid can substantially raise height of naps degree/Crypt depth ratio,
Illustrate belulinic acid Betulinic acid can improve small intestinal digest and assimilate and resist foreign substance invade function, be conducive to maintain the normal knot of tissue of small intestinal
Structure and physiological function.
Example 3:
Feeding experiment selects 90 21 age in days wean of same nursery house, Du × big × long ternary of body weight (5.73 ± 0.41) kg
Hybridization ablactational baby pig, is randomly divided into 3 experimental grouies, each experimental group 3 repetition, and wherein A group is blank group (base
Plinth daily ration, without any medicine), B group is belulinic acid Betulinic acid group (basal diet+0.2% belulinic acid Betulinic acid)；C group is antibiotic group (base
Plinth daily ration+50ppm oxytetracycline+50ppm chlortetracycline), experimental period 28d.Test first 1 week pig house is comprehensively cleaned,
Sterilization, before test, the anthelmintic of each group is once, uses closed management model management colony house.Early, middle and late three concentrations are divided to throw something and feed,
Having a small amount of feedstuff residue to be advisable with hopper, every day is stepped up.Use automatic drinking bowl freely to drink water, clear up feces in time, protect
Hold colony house well-ventilated.Ground keeps being dried, and every day checks feces.
The efficacy assessments result of the test of result of the test and example 2 is without significant difference, therefore does not do deposited stating at this.
Above embodiments of the invention are described in detail, but described content has been only presently preferred embodiments of the present invention, it is impossible to quilt
Think the practical range for limiting the present invention.All impartial changes made according to the present patent application scope and improvement etc., all should still return
Within belonging to the patent covering scope of the present invention.
Claims (5)
1. belulinic acid Betulinic acid application on the antidiarrhoea formulation of preparation domestic animal.
The belulinic acid Betulinic acid the most according to claim 1 application on the antidiarrhoea formulation of preparation domestic animal, it is special
Levying and be, described antidiarrhoea formulation can also improve the daily gain of domestic animal, reduces feedstuff-meat ratio.
The belulinic acid Betulinic acid the most according to claim 1 and 2 application on the antidiarrhoea formulation of preparation domestic animal,
It is characterized in that, described domestic animal is pig.
The belulinic acid Betulinic acid the most according to claim 3 application on the antidiarrhoea formulation of preparation domestic animal, it is special
Levying and be, described domestic animal is ablactational baby pig.
The belulinic acid Betulinic acid the most according to claim 1 and 2 application on the antidiarrhoea formulation of preparation domestic animal,
It is characterized in that, the preparation process of described belulinic acid Betulinic acid is: extract betulinol from Japanese birch bark,
After intermediate product betulonic acid is prepared in Jones reagent oxidation, synthesize belulinic acid Betulinic acid with sodium borohydride reduction.
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| CN108850614A (en) * | 2018-08-06 | 2018-11-23 | 杭州光启医疗科技发展有限公司 | A kind of disease-resistant fattening pannage of health care |
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| CN102960560B (en) * | 2012-11-23 | 2014-09-10 | 重庆优宝生物技术有限公司 | Composite acidifier for reducing post-weaning diarrhea rate |
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