CN104983821B - Application of the Aconitum kongboense in alleviating the damage of drosophila intestinal inflammatory - Google Patents
Application of the Aconitum kongboense in alleviating the damage of drosophila intestinal inflammatory Download PDFInfo
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- CN104983821B CN104983821B CN201510377387.7A CN201510377387A CN104983821B CN 104983821 B CN104983821 B CN 104983821B CN 201510377387 A CN201510377387 A CN 201510377387A CN 104983821 B CN104983821 B CN 104983821B
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Abstract
The present invention is application of the Aconitum kongboense extract in alleviating the damage of drosophila intestinal inflammatory.Aconitum kongboense extract is obtained using water extraction, drosophila intestinal inflammatory damage model reparative experiment is carried out to Aconitum kongboense extract, as a result, it has been found that Aconitum kongboense extract has good anti-inflammatory activity, it can be used for preparing anti-inflammatory drug, the drug of especially anti-intestinal inflammatory damage.
Description
Technical field
The present invention relates to application of the Aconitum kongboense extract in terms of alleviating drosophila intestinal inflammatory damage, and it is natural to be related to drosophila
Immunological technique field.
Background technology
Under normal circumstances, the normal flora in intestinal mucosal barrier and between host have certain balance.The undesirable external world
Environment can cause enteron aisle homeostasis to be lacked of proper care under the conditions of infection etc., and then body is induced to generate whole body or local inflammation reaction.Disease
After substance enters body by enteron aisle, induction enteron aisle epidermal cell activates a series of immune responses, anti-eventually by generating
Bacterium peptide and active oxygen kill pathogen.In drosophila, an intestinal stem cell (Intestinal stem cell, ISC) into
Row Asymmetric division can usually generate two cells, and one of cell keeps stem cell properties, another is not divided to be of short duration
Enteroblast (Enteroblast, EB), enteroblast can continue to be divided into absorption-type enterocyte (Enterocyte,
) or the endocrine cell of secreting type (Enteroendocrine cell, EE) EC.The collective effect of above-mentioned cell can realize intestines
The maintenance and update of road tissue.Compared with mammalian gut, enteron aisle structure and the cell composition of drosophila are relatively easy, and adjust
The coherent signal approach for saving function of intestinal canal is all similar to mammal.To sum up, drosophila intestine immunity regulation mechanism and enteron aisle are studied
Stem cell differentiation mechanism has important directive significance to the pathogenesis for disclosing human intestine's relevant disease.
Aconitum kongboense (Aconitum kongboense Lauener) is distributed in Tibet, China, and the ground such as western Sichuan, are normal
Use Tibetan medicine.Aconitum kongboense is toxic, has dispelling wind and eliminating dampness, analgesic and other effects, for treating wet arthralgia, traumatic injury, poisonous insect
It the illnesss such as bites.However, there is no the relevant report for being used to alleviate intestinal inflammatory damage to Aconitum kongboense extract at present.
Invention content
It is an object of the invention to disclose Aconitum kongboense extract to can be applied to alleviate intestinal inflammatory damage.
Above-mentioned purpose is achieved by following scheme：
Drosophila feeding toxic reagent (SDS and DSS) is given to establish drosophila intestinal inflammatory damage model first, by Aconitum kongboense
Extract carries out the experiments such as drosophila survival rate, intestinal epithelial cell quantity and immunostaining, Intestinal Tract Morphology observation, as a result, it has been found that
Aconitum kongboense extract has preferable anti-inflammatory activity, can be used for preparing anti-inflammatory drug, especially treats the drug of intestinal inflammatory.
Description of the drawings
Fig. 1 is influence of the Aconitum kongboense extract to drosophila survival rate after SDS and DSS processing；
Fig. 2 is influence of the Aconitum kongboense extract to drosophila intestinal stem cell and enteroblast quantity after SDS processing；
Fig. 3 is influence of the Aconitum kongboense extract to drosophila intestinal cell immunostaining results after SDS processing；
Fig. 4 is influence of the Aconitum kongboense extract to drosophila Intestinal Morphology after SDS processing；
Specific implementation mode
With reference to example, the invention will be further described：
Experimental animal：Wild type W1118Strain Drosophila melanogaster (Drosophila melanogaster) preserves for laboratory,
EsgGal4 UAS-GFP attack professor Rong Wen by Peking University and provide.Drosophila cultivation temperature is 25 DEG C, relative humidity 60%.
The preparation of Drosophila medium：Control group culture medium prescription is corn flour 14.2g, agar 1.1g, yeast powder 3.3g, big
Bean powder 1.9g, white sugar 17g, propionic acid 0.94mL, deionized water 200mL.It is added in experimental group isometric with control group deionized water
Aconitum kongboense extracting solution, a concentration of the 1% of Aconitum kongboense extract in experimental group.
Each experiment reagent that the present embodiment is related to is that commercially available analysis is pure：SDS and DSS etc..
The preparation of 1 Aconitum kongboense aqueous extract of embodiment
Aconitum kongboense powder 10g is weighed, 300mL deionized waters immersion 12h overnight is added, moderate heat boils 3h, filtering, filter residue
200mL deionized waters are added afterwards to be extracted 1 time with same method, 2 filtrates of merging are concentrated into 50mL and are to get to mass concentration
The Chinese herbal medicine aqueous extract of 200mg/mL (20%).4 DEG C of sealed storages are spare.
Influence of the 2 Aconitum kongboense extract of embodiment to drosophila survival rate after SDS and DSS processing
As a control group, the drosophila cultivated in Aconitum kongboense culture medium is as experiment for drosophila to be cultivated in ordinary culture medium
Group.Random collecting control group and experimental group have been sprouted wings 3~5 days drosophilas respectively, each 15 of female male drosophila in every group.Each group drosophila
It is transferred to after Nature enemy 2h in the drosophila pipe for putting the filter paper that is of five storeys, it includes 0.6% that 5% sucrose solution is separately added into filter paper
The pro-inflammatory cytokines such as SDS, 4%DSS make filter paper fully soak, and per filter paper of replacement for 24 hours and record drosophila death toll.
As a result it shows：SDS handles drosophila after 4 days, and control group survival rate drops to 26.6%, and the experiment of feeding Aconitum kongboense
Group survival rate is 57.1% (* * * P<0.001) 30.5% (Figure 1A), is improved compared with the control group.In addition, DSS handles drosophila 7
After it, the survival rate of control group is only 10%, and the experimental group survival rate of feeding Aconitum kongboense is respectively 50.9% (* * * P<
0.001), 40.9% (Figure 1B) is improved than control group.The above results illustrate that Aconitum kongboense extract can alleviate SDS and DSS is lured
The intestinal inflammatory damage led, considerably improves the survival rate of drosophila.
Influence of the 3 Aconitum kongboense extract of embodiment to drosophila intestinal stem cell and enteroblast quantity after SDS processing
The female drosophila feeding that random collecting control group and experimental group are sprouted wings 3~4 days contains SDS (0.6%) solution of 5% sucrose
16h, 10~15 drosophila enteron aisles of extraction are in formaldehyde under microscope, and room temperature is combined after fixing 30min with GFP antibody, 4 DEG C of mistakes
Night.DAPI dyes 10min after 2 anti-rat-FITC combinations 2h, finally uses glycerine mounting, in fluorescence microscopy microscopic observation and takes a picture.
As a result it shows：GFP positive cells in feeding Aconitum kongboense experimental group (precursor, including intestinal stem cell and at
Enterocyte) it counts compared with control group compared to significantly reducing, as a result see Fig. 2.The above result shows that Aconitum kongboense can preferably resist cause
The hyper-proliferative of stem cell and enteroblast after scorching factor treatment.
Influence of the 4 Aconitum kongboense extract of embodiment to drosophila intestinal cell immunostaining results after SDS processing
The female drosophila feeding that random collecting control group and experimental group are sprouted wings 3~4 days contains SDS (0.6%) solution of 5% sucrose
96h, for 10~15 drosophila enteron aisles of extraction in PBS, 7-AAD room temperatures dye 30min under microscope, and paraformaldehyde fixes 10min,
DAPI dyes 10min, and fluorescence microscopy under the microscope and is taken pictures after glycerine mounting.
As a result it shows：There is mortality in the intestinal epithelial cell of control group, and in the experimental group of feeding Aconitum kongboense
The The dead quantity of intestinal epithelial cell significantly reduces, and as a result sees Fig. 3.
Influence of 5 Aconitum kongboense of embodiment to drosophila Intestinal Morphology after SDS processing
The female drosophila to have sprouted wings 3~4 days is collected, 10~15 drosophila enteron aisles are extracted after SDS processing, under microscope in PBS
In, Intestinal Morphology variation is observed under natural light.
As a result it shows：The control group of SDS processing is due to intestinal epithelial cell mortality, and length is obviously shortened, Er Qie
There is apparent melanoma in middle hindgut position.When feeding Aconitum kongboense, enteron aisle length obviously increases compared with the control group,
And there is not melanoma, as a result see Fig. 4.
In conclusion by drosophila survival rate, intestinal stem cell and enteroblast quantity variation, intestinal cell dyeing and
The analysis of Intestinal Morphology, it is believed that Aconitum kongboense extract can alleviate the damage of drosophila intestinal inflammatory.
Claims (1)
1. Aconitum kongboense extract is applied in preparing alleviation drosophila intestinal inflammatory damage medicine, it is characterised in that:
Aconitum kongboense water extract can improve the survival rate after drosophila feeding SDS or DSS and reduce gut epithelium apoptosis cell, together
When alleviate the hyper-proliferative of enteron aisle melanoma and precursor caused by SDS.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103356724A (en) * | 2012-08-22 | 2013-10-23 | 东北林业大学 | Application of acanthopanax water extract in relieving intestinal inflammation of drosophila melanogaster |
| CN103599162A (en) * | 2013-11-22 | 2014-02-26 | 东北林业大学 | Application of Tibet rhodiola crenulata extract in preparing medicament for resisting intestinal inflammation |
| CN104017032A (en) * | 2014-05-12 | 2014-09-03 | 内蒙古农业大学 | Method for preparing lappaconitine tannate and uses thereof |
| CN104491035A (en) * | 2014-12-16 | 2015-04-08 | 甘肃中医学院 | Detoxification processing technology of aconitum sinomontanum nakai |
| CN105214046A (en) * | 2015-10-29 | 2016-01-06 | 青岛海之源智能技术有限公司 | Preparation for the treatment of Lateolabrax japonicus enteritis disease and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103356724A (en) * | 2012-08-22 | 2013-10-23 | 东北林业大学 | Application of acanthopanax water extract in relieving intestinal inflammation of drosophila melanogaster |
| CN103599162A (en) * | 2013-11-22 | 2014-02-26 | 东北林业大学 | Application of Tibet rhodiola crenulata extract in preparing medicament for resisting intestinal inflammation |
| CN104017032A (en) * | 2014-05-12 | 2014-09-03 | 内蒙古农业大学 | Method for preparing lappaconitine tannate and uses thereof |
| CN104491035A (en) * | 2014-12-16 | 2015-04-08 | 甘肃中医学院 | Detoxification processing technology of aconitum sinomontanum nakai |
| CN105214046A (en) * | 2015-10-29 | 2016-01-06 | 青岛海之源智能技术有限公司 | Preparation for the treatment of Lateolabrax japonicus enteritis disease and preparation method thereof |
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