CN115569144A - Application of slender dioscin in preparation of medicines for preventing inflammatory bowel diseases - Google Patents
Application of slender dioscin in preparation of medicines for preventing inflammatory bowel diseases Download PDFInfo
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- CN115569144A CN115569144A CN202211243568.7A CN202211243568A CN115569144A CN 115569144 A CN115569144 A CN 115569144A CN 202211243568 A CN202211243568 A CN 202211243568A CN 115569144 A CN115569144 A CN 115569144A
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- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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Abstract
The invention relates to the technical field of medicine, in particular to application of slender dioscin in preparation of a medicine for preventing inflammatory bowel diseases.
Description
Technical Field
The invention relates to the technical field of medicine, in particular to application of slender dioscin in preparation of a medicine for preventing inflammatory bowel disease.
Background
Inflammatory Bowel Disease (IBD), including ulcerative colitis and crohn's disease, is a chronic inflammatory condition that affects the gastrointestinal tract. The main clinical manifestations of the disease are persistent or recurrent diarrhea, abdominal pain, mucous bloody stool, etc. The IBD has a long course of disease, high recurrence rate and high cure difficulty, and patients often suffer long-term pain and directly affect the life quality of the patients, so that the IBD is listed as one of the most difficult diseases by the world health organization. At present, the common western medicines for treating IBD have short clinical remission time and great side effect. Although acute inflammation reaction is rapid and obvious when western medicine is used for treating acute inflammation, the traditional Chinese medicine is based on the scientific thinking of integral regulation and control, and aims at the pathological characteristics of IBD patients, so that chronic inflammation is more effectively treated. Therefore, the search for a traditional Chinese medicine with good therapeutic effect on IBD is of great significance for finding a novel therapeutic medicine for IBD.
The fine dioscin (Gracillin) is a natural steroid compound separated from dioscorea opposita and has the characteristics of tumor resistance, microorganism resistance, oxidation resistance, apoptosis resistance, inflammation resistance and the like. Gracillin is currently protective against atopic dermatitis and other inflammatory skin diseases. In addition, one study showed that Gracillin can reduce Lipopolysaccharide (LPS) -induced inflammatory injury of cardiomyocytes. However, there was no study of the effect of Gracillin on intestinal epithelial cell inflammation.
Disclosure of Invention
In view of the defects of the background art, the invention relates to the application of the slender dioscin in the preparation of medicines for preventing and treating inflammatory bowel diseases, discloses the potential effect of the slender dioscin on Caco-2 cells induced by LPS from a cellular level, and prompts that the slender dioscin can be regarded as an effective therapy for treating intestinal epithelium inflammation of patients with inflammatory bowel diseases.
The invention provides an application of slender dioscin in preparation of a medicine for preventing inflammatory bowel diseases.
The invention also provides application of the slender dioscin in preparation of a medicine for treating inflammatory bowel diseases.
Furthermore, the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
The invention also provides application of the slender dioscin in medicines for preventing inflammatory bowel diseases through bacteriostasis and sterilization.
The invention also provides application of the slender dioscin in preparation of treating the epithelial inflammation of the inflammatory bowel disease.
Furthermore, the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
The invention also provides application of the slender dioscin in preparation of a medicine for treating inflammatory bowel diseases induced by LPS.
Furthermore, the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
The invention has the main beneficial effects that:
1. the invention realizes the purpose of the slender dioscin in treating the intestinal epithelium inflammation;
2. the invention explores the function of the slender dioscin in inflammatory diseases, particularly the field of inflammatory bowel diseases, and provides a new medicine treatment mode for treating the inflammatory bowel diseases.
Drawings
FIG. 1: influence of different concentrations of dioscin (Gracillin) on LPS-induced cell viability of a Caco-2 inflammation model, *** p<0.001 vs. Control； # p<0.05， ### p<0.001 vs. LPS。
FIG. 2: influence of different concentrations of Gracillin on inflammation factors of a Caco-2 inflammation model induced by LPS, *** p<0.001 vs. Control； # p<0.05， ## p<0.01， ### p<0.001 vs. LPS。
FIG. 3: the influence of different concentrations of Gracillin on LPS-induced intestinal connexin in a Caco-2 inflammation model, *** p<0.001 vs. Control； ### p<0.001 vs. LPS。
FIG. 4 is a schematic view of: pairs of fine dioscin (Gracillin) with different concentrationsInfluence of LPS-induced Caco-2 inflammatory model barrier integrity, *** p<0.001 vs. Control； # p<0.05， ## p<0.01， ### p<0.001 vs. LPS。
FIG. 5 is a schematic view of: bacteriostatic action of Gracillin (Gracillin) with different concentrations on escherichia coli (e. ### p<0.001 vs.DMSO。
Detailed Description
While the embodiments of the present invention will be described and illustrated in detail with reference to the accompanying drawings, it is to be understood that the invention is not limited to the specific embodiments disclosed, but is intended to cover various modifications, equivalents, and alternatives falling within the scope of the invention as defined by the appended claims.
For the convenience of understanding the embodiments of the present invention, the following description will be further explained by taking specific embodiments as examples with reference to the drawings, and the embodiments are not to be construed as limiting the embodiments of the present invention.
Example 1 of the present invention relates to CCK-8 for detecting the effect of different concentrations of Gracillin (Gracillin) (20, 40 μmol/L) on cell viability when treated for 24 hours in LPS-induced intestinal epithelial cell Caco-2 inflammation model, the specific experimental method is as follows:
caco-2 was treated with 1. Mu.g/mL LPS alone for 24 hours or incubated with Gracillin (20, 40. Mu. Mol/L) for 24 hours. Thereafter, CCK-8 solution was added to each well and incubated for 4 hours. Finally, the absorbance was measured at 450nm by a microplate reader.
The results are shown in FIG. 1, gracillin significantly increases the LPS-inhibited cellular activity.
The embodiment 2 of the invention relates to a method for detecting the influence of different concentrations of Gracillin (Gracillin) (20 and 40 mu mol/L) on the expression of inflammatory factors (IL-6 and TNF-alpha) in a Caco-2 inflammation model induced by LPS by using a real-time quantitative polymerase chain reaction, and the specific experimental method comprises the following steps:
total RNA was isolated from each group of Caco-2 cells. Then, complementary DNA (cDNA) is synthesized by reverse transcription reaction using reverse transcriptase. qRT-PCR was performed using the Real-Time PCR system and the PCR Master Mix. GAPDH was used as a housekeeping gene. The relative expression level of each gene was quantified by the 2. DELTA.CT method.
The results are shown in FIG. 2, gracillin significantly inhibited LPS-promoted inflammatory factor expression.
The embodiment 3 of the invention relates to a Western blot method for detecting the influence of different concentrations of Gracillin (Gracillin) (20 and 40 mu mol/L) on the expression of intestinal connexin in a Caco-2 inflammation model induced by LPS, and the specific experimental method comprises the following steps:
total protein from Caco-2 cells was isolated by using lysis buffer and then protein concentration was determined by BCA protein kit. The separated proteins were transferred to polyvinylidene fluoride membranes. Thereafter, the membrane was treated with blocking buffer for 1 hour and then reacted with primary antibody at 4 ℃ overnight. The next day, incubation with secondary antibody was performed at room temperature for 2 hours. Protein signals were observed using an imaging system with enhanced chemiluminescent reagents.
The results are shown in FIG. 3, the expression of ZO-1, occludin and Claudin-1 is reduced by LPS treatment; gracillin increases ZO-1, occludin, claudin-1 expression inhibited by LPS.
Example 4 of the present invention relates to transepithelial electrical resistance (TEER) testing the effect of Gracillin on barrier integrity in LPS-induced Caco-2 inflammation model, the specific experimental method is as follows:
treated Caco-2 cells were seeded into 6-well plates each with a Transwell insert. Prior to measurement, cells were incubated in culture medium until fully differentiated. Epithelial transmembrane resistance meters were used to measure TEER at 0, 12, 24, 36 and 48 hours for each group.
The results are shown in FIG. 4, TEER is inhibited by LPS treatment and TEER inhibited by LPS is improved by Gracillin.
Embodiment 5 of the present invention relates to an analysis of bacteriostatic action of Gracillin on escherichia coli (e.coli), and the specific experimental method is as follows:
e, coli was cultured in LB medium for 24 hours, and inoculated into E.coli. Subsequently, gracillin (20, 40 μmol/L) treatment was performed, DMSO was used as a control group, incubation was performed at 37 ℃ for 24 hours, and bacterial liquid OD600 was measured by a full-wavelength plate reader to determine the Minimum Inhibitory Concentration (MIC) of tangerine peel essential oil to e.coli.
The results are shown in FIG. 5, gracillin inhibits growth and reproduction of E.
Table 1: bacterial cell density table of all groups
Therefore, the tenuous dioscin has bacteriostasis, and the bacteriostasis effect is increased along with the increase of the concentration of the tenuous dioscin.
Finally, it should be noted that: the above-mentioned embodiments are only specific embodiments of the present invention, which are used for illustrating the technical solutions of the present invention and not for limiting the same, and the protection scope of the present invention is not limited thereto, although the present invention is described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: any person skilled in the art can modify or easily conceive the technical solutions described in the foregoing embodiments or equivalent substitutes for some technical features within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the embodiments of the present invention, and they should be construed as being included therein. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (8)
1. Application of slender dioscin in preparation of medicines for preventing inflammatory bowel disease is provided.
2. Application of slender dioscin in preparation of medicines for treating inflammatory bowel diseases is provided.
3. The use of the fine dioscin according to claim 2 in the preparation of a medicament for the treatment of inflammatory bowel disease, wherein: the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
4. Application of slender dioscin in preparation of medicines for treating epithelial inflammation of inflammatory bowel disease is provided.
5. The use of a fine dioscin according to claim 4 in the preparation of a medicament for the treatment of epithelial inflammation in inflammatory bowel disease, wherein: the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
6. The application of slender dioscin in preparing medicines for preventing inflammatory bowel disease by inhibiting bacteria and killing bacteria.
7. Application of slender dioscin in preparation of medicine for treating inflammatory bowel diseases induced by LPS (lipopolysaccharide) is provided.
8. The use of the slender dioscin according to claim 7, in the preparation of a medicament for the treatment of inflammatory bowel disease induced by LPS, wherein: the concentration of the slender dioscin in the medicine prepared by the application is 20-40 mu mol/L.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211243568.7A CN115569144A (en) | 2022-10-12 | 2022-10-12 | Application of slender dioscin in preparation of medicines for preventing inflammatory bowel diseases |
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| CN202211243568.7A CN115569144A (en) | 2022-10-12 | 2022-10-12 | Application of slender dioscin in preparation of medicines for preventing inflammatory bowel diseases |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101257909A (en) * | 2005-07-06 | 2008-09-03 | 英国技术集团国际有限公司 | Core 2 glcnac-t inhibitors |
| CN101511369A (en) * | 2006-08-03 | 2009-08-19 | 肿瘤学研究国际有限公司 | Methods and compositions for inhibiting angiogenesis. |
| CN102617698A (en) * | 2012-03-14 | 2012-08-01 | 云南省药物研究所 | Method for preparing fine dioscin and application of fine dioscin |
| CN102784157A (en) * | 2011-05-18 | 2012-11-21 | 南方医科大学南方医院 | Application of slender dioscin and pharmaceutical composition containing the same |
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- 2022-10-12 CN CN202211243568.7A patent/CN115569144A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101257909A (en) * | 2005-07-06 | 2008-09-03 | 英国技术集团国际有限公司 | Core 2 glcnac-t inhibitors |
| CN101511369A (en) * | 2006-08-03 | 2009-08-19 | 肿瘤学研究国际有限公司 | Methods and compositions for inhibiting angiogenesis. |
| CN102784157A (en) * | 2011-05-18 | 2012-11-21 | 南方医科大学南方医院 | Application of slender dioscin and pharmaceutical composition containing the same |
| CN102617698A (en) * | 2012-03-14 | 2012-08-01 | 云南省药物研究所 | Method for preparing fine dioscin and application of fine dioscin |
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