CN117815262A - Application of herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis - Google Patents
Application of herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis Download PDFInfo
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- CN117815262A CN117815262A CN202410131550.0A CN202410131550A CN117815262A CN 117815262 A CN117815262 A CN 117815262A CN 202410131550 A CN202410131550 A CN 202410131550A CN 117815262 A CN117815262 A CN 117815262A
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- polysaccharide extract
- ulcerative colitis
- endive
- preventing
- mice
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Abstract
The application discloses application of a endive polysaccharide extract in preventing and treating ulcerative colitis, and belongs to the technical field of medicines. The endive polysaccharide extract can effectively treat or relieve the symptoms of ulcerative colitis: including weight change, colon shortening, DAI scoring, pathological injury, etc.; blood routine results show that the polysaccharide extract of the endives improves the blood routine parameters of mice with DSS induced enteritis; the result of the immune experiment shows that the polysaccharide extract of the endives can regulate the immune parameters; the 16S rRNA detection result shows that the intestinal microbiota has obvious difference in the composition of intestinal microbiota among groups after the treatment of the endive polysaccharide extract; the metabonomics research result shows that the endive polysaccharide extract can regulate the metabolite composition of enteritis mice. Therefore, the polysaccharide extract of Sonchus arvensis can be used as an effective drug for treating or preventing ulcerative colitis, and the protection effect on the ulcerative colitis of mice is possibly related to the diversity of intestinal microbiota and the composition of metabolites.
Description
Technical Field
The application relates to the technical field of medical medicines, in particular to application of a endive polysaccharide extract in preventing and treating ulcerative colitis.
Background
Digestive system diseases seriously affect the study, work and life of patients. Ulcerative Colitis (UC) is a chronic inflammatory disease of the gastrointestinal tract that is easily recurrent and difficult to treat. UC causes inflammation and digestive system disorders, and typical symptoms of UC are abdominal pain, diarrhea and hematochezia. Many reports indicate that the incidence and prevalence of global UC has increased over the last decades. In addition to developed countries, recent studies have shown that the prevalence of UC has also increased in asia, africa and south america. Epidemiological studies have shown that the severity and duration of UC also increases the risk of colon cancer in patients.
At present, the main pathological mechanism of UC is not clear, and the special etiology of UC is related to various factors such as environment, life style, immune function, genes, intestinal flora and the like. Many reports indicate that dysregulation of intestinal microbiota is associated with inflammatory bowel disease. Thus, dysregulation of intestinal microbiota is considered to be the primary pathological mechanism of UC. There is increasing evidence that the microbiota composition in UC is significantly altered compared to healthy people, with reduced biodiversity and species richness. Furthermore, changes in bacterial composition may lead to changes in metabolites, which may play a role in the etiology of UC. Metabonomic analysis found that there was a correlation between bacterial composition and metabolism.
Many studies have shown that traditional Chinese medicine can treat diseases by interfering with the homeostasis of the human intestinal flora. Herba Sonchi arvensis (Sonchus arvensis L.) of Sonchus of Compositae (Compositae) is dried whole herb, and can be used as medicine. The endive is perennial herb and is mainly used in the wild of the barren hillsides, beach and roadside of northwest, north China, northeast and other places in China. The endive has rich nutrition and contains rich calcium, magnesium, manganese, titanium, selenium, zinc and other elements. The endive has bitter and cold nature, has the effects of clearing heat and detoxicating, promoting blood circulation by removing blood stasis, promoting diuresis and expelling pus, cooling blood and stopping bleeding, removing blood stasis and detumescence, and is commonly used for treating acute pharyngitis, acute bacillary dysentery, haematuria, hemorrhoids swelling and pain, and the like, and is also eaten as wild vegetable in people. Polysaccharide is an important component of traditional Chinese medicine, it is known to extract over 300 polysaccharide compounds from traditional Chinese medicine, and many plant-derived drugs have been approved by the FDA in the united states. Therefore, research on the pharmacological activity of polysaccharide in traditional Chinese medicine plants has important significance for development and application of polysaccharide.
Many studies have shown that dietary fibers, polysaccharides, can regulate intestinal flora. Polysaccharides are one of four basic substances constituting life, and are natural biological macromolecules widely existing in microorganisms, plants and animals. It has been reported that many polysaccharides and their derivatives have important bioactive functions such as antioxidant, anti-aging, anti-inflammatory, antiviral, anti-radiation, antitumor, antithrombotic, hypolipidemic, immunoregulatory, and intestinal microbial composition regulating effects. Polysaccharide has been of interest to many researchers because of its remarkable pharmacological activity, small toxic and side effects, easy absorption, and the like. At present, no report on application of the polysaccharide extract of the endives in preventing and treating ulcerative colitis is found at home and abroad.
Disclosure of Invention
In view of the above-mentioned drawbacks or shortcomings in the prior art, it is desirable to provide an application of a herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis, and to prepare a medicament for effectively preventing or treating ulcerative colitis, and improving the conditions of weight loss, colonic tissue ulcer, colonic hemorrhage and the like caused by ulcerative colitis.
The application of the herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis comprises the application of the herba Sonchi arvensis polysaccharide extract in preparing medicines for preventing and treating ulcerative colitis.
Preferably, the extraction and preparation method of the endive polysaccharide extract comprises the following steps:
step one: cutting dried whole herb of Sonchus arvensis, weighing, soaking in ethanol water solution, and ultrasonic extracting for multiple times;
step two: volatilizing solvent, weighing the extracted endive, soaking in water, heating for extraction to obtain an extracting solution, extracting for multiple times, filtering after each extraction, centrifuging the supernatant, and finally combining multiple filtrates;
step three: concentrating the combined filtrate under reduced pressure to obtain concentrated solution, slowly adding 3-5 times of absolute ethyl alcohol into the concentrated solution, continuously stirring, standing at 4 ℃ overnight, filtering and centrifuging to take out precipitate;
step four: washing the precipitate with absolute ethanol, acetone and diethyl ether twice, and oven drying to obtain herba Sonchi arvensis polysaccharide extract.
Preferably, the concentration of ethanol in the ethanol water solution in the first step is 80-100%; the ultrasonic extraction times in the first step are 1-5 times, and the total extraction time is 0.5-2 hours.
Preferably, in the second step, the ratio of the extracted endives to water is 1:10 to 30 percent; the heating temperature in the second step is 90-100 ℃; the extraction times in the second step are 1-3 times, and the total extraction time is 1-3 hours.
Preferably, the application of the herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis is that the herba Sonchi arvensis polysaccharide extract can reduce one or more of ulcerative colitis symptoms, i.e. relieve or treat at least one of recognized ulcerative colitis symptoms such as weight loss, diarrhea, hematochezia, etc.
Preferably, the Sonchus arvensis polysaccharide extract can improve or reduce DAI score.
Preferably, the polysaccharide extract of Sonchus arvensis can improve colon length and morphology, and improve colon pathological conditions.
Preferably, the Sonchus arvensis polysaccharide extract can protect the colonic mucosa barrier and repair DSS induced colonic mucosa cell damage.
Preferably, the Sonchus arvensis polysaccharide extract is capable of modulating immune parameters of DSS-induced ulcerative colitis.
Preferably, the polysaccharide extract of endive can regulate the composition of intestinal flora, increase dominant bacteria and reduce harmful bacteria, and promote the recovery of intestinal flora.
Preferably, the chicory polysaccharide extract is capable of modulating the metabolite composition of enteritis mice.
Preferably, the medicine is prepared by taking the polysaccharide extract of the endives as an active ingredient, adding pharmaceutically acceptable auxiliary materials, auxiliary components or carriers into the active ingredient singly or in combination with other medicines.
Preferably, the dosage form of the medicament is powder, granules, capsules, tablets, pills, various oral liquid preparations, sustained release agents, injections or targeting preparations.
Preferably, the pharmaceutically acceptable carrier refers to a conventional pharmaceutical carrier in the pharmaceutical field.
Compared with the prior art, the beneficial effects of the application are as follows:
(1) The preparation and extraction process of the polysaccharide extract of the endives is standardized for the first time, the polysaccharide extract of the endives has high active ingredient content, reliable curative effect and less side effect, and is beneficial to further development and utilization of the plant resources of the endives;
(2) The invention discovers and proposes that the endive polysaccharide extract can be applied to the prevention and treatment of ulcerative colitis and the preparation of medicines for preventing and treating ulcerative colitis for the first time. Animal experiments show that the herba Sonchi arvensis polysaccharide extract can effectively improve weight loss, diarrhea and bloody stool caused by ulcerative colitis, improve colon length, regulate organ index and improve colon injury caused by ulcerative colitis; HE staining proves that the herba Sonchi arvensis polysaccharide extract can protect the epithelial tissue of the colonic mucosa, and effectively relieve the severe colonitis caused by DSS induction; the result of the immunological experiment shows that the polysaccharide extract of the endives can regulate the immune parameters; the intestinal flora is detected and analyzed through 16s rRNA sequencing, and the endive polysaccharide extract can regulate the dysbacteriosis phenomenon and promote the recovery of the intestinal flora; the metabonomics research result shows that the endive polysaccharide extract can regulate the metabolite composition of enteritis mice.
(3) The invention adopts the means of HE staining, immunological experiment, 16s rRNA sequencing, metabonomics and the like to respectively discuss the influence of the polysaccharide extract of the endives on the intestinal tissues, the mucous membrane barrier, the intestinal flora of the immune system and metabolism of the DSS-induced mouse colonitis model. Experimental results show that the endive polysaccharide extract has the effects of recovering the integrity of epithelial cells of colon tissues, healing mucous membrane barriers, protecting intestinal tracts and the like, thereby reducing the effect of DSS-induced colonitis in mice.
(4) The invention widens the application field of the herba Sonchi arvensis polysaccharide extract, provides experimental basis for the clinical application of the herba Sonchi arvensis polysaccharide extract in preparing medicines for treating ulcerative colitis, and has good application prospect.
It will be appreciated that pharmaceutically acceptable adjuvants, auxiliary ingredients or carriers in the present invention refer to diluents, adjuvants, excipients or vehicles that are co-administered with the active ingredient and which are suitable for contacting the tissues of humans and/or other animals without undue toxicity, irritation, allergic response, complications, or other problem, commensurate with a reasonable benefit/risk ratio, and within the scope of sound medical judgment.
It should be understood that, in the present invention, the term "treating" is used to include alleviation, inhibition or amelioration of symptoms or conditions of the disease; inhibiting the production of complications, improving or preventing potential metabolic syndrome; inhibiting the occurrence of a disease or condition, such as controlling the progression of a disease or condition; alleviating a disease or symptom; causing the disease or symptom to subside; alleviating complications caused by diseases or symptoms, or preventing or treating signs caused by diseases or symptoms. As used herein, administration may result in an improvement in a disease, symptom, or condition, particularly an improvement in severity, a delay in onset, a slowing in progression, or a reduction in duration of the condition.
It should be understood that the description in this summary is not intended to limit key or critical features of embodiments of the present application, nor is it intended to be used to limit the scope of the present application. Other features of the present application will become apparent from the description that follows.
Drawings
Other features, objects and advantages of the present application will become more apparent upon reading of the detailed description of non-limiting embodiments, made with reference to the following drawings, in which:
FIG. 1 is a graph showing the effect of the Sonchus arvensis polysaccharide extract on DSS-induced C57BL/6 mouse colitis in the examples of the present invention, wherein the data are expressed as mean.+ -. SD (n=5) and analyzed by student t-test, and the other groups are compared with the CL group, P<0.05 and P<0.001; the other groups are compared to the ML group and the AN group, △ P<0.05; the other groups are compared to the ML group, ## P<0.05; the other groups are compared to the AN group, ▽ P<0.05, a graph: schematic diagram of experimental design; b, drawing: body weight change profile for each group of mice; c, drawing: food intake change plots for each group of mice; d, drawing: water intake change plots for each group of mice; e, drawing: disease Activity Index (DAI) change profile for each group of mice; f and g: colon length map for each group of mice; h, drawing: colon HE staining pattern for each group of mice;
FIG. 2 is a graph showing the effect of the polysaccharide extract of Sonchus arvensis on the modulation of immune function parameters in DSS-induced mice in the examples of the present invention, wherein the data are expressed as mean.+ -. SD (n=5) and analyzed by student t-test, and the other groups are compared with the CL group, P<0.05 and P<0.01; the other groups are compared to the ML group, ## P<0.05; the other groups are compared to the AN group, ▽ P<panel a and b are graphs showing the effect on thymus index and spleen index in each group of mice; panels c, d and e are schematic illustrations of the effect on IgA, igG and IgM in the serum of mice of each group;
FIG. 3 is a schematic diagram showing the effect of the polysaccharide extract of Sonchus arvensis on the structure of DSS-induced intestinal flora in mice, wherein, in the embodiment of the invention, a is as follows: wien diagram; b, drawing: PCoA analysis chart; c, drawing: NMDS analysis chart; d, drawing: schematic of relative abundance at the gate level; e and f: schematic of the relative abundance of actionobacter and Verrucomicrobiota in the cecal microbiota of each group of mice; g diagram: MRPP and Amova analysis plots of intestinal flora for each group of mice;
FIG. 4 is a graph showing the clustering analysis of species abundance at the genus level of the effect of the Sonchus arvensis polysaccharide extract on DSS-induced intestinal flora of mice in the examples of the present invention.
FIG. 5 is a schematic diagram showing a comparative analysis of species differences of the polysaccharide extract of Sonchus arvensis to the intestinal flora of DSS-induced mice in the example of the present invention, wherein, a is as follows: CL group vs ML group; b, drawing: CL group vs PS group; c, drawing: ML group vs PS group; relative abundance of cecal content flora in CL, ML and PS mice: d, drawing: akkermansia; e, drawing: rikenella eae_Rc9_gun_group; f, drawing: rikenella; g diagram: unidentified_clostridium_vadinbb60_group; h, drawing: analysis of LEfSe of the polysaccharide extract of Sonchus arvensis on intestinal microbiota between groups in mice induced by DSS.
FIG. 6 is a schematic diagram showing the effect of the polysaccharide extract of Sonchus arvensis on DSS-induced intestinal metabolites of mice in the example of the present invention, wherein, figure a: PCA (principal component analysis) map; b, drawing: PLS-DA profile; c, drawing: differential metabolite thermogram; d and e: differential metabolites.
FIG. 7 is a graph of metabolic pathway enrichment profile analysis of differential metabolites via a KEGG database in an embodiment of the invention.
FIG. 8 is a graph showing the effect of the polysaccharide extract of Sonchus arvensis on DSS-induced intestinal metabolites of mice examined by pearson's correlation analysis in the examples of the present invention;
fig. 9 is an analysis chart of the relationship between intestinal microorganisms and metabolites examined by Spearman's correlation analysis in the example of the present invention, p <0.05, p <0.01.
Detailed Description
The present application is described in further detail below with reference to the drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the application and not limiting of the application. It should be noted that, for convenience of description, only the portions related to the application are shown in the drawings.
It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The test methods used in the examples are conventional methods unless otherwise specified; the materials, reagents and the like used, unless otherwise specified, are those commercially available. The present application will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.
Embodiment one: preparation of polysaccharide extract of Sonchus arvensis
Step one: cutting dried whole herb of Sonchus arvensis, weighing, soaking in 95% ethanol water solution, and ultrasonic extracting at 40deg.C for 3 times for 0.5 hr;
step two: volatilizing solvent, and mixing the materials according to a feed liquid ratio of 1:23 weighing the extracted endive, immersing the extracted endive in water and heating to 92 ℃ for extraction for 2 times, wherein the total extraction time is 1.81 hours, filtering the supernatant after each extraction, centrifuging the supernatant, and finally combining a plurality of filtrates;
step three: concentrating the combined filtrate in vacuum to obtain concentrated solution, slowly adding 4 times of absolute ethyl alcohol into the concentrated solution, standing overnight in a refrigerator at 4 ℃, filtering and centrifuging to take out precipitate;
step four: washing the precipitate with absolute ethanol, acetone and diethyl ether twice, and oven drying to obtain herba Sonchi arvensis polysaccharide extract.
Embodiment two: research on action of polysaccharide extract of endive on colon tissue of mice
1. Grouping animals
C57BL/6 mice, male, 8 week old, body weight 20+ -2 g, SPF grade, purchased from Peking Vitolith laboratory animal technologies Co. After 7 days of adaptive feeding, the mice were randomly divided into 4 groups of 5 mice each, which were respectively set as a control group (CL), a model group (ML), a drug treatment group (PS), and a positive control drug group (AN).
2. Dosing regimen
From the first day of the experiment, the drug treatment group irrigates the chicory polysaccharide extract every day, and the control group, the model group and the positive control drug group irrigate the stomach every day with an equal volume of physiological saline for 14 consecutive days; the ulcerative colitis model was established using DSS induction at day 15: the mice of the model group, the drug treatment group and the positive control drug group were perfused with 0.2mL of DSS solution (concentration of 0.6 g/mL) from day 15 for seven consecutive days, while the control group was perfused with an equal volume of physiological saline; the group for drug treatment is added with the polysaccharide extract of the endive herb of the intragastric irrigation every day, the mice of the positive control drug group are added with the aspirin of the intragastric irrigation, and the control group and the model group are added with the physiological saline with the same volume of the intragastric irrigation.
3. Experimental method and data processing
3.1 disease Activity index (Disease activity index, DAI) score
Mice were observed daily and recorded for body weight, diet, stool consistency, and hematochezia. Scoring according to DAI scoring criteria, comprehensively scoring according to the weight reduction percentage of animals (the weight is not reduced by 0, the reduction is less than or equal to 5 percent and is more than 0 and is 1 minute, the reduction is less than or equal to 10 percent and is more than 5 percent and is 2 minutes, the reduction is less than or equal to 15 percent and is more than 10 percent and is 3 minutes, the reduction is more than 15 percent and is 4 minutes), the stool viscosity (normal 0, loose stool is 2 minutes and diarrhea is 4 minutes) and the stool bleeding (normal 0 minutes, occult blood positive is 2 minutes and dominant bleeding is 4 minutes), and dividing the total score of 3 results by 3 to obtain the DAI.
Dai= (body mass index + stool consistency index + bleeding index)/3.
3.2 colon Length
After the experiment, each group of mice is sacrificed by cervical dislocation, the whole colon tissue is separated, the colon length of each group of mice is measured by a ruler, and statistical analysis is performed.
3.3 pathological histological observation of the colon
Colon tissue of each group of mice was paraffin-embedded, sectioned and HE stained. And then observed by an optical microscope.
3.4 results
During the course of the experiment, mice were monitored daily for weight loss, diarrhea and bloody stool. The ML, PS and AN mice significantly reduced body weight compared to the control CL group, especially the ML and AN groups (fig. 1 b), whereas the PS mice significantly reduced body weight compared to the ML and AN groups (fig. 1 b).
To investigate whether weight loss is related to food or water intake, water intake and food intake were examined: the water intake and food intake were slightly reduced in the PS group compared to the CL group (fig. 1c and 1 d), while those in the ML and AN groups were significantly reduced (fig. 1c and 1 d). These results indicate that the polysaccharide extract of Sonchus arvensis significantly improves DSS-induced weight loss in UC mice.
The DAI scores were significantly higher in ML, FE and AN mice than in the control CL, while the DAI scores were significantly lower in PS mice than in ML and AN (fig. 1 e).
The colon changes in ML and AN mice showed a significant reduction, while those in PS mice showed a significant improvement over those in ML and AN (fig. 1f and 1 g).
The severity of colonic inflammation was further assessed by HE analysis and the pathological changes were significantly improved after administration of the chicory polysaccharide extract to DSS-induced UC mice (fig. 1 h).
These results indicate that the polysaccharide extract of Sonchus arvensis can significantly alleviate DSS-induced colitis in mice, and the therapeutic effect of the polysaccharide extract of Sonchus arvensis is superior to that of aspirin.
Embodiment III:
1. blood routine detection:
at the end of the experiment, blood samples of each group of mice are collected, after anticoagulation treatment, conventional blood detection is carried out by using a full-automatic blood analyzer, and relevant cells and relevant parameters in the blood are recorded.
2. Analysis of results
Blood routine test results are shown in table 1, and the results show that almost all blood parameters of ML group and AN group mice are significantly different compared with CL group, whereas almost all blood parameters of PS group mice are not significantly different compared with CL group. The result analysis shows that the polysaccharide extract of the endives significantly improves the blood routine parameters of the UC mice induced by DSS.
Embodiment four: immune index modulation
1. Immune organ index determination
Each group of mice was weighed at the end of the experiment, thymus and spleen were stripped and weighed separately. The organ index formula is as follows:
immune organ index (mg/g) =immune organ index body weight (mg)/body weight (g)
ELISA test
At the end of the experiment, the mice were bled from their orbit and then allowed to stand at room temperature for 2h. Serum was obtained by centrifugation according to ELISA kit instructions. IgA, igG, igM levels were detected using an ELISA kit.
3. Results
Fig. 2a shows that there is no significant difference in thymus index between the CL and PS mice, but that the thymus index is significantly reduced in the ML and AN mice compared to the CL and PS mice.
Fig. 2b shows that in terms of spleen index, the spleen index of ML group, AN group, PS group mice is significantly increased compared to CL group, wherein the spleen index of ML group and AN group mice is significantly increased.
Immunoglobulins play an important role in neutralizing pathogens and toxins. The effect of the polysaccharide extract of Sonchus arvensis on serum immunoglobulin is shown in FIG. 2c, FIG. 2d and FIG. 2e, respectively. Figure 2d shows a significant reduction in IgG production in AN group AN mice following aspirin treatment, suggesting that aspirin inhibits immunoglobulin IgG secretion. FIG. 2e shows that the IgM levels were significantly higher in the ML group than in the CL, AN and PS groups, indicating that DSS enhanced secretion of immunoglobulin IgM. These results indicate that the polysaccharide extract of Sonchus arvensis plays an important role in immunomodulation.
Fifth embodiment: intestinal microbiota analysis
1. Fecal sample collection and treatment
After the experiment is finished, the mice are sacrificed, the abdominal cavity is dissected, the anus is taken out to complete colon tissues of the cecum and placed on an ice tray, the faeces in the cecum are taken out and collected in a freezing tube, and the faeces are immediately placed in liquid nitrogen for preservation, so that bacterial DNA in the faeces is extracted.
2. Extraction and sequencing of bacterial DNA in feces
The cecal content was collected and total genomic DNA was extracted from all samples using CTAB method. The 16S rRNA/18SrRNA/ITS genes of the different regions were amplified. The product was then purified and quantified. Sequence analysis was performed using Uparse software. Sequences with a similarity of 97% or more were assigned to the same OTUs. The representative sequence of each OTU was screened for further annotation. OTUs abundance information is normalized using the sequence number standard corresponding to the sample with the least sequence. Subsequent alpha diversity and beta diversity analyses were all analyzed based on the normalized data.
3. Results
The wien diagram of fig. 3a shows that the CL, ML and PS groups share 536 OTUs, 734 OTUs between the CL and ML groups, 568 OTUs between the CL and PS groups, 734 OTUs between the ML and PS groups.
The PCoA and NMDS analyses of FIG. 3b and 3c indicated that the microbiota composition of the CL, ML and PS groups had a distinct cluster, and the results indicated that the Sonchus arvensis polysaccharide extract induced a significant change in the composition of the intestinal microbiota.
Fig. 3g shows that MRPP and Amova results indicate significant differences in gut microbiota composition between the three groups (P < 0.05).
FIG. 3d shows that the top 10 species with the highest abundance of gate levels were analyzed, with the thick-walled and Bacteroides being the most abundant two gates in all groups.
Notably, in the DSS-induced UC mice group, the abundance of Verrucomicrobiota flora increased significantly, while in the PS group of mice this change was reversed significantly (fig. 3 f). Furthermore, the abundance of actionobacter in PS group mice was significantly enhanced (fig. 3 e). FIG. 3g shows that these findings indicate that the polysaccharide extract of Sonchus arvensis can alter the abundance of intestinal flora.
As shown in fig. 4, analyzing species abundance information at the genus level for all group samples, the species abundance cluster map showed that CL group and PS group were mainly clustered together at the genus level, clearly separated from ML group, suggesting that the intestinal flora composition of PS group mice was closer to CL group.
Further t-tests were performed at the genus level, and figures 5 a-c indicated that there was a significant difference in the intestinal flora of mice between groups at the genus level, whereas the abundance of the intestinal flora of the ML group was significantly reversed after treatment with the chicory polysaccharide extract (figures 5 d-g).
Further LEfSe analysis, DSS-induced UC mice were altered in biomarkers following treatment with the Sonchus arvensis polysaccharide extract (FIG. 5 h), which was similar to the t-test results.
In general, DSS induction results in dysbacteriosis of the intestinal microbiota of mice, whereas the chicory polysaccharide extract can regulate dysbacteriosis of the intestinal microbiota and restore the intestinal homeostasis environment.
Example six: intestinal metabolite analysis
1. Fecal sample collection and treatment
After the experiment is finished, the mice are sacrificed, the abdominal cavity is dissected, the anus is taken out to complete colon tissues of the cecum and placed on an ice tray, the faeces in the cecum are taken out and collected in a freezing tube, and the faeces are immediately placed in liquid nitrogen for storage for later use.
2. Non-targeted metabolite assay
The cecal content was assayed for non-targeted metabolites by UHPLC system by appropriate treatment. Chromatographic data were analyzed using MSDIAL3.82 software.
3. Results
Analysis of the PCA plot of FIG. 6a and the PLS-DA plot of FIG. 6b indicated that the composition of the non-targeted metabolites was significantly different for the CL, ML and PS groups, with 97 different metabolites between each group (FIG. 6 c).
Further data analysis found that DSS-induced intestinal metabolite abundance in UC mice was significantly reversed following treatment with the chicory polysaccharide extract (fig. 6d and e). These results indicate that the chicory polysaccharide extract can affect intestinal metabolites of UC mice.
As shown in FIG. 7, the analysis of metabolic pathway enrichment pattern of differential metabolites by KEGG database revealed that biosynthesis of main unsaturated fatty acids and linoleic acid metabolic pathway were affected after treatment of Sonchus arvensis polysaccharide extract.
In order to investigate the effect of the polysaccharide extract of Sonchus arvensis on the metabolites, pearson's correlation analysis was performed on the metabolites, indicating the correlation between the metabolites, as shown in FIG. 8, and further, the correlation between the intestinal microbiota and the metabolites was examined using the Spearman's correlation analysis, as shown in FIG. 9, to elucidate the correlation between the metabolite correlation and the intestinal microbiota.
TABLE 1 routine blood test results for each group
Note that: * P (P)<0.05,**P<0.01,***P<0.001vs CL； # P<0.05, ## P<0.01, ### P<0.001vsML； ▲ p＜0.05vs AN
The effect of the invention was evaluated by establishing a DSS-induced acute mouse colitis model. The animal model is commonly used for researching pathological mechanism of ulcerative colitis and evaluating curative effect of medicines. Through a DSS-induced colitis mouse model, we analyzed the general weight profile, DAI score, colon length, colon histopathological changes of mice. The experimental results show that: the polysaccharide extract of the endive can effectively treat and relieve the symptoms of ulcerative colitis: including weight loss, diarrhea, and hematochezia; can further protect the mucosal barrier and repair colon lesions; can regulate intestinal flora composition, promote flora recovery, and regulate intestinal health; can regulate metabolite composition of enteritis mice. Therefore, the herba Sonchi arvensis polysaccharide extract can be used as an effective therapeutic drug for ulcerative colitis, and can be prepared into various dosage forms or pharmaceutical compositions for treating ulcerative colitis.
In the description of the present specification, the terms "one embodiment," "some embodiments," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing is merely a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and variations may be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.
Claims (10)
1. The application of the herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis is characterized by comprising the application of the herba Sonchi arvensis polysaccharide extract in preparing medicines for preventing and treating ulcerative colitis.
2. The use of the polysaccharide extract of endive according to claim 1 for preventing and treating ulcerative colitis, wherein the method for preparing the polysaccharide extract of endive comprises the following steps:
step one: cutting dried whole herb of Sonchus arvensis, weighing, soaking in ethanol water solution, and ultrasonic extracting for multiple times;
step two: volatilizing solvent, weighing the extracted endive, soaking in water, heating for extraction to obtain an extracting solution, extracting for multiple times, filtering after each extraction, centrifuging the supernatant, and finally combining multiple filtrates;
step three: concentrating the combined filtrate under reduced pressure to obtain concentrated solution, slowly adding 3-5 times of absolute ethyl alcohol into the concentrated solution, continuously stirring, standing at 4 ℃ overnight, filtering and centrifuging to obtain precipitate;
step four: washing the precipitate with absolute ethanol, acetone and diethyl ether twice respectively, and drying in an oven to obtain the herba Sonchi arvensis polysaccharide extract.
3. The use of the endive polysaccharide extract according to claim 2 for preventing and treating ulcerative colitis, wherein the concentration of ethanol in the aqueous ethanol solution in the first step is 80-100%; the ultrasonic extraction times in the first step are 1-5 times, and the total extraction time is 0.5-2 hours.
4. The application of the endive polysaccharide extract according to claim 3 in preventing and treating ulcerative colitis, wherein the feed liquid ratio of the extracted endive to water in the second step is 1:10 to 30 percent; the heating temperature in the second step is 90-100 ℃; the extraction times in the second step are 1-3 times, and the total extraction time is 1-3 hours.
5. The use of the endive polysaccharide extract according to claim 1 for preventing and treating ulcerative colitis, wherein the endive polysaccharide extract is capable of improving or reducing DAI score; the herba Sonchi arvensis polysaccharide extract can reduce more than one of ulcerative colitis symptoms, i.e. relieve or treat at least one of weight loss, diarrhea and hematochezia-recognized ulcerative colitis symptoms.
6. The use of the polysaccharide extract of endive according to claim 1 for preventing and treating ulcerative colitis, wherein the polysaccharide extract of endive is capable of improving colon length and morphology and improving colon pathological conditions.
7. The use of the herba Sonchi arvensis polysaccharide extract according to claim 1 for preventing and treating ulcerative colitis, wherein the herba Sonchi arvensis polysaccharide extract is capable of protecting the colonic mucosa barrier and repairing DSS-induced colonic mucosa cell damage.
8. The use of the endive polysaccharide extract according to claim 1 for preventing and treating ulcerative colitis, wherein the endive polysaccharide extract is capable of regulating immune parameters of ulcerative colitis;
the polysaccharide extract of the endive can regulate the composition of intestinal flora, increase dominant bacteria and reduce harmful bacteria, and promote the recovery of the intestinal flora;
the herba Sonchi arvensis polysaccharide extract can regulate the metabolite composition of enteritis.
9. The application of the herba Sonchi arvensis polysaccharide extract according to claim 1 in preventing and treating ulcerative colitis, wherein the medicament is prepared into a pharmaceutically acceptable dosage form by taking the herba Sonchi arvensis polysaccharide extract as an active ingredient and adding pharmaceutically acceptable auxiliary materials, auxiliary components or carriers, alone or in combination with other medicaments.
10. The use of the endive polysaccharide extract according to claim 9 for preventing and treating ulcerative colitis, wherein the medicament is in the form of powder, granules, capsules, tablets, pills, oral liquid, sustained release preparation, injection or targeting preparation.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310053754.2A CN116270705A (en) | 2023-02-03 | 2023-02-03 | Application of herba Sonchi arvensis polysaccharide extract in preventing and treating ulcerative colitis |
| CN2023100537542 | 2023-02-03 |
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