Application of rice worm in preparation of medicine for preventing gastric ulcer

Abstract

The invention provides application of rice worm in preparation of a medicine for preventing gastric ulcer, and relates to the field of biological medicine. According to the application, a mouse alcoholic gastric ulcer model is constructed, the prevention effect of the rice worm on absolute ethyl alcohol induced gastric ulcer is researched, the research result proves that the rice worm has a good prevention effect on the mouse alcoholic gastric ulcer, and the action mechanism of the rice worm is related to MAPK14 gene expression increase.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of P. oryzae in preparing medicine for preventing gastric ulcer. Background Art

[0002] The active protein of the rice weevils contains unique antimicrobial peptides. When the body is damaged or invaded by pathogenic microorganisms, the antimicrobial peptides quickly track and kill the invaders. The antimicrobial peptides are like immune special police, using sharp knives to attack the "enemy", chiseling ion channels on the cell membrane structure of bacteria and viruses, destroying the bacterial cell membrane structure, causing the outflow of water-soluble substances in the cells, and thus completely killing the bacteria. The active protein of the rice weevils is rich in chitin. Chitin is one of the important pillars of all biological life and is known as the "sixth life nutrient element" after protein, sugar, fat, vitamins, and minerals. It is like a hard-working gardener, playing the role of three adjustments (bidirectional immune regulation, pH regulation, hormone regulation) and three eliminations (eliminating harmful substances in cells and body fluids, eliminating heavy metal ions, and eliminating oxygen toxins) in the immune system of organisms, and constantly maintaining the internal environment of organisms. The active protein of the rice weed also contains exogenous lectins, which can promote cell adhesion and inhibit their proliferation. It can not only make normal cells more active, but also kill mutant cells, resist the spread of viruses and activate immunity.

[0003] At present, there is no research on using Diplocaulidae as a drug for preventing gastric ulcer in mice, but the present invention confirms that Diplocaulidae can be used as a drug for preventing gastric ulcer in mice. Summary of the invention

[0004] In view of this, the present invention provides the use of Diplococephala as a drug for preventing gastric ulcer, and the use confirms that Diplococephala has a good preventive effect on alcoholic gastric ulcer.

[0005] The present invention is achieved by adopting the following technical solutions:

[0006] The present invention constructs a mouse alcoholic gastric ulcer model to study the preventive effect of Dioscorea oryzae on anhydrous ethanol-induced gastric ulcer. The model construction and drug administration are as follows:

[0007] Model construction: The mice were fasted and deprived of water for 24 hours to ensure gastric emptying. After fasting and deprivation of water for 24 hours, each mouse was given 0.2mL / 20g anhydrous ethanol by gavage, and 1 hour later, the mice were killed by cervical dislocation.

[0008] The invention administers medicine to mice before model building, 1-2 times a day, with a dosage of 2.5g / kg-3g / kg, for 7-14 consecutive days, by gavage of P. oryzae.

[0009] Compared with the prior art, the present invention has the following beneficial effects:

[0010] The invention confirms that P. oryzae has a good preventive effect on alcoholic gastric ulcer in mice, and its mechanism of action is related to the increased expression of MAPK14 gene. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 The figure is a bar graph comparing the gastric ulcer areas of mice in Comparative Example 1, Comparative Example 2 and Example 1;

[0012] Figure 2 The anatomical diagram of the gastric mucosa of mice of Comparative Example 1, Comparative Example 2 and Example 1;

[0013] Figure 3 The figure is a comparative bar graph of gastric ulcer index of mice in comparative example 1, comparative example 2 and embodiment 1;

[0014] Figure 4 The figure is a comparative bar graph of gastric ulcer scores in mice of Comparative Example 1, Comparative Example 2 and Example 1;

[0015] Figure 5 HE staining results of pathological sections of gastric ulcer in mice of Comparative Example 1, Comparative Example 2 and Example 1;

[0016] Figure 6 The results of QPCR detection of the gene expression of MAPK1, MAPK8, MAPK14 and RELA in the gastric tissue of mice in Comparative Example 1, Comparative Example 2 and Example 1; DETAILED DESCRIPTION

[0017] The present invention will be further described below in conjunction with the embodiments.

[0018] The mice used in Example 1 of the present invention, Comparative Example 1 and Comparative Example 2 were all SPF-grade BABL / c mice, 6-8 weeks old, and 5 males.

[0019] Example 1

[0020] Before the model was established, 5 normally fed mice were gavaged with 2.5 g / kg of Diplococephalus once a day for 7 consecutive days. After the last administration, they were deprived of food and water for 24 hours to ensure gastric emptying. After fasting and deprivation of water for 24 hours, each mouse was gavaged with 0.2 mL / 20 g of anhydrous ethanol and killed by cervical dislocation 1 hour later.

[0021] Comparative Example 1

[0022] Five normally fed mice were fasted and deprived of water for 24 h to ensure gastric emptying. After fasting and deprivation of water for 24 h, each mouse was gavaged with 0.2 mL / 20 g normal saline and killed by cervical dislocation 1 h later.

[0023] Comparative Example 2

[0024] Five normally fed mice were fasted and deprived of water for 24 hours to ensure gastric emptying. After fasting and deprivation of water for 24 hours, each mouse was given 0.2 mL / 20 g anhydrous ethanol by gavage and was killed by cervical dislocation 1 hour later.

[0025] The test indicators of Example 1, Comparative Example 1 and Comparative Example 2 are as follows:

[0026] 1. Gastric ulcer area

[0027] After taking pictures using a stereo microscope, image J was used to calculate the gastric ulcer area.

[0028] The statistics of gastric ulcer areas of Comparative Example 1, Comparative Example 2 and Example 1 are shown in Table 1.

[0029] Table 1

[0030]

[0031] according to Figure 1 Comparative Example 1, Comparative Example 2 and Example 1 in the mouse gastric ulcer area comparison bar graph and Figure 2 1, Comparative Example 2 and Example 1 of the mouse gastric mucosa anatomical diagram, in conjunction with Table 1, it can be known that, compared with Comparative Example 2, the mouse gastric ulcer area of ​​Comparative Example 1 and Example 1 presents a very significant difference. This difference is not only significant, but also extremely significant, reaching the statistical P < 0.0001 level. This data result fully proves that the experimental conditions have a high degree of reliability and importance for the impact of gastric ulcer area. This difference is not only reflected in the numerical value, but also reveals the potential impact of the treatment method or drug dosage on the gastric ulcer area in a deeper level. Very obviously reveals the very significant effect of Dioscorea sphaerocephala in treating gastric ulcer.

[0032] 2. Gastric ulcer index

[0033] Calculation method of ulcer index: 1 point for ulcer length less than 1 mm, 2 points for ulcer length between 1 and 2 mm, 3 points for ulcer length greater than 2 mm, and so on. The index is doubled for ulcer width greater than 1 mm.

[0034] See Table 2 for the gastric ulcer index scoring results of mice in Comparative Example 1, Comparative Example 2 and Example 1.

[0035] Table 2

[0036]

[0037] The comparison bar graph of the gastric ulcer index of mice in Example 1, Comparative Example 2 and Example 1 (as shown in FIG. Figure 3As shown in Table 2, the difference between Comparative Example 1 and Comparative Example 2 is extremely significant, which is clearly verified by statistical analysis (P<0.0001). Similarly, the difference between Example 1 and Comparative Example 2 is also highly significant, and the statistical data show that its P value is 0.0011. It is worth noting that the gastric ulcer index of mice in Example 1 is significantly lower than that in Comparative Example 2, and this finding further confirms the effectiveness of Example 1 in reducing the gastric ulcer index.

[0038] Specifically, when comparing Comparative Example 1 with Comparative Example 2, it was found that there was a significant difference between the two. This difference was not only reflected in the numerical value, but also strictly verified by statistical analysis (P<0.0001). Similarly, the comparison between Example 1 and Comparative Example 2 also showed similar results, with a P value of 0.0011, indicating that the difference between the two was also extremely significant.

[0039] Moreover, Example 1 showed a significant effect in reducing the gastric ulcer index of mice. Compared with Comparative Example 2, the gastric ulcer index of mice in Example 1 was significantly smaller. This finding not only verifies the effectiveness of Example 1, but also provides a new idea for further exploring the treatment of gastric ulcer.

[0040] 3. Gastric ulcer scoring

[0041] The ulcer condition of the mice was scored, with partial redness and congestion being scored as 1 point, punctate bleeding or erosion being scored as 1 point each, and linear erosion being scored as 3 points. The total score was calculated as the scoring index.

[0042] See Table 3 for the gastric ulcer scoring index results of Comparative Example 1, Comparative Example 2 and Example 1.

[0043] Table 3

[0044]

[0045] The mouse gastric ulcer score index comparison bar graph (such as Figure 4 As shown in Table 3, it can be seen that there are significant differences among Comparative Example 1, Comparative Example 2 and Example 1 in the gastric ulcer scoring index of mice. Figure 4 The effects of each experimental group on treating gastric ulcers are shown in an intuitive way. Upon careful observation, it can be seen that the bar graph of Comparative Example 1 is significantly lower than that of Comparative Example 2, which indicates that Comparative Example 1 has a significant advantage in treating gastric ulcers in mice, and statistical data show that this advantage is extremely significant (P<0.0001), which undoubtedly provides strong evidence for the superiority of Comparative Example 1.

[0046] At the same time, compared with comparative example 2, the histogram of example 1 also showed a significant downward trend, indicating that example 1 had a significant effect in reducing the gastric ulcer score index of mice. Moreover, this difference also reached a very significant level in statistics (P=0.0007), further proving the reliability and effectiveness of example 1 in terms of therapeutic effect.

[0047] Further analysis revealed that the gastric ulcer score index of mice in Example 1 was significantly lower than that in Comparative Example 2. This result fully demonstrated the excellent performance of Example 1 in reducing the degree of gastric ulcer. By comparing the data of Comparative Example 2 and Example 1, it can be clearly seen that Example 1 significantly improved the efficacy, which provides a new and effective treatment plan for future research on the prevention of gastric ulcers.

[0048] In summary, through Figure 4 Based on the in-depth analysis and the data in Table 3, it can be confirmed that Example 1 has shown significant advantages in treating gastric ulcer in mice. Its therapeutic effect is not only significant but also highly reliable, which provides a new direction and idea for our further research on the treatment of gastric ulcer.

[0049] 4. Pathological sections

[0050] The gastric tissue was immersed in formalin, embedded in paraffin, sliced, and stained with hematoxylin-eosin (HE) to observe the damage of gastric mucosa.

[0051] The results of HE staining of the mouse gastric ulcer pathological sections of Comparative Example 1, Comparative Example 2 and Example 1 are shown in Figure 5 According to the results of pathological sections, the gastric mucosa of the mice in Comparative Example 1 was intact and there was no bleeding; the gastric mucosa of the mice in Comparative Example 2 was obviously damaged and had a large number of bleeding spots; the gastric mucosal damage of the mice in Example 1 was relatively mild, and there was a small amount of bleeding.

[0052] 5. mRNA detection

[0053] Real-time fluorescence quantitative nucleic acid amplification detection (QPCR detection) of MAPK1, MAPK8, MAPK14, and RELA gene expression in mouse gastric tissue.

[0054] The results of QPCR detection of MAPK1, MAPK8, MAPK14, and RELA gene expression in the gastric tissues of mice in Comparative Example 1, Comparative Example 2, and Example 1 are shown in Table 4.

[0055] Table 4

[0056]

[0057]

[0058] according to Figure 6The results of QPCR detection of MAPK1, MAPK8, MAPK14 and RELA gene expression in the mouse gastric tissue of Comparative Example 1, Comparative Example 2 and Example 1, combined with Table 4, can be seen that these genes play a key role in cell signal transduction and regulation. Therefore, understanding their expression differences in mouse gastric tissue is of great significance for in-depth research on the function and pathological changes of gastric tissue.

[0059] Expression of MAPK1 gene in mouse gastric tissue: Compared with Comparative Example 2, the expression level of MAPK1 gene in Comparative Example 1 showed a very significant difference (P=0.0006). This extremely low P value means that the difference is very likely due to differences in experimental conditions rather than random errors. This very significant difference may reflect that the MAPK1 gene is strongly regulated or affected under specific experimental conditions, which may be related to certain specific biological processes or pathological changes in mouse gastric tissue.

[0060] Expression of MAPK8 gene in mouse gastric tissue: In Comparative Example 1, Comparative Example 2 and Example 1, the expression of MAPK8 gene did not show significant differences. This indicates that in mouse gastric tissue, the expression of MAPK8 gene may be relatively stable and not easily disturbed by external factors. This stability of the MAPK8 gene means that its function in gastric tissue is relatively basic or universal, and is not specific to certain biological processes or pathological states.

[0061] Expression of MAPK14 gene in mouse gastric tissue: Compared with Comparative Example 2, the expression level of MAPK14 gene in Comparative Example 1 showed a significant difference (P=0.0210), while the expression in Example 1 showed an extremely significant difference (P<0.0001). This significant and extremely significant difference indicates that the MAPK14 gene plays an important role in mouse gastric tissue, and its expression may be regulated by multiple factors. The extremely significant difference in the expression of MAPK14 gene in Example 1 means that under specific experimental conditions, the expression of this gene is particularly strongly regulated or affected, involving key biological processes or pathological changes in gastric tissue.

[0062] Expression of RELA gene in mouse gastric tissue: Unlike MAPK1 and MAPK14, RELA gene showed no significant difference in Comparative Example 1, Comparative Example 2 and Example 1. This indicates that in mouse gastric tissue, the expression of RELA gene is relatively stable and is not easily affected by experimental conditions or treatment methods. This stability of RELA gene means that its function in gastric tissue is relatively fixed or universal, and is not specific to certain biological processes or pathological states.

[0063] In summary, by analyzing the expression of the four genes MAPK1, MAPK8, MAPK14 and RELA in mouse gastric tissue in detail, we can clearly see the significance and stability of the expression of these genes. The significant and extremely significant differences in the MAPK1 and MAPK14 genes in some experimental groups reveal the important role of these genes in specific biological processes or pathological changes. The stable expression of the MAPK8 and RELA genes reflects their basic or universal functions in gastric tissue. These results not only provide important clues for us to further study the mechanism of action of these genes in gastric tissue, but also provide a new perspective for understanding the physiological functions and pathological changes of gastric tissue.

[0064] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of Dioscorea opposita in the preparation of medicines for preventing gastric ulcers.

2. The use according to claim 1, characterized in that: The medicament also includes medically acceptable adjuvants.

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