A composition for the treatment of radiation-induced intestinal injury and uses thereof

Abstract

The application discloses a kind of for radiation intestinal injury treatment composition and its application, it is related to medical technical field.The composition includes the following components: aesculetin, fraxinol, fraxin and baicalein.The composition of the application can reduce intestinal villus injury by regulating the NF-κB pathway and Caspase-9 / 3 pathway of intestinal epithelial cells, reduce the infiltration of intestinal inflammatory cells, reduce intestinal cell apoptosis, to relieve intestinal injury, and then multi-component, multi-pathway jointly play the efficacy of treating radiation intestinal injury.The application solves the problem that there is no effective drug for treating radiation intestinal injury in the prior art.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for treating radiation-induced intestinal injury and an application thereof. Background Art

[0002] Radiation-induced bowel injury (RII) is a radiation-induced intestinal injury that can affect the small intestine, colon, and rectum. Symptoms include abdominal pain, diarrhea, and bloody stools. Currently, my country faces a large number of patients with RII and a shortage of therapeutic drugs. Furthermore, the complex mechanisms of RII hinder the development of new anti-RI drugs. Therefore, developing low-toxic, effective drugs with clear mechanisms of action and formulating more rational treatment plans are crucial strategies for addressing the treatment of RII. Summary of the Invention

[0003] In order to solve the above technical problems, the purpose of the present invention is to provide a composition for treating radiation-induced intestinal injury and its application, so as to solve the problem of lack of effective drugs for treating radiation-induced intestinal injury in the prior art.

[0004] The technical solution of the present invention to solve the above technical problems is as follows: providing a composition for treating radiation-induced intestinal injury, comprising the following components: aesculin, aesculetin, aesculetin and baicalein.

[0005] On the basis of the above technical solution, the present invention can also be improved as follows:

[0006] Furthermore, the molar ratio of aesculetin, aesculetin, aesculetin and baicalein is (1-100):(10-150):(0.1-30):(0.1-50).

[0007] The beneficial effect of adopting the above further technical solution is: adopting the above molar ratio can play a role in treating radiation-induced intestinal damage.

[0008] Furthermore, the molar ratio of aesculetin, aesculetin, aesculetin and baicalein is 50:70:5:10.

[0009] Furthermore, the molar ratio of aesculetin, aesculetin, aesculetin and baicalein is 1:10:0.1:0.1.

[0010] Furthermore, the molar ratio of aesculetin, aesculetin, aesculetin and baicalein is 100:150:30:50.

[0011] The present invention also provides use of the above composition in preparing medicine for treating radiation-induced intestinal damage.

[0012] Furthermore, radiation-induced intestinal injury includes acute radiation-induced intestinal injury, chronic radiation-induced intestinal injury and ulcerative colitis.

[0013] The present invention also provides a medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0014] Furthermore, the dosage form of the drug is an injection, enema preparation, powder, pill, tablet, microcapsule, soft capsule, film, ointment, tincture, granule or aerosol allowed in pharmaceutical science.

[0015] The present invention has the following beneficial effects:

[0016] 1. The present invention proposes a composition for treating radiation-induced intestinal injury. The composition can improve the shedding of intestinal villi, reduce the inflammatory infiltration of intestinal epithelial cells, and maintain the intestinal barrier function through the NF-κB pathway of intestinal epithelial cells; it can reduce the apoptosis of intestinal epithelial cells and alleviate intestinal damage through the Caspase-9 / -3 pathway of intestinal epithelial cells; the radiation-induced intestinal injuries that can be treated include acute radiation-induced intestinal injury, chronic radiation-induced intestinal injury, and ulcerative colitis.

[0017] 2. The composition of the present invention can reduce intestinal villus damage, reduce infiltration of intestinal inflammatory cells, and alleviate intestinal cell apoptosis by regulating the NF-κB pathway and Caspase-9 / -3 pathway of intestinal epithelial cells, thereby alleviating intestinal damage, and thus multiple components and multiple pathways can jointly exert the efficacy of treating radiation-induced intestinal damage. DETAILED DESCRIPTION

[0018] The following is a description of the principles and features of the present invention. The examples are intended only to explain the present invention and are not intended to limit the scope of the present invention. In the examples, if no specific conditions are specified, the methods were performed according to conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, if the manufacturer is not specified, are conventional products that can be purchased commercially.

[0019] Example 1:

[0020] A composition for treating radiation-induced intestinal injury comprises the following components: aesculin, aesculetin, aesculetin and baicalein; the molar ratio of aesculin, aesculetin, aesculetin and baicalein is 50:70:5:10.

[0021] A medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0022] Example 2:

[0023] A composition for treating radiation-induced intestinal injury comprises the following components: aesculin, aesculetin, aesculetin and baicalein; the molar ratio of aesculin, aesculetin, aesculetin and baicalein is 1:10:0.1:0.1.

[0024] A medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0025] Example 3:

[0026] A composition for treating radiation-induced intestinal injury comprises the following components: aesculin, aesculetin, aesculetin and baicalein; the molar ratio of aesculin, aesculetin, aesculetin and baicalein is 100:150:30:50.

[0027] A drug for treating radiation-induced intestinal injury, comprising the above-mentioned composition for treating radiation-induced intestinal injury.

[0028] A composition for treating radiation-induced intestinal injury comprises the following components: aesculetin; the molar number of aesculetin is 70.

[0029] A medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0030] Comparative Example 2:

[0031] A composition for treating radiation-induced intestinal injury comprises the following components: aesculin and aesculetin; the molar numbers of aesculin and aesculetin are 50 and 70 respectively.

[0032] A medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0033] Comparative Example 3:

[0034] A composition for treating radiation-induced intestinal injury comprises the following components: aesculin, aesculetin and aesculetin; the molar numbers of aesculin, aesculetin and aesculetin are 50, 70 and 5 respectively.

[0035] A medicine for treating radiation-induced intestinal injury, comprising the above composition for treating radiation-induced intestinal injury.

[0036] Test example

[0037] 1. Pathological injury evaluation

[0038] The compositions obtained in Example 1 (at molar ratios of 50, 70, 5, and 10, respectively) and Comparative Examples 1-3 were administered orally to mice for evaluation of intestinal pathological damage. The mice were divided into a normal group, a model group, and the four groups treated with the aforementioned drugs. Following X-ray irradiation, the mice were gavaged for seven consecutive days. On the eighth day, H&E results of the mouse intestinal tissue were obtained, as shown in Table 1.

[0039] Table 1 Chinese medicine and drug pairs alleviate intestinal pathological damage in RII mice

[0040]

[0041] Experimental results: As shown in Table 1, the H&E results of mouse intestinal tissue showed that the intestinal villi and intestinal crypt structures of the normal group of mice were intact, without villus breakage and inflammatory cell infiltration; after irradiation, the intestinal structure of the mice was destroyed, inflammatory cells infiltrated, intestinal villi were broken, intestinal crypts were enlarged, the number of crypts decreased, and the Chiu's score increased; after drug treatment, the pathological damage of intestinal tissue of the radiation-damaged mice model was alleviated, among which the effect of aesculin A + aesculin E + aesculin A + baicalein was better than that of other groups.

[0042] 2. Pathological Damage Evaluation

[0043] The composition of Example 1 was administered orally to conduct an experiment to evaluate the pathological damage of intestinal tissue in mice. The implementation method was as follows: after X-ray irradiation, the mice were weighed every day for 8 consecutive days; the mice were divided into normal group, model group, 90 μmol / kg of aesculin + 126 μmol / kg of aesculin + 9 μmol / kg of aesculin + 18 μmol / kg of baicalein, with the low and medium doses decreasing by 2 times in sequence; samples were collected on the 8th day, and the H&E results of the mouse intestinal tissue are shown in Table 2.

[0044] Table 2 Villus length, crypt number and Chiu's score of mouse intestinal tissue

[0045]

[0046] ## P < 0.01 compared with the Control group; ** P < 0.01 compared with the RII group; * P<0.05 compared with RII group

[0047] Experimental results: After irradiation, the body weight of the mice in the model group decreased significantly; administration of low, medium and high doses of the composition can inhibit the loss of body weight of the mice. On the 8th day, the experimental results suggested that the composition of the present invention can inhibit the weight loss of mice after irradiation.

[0048] As shown in Table 2, there was no villus rupture in the intestines of the mice in the normal group, and the intestinal villi and intestinal crypt structures of the mice were intact; after irradiation, the intestinal structure of the mice was destroyed, the intestinal villi were ruptured, the intestinal crypts were enlarged, the number of crypts was reduced, and the Chiu's score was increased; after treatment with the composition, the villus rupture and crypt enlargement were reduced, and the composition can reduce the pathological damage of the intestinal tissue of the radiation damage model mice. After treatment with the composition, the atrophy of the intestinal mucosa and the shedding of intestinal villi were significantly reduced, the inflammatory infiltration of intestinal epithelial cells was reduced, and the Chiu's score of intestinal tissue damage was reduced.

[0049] 3. Inflammatory Factor Detection

[0050] Based on the above, intestinal samples were taken from each group of mice, and the levels of inflammatory factors (IL-6, IL-10, IL-1β, TNF-α) were detected according to the operating instructions of the ELISA kit. After irradiation, the intestinal inflammatory response of mice was an important pathological process. GraphPad Prism 8.4.0 software was used to analyze and statistically analyze the results. The results are shown in Table 3.

[0051] Table 3 Levels of inflammatory factors in the intestine of mice

[0052]

[0053] ## P < 0.01 compared with the Control group; ** P < 0.01 compared with the RII group; * P<0.05 compared with RII group

[0054] Table 3 shows that compared with the normal control group, radiation caused a significant increase in the pro-inflammatory factors TNF-α, IL-1β, and IL-6, and a decrease in the anti-inflammatory factor IL-10. However, administration of the composition reduced the expression of pro-inflammatory factors and increased the expression of anti-inflammatory factors in a dose-dependent manner. This indicates that the composition of the present invention can reduce the levels of intestinal inflammatory factors in mice with radiation-induced intestinal injury.

[0055] 4. Inflammatory Factor Detection

[0056] HIEC-6 cells were used and grouped into normal group, model group, and combination group (15 μmol / L + aesculetin 21 μmol / L + aesculetin 1.5 μmol / L + baicalein 3 μmol / L). The low and medium doses were decreased by 2-fold in sequence. After X-ray irradiation, the levels of inflammatory factors (IL-6, IL-10, IL-1β, TNF-α) were detected according to the instructions of the ELISA kit. The results are shown in Table 4.

[0057] Table 4 Levels of inflammatory factors in HIEC-6 cells

[0058]

[0059] ## P < 0.01 compared with the Control group; ** P < 0.01 compared with the RII group; * P<0.05 compared with RII group

[0060] As shown in Table 4, compared with the normal control group, radiation caused increased expression of proinflammatory factors TNF-α, IL-1β, and IL-6, and decreased expression of the anti-inflammatory factor IL-10. However, after administration of the composition, the expression of proinflammatory factors decreased and the expression of anti-inflammatory factors increased in a dose-dependent manner. This indicates that the composition of the present invention can reduce the levels of inflammatory factors in HIEC-6 cells induced by radiation.

[0061] 5. Cell Apoptosis

[0062] HIEC-6 cells were used and divided into normal group, model group, and combination group: aesculin 15 μmol / L + aesculin 21 μmol / L + aesculin 1.5 μmol / L + baicalein 3 μmol / L. The low and medium doses were decreased by 2-fold in sequence. After X-ray irradiation, cell apoptosis was detected by TUNEL kit. Apoptosis is an important pathogenesis of radiation-induced intestinal injury. The results are shown in Table 5.

[0063] Table 5 TUNEL levels in HIEC-6 cells

[0064]

[0065]

[0066] ## P < 0.01 compared with the Control group; ** P < 0.01 compared with the RII group; * P<0.05 compared with RII group

[0067] As shown in Table 5, compared with the normal group, the apoptosis of HIEC-6 cells after irradiation was significantly increased. The administration of the composition can dose-dependently inhibit the apoptosis of HIEC-6 cells induced by X-rays, indicating that the composition of the present invention can inhibit the apoptosis of HIEC-6 cells after irradiation.

[0068] 6. Effects on the Expression of NF-κB Pathway Proteins

[0069] The effect of the composition of Example 1 on the expression of NF-κB pathway proteins was evaluated using normal human intestinal epithelial cells (HIEC-6) (ATCC Cell Bank, USA). The NF-κB inhibitor HY-18738 (MedChemExpress, USA) was used as a control. Conventional Western blot analysis was performed. The following procedures were used: a radiation model was established, wherein HIEC-6 cells were placed in a carbon dioxide incubator (5% CO2, 37°C) and cultured in DMEM / HIGH GLUCOSE medium supplemented with 10% fetal bovine serum. Groups were divided into: a normal group (Con), a model group (RII), a composition group, an inhibitor group (HY-18738), and a composition + inhibitor group (composition + HY-18738). The composition dosages were 15 μmol / L aesculin, 21 μmol / L aesculin, 1.5 μmol / L aesculin, and 3 μmol / L baicalein. The inhibitor dosage was 100 μM. The cells were irradiated with 4 Gy of X-rays and then given corresponding drugs. The results are shown in Tables 6-7.

[0070] Table 6 Levels of inflammatory factors in HIEC-6 cells

[0071]

[0072] ## P < 0.01 compared with the Control group; ** P<0.01 compared with RII group

[0073] Table 7 Effects of the composition on the expression of key proteins in the NF-κB pathway

[0074]

[0075] ## P < 0.01 compared with the Control group; ** P<0.01 compared with RII group

[0076] As shown in Tables 6-7, after irradiation, the phosphorylation level of P65 was significantly increased, while the phosphorylation expression of P65 in the combination group, HY-18738 group, and combination + HY-18738 group was significantly decreased, indicating that the composition of the present invention can significantly reduce the phosphorylation level of P65, thereby alleviating inflammation in HIEC-6 intestinal epithelial cells.

[0077] VII. Effects on the Expression of Caspase-9 / -3 Pathway Proteins

[0078] The effect of the composition of Example 1 on the expression of Caspase-9 / -3 pathway proteins was evaluated using normal human intestinal epithelial cells (HIEC-6) (ATCC Cell Bank, USA). The Caspase-9 / -3 inhibitor HY-12305 (MedChemExpress, USA) was used as a control. Conventional Western blot analysis was performed. The following procedures were performed: First, a radiation model was established: HIEC-6 cells were placed in a carbon dioxide incubator (5% CO2, 37°C) and cultured in DMEM / HIGH GLUCOSE medium supplemented with 10% fetal bovine serum. A normal group (Con), a model group (RII), a composition group, an inhibitor group (HY-12305), and a composition + inhibitor group (composition + HY-12305) were administered. The composition dosages were 15 μmol / L aesculin, 21 μmol / L aesculetin, 1.5 μmol / L aesculin, and 3 μmol / L baicalein. The inhibitor dosage was 50 nM. The cells were irradiated with 4 Gy of X-rays. The results are shown in Table 8.

[0079] Table 8 Effects of the composition on the expression of key proteins in the Caspase-9 / -3 pathway

[0080]

[0081] ## P < 0.01 compared with the Control group; ** P<0.01 compared with RII group

[0082] As shown in Table 8, after irradiation, the expression of Caspase-9, Caspase-3, Cle-Caspase-9 and Cle-Caspase-3 was significantly increased, while the expression of Caspase-9, Caspase-3, Cle-Caspase-9 and Cle-Caspase-3 in the combination group, HY-12305 group and combination + HY-12305 group was significantly decreased, indicating that the composition of the present invention can reduce the expression of Caspase-9, Caspase-3, Cle-Caspase-9 and Cle-Caspase-3, thereby alleviating the apoptosis of HIEC-6 intestinal epithelial cells.

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition for treating radiation-induced intestinal injury, characterized in that: The active ingredients are: aesculin, aesculetin, aesculetin and baicalein; the molar ratio of aesculin, aesculetin, aesculetin and baicalein is (1-100): (10-150): (0.1-30): (0.1-50).

2. The composition for treating radiation-induced intestinal injury according to claim 1, characterized in that: The molar ratio of aesculetin, aesculetin, aesculetin and baicalein is 50:70:5:

10.

3. The composition for treating radiation-induced intestinal injury according to claim 1, characterized in that: The molar ratio of aesculetin, aesculetin, aesculetin and baicalein is 1: 10：0.1：0.1。 4. Use of the composition for treating radiation-induced intestinal injury according to any one of claims 1 to 3 in the preparation of a drug for treating radiation-induced intestinal injury.

5. Use of the composition for treating radiation-induced intestinal injury according to claim 4 in preparing a drug for treating radiation-induced intestinal injury, characterized in that: The radiation-induced intestinal injury includes acute radiation-induced intestinal injury, chronic radiation-induced intestinal injury and ulcerative colitis.

6. A drug for treating radiation-induced intestinal injury, characterized in that: The composition for treating radiation-induced intestinal injury comprises the composition according to any one of claims 1 to 3.

7. The drug for treating radiation-induced intestinal injury according to claim 6, characterized in that: The dosage form of the drug is an injection, enema preparation, powder, pill, tablet, microcapsule, soft capsule, film, ointment, tincture, granule or aerosol allowed in pharmacy.

Images