A composition for treating radiation-induced intestinal injury and use thereof

Abstract

The application discloses a kind of compositions for treating radiation enteropathy and application thereof, belong to medical technology field.The composition includes the following components: anemarrhena asphodeloides B4, obakunine, berberine, aesculetin, fraxinellone and fraxinellin.The composition has no toxic side effects, has good safety, can reduce the inflammatory infiltration of intestinal epithelial cells, reduce intestinal epithelial cell apoptosis, protect intestinal barrier and protect intestinal oxidative damage by regulating NF-κB, Caspase 9 / 3 and NRF2 pathway, and then multi-component, multi-pathway jointly play the efficacy of treating radiation enteropathy.The problem that there is no effective drug for treating radiation enteropathy in the prior art is solved.

Description

Technical Field

[0001] This invention relates to the field of medical technology, and more specifically to a composition for treating radiation-induced intestinal injury and its application. Background Technology

[0002] Ionizing radiation is widely used in medicine, military, energy, agriculture, and many other fields. While benefiting human life, it also poses potential safety hazards. Radiation enteropathy is an intestinal disease caused by long-term or high-dose exposure to radiation. This condition usually occurs after radiation therapy or accidental radiation exposure. Main symptoms include abdominal pain, diarrhea, rectal bleeding, and tenesmus, which can be life-threatening and severely reduce the patient's quality of life. The mechanism of radiation enteropathy is complex, and there is currently no effective treatment. Therefore, the development of a drug to treat radiation enteropathy is urgently needed. Summary of the Invention

[0003] In order to solve the above-mentioned technical problems, the object 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 the lack of effective drugs for treating radiation-induced intestinal disease.

[0004] The technical solution of the present invention to solve the above-mentioned technical problems is as follows:

[0005] A composition for treating radiation-induced intestinal injury comprises the following components: Pulsatilla saponin B4, berberine, berberine, fraxetin, fraxetin A, and fraxetin B.

[0006] The beneficial effects of this invention are as follows: This invention employs a combination of multiple drugs, targeting the long course and complex pathogenesis of radiation-induced intestinal injury through multi-component and multi-target action, achieving good clinical results. This composition has no toxic side effects, good safety, and can significantly reduce radiation-induced intestinal injury, improve the antioxidant capacity of intestinal tissue, and inhibit radiation-induced intestinal inflammatory responses. It can also reduce inflammatory infiltration of intestinal epithelial cells, reduce intestinal epithelial cell apoptosis, protect the intestinal barrier, and protect against intestinal oxidative damage by regulating the NF-κB, Caspase 9 / 3, and NRF2 pathways. Thus, multiple components and multiple pathways work together to treat radiation-induced intestinal disease.

[0007] Furthermore, the molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is (1-100):(1-150):(1-50):(1-20):(1-60):(1-60).

[0008] Furthermore, the molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is ((10-30):(5-20):(5-20):(1-5):(20-40):(1-10).

[0009] Furthermore, the molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is 15:10:8:2:30:4.

[0010] The above-mentioned composition for treating radiation-induced intestinal injury is used in the preparation of a medicament for treating radiation-induced intestinal disease.

[0011] Furthermore, radiation enteropathy includes acute radiation enteropathy, chronic radiation enteropathy, ulcerative colitis, and intestinal obstruction.

[0012] A medicine for treating radiation enteropathy, the active ingredient comprising the above-described composition for treating radiation enteropathy.

[0013] Furthermore, the dosage form of the drug is injection, powder, pill, microcapsule, soft capsule, film, ointment, tincture, granule or aerosol.

[0014] The present invention has the following beneficial effects:

[0015] Radiation-induced intestinal injury is characterized by a long course and complex pathogenesis. This invention employs a combination of multiple drugs, achieving good clinical results through multi-component and multi-target action. Experiments have demonstrated that this composition has no toxic side effects and good safety. It can significantly reduce intestinal mucosal atrophy and villus shedding, lower Chiu's score for intestinal tissue damage, increase villus length and crypt number, enhance the antioxidant capacity of intestinal tissue, and inhibit radiation-induced intestinal inflammatory response. Furthermore, it can reduce inflammatory infiltration of intestinal epithelial cells, reduce intestinal epithelial cell apoptosis, protect the intestinal barrier, and protect against intestinal oxidative damage by regulating the NF-κB, Caspase 9 / 3, and NRF2 pathways. Thus, the multi-component and multi-pathway approach works synergistically to treat radiation-induced intestinal disease. Detailed Implementation

[0016] The principles and features of the present invention are described below with reference to embodiments. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, specific conditions or manufacturer-recommended conditions should be followed in the embodiments. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0017] Example 1:

[0018] A composition for treating radiation-induced intestinal injury, comprising the following components:

[0019] Pulsatilla saponin B4, berberine, fraxin, fraxin A and fraxin B;

[0020] The molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is 15:10:8:2:30:4.

[0021] Example 2:

[0022] A composition for treating radiation-induced intestinal injury, comprising the following components:

[0023] Pulsatilla saponin B4, berberine, fraxin, fraxin A and fraxin B;

[0024] The molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is 1:1:1:1:1:1.

[0025] Example 3:

[0026] A composition for treating radiation-induced intestinal injury, comprising the following components:

[0027] Pulsatilla saponin B4, berberine, fraxin, fraxin A and fraxin B;

[0028] The molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is 100:150:50:20:60:60.

[0029] Experimental Example 1: Evaluation Experiment of Intestinal Pathological Damage

[0030] An intestinal pathological damage evaluation experiment was conducted on the composition used to treat radiation-induced intestinal injury via gavage.

[0031] The experimental method is as follows:

[0032] Adult, healthy male C57BL / 6J mice (7-8 weeks old, weighing 18-20g) were acclimatized for one week. Before irradiation, each mouse received an abdominal injection of 0.2 mL of sodium pentobarbital (0.3% concentration) to ensure anesthesia during the irradiation process. A mouse model of radiation enteropathy was established by a single whole-body irradiation of 10 Gy using a medical linear accelerator (CLINAC 23EX-SN297) in 6MV X-ray mode, with a source-to-skin distance of 100 cm and a dose rate of 400 MU / min. The C57BL / 6J male mice were randomly assigned to 6 groups, with 6 mice in each group. The study included a normal group, a model group, and low, medium, and high dose groups of the combined composition. The high-dose group consisted of: Pulsatilla saponin B4 30 μM / kg + Phellodendron chinense 20 μM / kg + Berberine 16 μM / kg + Fraxinus 4 μM / kg + Fraxinus A 60 μM / kg + Fraxinus B 8 μM / kg. The medium and low-dose groups were diluted twofold. After modeling, the patients were administered the medication by gavage for 7 consecutive days, followed by sacrifice and tissue sampling. Hematoxylin and eosin (H&E) staining was then performed. Chiu's score was used to evaluate intestinal tissue damage, and GraphPadPrism 8.4.0 software was used for analysis and statistical data collection.

[0033] The experimental results are shown in Table 1. Among them, ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group; * P<0.05 compared with the model group.

[0034] Table 1. Statistics of Chiu's score, villus length, and crypt number in different dosage groups of the composition.

[0035] Grouping Chiu's rating Fiber length (μmol / L) Number of crypts normal group 0.50±0.54 397.66±55.48 29.83±2.92 Model group <![CDATA[4.20±0.83 ## ]]> <![CDATA[198.00±18.13 ## ]]> <![CDATA[13.00±1.58 ## ]]> Low-dose group of the composition 3.00±0.70 <![CDATA[253.50±14.37 * ]]> 16.20±2.58 Dosage group in the composition <![CDATA[2.33±0.51 ** ]]> <![CDATA[284.50±27.86 ** ]]> <![CDATA[21.33±1.63 ** ]]> High-dose group of the composition <![CDATA[2.00±0.70 ** ]]> <![CDATA[376.66±15.38 ** ]]> <![CDATA[25.16±1.47 ** ]]>

[0036] Experimental results showed that the composition significantly reduced intestinal mucosal atrophy, villus loss, and Chiu's score for intestinal tissue damage after treatment, and increased villus length and crypt number. The results indicate that the composition can effectively protect against intestinal pathological damage and reduce intestinal bleeding.

[0037] Experiment Example 2: Evaluation of Anti-inflammatory and Antioxidant Capabilities

[0038] Based on Experiment 1, intestinal tissue samples were taken from mice in each group and tested according to the instructions of ELISA kit, SOD kit and MDA kit to detect changes in the levels of antioxidant enzyme (SOD), malondialdehyde (MDA) and inflammatory factors (IL-6, IL-10, IL-1β and TNF-α).

[0039] The experimental results are shown in Tables 2 and 3. Among them, ## P<0.01 compared with the normal group; **P<0.01 compared with the model group; * P<0.05 compared with the model group.

[0040] Table 2 Antioxidant capacity of different dosages of the composition

[0041] Grouping SOD MDA normal group 132.40±4.07 8.26±1.00 Model group <![CDATA[86.90±4.23 ## ]]> <![CDATA[19.69±2.31 ## ]]> Low-dose group of the composition <![CDATA[97.89±4.92 ** ]]> 17.34±2.02 Dosage group in the composition <![CDATA[105.50±4.69 ** ]]> <![CDATA[13.74±1.53 ** ]]> High-dose group of the composition <![CDATA[112.20±4.48 ** ]]> <![CDATA[11.16±1.46 ** ]]>

[0042] Table 3. Anti-inflammatory efficacy of different dosages of the composition.

[0043] Grouping TNF-α IL-1β IL-6 IL-10 normal group 58.72±1.52 14.82±1.47 52.71±1.46 87.48±1.94 Model group <![CDATA[76.90±1.51 ## ]]> <![CDATA[34.59±0.94 ## ]]> <![CDATA[96.80±1.40 ## ]]> <![CDATA[57.93±1.67 ## ]]> Low-dose group of the composition <![CDATA[64.25±1.02 ** ]]> <![CDATA[27.81±0.76 ** ]]> <![CDATA[87.72±1.87 ** ]]> <![CDATA[61.88±0.93 * ]]> Dosage group in the composition <![CDATA[61.55±0.88 ** ]]> <![CDATA[24.58±0.86 ** ]]> <![CDATA[78.92±1.30 ** ]]> <![CDATA[65.45±1.29 ** ]]> High-dose group of the composition <![CDATA[59.40±1.07 ** ]]> <![CDATA[19.62±1.11 ** ]]> <![CDATA[71.24±1.79 ** ]]> <![CDATA[75.31±1.34 ** ]]>

[0044] The experimental results showed that, compared with the normal group, the model group mice exhibited significantly decreased activity of the intestinal antioxidant enzyme SOD, significantly increased levels of the lipid peroxide MDA, elevated levels of pro-inflammatory factors TNF-α, IL-1β, and IL-6, and decreased levels of the anti-inflammatory factor IL-10. The composition dose-dependently inhibited the levels of pro-inflammatory factors TNF-α, IL-1β, and IL-6, promoted the secretion of the anti-inflammatory factor IL-10, and alleviated intestinal inflammation. The composition dose-dependently increased SOD activity and decreased MDA content, thereby enhancing the antioxidant capacity of the intestine. These results indicate that the composition can improve the antioxidant capacity of the intestinal tissue of irradiated mice and inhibit radiation-induced intestinal inflammatory responses.

[0045] Experimental Example 3: Effect of the Composition on the Expression of NF-κB Pathway Proteins

[0046] The prepared composition was used to conduct experiments to evaluate its effect on the expression of NF-κB pathway proteins. The cells used in the experiment were normal human intestinal epithelial cells HIEC-6 (ATCC cell bank, USA), with HY-18738 inhibitor as a control. The conventional Western blot method was used for testing.

[0047] The implementation method was as follows: A cell radiation damage model was constructed using X-rays at a dose of 4 Gy. The cell experiments were divided into several groups: normal group, model group, high-dose combination group, inhibitor group, and high-dose combination + inhibitor group. The high-dose combination group consisted of: Pulsatilla saponin B4 2.3 μmol / L + Phellodendron chinense 1.53 μmol / L + Berberine 1.23 μmol / L + Fraxinus chinensis 0.31 μmol / L + Fraxinus chinensis A 4.60 μmol / L + Fraxinus chinensis B 0.61 μmol / L. The inhibitor group consisted of: 100 μmol / L. After 24 hours of treatment in each group, samples were collected and analyzed.

[0048] The experimental results are shown in Table 4. Among them, ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.

[0049] Table 4. Effects of the composition on NF-κB pathway protein expression

[0050] Grouping pP-65 / P-65 normal group 0.18±0.02 Model group <![CDATA[0.79±0.05 ## ]]> High-dose group of the composition <![CDATA[0.39±0.04 ** ]]> Inhibitor group <![CDATA[0.32±0.05 ** ]]> High-dose combination + inhibitor group <![CDATA[0.40±0.04 ** ]]>

[0051] The experimental results showed that the expression of p-P65 protein was significantly increased in the model group, and significantly decreased after treatment with the composition, indicating that the composition can exert anti-inflammatory effects by inhibiting the NF-κB pathway.

[0052] Experimental Example 4: Effect of the Composition on the Expression of Caspase 9 / 3 Pathway Proteins

[0053] The prepared composition was used to conduct experiments to evaluate its effect on the expression of Caspase 9 / 3 pathway proteins. The cells used in the experiment were normal human intestinal epithelial cells HIEC-6 (ATCC cell bank, USA), with HY-12305 inhibitor as a control. The conventional Western blot method was used for testing.

[0054] The implementation method was as follows: A cell radiation damage model was constructed using X-rays at a dose of 4 Gy. The cell experiments were divided into several groups: normal group, model group, high-dose combination group, inhibitor group, and high-dose combination + inhibitor group. The high-dose combination group consisted of: Pulsatilla saponin B4 2.3 μmol / L + Phellodendron chinense 1.53 μmol / L + Berberine 1.23 μmol / L + Fraxinus chinensis 0.31 μmol / L + Fraxinus chinensis A 4.60 μmol / L + Fraxinus chinensis B 0.61 μmol / L. The inhibitor group was treated with 400 nM. After 24 hours of treatment in each group, samples were collected and analyzed.

[0055] The experimental results are shown in Table 5. Among them, ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.

[0056] Table 5. Effects of the composition on the expression of proteins in the Caspase 9 / 3 pathway.

[0057]

[0058] The experimental results showed that the expression of Cle-Caspase 9 / 3 protein was significantly increased in the model group, and the expression of Cle-Caspase 9 / 3 protein was significantly decreased after treatment with the composition, indicating that the composition can exert an anti-apoptotic effect by inhibiting the Caspase 9 / 3 pathway and protect the intestinal barrier.

[0059] Experimental Example 5: Effect of the Composition on NRF2 Pathway Protein Expression

[0060] The composition prepared in Example 1 was used to evaluate the effect of the composition on the expression of NRF2 pathway proteins. The cells used in the experiment were normal human intestinal epithelial cells HIEC-6 (ATCC cell bank, USA), and the HY-112675 inhibitor was used as a control. The conventional Western blot method was used for testing.

[0061] The implementation method was as follows: A cell radiation damage model was constructed using X-rays at a dose of 4 Gy. The cell experiments were divided into several groups: normal group, model group, high-dose combination group, agonist group, and high-dose combination + agonist group. The high-dose combination group consisted of: Pulsatilla saponin B4 2.3 μmol / L + Phellodendron chinense 1.53 μmol / L + Berberine 1.23 μmol / L + Fraxinus chinensis 0.31 μmol / L + Fraxinus chinensis A 4.60 μmol / L + Fraxinus chinensis B 0.61 μmol / L. The agonist group consisted of: 30 μmol / L. After 24 hours of drug intervention in each group, samples were collected and analyzed.

[0062] The experimental results are shown in Table 6. Among them, ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.

[0063] Table 6. Effects of the composition on NRF2 pathway protein expression

[0064] Grouping NRF2 / GAPDH normal group 0.81±0.03 Model group <![CDATA[0.37±0.03 ## ]]> High-dose group of the composition <![CDATA[1.02±0.10 ** ]]> agonist group <![CDATA[0.93±0.06 ** ]]> High-dose combination + agonist group <![CDATA[0.98±0.07 ** ]]>

[0065] The experimental results showed that NRF2 protein expression was significantly decreased in the model group, and significantly increased after treatment with the composition, indicating that the composition can exert an antioxidant effect by inhibiting the NRF2 pathway and protecting the intestinal barrier.

[0066] 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 within the protection scope of the present invention.

Claims

1. A composition for treating radiation-induced intestinal injury, characterized in that, It is composed of the following components: Pulsatilla saponin B4, berberine, fraxin, fraxin A and fraxin B; The molar ratio of Pulsatilla saponin B4, berberine, fraxetin, fraxetin A, and fraxetin B is (10~30):(5~20):(5~20):(1~5):(20~40):(1~10).

2. The composition for treating radiation-induced intestinal injury according to claim 1, characterized in that, The molar ratio of Pulsatilla saponin B4, berberine, fraxin, fraxin A, and fraxin B is 15:10:8:2:30:

4.

3. The use of the composition for treating radiation-induced intestinal injury as described in claim 1 or 2 in the preparation of a medicament for treating radiation enteropathy.

4. The application according to claim 3, characterized in that, The radiation enteropathy includes acute radiation enteropathy and chronic radiation enteropathy.

5. A drug for treating radiation-induced enteropathy, characterized in that, The active ingredient includes the composition for treating radiation-induced intestinal injury as described in claim 1 or 2.

6. The medicament for treating radiation enteropathy according to claim 5, characterized in that, The dosage form of the drug is injection, powder, pill, microcapsule, soft capsule, film, ointment, tincture, granule or aerosol.

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