A composition for treating radiation enteropathy and uses thereof
By using a combination of puerarin, baicalin, berberine, and glycyrrhetinic acid to regulate multiple signaling pathways, the problem of the lack of effective treatments for radiation enteropathy in existing technologies has been solved, and the effect of significantly reducing intestinal lesions caused by radiation enteropathy has been achieved.
Patent Information
- Application Number
- CN202410879634.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-07-02
- Publication Date
- 2025-12-16
- Estimated Expiration
- 2044-07-02
AI Technical Summary
There is a lack of effective drugs for treating radiation enteropathy in the current technology, especially due to the serious side effects and narrow treatment window of existing drugs such as Amifostine, which limits their clinical application.
A combination of puerarin, baicalin, berberine, and glycyrrhetinic acid was used to promote the proliferation of intestinal cells after radiation damage, reduce intestinal tissue inflammation, reduce cell apoptosis, and improve radiation damage to intestinal epithelial cells by regulating the Wnt/β-catenin, NF-κB, and Caspase-9/-3 signaling pathways.
This composition significantly reduces the shortening of intestinal villi caused by radiation enteropathy, increases the number of crypts, lowers Chiu's score, alleviates intestinal pathological damage, promotes cell proliferation, and reduces apoptosis and inflammatory response, demonstrating a significant therapeutic effect on radiation enteropathy.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological medicine, in particular to a composition for treating radiation enteropathy and application thereof. BACKGROUND
[0002] Radiation-induced intestinal injury (RII) is an intestinal injury caused by ionizing radiation, and the ionizing radiation sources include x-rays, gamma rays, beta rays and neutron rays. The pathogenesis of RII involves apoptosis and senescence of intestinal epithelial cells, activation of intestinal inflammation, activation of oxidative stress, changes in intestinal flora and intestinal fibrosis, and other factors. The pathological mechanism is complex, and the pathological process is diverse. At present, there is no ideal treatment measure.
[0003] In clinical treatment of RII, anti-free radical oxidative damage drugs such as vitamin C and vitamin E are often used. Amifostine, as a free radical scavenger, can reduce the side effects of radiotherapy and is the only drug approved for clinical anti-RII. However, its serious side effects and narrow treatment time window seriously limit its clinical use. Therefore, it is of great clinical value to further search for possible effective treatment drugs for RII. SUMMARY
[0004] In order to solve the above technical problems, the purpose of the present application is to provide a composition for treating radiation enteropathy and application thereof, so as to solve the problem that there is no effective treatment drug for radiation-induced intestinal injury.
[0005] The technical scheme for solving the above technical problems of the present application is as follows:
[0006] A composition for treating radiation enteropathy, the composition comprising puerarin, baicalein, berberine and glycyrrhetinic acid.
[0007] The present application provides a composition for treating radiation enteropathy, which can effectively reduce the shortening degree of intestinal villi caused by radiation enteropathy, increase the number of crypts, reduce Chiu's score, and reduce the pathological damage of the intestinal tract caused by radiation. The composition activates the Wnt / β-catenin signaling pathway to promote the proliferation of HIEC-6 cells after radiation damage, inhibits the Caspase-9 / -3 signaling pathway to reduce the apoptosis of HIEC-6 cells, and affects the NF-κB signaling pathway to reduce the inflammatory response of cells caused by radiation damage, thereby playing a significant role in treating radiation enteropathy.
[0008] Further, the molar ratio of puerarin, baicalein, berberine and glycyrrhetinic acid in the composition is (180-260):(100-200):(5-30):(1-10).
[0009] Further, the molar ratio of puerarin, baicalein, berberine and glycyrrhetinic acid in the composition is 220:150:15:5.
[0010] The composition for treating radiation enteropathy is used in the preparation of a medicament for treating radiation enteropathy.
[0011] Further, the radiation enteropathy includes acute / chronic enteropathy caused by at least one of x-rays, gamma rays, alpha rays, beta rays and neutron rays.
[0012] Further, the acute / chronic enteropathy includes acute / chronic enteritis, intestinal obstruction, intestinal hemorrhage and intestinal ulcer.
[0013] A medicament for treating radiation enteropathy, comprising the composition.
[0014] Further, the dosage form of the medicament includes oral preparations or enema preparations.
[0015] The present application has the following beneficial effects:
[0016] The present application provides a composition for treating radiation enteropathy, which can promote the proliferation of intestinal cells after radiation damage, reduce the inflammation level of intestinal tissue after radiation, reduce the apoptosis of intestinal cells after radiation, etc., improve the radiation damage of intestinal epithelial cells, play an anti-radiation intestinal damage role, and has a significant effect on treating radiation enteropathy by regulating Wnt / β-catenin, NF-κB and Caspase-9 / -3 signaling pathways, etc. DETAILED DESCRIPTION
[0017] The principles and features of the present application are described below, and the examples are only used to explain the present application and are not used to limit the scope of the present application. If the specific conditions are not specified in the examples, the conventional conditions or the conditions recommended by the manufacturer are used. If the reagents or instruments used are not specified by the manufacturer, they are all conventional products that can be purchased on the market.
[0018] Example 1: Evaluation experiment of radiation intestinal mucosa damage
[0019] The evaluation experiment of radiation intestinal mucosa damage was performed by gavage.
[0020] (1) Experimental method
[0021] C57 BL / 6J mice were taken, X-ray 6MV mode was used, source-skin distance was 100 cm, dose rate was 400 MU / min, mice were irradiated once, and a mouse radiation damage model was constructed to simulate radiation enteropathy.
[0022] The mice were divided into a normal group, a model group, a composition low-dose group, a composition medium-dose group and a composition high-dose group.
[0023] The composition low-dose group was administered in an amount of 2.16 mg / kg of puerarin, 0.9 mg / kg of baicalein, 0.18 mg / kg of berberine and 0.04 mg / kg of glycyrrhetinic acid;
[0024] The composition medium-dose group was administered in an amount of 4.32 mg / kg of puerarin, 1.8 mg / kg of baicalein, 0.36 mg / kg of berberine and 0.08 mg / kg of glycyrrhetinic acid;
[0025] The composition high-dose group was administered in an amount of 8.64 mg / kg of puerarin, 3.6 mg / kg of baicalein, 0.72 mg / kg of berberine and 0.16 mg / kg of glycyrrhetinic acid.
[0026] After the modeling was completed, the mice were continuously administered intragastrically for 7 days and then sacrificed, followed by H&E staining, and the damage to the intestinal tissue was evaluated by Chiu's score, villus length and intestinal crypt number.
[0027] (2) Experimental results
[0028] The experimental results are shown in Table 1. ## P<0.01 compared with the normal group; ** P<0.01, * P<0.05 compared with the model group.
[0029] Table 1: Evaluation of the pathological damage to the RII intestinal mucosa by the composition
[0030] Group Chiu's score Villus length (pm) Crypt number (number) Normal group 0.50±0.55 592.59±10.94 29.33±2.73 Model group 4.67 ± 0.52 ## ]] 279.66 ± 12.51 ## ]] 13.5 ± 1.05 ## ]] Composition low-dose group 4.0±0.63* 451.07±20.79** 18.67±1.37** Composition medium-dose group 3.5±0.55** 510.64±13.04** 22.33±1.63** Composition high-dose group 3.0±0.63** 558.20±6.44** 24.5±1.52**
[0031] The results show that, compared with the normal control group, the intestinal villi of the radiation model group mice were defoliated and shortened, the intestinal villus epithelial cells were denatured and necrotic and fell off, the crypt number was reduced, and the Chiu's score was significantly increased (P<0.01). Compared with the model group, the degree of shortening of the intestinal villi of the composition group was reduced, the crypt number was increased, and the Chiu's score was reduced, indicating that the damage to the intestinal tissue was reduced, and indicating that the composition can significantly reduce the pathological damage to the intestinal tract caused by radiation.
[0032] Example 2: Evaluation of the radiation cell proliferation level
[0033] The evaluation of the radiation cell proliferation level was performed by an in vitro cell experiment.
[0034] (1) Experimental method
[0035] Human intestinal epithelial cells HIEC-6 were inoculated in the hole plate, and a cell radiation injury model was constructed by X-ray at a dose of 4 Gy. The cell experiment was divided into a normal group, a model group, a composition group, a HY-148055A (Wnt / β-catenin agonist, purchased from MedChemExpres company) group, and a composition + HY-148055A (Wnt / β-catenin agonist) group.
[0036] In the composition group and the composition + HY-148055A group, the administration amount of the composition was 6.043 μg / mL of puerarin + 2.34 μg / mL of baicalein + 0.35 μg / mL of berberine + 0.031 μg / mL of glycyrrhetinic acid; after 24 hours of administration intervention, the expression of Ki67 was detected by immunofluorescence method.
[0037] (2) Experimental results
[0038] The experimental results are shown in Table 2. ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.
[0039] Table 2 Effect of the composition on the proliferation of RII intestinal epithelial cells
[0040] Group Relative proliferation rate (%) Normal group 45.61±3.39 Model group 5.23 ± 0.73 ## ]] Composition group 26.76±3.88** HY-148055A group 29.52±2.40** Composition + HY-148005A group 30.19±3.74**
[0041] Ki67 is a kind of nuclear protein closely related to the activity of cell proliferation, and can be used as a marker of cell proliferation. The results in the above table show that compared with the normal group, the expression of Ki67 in the cells of the model group after radiation was significantly reduced (P<0.01), and the cell proliferation was significantly inhibited. Compared with the model group, the composition group can promote the proliferation of cells, which is manifested by the significant increase of the expression of lipoprotein Ki67 (P<0.01). It is shown that the composition can promote the proliferation of HIEC-cells after radiation injury by activating the Wnt / β-catenin signaling pathway.
[0042] Example 3: Evaluation experiment of radiation cell apoptosis level
[0043] The evaluation experiment of radiation cell apoptosis level was carried out by in vitro cell experiment.
[0044] (1) Experimental method
[0045] Human intestinal epithelial cells HIEC-6 were inoculated in the hole plate, and a cell radiation injury model was constructed by X-ray at a dose of 4 Gy. The cell experiment was divided into a normal group, a model group, a composition group, a HY-12305 (Caspase-9 / -3 inhibitor, purchased from MedChemExpress company) group, and a composition + HY-12305 (Caspase-9 / -3 inhibitor) group.
[0046] The dosages of the composition in the composition group and the composition + HY-12305 group are: puerarin 6.043 μg / mL + baicalein 2.34 μg / mL + berberine 0.35 μg / mL + glycyrrhetinic acid 0.031 μg / mL; after 24 hours of drug intervention, the apoptosis level of each group is detected by TUNEL experiment.
[0047] (2) Experimental results
[0048] The experimental results are shown in Table 3. ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.
[0049] Table 3 Effect of the composition on apoptosis of RII enterocyte
[0050] Group Relative proliferation rate (%) Normal group 3.33±0.68 Model group 18.45 ± 2.75 ## ]] Composition group 7.98±0.47** HY-12305 group 6.58±1.41** Composition + HY-12305 group 5.80±0.85**
[0051] The results show that compared with the normal group, the number of apoptotic cells in the model group after radiation increases significantly (P<0.01), and the cell damage is serious. Compared with the model group, the number of apoptotic cells in the composition group, the HY-12305 group and the composition + HY-12305 group decreases significantly (P<0.01), indicating that the composition can reduce the apoptosis of HIEC-6 cells by inhibiting the Caspase-9 / -3 signaling pathway.
[0052] Example 4: Evaluation experiment of inflammatory factor level secreted by radiation cells
[0053] The evaluation experiment of inflammatory factor level secreted by radiation cells is carried out by in vitro cell experiment.
[0054] (1) Experimental method
[0055] Human enterocyte HIEC-6 is inoculated in the well plate, and a cell radiation damage model is constructed by using X-ray 4Gy dose. The cell experiment is divided into a normal group, a model group, a composition group, a HY-18738 (NF-κB inhibitor, purchased from MedChemExpress company) group and a composition + HY-18738 (NF-κB inhibitor) group.
[0056] The dosages of the composition in the composition group and the composition + HY-18738 group are: puerarin 6.043 μg / mL + baicalein 2.34 μg / mL + berberine 0.35 μg / mL + glycyrrhetinic acid 0.031 μg / mL; after 24 hours of drug intervention, the factor level of each group is detected by ELISA kit.
[0057] (2) Experimental results
[0058] The experimental results are shown in Table 4. ## P<0.01 compared with the normal group; ** P<0.01, * P<0.05 compared with the model group.
[0059] Table 4 Effect of the composition on secretion of inflammatory factors of RII intestinal epithelial cells
[0060]
[0061] The results show that, compared with the normal group, the model group after radiation can cause the increase of the levels of pro-inflammatory factors TNF-α, IL-1β and IL-6, and the decrease of the level of anti-inflammatory factor IL-10. Compared with the model group, the composition and the KF-κB signal pathway inhibitor can both alleviate the inflammatory response of the cells to a certain extent, and the anti-inflammatory levels are equivalent, indicating that the composition can alleviate the inflammatory response of the cells caused by radiation damage by affecting the KF-κB signal pathway.
[0062] Example 5: Evaluation experiment of expression levels of key proteins of various signal pathways
[0063] On the basis of Examples 2-4, the proteins of each group were extracted, and the changes in the expression levels of key proteins of the Wnt / β-catenin signal pathway, the Caspase-9 / -3 signal pathway and the NF-κB signal pathway were detected by the immunoblotting method.
[0064] The experimental results are shown in Table 5. ## P<0.01 compared with the normal group; ** P<0.01 compared with the model group.
[0065] Table 5 Effect of the composition on proteins of related signal pathways of RII intestinal epithelial cells
[0066]
[0067] The activation of Wnt / β-catenin signaling pathway is due to the increased transfer of β-catenin to the nucleus, and in the nucleus, β-catenin binds to the transcription factor family TCF / LEFs, which can promote the expression of proteins such as CyclinD1 (CCND1) and MYC, and promote cell proliferation. The Western blot detection results show that the composition significantly increases the expression of β-catenin in the nucleus, indicating that the composition activates the Wnt / β-catenin pathway by promoting the nuclear translocation of β-catenin. There is no significant difference in the expression level of nuclear β-catenin between the composition + HY-148055A group and the HY-148055A group. The above results show that the composition promotes the proliferation of intestinal epithelial cells by activating the Wnt / β-catenin signaling pathway.
[0068] Compared with the normal group, the composition group and the composition + HY-12305 group significantly inhibited the expression levels of Caspase-3, Caspase-9, Cleaved-Caspase-3 and Cleaved-Caspase-9 proteins. The above results show that the composition plays a role in resisting radiation-induced cell apoptosis by inhibiting the activation of Caspase-9 / -3 signaling pathway.
[0069] Compared with the model group, the composition group, the HY-18738 group and the composition + HY-18738 group significantly inhibited the expression level of phosphorylated P65 protein, and the composition + HY-18738 group and the HY-18738 group had no significant difference in the expression level of phosphorylated P65 protein, and had no significant effect on the expression of P65. The above results show that the composition can inhibit the activation of NF-κB signaling pathway by inhibiting the phosphorylation level of P65 protein, thereby playing a role in resisting radiation-induced intestinal injury inflammatory response.
[0070] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. A composition for treating radiation enteropathy, characterized in that, The composition consists of puerarin, baicalein, berberine and glycyrrhetinic acid; the molar ratio of puerarin, baicalein, berberine and glycyrrhetinic acid is (180-260):(100-200):(5-30):(1-10).
2. The composition for treating radiation enteropathy according to claim 1, characterized by, The molar ratio of puerarin, baicalein, berberine and glycyrrhetinic acid in the composition is 220:150:15:
5.
3. Use of the composition for treating radiation enteropathy according to claim 1 or 2 in the preparation of a medicament for treating radiation enteropathy.
4. Use according to claim 3, characterized in that, The radiation enteropathy includes acute / chronic enteropathy caused by at least one of x-ray, gamma-ray, alpha-ray, beta-ray and neutron ray.
5. Use according to claim 4, characterized in that, The acute / chronic enteropathy includes acute / chronic enteritis, intestinal obstruction, intestinal hemorrhage and intestinal ulcer.
6. A medicament for treating radiation enteropathy, characterized by, The composition according to claim 1 or 2 is included.
7. The medicament for treating radiation enteropathy according to claim 6, characterized in that, The dosage form of the medicament includes oral preparation or enema preparation.
Citation Information
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