Application of biliverdin in preparation of medicine for preventing and treating lung injury caused by radon exposure

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of biliverdin in preparation of medicine for preventing and treating lung injury. By constructing a radon inhalation-induced lung injury mouse model, after exposure at different radon levels, the weight of a mouse is reduced, the lung coefficient is increased, the tidal volume of the lung, the ventilation volume per minute and the maximum inspiration flow are reduced, the respiratory function is inhibited, the alveolar cavity of the mouse is abnormally expanded, the pulmonary septum is thickened and fractured, and more fibrin exudation is accompanied; obvious inflammatory infiltration occurs beside bronchial tubes, extravasated blood occurs in small blood vessels, lung tissue collagen deposition occurs, pulmonary fibrosis can be caused, lung injury can be caused, and the phenomena can be effectively inhibited through gavage of biliverdin. Therefore, biliverdin can be used for preventing and treating lung injury caused by radon exposure.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of biliverdin in preparing medicine for preventing and treating lung damage caused by radon exposure. Background Art

[0002] Indoor radon is considered to be the main source of radiation exposure to humans. The inhaled dose caused by radon and its progeny accounts for about 48% of the average annual dose of natural background radiation for humans. Due to the special mode of action of radon and its progeny, bronchial epithelial basal cells, mucus cells and lung epithelial cells become their important targets. Long-term radon exposure can cause oxidative stress, DNA damage and inflammatory response in lung and bronchial epithelial cells, and lead to chronic lung diseases, including pulmonary fibrosis and lung cancer. At present, there is no effective treatment for pulmonary fibrosis caused by radon exposure in clinical practice. Therefore, actively exploring effective targets and preparations for the prevention and treatment of pulmonary fibrosis caused by radon exposure has important clinical value. Summary of the invention

[0003] The purpose of the present invention is to develop a drug for preventing and treating lung damage caused by radon exposure and effectively inhibiting lung collagen deposition and fibrosis.

[0004] In order to achieve the above object, the present invention provides the use of biliverdin in preparing a product for preventing and treating lung damage caused by radon exposure.

[0005] Preferably, the lung damage caused by radon exposure includes lung respiratory function damage caused by radon exposure.

[0006] Preferably, the radon exposure-induced pulmonary respiratory function damage includes radon exposure-induced decrease in minute ventilation and / or maximum inspiratory flow.

[0007] Preferably, the lung damage caused by radon exposure includes pulmonary fibrosis caused by radon exposure.

[0008] Preferably, the lung damage caused by radon exposure includes pulmonary collagen deposition caused by radon exposure.

[0009] Preferably, the lung damage caused by radon exposure includes an increase in the lung coefficient caused by radon exposure.

[0010] Preferably, the relative cumulative exposure dose of radon in the radon exposure is 30 to 120 WLM.

[0011] Preferably, the radon exposure concentration is 100000 Bq / m 3 , time is 189~755h.

[0012] The present invention also provides a medicine for preventing and treating lung damage caused by radon exposure, wherein the effective ingredient of the medicine includes biliverdin.

[0013] Preferably, the drug is formulated as a single-use dosage form with 35 mg biliverdin per dose.

[0014] Beneficial effects:

[0015] The present invention found that biliverdin can inhibit weight loss and increase of lung coefficient in mice, improve respiratory function, restore the decrease of lung tidal volume, minute ventilation and maximum inspiratory flow caused by lung injury, inhibit lung collagen deposition and fibrosis, and can be used to prevent and treat lung injury caused by radon exposure. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0017] Figure 1 This is a graph showing the changes in weight gain of mice after inhalation of different doses of radon in Example 1;

[0018] Figure 2 This is a graph showing the change in lung coefficient of mice after inhalation of different doses of radon in Example 1;

[0019] Figure 3 This is a graph showing the tidal volume test results of mice after inhalation of different doses of radon in Example 1;

[0020] Figure 4 This is a graph showing the minute ventilation test results of mice after inhalation of different doses of radon in Example 1;

[0021] Figure 5 This is a graph showing the maximum inspiratory flow rate test results of mice after inhalation of different doses of radon in Example 1;

[0022] Figure 6 The results of H&E staining of mouse lung tissue after inhalation of different doses of radon in Example 1; wherein the scale is 250 μm;

[0023] Figure 7 This is a diagram showing the H&E scoring results of mouse lung tissue after inhalation of different doses of radon in Example 1;

[0024] Figure 8 The Masson staining results of mouse lung tissue after inhalation of different doses of radon in Example 1; wherein the scale is 250 μm;

[0025] Fig. 9 This is a diagram showing the results of the Masson quantitative analysis of mouse lung tissue after inhalation of different doses of radon in Example 1;

[0026] Fig.10 This is a graph showing the changes in weight gain of mice after inhalation of different doses of radon in Example 2;

[0027] Fig.11This is a graph showing the change in lung coefficient of mice after inhalation of different doses of radon in Example 2;

[0028] Fig.12 This is a graph showing the tidal volume test results of mice after inhalation of different doses of radon in Example 2;

[0029] Fig.13 This is a graph showing the minute ventilation test results of mice after inhalation of different doses of radon in Example 2;

[0030] Fig.14 This is a graph showing the maximum inspiratory flow rate test results of mice after inhalation of different doses of radon in Example 2;

[0031] Fig.15 This is the H&E staining result of mouse lung tissue after inhalation of different doses of radon in Example 2; wherein the scale is 250 μm;

[0032] Fig.16 This is a diagram showing the H&E scoring results of mouse lung tissue after inhalation of different doses of radon in Example 2;

[0033] Fig.17 The Masson staining results of mouse lung tissue after inhalation of different doses of radon in Example 2; wherein the scale is 250 μm;

[0034] Fig.18 This is a diagram showing the results of the Masson quantitative analysis of mouse lung tissue after inhalation of different doses of radon in Example 2;

[0035] In the figure, ns means no statistical significance, **** means p < 0.0001, *** means p < 0.001, ** means p < 0.01, and * means p < 0.05. DETAILED DESCRIPTION

[0036] The invention provides application of biliverdin in preparing products for preventing and treating lung damage caused by radon exposure.

[0037] As an embodiment, the product of the present invention is a drug or reagent; as an embodiment, the product of the present invention is a drug. As an embodiment, the radon relative cumulative exposure dose of radon in the radon exposure of the present invention is 30 to 120 WLM; as another embodiment, the radon relative cumulative exposure dose of radon in the radon exposure of the present invention is 60 to 120 WLM. As an embodiment, the concentration of radon exposure in the present invention is 100000 Bq / m 3 , time is 189~755h; As another embodiment, the concentration of radon exposure in the present invention is 100000Bq / m 3 , time is 377~755h.

[0038] The present invention constructs a mouse model of lung injury caused by radon inhalation and finds that after exposure to different radon levels, the mice lose weight, increase the lung coefficient, reduce the lung tidal volume, minute ventilation and maximum inspiratory flow, and inhibit respiratory function. The alveolar cavity of the mice is abnormally expanded, the lung septum is thickened and broken, accompanied by more fibrin exudation, obvious inflammatory infiltration appears around the bronchus, congestion appears in small blood vessels, and collagen deposition in lung tissue can lead to the occurrence of pulmonary fibrosis and lung injury. Oral administration of biliverdin can effectively inhibit the above phenomenon. Therefore, biliverdin can be used to prevent and treat lung injury caused by radon exposure.

[0039] The present invention also provides a medicine for preventing and treating lung damage caused by radon exposure, wherein the effective ingredient of the medicine includes biliverdin.

[0040] As an embodiment, the drug of the present invention is formulated as a single-use dosage form with 35 mg of biliverdin per dose.

[0041] The present invention also provides a method for preventing and treating lung injury, which comprises intragastric administration of the drug described in the above technical solution.

[0042] To further illustrate the present invention, the application of biliverdin provided by the present invention in the preparation of a drug for preventing and treating lung damage caused by radon exposure is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1

[0044] 1. Experimental Materials

[0045] (1) Experimental animals: 18 C57 mice purchased from SpEffect Biotechnology.

[0046] (2) Experimental reagents: H&E staining kit, purchased from Solebro; Masson staining kit, purchased from Solebro.

[0047] 2. Model Construction

[0048] The experimental mice were randomly divided into a control group (NC) and a radon exposure group (Rn), with 9 mice in each group. The mice in the control group were placed in an environment with a background radon concentration of 8.20 Bq / m 3 The mice in the radon exposure group were exposed to the HD-3 multifunctional radon chamber with a radon exposure concentration of 100,000 Bq / m 3 The exposure time of the control group and radon exposure group were 189, 377, and 755 h, respectively, and the relative cumulative exposure dose was 30, 60, and 120 working level months (WLM).

[0049] 3. Experimental testing and conclusions

[0050] (1) The dynamic changes of weight gain and lung coefficient of mice in different treatment groups after radon inhalation were detected; the weight gain was the weight after exposure (g) minus the weight before exposure (g); the lung coefficient was obtained by dividing the lung wet weight (mg) by the body weight (g). The test results are as follows: Figure 1 and Figure 2 As shown. Figure 1 and Figure 2 It can be seen that compared with the control group, the mice in the radon exposure group lost weight and increased lung coefficient after exposure to different radon levels. Radon exposure can cause pulmonary fibrosis and lung damage.

[0051] (2) Non-invasive small animal respirometer was used to measure the tidal volume (TV, mL), minute ventilation (MV, mL), maximum inspiratory flow rate (PIF, mL / s) and other parameters of the mice in the same treatment group after radon inhalation poisoning to reflect the lung function level of the mice. The results are as follows: Figures 3 to 5 As shown. Figures 3 to 5 It can be seen that exposure to different radon levels can reduce the tidal volume, minute ventilation and maximum inspiratory flow of mice, indicating that the lung respiratory function of mice is inhibited, which is common in restrictive lung diseases such as pulmonary fibrosis.

[0052] (3) Lung tissue sections of mice in different treatment groups were subjected to H&E staining. Semi-quantitative lung pathology scoring was performed according to the method of Szapiel et al. (doi: 10.1164 / arrd.1979.120.4.89; doi: 10.1136 / jcp.41.4.467) to quantify the degree of lung injury. The scores of three sections of each mouse were averaged, and the average of three mice in each group was again taken to finally obtain the lung injury score of the mice in that group. The results are shown in Table 1. Figure 6 and Figure 7 As shown. Figure 6 and Figure 7 It can be seen that radon exposure caused abnormal expansion of the alveolar cavity of mice, thickening and rupture of the lung septum, accompanied by more fibrin exudation, obvious inflammatory infiltration around the bronchus, and congestion in small blood vessels. As the exposure dose accumulated, the degree of lung damage worsened.

[0053] (4) Masson staining was performed on lung tissue sections of mice in different treatment groups, and the collagen content was quantitatively analyzed using Image J software. The average value of three sections of each mouse was taken, and the average value of three mice in each group was taken again. Finally, the quantitative analysis results of collagen in the lung tissue of this group of mice were obtained. The results are shown in Figure 8 and Fig. 9 As shown. Figure 8 and Fig. 9 It can be seen that exposure to different radon levels promoted collagen deposition in the lung tissue of mice to varying degrees.

[0054] Example 2

[0055] 1. Experimental Materials

[0056] (1) Experimental animals: 24 C57 mice purchased from SpEffect Biotechnology.

[0057] (2) Experimental reagents: biliverdin, CAS number: 114-25-0;

[0058] Vehicle solvent: 0.2NNaOH, pH adjusted to 7.4 with 1N HCl;

[0059] H&E staining kit, purchased from Solebol;

[0060] Masson staining kit was purchased from Solebro.

[0061] 2. Model Construction

[0062] The experimental mice were randomly divided into a solvent control group (Rn+Vehicle) and a metabolite intervention group (Rn+BV), with 12 mice in each group. They were exposed to a HD-3 multifunctional radon chamber with a radon exposure concentration of 100,000 Bq / m 3 The exposure time was 377 and 755 h, respectively, and the relative cumulative exposure dose was 60 and 120 working level months (WLM); among them, the mice in the metabolite intervention group were intraperitoneally injected with biliverdin (dissolved in Vehicle solvent) at a dose of 35 mg / kg 24 h and 2 h before radon exposure to construct a metabolite intervention mouse model; the mice in the solvent control group were intraperitoneally injected with the same dose of Vehicle solvent as that in the metabolite intervention group 24 h and 2 h before radon exposure to construct a solvent control intervention mouse model.

[0063] 3. Experimental testing and conclusions

[0064] (1) The dynamic changes of weight gain and lung coefficient of mice in different treatment groups after radon inhalation were detected; the weight gain was the weight after exposure (g) minus the weight before exposure (g); the lung coefficient was obtained by dividing the lung wet weight (mg) by the body weight (g). The test results are as follows: Fig.10 and Fig.11 As shown. Fig.10 and Fig.11 It can be seen that biliverdin can effectively inhibit the weight loss and increase of lung coefficient in radon-exposed mice, inhibit the occurrence of pulmonary fibrosis, and prevent and treat lung damage.

[0065] (2) Non-invasive small animal respirometer was used to measure the tidal volume (TV, mL), minute ventilation (MV, mL), maximum inspiratory flow rate (PIF, mL / s) and other parameters of the mice in the same treatment group after radon inhalation poisoning to reflect the lung function level of the mice. The results are as follows: Figures 12-14As shown. Figures 12-14 It can be seen that biliverdin can significantly improve the decrease in minute ventilation and maximum inspiratory flow of mice caused by radon exposure, improve lung function, and improve the lung function of mice after radon exposure.

[0066] (3) Lung tissue sections of mice in different treatment groups were subjected to H&E staining. Semi-quantitative lung pathology scoring was performed according to the method of Szapiel et al. (doi: 10.1164 / arrd.1979.120.4.89; doi: 10.1136 / jcp.41.4.467) to quantify the degree of lung injury. The scores of three sections of each mouse were averaged, and the average of six mice in each group was taken to finally obtain the lung injury score of the group of mice. The results are shown in the figure. Fig.15 and Fig.16 As shown. Fig.15 and Fig.16 It can be seen that biliverdin can significantly improve lung damage caused by radon exposure.

[0067] (4) Masson staining was performed on lung tissue sections of mice in different treatment groups, and the collagen content was quantitatively analyzed using Image J software. The average value of three sections for each mouse and the average value of six mice in each group were taken to finally obtain the quantitative analysis results of collagen in the lung tissue of the mice in this group. The results are shown in Figure 2. Fig.17 and Fig.18 As shown. Fig.17 and Fig.18 It can be seen that biliverdin can significantly improve collagen deposition and lung damage caused by radon exposure.

[0068] Based on the above content, it can be seen that biliverdin can prevent and treat lung damage, especially lung damage caused by radon exposure.

[0069] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of biliverdin in the preparation of products for preventing and treating lung damage caused by radon exposure.

2. The use according to claim 1, characterized in that: The lung damage caused by radon exposure includes lung respiratory function damage caused by radon exposure.

3. The use according to claim 2, characterized in that: The radon exposure-induced pulmonary respiratory function damage includes radon exposure-induced decrease in minute ventilation and / or maximum inspiratory flow.

4. The use according to claim 1, characterized in that: The radon exposure-induced lung damage includes radon exposure-induced pulmonary fibrosis.

5. The use according to claim 1, characterized in that: The radon exposure-induced lung damage includes radon exposure-induced lung collagen deposition.

6. The use according to claim 1, characterized in that: The radon exposure-induced lung damage includes radon exposure-induced lung coefficient increase.

7. The use according to any one of claims 1 to 6, characterized in that: The relative cumulative exposure dose of radon in the radon exposure is 30 to 120 WLM.

8. The use according to claim 7, characterized in that: The radon exposure concentration is 100000Bq / m 3 , time is 189~755h.

9. A drug for preventing and treating lung damage caused by radon exposure, characterized in that: The active ingredient of the drug includes biliverdin.

10. The drug according to claim 9, characterized in that The drug is formulated as a single-use dosage form with 35 mg biliverdin per dose.

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