Application of astilbe chinensis in preparation of medicine for preventing and treating acute lung injury

Abstract

The invention discloses application of astilbe chinensis in preparation of a medicine for preventing and treating acute lung injury, and belongs to the field of biological pharmacy. According to the astilbe chinensis aqueous extract prepared by the invention, through the intervention effect on an acute lung injury mouse model, the astilbe chinensis has the effect of inhibiting the acute lung injury, a new medicine application is provided for the astilbe chinensis aqueous extract, and the astilbe chinensis aqueous extract has important clinical significance.

Description

Technical Field

[0001] The invention belongs to the field of biopharmaceuticals, and particularly relates to the application of Astilbe chinensis in preparing a medicine for preventing and treating acute lung injury. Background Art

[0002] Acute lung injury (ALI) is a respiratory emergency with pathological manifestations of acute hypoxic insufficiency or respiratory failure, increased alveolar permeability, and severe alveolar edema. In severe cases, acute respiratory distress syndrome (ARDS) may occur, which is characterized by severe lung inflammation, destruction of the alveolar capillary barrier, and pulmonary edema. ALI can lead to the development of chronic diseases, such as idiopathic pulmonary fibrosis (IPF) [Kligerman S. Pathogenesis, Imaging, and Evolution of Acute Lung Injury. Radiol Clin North Am. 2022 Nov; 60(6): 925-939.], chronic obstructive pulmonary disease (COPD), asthma, lung cancer, etc. [Ayilya BL, Balde A, Ramya M, Benjakul S, Kim SK, Nazeer RA. Insights on the mechanism of bleomycin to induce lung injury and associated in vivo models: A review. Int Immunopharmacol. 2023 Aug; 121: 110493.]. ALI is a common clinical emergency with a mortality rate of up to 40%. However, there is currently no specific treatment strategy for ALI in clinical practice.

[0003] Astilbe chinensis, scientific name Astilbe chinensis (Maxim.) Franch. et Savat., belongs to the genus Astilbe of the Saxifragaceae family. In Zhejiang, Astilbe chinensis is a commonly used medicinal material. It has the effects of clearing away heat and relieving exterior symptoms, relieving cough, removing phlegm and relieving asthma. It is widely used in the treatment of colds, coughs and other diseases. Astilbe contains a variety of active ingredients, such as bergenin, which exhibit multiple pharmacological effects such as anti-inflammatory, antibacterial, antiviral, expectorant and antitussive effects [Tang Q, Wang Q, Sun Z, et al. Bergenin Monohydrate Attenuates Inflammatory Response via MAPK and NF-κB Pathways Against Klebsiella pneumonia Infection. Front Pharmacol. 2021 May 4; 12: 651664.][Yang L, Zheng Y, Miao YM, et al. Bergenin, a PPARγagonist, inhibits Th17 differentiation and subsequent neutrophilic asthma by preventing GLS1-dependent glutaminolysis. Acta Pharmacol Sin. 2022 Apr; 43(4): 963-976.]. These mechanisms of action provide theoretical support for the use of Astilbe in the treatment of lung inflammation. Pharmacodynamic tests show that the water extract of Astilbe can significantly inhibit the swelling of the auricle of mice and counteract the swelling of the plantars of mice caused by Chondrus [Jiang Shipeng, Lai Pingfan, Yang Yiling. Experimental study on the antipyretic, anti-inflammatory, antibacterial and anti-endotoxin effects of Astilbe root [J]. Chinese Medicinal Materials, 2007, (01): 77-80.], indicating that it has a significant anti-inflammatory effect. In addition, the "Chinese Pharmacopoeia" points out that bergenin in Astilbe is an effective drug for treating bronchitis. Experimental studies have confirmed that Astilbe can significantly prolong the cough latency of mice, reduce the number of coughs, and increase the excretion of phenol red, indicating that it has a good expectorant and antitussive effect [Lai Pingfan, Jiang Shipeng, Yang Yiling. Research on the analgesic, blood-activating, antitussive and expectorant effects of Astilbe root. Chinese Journal of Traditional Chinese Medicine. 24 (2006): 2061-2064.]. This has a significant effect on relieving symptoms such as cough and expectoration caused by lung inflammation. In summary, Astilbe chinensis has anti-inflammatory, analgesic, antitussive and expectorant effects, and can treat bronchitis, but there are no reports on the effects of Astilbe chinensis granules on lung inflammation in acute lung injury, and there are no reports on the study of the mechanism of action of Astilbe chinensis in the direct treatment of ALI. Summary of the invention

[0004] Based on the deficiencies in the prior art, the purpose of the present invention is to provide a new pharmaceutical use of Astilbe chinensis in treating, alleviating or preventing ALI, namely, the use of Astilbe chinensis in the preparation of a drug for preventing and treating acute lung injury.

[0005] In order to achieve the above-mentioned purpose, the specific technical scheme of the present invention is as follows:

[0006] The invention provides application of Astilbe chinensis in preparing medicine for preventing and treating acute lung injury.

[0007] Specifically, the Astilbe truncatum is an aqueous extract of Astilbe truncatum, and the preparation method is as follows: boiling Astilbe truncatum with water to obtain the aqueous extract of Astilbe truncatum.

[0008] Preferably, the Astilbe chinensis water extract contains 5 g of crude drug per milliliter.

[0009] In some embodiments of the present invention, the dosage form of the drug is liquid, powder injection, tablet, capsule, powder, pill, paste, granule or dressing.

[0010] Preferably, when the drug is in the form of a liquid, the dosage of the drug is 1.56 to 15.6 g / kg. The drug is administered continuously for 7 days.

[0011] The mice were randomly divided into a normal group, a model group, a dexamethasone group (DXM), a high-dose Astilbe nutans group (LXF H), a medium-dose Astilbe nutans group (LXF M), and a low-dose Astilbe nutans group (LXF L). The drug-administered groups were given preventive medication for 7 consecutive days. After 7 days of medication, the model group and the four drug-administered groups were respectively given LPS for tracheal instillation to establish models.

[0012] After LPS modeling, mice were anesthetized, intubated, and placed in the forced lung function instrument chamber for lung function testing. Astilbe chinensis can reduce the functional residual capacity (FRC) of ALI mice and alleviate lung function damage.

[0013] After the mice were killed, part of the lung tissue was immediately removed and added to 10% serum culture medium to process into a single cell suspension, and the polarization of macrophages in the lung tissue cells was detected. High doses of Astilbe chinensis can significantly increase the proportion of M2 cells in the lung tissue of ALI mice and promote the repair of lung damage.

[0014] After the mice were killed, part of the lung tissue was taken and added to PBS to prepare a 10% homogenate, and the supernatant was centrifuged to detect IL-6, IL-1β, and TNFα inflammatory factors. Medium and low doses of Astilbe chinensis can significantly reduce the content of inflammatory factors in lung tissue homogenate.

[0015] Part of the lung tissue was fixed in neutral formalin, embedded in paraffin, sliced, and stained with HE to observe the changes in the pathological morphology of the lung tissue of each group of mice. The high-dose and medium-dose groups of Astilbe chinensis can improve the infiltration of bronchial inflammatory cells in the lungs of ALI mice, reduce the thickness of the airway wall, and relieve airway inflammation.

[0016] Part of the lung tissue was quickly frozen in liquid nitrogen after being removed. The total RNA in the lung tissue was extracted using a rapid RNA extraction kit, and then reverse transcribed into cDNA. The relative expression of genes of CCR1 and CCR5 targets in the chemokine signaling pathway was analyzed using qRT-PCR. GAPDH was selected as an endogenous reference. Each dose group of Astilbe chinensis can significantly reduce the relative expression of CCR1 and CCR5 genes in the lung tissue of ALI mice.

[0017] Animal model studies have found that Astilbe chinensis can significantly increase the M2 ratio, reduce the animal's forced lung function FRC index, alleviate pathological changes of lung inflammation, reduce the inflammatory factors TNF-α and IL-6 in lung tissue, and reduce the expression of CCR1 and CCR5 genes.

[0018] The present invention also provides a medicine for preventing and treating acute lung injury, which is Astilbe truncatum itself or a water extract of Astilbe truncatum, and a pharmaceutically acceptable salt thereof.

[0019] Preferably, the drug is a liquid, the dosage of the drug is 1.56 to 15.6 g / kg, and the drug is administered continuously for 7 days.

[0020] Beneficial effects of the present invention:

[0021] The present invention prepares the Astilbe chinensis water extract, and through the intervention effect on the ALI mouse model, clarifies that Astilbe chinensis has the effect of inhibiting ALI, provides a new drug use for the Astilbe chinensis water extract, and has important clinical significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The effect of Astilbe chinensis on lung function of ALI mice. Compared with the normal group, "Δ" p<0.05, "ΔΔ" p<0.01; compared with the model group, "*" p<0.05.

[0023] Figure 2 The effect of Astilbe chinensis on inflammatory factors (IL-6 and TNFα) in lung tissue of ALI mice. Compared with the normal group, "Δ" p<0.05, "ΔΔ" p<0.01; compared with the model group, "*" p<0.05.

[0024] Figure 3The effect of Astilbe chinensis on the pathological structural changes of the lung airway tissue in ALI mice; A is the airway inflammation score; B is the airway wall thickness (μm); C is the normal group, D is the model group, E is the dexamethasone group, F is the Astilbe chinensis high-dose group, G is the Astilbe chinensis medium-dose group, and H is the Astilbe chinensis low-dose group. The scale bar is 100μm. Compared with the normal group, "Δ" p < 0.05, "ΔΔ" p < 0.01; compared with the model group, "*" p < 0.05, "**" p < 0.01.

[0025] Figure 4 The effect of Astilbe chinensis on the polarization of macrophages in lung tissue of ALI mice. "ΔΔ" compared with the normal group, p < 0.01; "**" compared with the model group, p < 0.01.

[0026] Figure 5 The effect of Astilbe chinensis on the expression of CCR1 and CCR5 genes in the lung tissue of ALI mice. Compared with the normal group, "Δ" p<0.05, "ΔΔ" p<0.01; compared with the model group, "*" p<0.05, "**" p<0.01. DETAILED DESCRIPTION

[0027] Example 1: Preparation method of Astilbe chinensis medicine

[0028] Take 5000g of Astilbe chinensis slices, grind them, add 5 times the amount of water as the medicinal material, soak for 30 minutes, boil at 100℃ for 1 hour, add water and boil, filter the boiled solution to obtain a filtrate, concentrate the filtrate to a clear paste, and prepare a water extract containing 5g of the crude drug per milliliter.

[0029] Example 2 Experiment on the treatment of acute lung injury in mice with Astilbe chinensis

[0030] 1 Experimental methods

[0031] (1) Animal grouping

[0032] The mice were randomly divided into normal group, model group, dexamethasone group (DXM), LXF high-dose group (LXF H), LXF medium-dose group (LXF M), and LXF low-dose group (LXF L), with 10 mice in each group.

[0033] (2) Animal modeling and drug dosage

[0034] LPS (lipopolysaccharide) modeling: 4 mg / kg LPS was instilled into the mouse trachea, with an LPS concentration of 4 mg / ml.

[0035] Dosage: Dexamethasone is administered by intraperitoneal injection at a dose of 5 mg / kg. Astilbe chinensis is administered by oral gavage, with a low dose of 1.56 g / kg according to the clinical dose, a medium dose of 7.8 g / kg, 5 times the clinical dose, and a high dose of 15.6 g / kg, 10 times the clinical dose.

[0036] (3) Animal modeling and drug administration

[0037] Except for the normal group and the model group, the mice were given preventive medication for 7 days. After 7 days of medication, the model group and the four medication groups were given LPS by tracheal instillation to establish models.

[0038] (4) Mouse forced lung function test

[0039] After LPS modeling of mice, the mice were anesthetized with 1% sodium pentobarbital at a dose of 50 mg / kg, tracheal intubation was performed, and the mice were placed in the cavity of a forced lung function instrument for lung function testing.

[0040] (5) Flow cytometry detection of lung tissue macrophage polarization

[0041] After the mice were killed, part of the lung tissue was immediately removed and added to 10% serum medium to make a single cell suspension. The single cell suspension was centrifuged (300g / 5min), the supernatant was discarded, and a certain amount of PBS was added to wash 1-2 times. The supernatant was discarded, the cells were flicked and resuspended to 5×10 6 -10×10 6 Take 500 μL of PBS to resuspend the cells in an EP tube or flow tube, take 100 μL in each tube, and add 5 μL of surface staining antibodies. Add CD11b, CD86, CD206, and F4-80 antibodies respectively and mix well to detect the polarization of macrophages in lung tissue cells.

[0042] (6) ELISA detection of inflammatory factors in lung tissue

[0043] After the mice were killed, part of the lung tissue was taken and added to PBS to prepare a 10% homogenate, and the supernatant was centrifuged to detect IL-6, IL-1β, and TNFα inflammatory factors.

[0044] (7) Pathological morphological changes of lung tissue

[0045] Part of the lung tissue was fixed in neutral formalin, embedded in paraffin, sectioned, and stained with HE to observe the changes in the pathological morphology of the lung tissue of mice in each group.

[0046] (8) Target gene expression in the chemokine signaling pathway of lung tissue

[0047] Part of the lung tissue was quickly frozen in liquid nitrogen after being removed. The total RNA in the lung tissue was extracted using a rapid RNA extraction kit, and then reverse transcribed into cDNA. The relative expression of genes of CCR1 and CCR5 targets in the chemokine signaling pathway was analyzed using qRT-PCR. GAPDH was selected as an endogenous reference. The primer sequences of the target genes are shown in Table 1. The relative expression of the target genes was calculated using the 2-Δct method.

[0048] Table 1 Primer sequences

[0049]

[0050]

[0051] 2 head test results

[0052] (1) Effect of Astilbe chinensis on lung function of ALI mice: After LPS modeling, the Force Vital Capacity (FVC) of ALI mice was significantly reduced (p < 0.05), while the Functional Residual Capacity (FRC) was significantly increased (p < 0.05). The positive drug, high-dose and medium-dose Astilbe chinensis groups could significantly reduce FRC (p < 0.05). The results are shown in Figure 1 .

[0053] (2) Effect of Astilbe chinensis on inflammatory factors in lung tissue of ALI mice: After LPS modeling, the levels of inflammatory factors IL-6 and TNFα in lung tissue homogenate of ALI mice were significantly increased. Medium and low doses of Astilbe chinensis could significantly reduce the levels of inflammatory factors in lung tissue homogenate (p < 0.05). Figure 2 .

[0054] (3) Observation on the pathological morphology of lung tissue of ALI mice with Astilbe chinensis: After LPS modeling, the infiltration of inflammatory cells around the bronchi in the lung tissue of ALI mice was obvious, and the airway wall was significantly thickened. The high-dose and medium-dose groups of Astilbe chinensis could improve the infiltration of inflammatory cells around the bronchi in the lungs of ALI mice, reduce the thickness of the airway wall, and relieve airway inflammation (p < 0.05). The results are shown in Figure 3 .

[0055] (4) Effect of Astilbe chinensis on polarization of macrophages in lung tissue of ALI mice: After LPS modeling, the number of M2 cells in ALI mice decreased significantly (p < 0.01), while the number of M1 cells increased. However, there was no significant difference between the model group and the normal group. High doses of Astilbe chinensis can significantly increase the proportion of M2 cells (p < 0.01). Figure 4 .

[0056] (5) Effects of Astilbe chinensis on genes related to chemokine signaling pathways in lung tissue: Results Figure 5As shown, administration of Astilbe chinensis can significantly reduce the gene expression levels of chemokine receptors CCR1 and CCR5 (p < 0.01).

[0057] 3 Conclusion

[0058] Astilbe chinensis can protect the lung function of mice with acute lung injury, reduce the levels of inflammatory factors IL-6 and TNFα in the lung tissue of mice with acute lung injury, promote the differentiation ratio of anti-inflammatory macrophages M2 in lung tissue, alleviate the infiltration of inflammatory cells in the pathological structure of lung tissue and reduce airway wall thickness. Astilbe chinensis can significantly reduce the gene expression levels of chemokines CCR1 and CCR5, suggesting that the effect of Astilbe chinensis in treating acute lung injury may be through chemokine receptors acting on the chemokine signaling pathway, regulating the chemotaxis of inflammatory cells to alleviate lung inflammation.

Claims

1. Application of Astilbe chinensis in the preparation of medicines for preventing and treating acute lung injury.

2. The use according to claim 1, characterized in that The Astilbe truncatum is an aqueous extract of Astilbe truncatum, and the preparation method is as follows: Astilbe truncatum is decocted with water to obtain the aqueous extract of Astilbe truncatum.

3. The use according to claim 2, characterized in that Each milliliter of the Astilbe chinensis water extract contains 5 g of the crude drug.

4. The use according to claim 1, characterized in that The dosage form of the medicine is liquid, powder injection, tablet, capsule, powder, pill, paste, granule or dressing.

5. The use according to claim 4, characterized in that When the dosage form of the drug is a liquid, the dosage of the drug is 1.56 to 15.6 g / kg.

6. The use according to claim 5, characterized in that The drug was administered continuously for 7 days.

7. A drug for preventing and treating acute lung injury, characterized in that: The medicine is Astilbe truncatum itself or the water extract of Astilbe truncatum, and pharmaceutically acceptable salts thereof.

8. The drug for preventing and treating acute lung injury according to claim 7, characterized in that: The medicine is a liquid.

9. The drug for preventing and treating acute lung injury according to claim 8, characterized in that: The dosage of the drug is 1.56 to 15.6 g / kg.

10. The drug for preventing and treating acute lung injury according to claim 9, characterized in that: The drug was administered continuously for 7 days.

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