阅读说明：本技术 羊齿烯醇用于治疗肺纤维化的用途 (Application of fernasenol in treating pulmonary fibrosis ) 是由 于海涛 杨晓旭 于 2020-07-13 设计创作，主要内容包括：本发明公开了羊齿烯醇用于治疗肺纤维化的用途。研究发现,羊齿烯醇可以有效抑制人胚肺成纤维细胞株MRC-5的增殖,还可以有效促进人胚肺成纤维细胞株MRC-5的凋亡,因此羊齿烯醇具有开发成抗肺纤维化药物的前景。(The invention discloses application of ferenol in treating pulmonary fibrosis. Researches show that the ferenol can effectively inhibit the proliferation of a human embryonic lung fibroblast cell strain MRC-5 and can effectively promote the apoptosis of the human embryonic lung fibroblast cell strain MRC-5, so the ferenol has the prospect of developing anti-pulmonary fibrosis drugs.)

1. Application of fernasenol in preparing medicines for treating pulmonary fibrosis is disclosed.

2. A medicament for treating pulmonary fibrosis, which is characterized in that: takes the fersenol as an active component and is prepared into a pharmaceutically acceptable preparation formulation by using pharmaceutically acceptable auxiliary materials.

3. The medicament of claim 2, wherein: the auxiliary material can be solid auxiliary material or liquid auxiliary material.

4. The medicament of claim 2, wherein: the preparation can be tablet, capsule, granule, and injection.

Technical Field

The invention relates to medical application of fersenol, in particular to application of fersenol in treating pulmonary fibrosis.

Background

Pulmonary fibrosis is caused by various pulmonary interstitial diseases, progressive dyspnea occurs after the pulmonary fibrosis is serious, and patients die due to respiratory failure and exhaustion, and no particularly effective medicine is used for treating the pulmonary fibrosis at present.

The lung fibroblasts are the most important cells in the formation process of pulmonary fibrosis, and on one hand, the cells can continuously proliferate and synthesize a large amount of collagen to participate in the pulmonary fibrosis, and on the other hand, the cells can secrete a plurality of cytokines to promote the progression of the pulmonary fibrosis. Therefore, inhibiting the proliferation of lung fibroblasts can effectively inhibit the progression of pulmonary fibrosis.

Disclosure of Invention

The invention aims to provide the application of the fersenol in treating pulmonary fibrosis.

The technical scheme for realizing the purpose is as follows:

application of fernasenol in preparing medicines for treating pulmonary fibrosis is disclosed.

A medicine for treating pulmonary fibrosis is prepared from fersenol as active component and pharmacologically acceptable auxiliaries through proportional mixing.

Preferably, the adjuvant may be a solid adjuvant or a liquid adjuvant.

Preferably, the dosage form can be tablets, capsules, granules and injections.

Has the advantages that:

researches show that the ferenol can effectively inhibit the proliferation of a human embryonic lung fibroblast cell strain MRC-5 and can effectively promote the apoptosis of the human embryonic lung fibroblast cell strain MRC-5, so the ferenol has the prospect of developing anti-pulmonary fibrosis drugs.

Drawings

FIG. 1 shows the proliferation inhibition rate of human embryonic lung fibroblast cell line MRC-5 after different concentrations of drug intervention for 48 h.

FIG. 2 shows the effect of the drug on the expression of the apoptosis proteins Bcl-2 and Bax in human embryonic lung fibroblast strain MRC-5, wherein B is a control group, Y is ferylenol group, and P is taraxanol group.

Detailed Description

First, test materials

Human embryonic lung fibroblast line MRC-5 was purchased from Yaji biology, Shanghai, and frozen in liquid nitrogen.

The purity of the prenyl alcohol and the taraxacin is not less than 98 percent, and the prenyl alcohol and the taraxacin are prepared into a medicinal solution for later use.

RPMI1640 medium, fetal bovine serum was purchased from Gibco.

Penicillin, streptomycin was purchased from Sigma.

The CCK8 test kit was purchased from Nanjing Binyan Yuntan.

Second, test method

1. Cell culture

Recovering human embryonic lung fibroblast strain MRC-5 frozen in liquid nitrogen according to conventional method, suspension culturing in RPMI1640 culture solution containing 10% fetal calf serum, 100U/ml penicillin and 100 μ g/ml streptomycin, at 37 deg.C and 5% CO₂And culturing in a constant-temperature incubator with saturated humidity, changing liquid for passage every 2-3 d, and taking cells in logarithmic growth phase for experiment.

2. CCK8 method for determining proliferation inhibition effect of drug on human embryonic lung fibroblast cell line MRC-5

Taking human embryonic lung fibroblast cell line MRC-5 in logarithmic growth phase, digesting and re-suspending to prepare the human embryonic lung fibroblast cell line MRC-5 with the density of 5 × 10⁴The cell suspension/mL was inoculated into a 96-well plate at an inoculum size of 200. mu.L per well, acclimatized for 24 hours, the medium was replaced with a complete medium containing 2, 5, 10. mu.M of prenol or taraxyl alcohol at 3 duplicate wells per concentration, and a cell-free medium was placed in a blank set at 37 ℃ in 5% CO₂And after the culture is continued for 48 hours in the constant temperature incubator with saturated humidity, 10 mu L of CCK8 reagent is added into each hole, the OD450 value of each hole is measured at the wavelength of 450nm of an enzyme linked immunosorbent assay (ELISA) detector after the incubation is carried out for 4 hours, the average value of 3 holes is taken, and the proliferation inhibition rate is calculated.

The proliferation inhibition ratio (%) was [ 1- (OD drug group-OD blank)/(OD control group-OD blank) ] × 100%.

3. WB method for determining influence of drug on expression of apoptosis proteins Bcl-2 and Bax in human embryonic lung fibroblast cell line MRC-5

Taking human embryonic lung fibroblast cell line MRC-5 in logarithmic growth phase, digesting and re-suspending to prepare the human embryonic lung fibroblast cell line MRC-5 with the density of 1 × 10⁵The cell suspension/mL, 2mL per well was inoculated into 6-well plate, after adaptive culture for 24h, the medium was replaced with complete medium containing 5. mu.M of ferocinolol or taraxyl alcohol, and cells cultured without drug were used as a control group and placed at 37 ℃ in 5% CO₂And continuously culturing for 48 hours in a constant-temperature incubator with saturated humidity, collecting cells, washing by PBS, cracking by lysate, determining the protein concentration, performing SDS-PAGE electrophoresis on each group of 30 mu g of protein, transferring membranes, sealing by 5% of skimmed milk powder, adding Bcl-2, Bax and GAPDH primary antibodies, incubating overnight at 4 ℃, washing the membranes, adding horseradish peroxidase-labeled secondary antibodies, incubating for 2 hours at room temperature, developing, and performing gray scanning analysis.

4. Statistical analysis

Software SPSS17.0 is adopted for statistical analysis processing, the metering data is expressed by mean +/-standard deviation, t test is adopted for comparison among groups, variance analysis is adopted for comparison among groups, and the difference P < 0.05 has statistical significance.

Third, test results

1. Proliferation inhibiting effect on human embryonic lung fibroblast cell line MRC-5

The proliferation inhibition rates of the prenyl alcohol and the taraxerol with different concentrations on the human embryonic lung fibroblast strain MRC-5 in 48h are shown in table 1 and figure 1, and the prenyl alcohol and the taraxerol can obviously inhibit the proliferation of the human embryonic lung fibroblast strain MRC-5 and have obvious dose-dependent effect.

TABLE 1 inhibition rate of different concentrations of drug intervention on proliferation of human embryonic lung fibroblast cell line MRC-5 for 48h

2. Apoptosis promoting effect on human embryonic lung fibroblast strain MRC-5

FIG. 2 shows the effect of prenyl alcohol and taraxerol on the expression of apoptosis proteins Bcl-2 and Bax in human embryonic lung fibroblast MRC-5 (B is a control group, Y is prenyl alcohol group, and P is taraxerol group). Bcl-2 and Bax are the most important cytokines for regulating apoptosis, Bcl-2 inhibits apoptosis, and Bax promotes apoptosis. It can be found that the expression of Bcl-2 protein is obviously reduced and the expression of Bax protein is up-regulated by both the fernasenol and the taraxerol, and the influence of the taraxerol on the expression levels of the Bcl-2 and Bax protein is stronger than that of the fernasenol, and both the fernasenol and the taraxerol have obvious effects of promoting the apoptosis of the human embryonic lung fibroblast strain MRC-5.