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Application of flavone compound in treating lung fibrosis

A technology of pulmonary fibrosis and composition, which is applied in the application field of neohesperidin in the treatment of pulmonary fibrosis, and can solve problems such as no neohesperidin

Inactive Publication Date: 2017-02-01
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] So far, there is no report that neohesperidin can inhibit TGF-β signaling pathway and slow down pulmonary fibrosis

Method used

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  • Application of flavone compound in treating lung fibrosis
  • Application of flavone compound in treating lung fibrosis
  • Application of flavone compound in treating lung fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Neohesperidin inhibits TGF-β1 signaling in a dose-dependent manner

[0022] The cells used in this example are NIH3T3 mouse fibroblasts stably transfected with the CAGA-luciferase reporter gene.

[0023] The NIH3T3 cells in the logarithmic growth phase were planted in a 96-well plate, and after growing to 80%, they were treated with 0.1% serum for 24 hours, and a final concentration of 5 ng / ml TGF-β1 pure protein and different concentrations of neohesperidin were added. The final concentrations were 0.1 μmol / L, 1 μmol / L, 5 μmol / L, and 10 μmol / L (three replicate wells were set for one compound), and cultured at 37°C for 24 hours. The cells in the 96-well plate were washed twice with PBS buffer. Add 50 μL of 1×passive (Passive) lysis buffer, place the 96-well plate on a horizontal shaker at 200 rpm / min, and lyse the cells at room temperature for 30 min. Take 40 μL of cell lysate into a 96-well plate for measurement, add 35 μL of substrate, and perform lucifera...

Embodiment 2

[0027] Example 2 Neohesperidin inhibits TGF-β1-induced myofibroblast differentiation

[0028]The cells used in this example were the mouse lung fibroblast cell line Mlg obtained from the American Type Culture Collection (ATCC). After Mlg fibroblasts were overgrown, 0.1% serum was treated for 24 hours, and 5 ng / mL LTGF-β1 protein was added with a final concentration of 5 μmol / L neohesperidin or hesperidin (as a positive control). After 12 hours of treatment, RNA was collected for real-time fluorescent quantitative PCR to detect α-SMA; after 24 hours of treatment, cells were collected and Western Blotting was used to detect the expression of α-SMA protein. Real-time fluorescent quantitative PCR refers to preparing total RNA from cells using Trizol (Invitrogen), synthesizing cDNA using ThermoScript RT-PCR kit (Invitrogen), and detecting the expression of α-SMA using SYBR Green kit (Roche). Relative gene expression using 2 -Δ(ΔCT) method, and β-actin was used as an internal refe...

Embodiment 3

[0035] Example 3 Neohesperidin slows down bleomycin-induced pulmonary fibrosis

[0036] Pulmonary fibrosis animal model preparation refers to male C57BL / 6J (age 8-10 weeks) wild-type mice, anesthetized mice with 0.6mL / 100g intraperitoneal (I.P.) injection of 7.5% chloral hydrate, intratracheal injection of Bray Mycin 2.5U / kg. The specific scheme is as follows: after weighing and recording the body weight, fix the mouse on the operating table, disinfect the neck with 70% alcohol, use a scalpel to cut vertically about 1 cm in the neck of the mouse, use micro-tweezers to separate the tissue to expose the trachea, Insert the syringe into the trachea through the gap between the tracheal cartilage rings towards the heart, then slowly inject 2.5 U / kg of bleomycin saline solution in a volume appropriate to its body weight, and immediately turn the animal upright and rotate left and right to make the liquid medicine Evenly distributed in the lungs.

[0037] Neohesperidin treatment re...

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PUM

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Abstract

The invention relates to application of a flavone compound in treating lung fibrosis, particularly relates to application of neohesperidin in treating lung fibrosis, particularly relates to restraining of the neohesperidin on expression of a CAGA-luciferase reporter gene induced by TGF-beta1 by using dose dependency, particularly relates to application of the neohesperidin in restraining in muscle fiber mother cell differentiation induced by TGF-beta1, and particularly relates to application of the neohesperidin in alleviating murine lung fibrosis induced by bleomycin.

Description

technical field [0001] The present invention relates to a new application of neohesperidin (Neohesperidin), in particular to the application of neohesperidin in treating pulmonary fibrosis. Background technique [0002] Based on the support of the National Natural Science Foundation of China (81430001 31471373 31271559) and the Tianjin Applied Basic and Frontier Technology Research Project (15JCYBJC29100), the present invention studies the material basis of the treatment of pulmonary fibrosis. [0003] The occurrence of pulmonary fibrosis is related to many factors, including smoking, air pollution, autoimmune diseases, viruses, inappropriate treatment methods, etc. For example, SARS virus can cause fibrosis in some patients, and the use of a large amount of antibiotics during treatment can accelerate the process of fibrosis, leading to serious sequelae. Idiopathic pulmonary fibrosis (IPF) is the most common, most serious, and most refractory disease among pulmonary fibrosi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61P11/00A23L33/00
CPCA61K31/7048A23V2002/00A23V2200/30
Inventor 宁文方银善
Owner NANKAI UNIV
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