Cryptotanshinone for preventing and treating pulmonary fibrosis and application thereof
A technology for cryptotanshinone and pulmonary fibrosis, which is applied in the field of medicine and can solve problems such as the absence of cryptotanshinone
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experiment example 1
[0025] Experimental example 1 The effect of cryptotanshinone on the proliferation of human embryonic lung fibroblast (HLF) cells:
[0026] (1) Experimental materials: HLF cells were purchased from the Experimental Animal Center of Sun Yat-sen University. DMEM medium (Gibco, USA), fetal bovine serum-like (Hyclone, USA), MTT (Sigma);
[0027] (2) Experiment method: 5.0×10 3 The HLF cells were added to the wells of a 96-well plate and cultured for 24 h. Aspirate the original culture solution, add the complete culture solution containing different concentrations (2.5 μM, 5 μM, 10 μM, 20 μM) of cryptotanshinone, and add the complete culture solution without cryptotanshinone as a normal control. Continue to culture for 48 h, add 20 μL of phosphate-buffered saline containing 5 mg / mL MTT to each well, after 4 h of culture, aspirate the culture medium in each well, and add 200 μL of dimethyl sulfoxide (DMSO ). Shake for 15 min, at a wavelength of 490 nm, detect the absorbance value...
experiment example 2
[0031] Experimental Example 2 The effect of cryptotanshinone on the abnormal activation of human embryonic lung fibroblast (HLF) cells stimulated by TGF-β1:
[0032] (1) Experimental materials: HLF cells were purchased from the Experimental Animal Center of Sun Yat-sen University. DMEM medium (Gibco, USA), fetal bovine serum-like (Hyclone, USA), human transforming growth factor-β1 (HumanTGF-β1, PeproTech Inc);
[0033] (2) Experimental method: 5.0×103 HLF cells were added to the wells of a 96-well plate and cultured for 24 hours. Divide lung fibroblasts into:
[0034] TGF-β1 treatment group (5 ng / mL action time 48 h); TGF-β1 treatment group (5 ng / mL action time 48 h) + cryptotanshinone 2.5 μM; TGF-β1 treatment group (5 ng / mL action time 48 h) + Cryptotanshinone 5 μM; TGF-β1 treatment group (5 ng / mL for 48 h) + cryptotanshinone 10 μM. TGF-β1 treatment group (5 ng / mL for 48 h) + cryptotanshinone 20 μM. After 48h, BrdU was used to detect cell proliferation. Taking the TGF-β1...
experiment example 3
[0040] Experimental Example 3 The effect of cryptotanshinone on the expression of fibrin in lung fibroblasts stimulated by TGF-β1:
[0041] (1) Materials and methods: HLF cells were purchased from the Experimental Animal Center of Sun Yat-sen University;
[0042] DMEM medium (Gibco, USA), human transforming growth factor-β1 (HumanTGF-β1, PeproTech Inc), α-SMA antibody (Boster Company), mouse secondary antibody (Thermo, USA); trypsin, α-tubulin antibody (Sigma USA ), rabbit secondary antibody (Promega, USA);
[0043] (2) Test method: cells were seeded in 6-well plates and cultured in a cell incubator. When the cells grew confluent to 70-80%, they were starved for 24 h with serum-free medium, and replaced with fresh medium, and the passaged HLF cells were divided into: control group, cryptotanshinone group, cryptotanshinone 25 μM, and the action time was 48 h TGF-β1 Treatment group (5 ng / mL action time 48 h); TGF-β1 treatment group (5 ng / mL action time 48 h) + cryptotanshinone...
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