Résumé
Objective To investigate the effect of artesunate on the expression of heat shock protein 47(Hsp47)in pulmonary fi-brosis in rats.Methods Thirty SD male rats were randomly divided into control group,model group and artesunate intervention group (intervention group).Rats in control group were intrachacheally instilled with saline (0.4 mL),while rats in other two groups 5 mg/kg bleomycin.Rats in both control group and model group were intraperitoneal injected with 1 mL saline per day sub-sequently.And the rats in intervented group were intraperitoneal injected with 10 mg/100 g artesunate per day.The body weight of day 0,day 14 and day 28 were recorded.All the rats were sacrificed on day 28.The pulmonary coefficient and the hydroxyproline content were observed.The expression of Hsp47mRNA and CollagenⅠmRNA were evaluated by RT-PCR.The Hsp47 expression was also detected by Western Blot.The pathological changes were analyzed by hamatoxylin-eosin (HE)stain and Masson stain.Re-sults (1)The body weight of control group grew much faster than other groups (P intervention group (8.54±0.67)mg/g>control group(4.81±0.38)mg/g (P control group (3.08±0.56),model group(5.70± 1.09)>intervention group (4.01 ± 1.25).There was no significant difference between control group and intervention group(P >0.05).(4)Hydroxyproline content:model group (620.33±66.16)μg/g>control group(379.00±35.51)μg/g,model group (620.33±66.16)μg/g >intervention group(429.00± 36.51)μg/g (P control group (P 0.05).CollagenⅠmRNA:model group > intervention group>control group (P <0.05).(6)Western blot showed that Hsp47 ex-pression of model group was the highest in all three groups.Conclusion These results indicate that artesunate might inhibit pulmo-nary fibrosis and depress CollagenⅠproduction,and it′s probably a result of Hsp47 down regulation.
Résumé
This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.