| Objective: To observe the effect of tetrandine on genetic experession of collagenI/III、 transformation growth factor-β1and suppression effect of tetrandine onhypertrophic scar of rabbits’ ear, so as to provide theoretical support for its furtherclinical application.Methods: Ten big-eared Japanese rabbits were used to establish animal model forhypertrophic scars. Six square wounds sized1.0cm2on rabbits ears were made,full-thickness skin along with perichondrium was removed and cartilage was kept.Wound interval was kept with1.0cm.24days after injury, seventy-two scars’samples were divided into four groups randomly: The samples of group A were gevennormal saline, group B prednisolone, group C low-concentration tetrandine1.0mg/ml,group D high-concentration tetrandine7.5mg/ml. The bases of scar tissues in everygroup were injected into the above corresponding liquid20ul, every3days one time,totally for10times. Morphological changes of wound and scar were dynamicallyobserved; The number of fibroblasts and expression and arrangement changes ofcollagen were observed by HE and Masson staining; The expression of collagen I/IIIand transformation growth factor-β1were observed by ImmunoHistochemical andimage analysis. The mRNA expression of collagen I/III and transformation growthfactor-β1and signal factor Smad3were detected by RT-PCR.Results:(1) Wound bengan scabby after3-5days, completely scabby after6days,most of the distal wound healed in15days.24days after injury, all woundepidermized and formed tough, dark-redded hyperplasia tissues outstanding leatherface. HE and Masson staining shows: In scar hyperplasia tissues, the number offibroblasts and the density of collagen were significantly higher than normal skin. Inaddition, the arrangement of collagen is disorder and vortex.(2) Compared withsaline, the degree of congestion, hardness, and thicknesson in scar in tetrandrine andprednisolone groups is reduced. The morphological improvement of tetrandrine groupis better than that of prednisolone group, and high-concentration tetrandrine groupshowed the best improvement.3) HE and Masson staining shows: Compared with saline group, the number of fibroblasts and density of collagen in tetrandrine andprednisolone group was significantly reduced (P <0.01), and the arrangement ofcollagen was sparse, no vortex and nodules were found. High-concentrationtetrandrine group show the most obvious reduction in fibroblast quantity and collagendensity.(4) Immunohistochemical shows: In each group, collagen I/III and TGF-β1were positively expressed; The surface density of collagen type I and III and thepositive expression rate of TGF-β1in saline group, prednisolone group, low-concentration Tet group and high-concentration Tet group are in turn decrease (P <0.01); Another collagen type I and III surface density ratio also declined in turn:saline groups> low concentration Tet group> high concentration Tet group andprednisolone group.(5) RT-PCR detection results: the amplification bands brightnessof collagen I/III and TGF-β1and signal molecular Smad3mRNA in scar tissueshowed significant difference between groups; Compared with saline group, collagenI/III and TGF-β1and Smad3mRNA expressed lower in tetrandine group andprednisolone group (P <0.01); The expression of type I collagen and TGF-β1andSmad3mRNA showed the most obvious reduction in high-concentration Tet group.Conclusion: Tetrandine can significantly suppress the expression of collagen I/III andTGF-β1and signal factor Smad3mRNA, and effectively inhibit the proliferation offibroblasts. The reduction in the content and ratio of collagen type I and III, reduceTGF β1expression. It’s may be one of the important mechanism for tetrandineinhibit scar hyperplasia. |
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