Zhou Xiao-Liang, Liu De-Wu, Mao Yuan-Gui, Lü Jing
Zhonghua Zheng Xing Wai Ke Za Zhi. 2013 Nov;29(6):406-12.
To observe the effect of tetrandine on gene expression of collagen type I, collagen type III, transformation growth factor-beta1 and to investigate the inhibitory effect of tetrandine on the scar tissue hyperplasia in rabbits' ears.
After the scar model was formed on the rabbits' ears, the rabbits were divided into 4 groups to receive intro-lesion injection with saline, or prednisolone (Pre) or tetrandrine in low concentration (L-Tet, 1.0 mg/ml) or tetrandrine in high concentration (H-Tet, 7.5 mg/ml). The morphological changes of scar tissue were observed. The changes of fibroblasts quantity and collagen expression were observed with HE and Masson staining. Immunohistochemical study was used to observe the expression level of collagen type I and collagen type III and TGF-beta1. Collagen type I and collagen type III and TGF-beta1, and signal factor Smad 3 mRNA were detected with RT-PCR.
(1) 24 days after injury, all the wounds healed completely with formation of red, tough and hypertrophic scar. HE and Masson staining showed significant increase of fibroblasts and collagen density with irregularly arrangement. (2) Compared with that in saline group, the scar in other groups became softer, lighter and thinner, especially in H-Tet group. (3) HE and Masson staining shows the scar in Tet and Pre groups contained less fibroblasts and lower collagen dentsity with comparatively regular arrangement than that in saline group (P < 0.01), especially in H-Tet group. (4) According to the immunohistochemical study, the expression of collage type I and III and TGF-beta was positive in all the groups, but the positive rate and the ratio of collagen density I to III decreased in the order of saline, L-Tet, H-Tet and Pre groups (P < 0.01). (5) PT-PCR detection results showed that the amplification bands brightness of collagen type I and III and TGF-beta1 and signal molecular Smad 3 mRNA in scar tissue were obviously different. Compared with that in saline group, the expression of collagen type I and III and TGF-beta1 and Smad 3 mRNA decreased in Tet and Pre groups (P < 0.01). H-Tet group showed the most obvious reduce in the expression of type I collagen and TGF-beta1 and Smad 3 mRNA. Conclusions Tetrandine can significantly suppress the expression of collagen type I and collagen type III and TGF-beta1 on hypertrophic scar of rabbit ears, and reduce signal factor Smad 3 mRNA' s expression. It may be one of the important mechanism for its inhibitory effect on scar hyperplasia.
观察粉防己碱对Ⅰ型胶原、Ⅲ型胶原、转化生长因子-β1基因表达的影响，探讨粉防己碱对兔耳瘢痕组织增生的抑制作用。
兔耳瘢痕模型形成后，将兔分为4组，分别于瘢痕内注射生理盐水、泼尼松龙（Pre）、低浓度粉防己碱（L-Tet，1.0mg/ml）或高浓度粉防己碱（H-Tet，7.5mg/ml）。观察瘢痕组织的形态学变化。采用HE和Masson染色观察成纤维细胞数量及胶原表达的变化。采用免疫组织化学法观察Ⅰ型胶原、Ⅲ型胶原及TGF-β1的表达水平。采用RT-PCR检测Ⅰ型胶原、Ⅲ型胶原、TGF-β1及信号因子Smad 3 mRNA。
（1）伤后24天，所有创面均完全愈合，形成红色、坚韧、肥厚的瘢痕。HE和Masson染色显示成纤维细胞和胶原密度显著增加，排列不规则。（2）与生理盐水组相比，其他组瘢痕变软、变浅、变薄，尤其是H-Tet组。（3）HE和Masson染色显示，Tet组和Pre组瘢痕中成纤维细胞较少，胶原密度较低，排列较生理盐水组规则（P<0.01），尤其是H-Tet组。（4）免疫组织化学研究显示，所有组Ⅰ型胶原、Ⅲ型胶原及TGF-β表达均为阳性，但阳性率及胶原密度Ⅰ/Ⅲ比值按生理盐水组、L-Tet组、H-Tet组和Pre组顺序降低（P<0.01）。（5）PT-PCR检测结果显示，瘢痕组织中Ⅰ型胶原、Ⅲ型胶原、TGF-β1及信号分子Smad 3 mRNA的扩增条带亮度明显不同。与生理盐水组相比，Tet组和Pre组Ⅰ型胶原、Ⅲ型胶原、TGF-β1及Smad 3 mRNA表达降低（P<0.01）。H-Tet组Ⅰ型胶原、TGF-β1及Smad 3 mRNA表达降低最明显。结论粉防己碱可显著抑制兔耳增生性瘢痕中Ⅰ型胶原、Ⅲ型胶原及TGF-β1 的表达，降低信号因子Smad 3 mRNA的表达。这可能是其抑制瘢痕增生的重要机制之一。
