阳和解凝膏联合梅花针对局限性硬皮病小鼠TGF-β_(1)/Smad3信号通路的影响

The Effect of Yanghe Jiening Gao(阳和解凝膏)Combined with Plum-blossom Needle on TGF-β_(1)/Smad3 Signaling Pathway in Mice with Localized Scleroderma

目的:观察阳和解凝膏联合梅花针对局限性硬皮病小鼠模型转化生长因子-β_(1)(TGF-β_(1))/Smad同源物3(Smad3)信号通路的影响。方法:将36只雌性BALB/c小鼠随机分为空白对照组、模型对照组、肝素钠组、阳和解凝膏组、梅花针组、阳和解凝膏联合梅花针组(联合治疗组),每组6只。除空白对照组外,其余各组小鼠均皮下注射博莱霉素建立局限性硬皮病小鼠模型。治疗4周后,处死小鼠取皮肤组织,采用Rodnan评分观察皮肤受累程度,HE染色观察皮肤组织病理学变化,偏振光皮肤镜观察皮肤组织表面血管和毛发的变化情况,免疫组织化学染色检测皮肤组织中TGF-β_(1)、白细胞介素-4(IL-4)、Smad3、Smad7的表达;Masson特殊染色检测皮肤胶原纤维的面积。结果:阳和解凝膏组、梅花针组、联合治疗组小鼠治疗后皮肤组织表面血管和毛发恢复生长;HE染色观察真皮变薄;Masson特殊染色胶原纤维面积减少;TGF-β_(1)、IL-4、Smad3表达降低,Smad7表达升高,且以联合治疗组效果最佳,差异有统计学意义(P<0.05)。结论:阳和解凝膏、梅花针可以通过抑制TGF-β_(1)/Smad3信号通路的表达干预局限性硬皮病小鼠皮肤组织纤维化。阳和解凝膏联合梅花针治疗局限性硬皮病小鼠可以起到协同增效的作用。

Objective:To observe the effect of Yanghe Jiening Gao(阳和解凝膏)combined with plum-blossom needle on TGF-β_(1)/Smad3 signaling pathway in mice with localized scleroderma.Methods:Thirty-six female BALB/c mice were randomly divided into a blank control group,a model control group,a heparin sodium group,a Yanghe Jiening Gao group,a plum-blossom needle group,and a combination of Yanghe Jiening Gao and plum-blossom needle group(the combined treatment group),with 6 mice in each group.Except the blank control group,all other groups of mice were subcutaneously injected with Bleomycin to establish mouse models of localized scleroderma.After 4 weeks of treatment,the mice were euthanized and skin tissue was taken,and the degree of skin involvement was observed using the Rodnan skin thickness score;HE staining was used to observe pathological changes in skin tissue;changes in blood vessels and hair on the surface of skin tissue were observed using polarized light dermatoscopy;immunohistochemical staining was used for detecting the expression of TGF-β_(1),IL-4,Smad3,and Smad7 in the skin tissue;Masson special staining was used to detect the area of collagen fibers in the skin.Results:After treatment,the blood vessels and hair on the surface of the skin tissue of mice in the Yanghe Jiening Gao group,the plum-blossom needle group,and the combined treatment group resumed growth;HE staining observed thinning of the dermis;Masson's special staining reduced the area of collagen fibers;the expression of TGF-β_(1),IL-4 and Smad3 decreased,while the expression of Smad7 increased,and the combined treatment group had the most significant therapeutic effect,with statistical significance(P<0.05).Conclusion:Yanghe Jiening Gao and plum-blossom needle can inhibit the expression of TGF-β_(1)/Smad3 signaling pathway to intervene the skin tissue fibrosis in mice with localized scleroderma.The combination of them have a synergistic effect on the treatment of localized scleroderma in mice.