Yao N, Xia J-X, Liu X-M, Wang N, Mmi X-G, Wang Y-F, Guan L-L, Yang J, Dong Y-Y, Wang F-W, Li H-Y, Li X-K
Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agricultural University, Changchun, China.
Eur Rev Med Pharmacol Sci. 2014;18(7):1085-91.
Psoriasis is a chronic inflammatory skin disease characterized by excessive proliferation of keratinocytes. Fibroblast growth factor 10 (FGF10) acts as a growth factor for keratinocyte proliferation. The aim of this study is to investigate whether FGF10 blockage, a new monoclonal antibody against FGF10 we generated, could mitigate topical propranolol-induced psoriasis-like lesions in guinea pigs.
The monoclonal anti-FGF10 was generated by a routine method and purified by affinity chromatography. The effect of FGF10 and anti-FGF10 on human keratinocyte HaCaT cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The back of the ears of individual guinea pigs was topically exposed to 5% propranolol emulsion to induce psoriasis-like lesions and randomly treated topically with phosphate buffered saline (PBS), hydrocortisone butyrate, or different doses of anti-FGF10. The pathologic changes and the degrees of inflammation in the auricular areas of individual animals were examined histologically.
Characterization revealed that anti-FGF10 had a purity of 90% and a titer of 1:12800. We found that FGF10 stimulated HaCaT cell proliferation while treatment with different doses of anti-FGF10 inhibited FGF10-induced cell proliferation in a dose-dependent manner (100, 200 ng/ml, p < 0.05 vs. control; 400, 800, 1600 ng/ml, p < 0.01 vs. control). Compared to PBS-treated psoriatic animals, treatment with anti-FGF10, like hydrocortisone butyrate, greatly inhibited the severity of psoriasis-like lesions by reducing the Baker's scores, the thickness of epidermis, and the numbers of monocyte infiltrates in the dermis of animals.
The newly generated anti-FGF10 monoclonal antibody inhibited the proliferation of human keratinocytes in vitro and mitigated inflammation and pathogenic changes in propranolol-induced psoriasis-like lesions in animals. Therefore, these findings may provide a proof of principle that blockage of FGF-10 may inhibit psoriasis-related inflammation.
银屑病是一种以角质形成细胞过度增殖为特征的慢性炎症性皮肤病。成纤维细胞生长因子10（FGF10）作为角质形成细胞增殖的生长因子。本研究的目的是调查我们新生成的一种针对FGF10的单克隆抗体阻断FGF10是否能减轻局部应用普萘洛尔诱导的豚鼠银屑病样病变。
采用常规方法制备抗FGF10单克隆抗体，并通过亲和层析进行纯化。通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法测定FGF10和抗FGF10对人角质形成细胞HaCaT细胞增殖的影响。将豚鼠个体耳后局部暴露于5%普萘洛尔乳剂以诱导银屑病样病变，并随机局部给予磷酸盐缓冲盐水（PBS）、丁酸氢化可的松或不同剂量的抗FGF10。对各动物耳部区域的病理变化和炎症程度进行组织学检查。
鉴定显示抗FGF10的纯度为90%，效价为1:12800。我们发现FGF10刺激HaCaT细胞增殖，而用不同剂量的抗FGF10处理以剂量依赖方式抑制FGF10诱导的细胞增殖（100、200 ng/ml，与对照组相比p<0.05；400、800、1600 ng/ml，与对照组相比p<0.01）。与PBS处理的银屑病动物相比，抗FGF10处理与丁酸氢化可的松一样，通过降低动物的贝克评分、表皮厚度和真皮中单核细胞浸润数量，极大地抑制了银屑病样病变的严重程度。
新生成的抗FGF10单克隆抗体在体外抑制人角质形成细胞增殖，并减轻普萘洛尔诱导的动物银屑病样病变中的炎症和病理变化。因此，这些发现可能提供了一个原理证明，即阻断FGF-10可能抑制银屑病相关炎症。
