绵羊痒螨重组丝氨酸蛋白酶抑制剂对小鼠银屑病样皮肤炎症的影响

doi:10.11843/j.issn.0366-6964.2024.12.039

摘要/Abstract

摘要：

课题组前期发现绵羊痒螨的重组丝氨酸蛋白酶抑制剂4和5(recombinant serine protease inhibitors 4 and 5, rPsoSP4和rPsoSP5)可体外抑制糜蛋白酶、胰蛋白酶和弹性蛋白酶的活性，因此推测其可能具有抗炎活性，为证实上述推测，评价了rPsoSP4和rPsoSP5是否可以改善小鼠银屑病样皮肤炎症。BALB/c小鼠随机分为5组，用凡士林或咪喹莫特乳膏(imiquimod cream, IMQ)连续涂抹背部皮肤6 d, 建立空白对照组和银屑病模型，期间模型组每日皮内注射PBS(IMQ组)或pET32a(+)载体蛋白(IMQ+pET32a(+)组)或rPsoSP4(IMQ+rPsoSP4组)或rPsoSP5(IMQ+rPsoSP5组)，每天观察小鼠银屑病样皮损区变化，于第7天处死小鼠，比较各组皮损组织病理变化和皮损组织中IL-1β、IL-6和TNF-α的转录水平。结果表明：IMQ造模成功，小鼠呈现典型的银屑病样皮损；与模型组(IMQ组)相比，IMQ+rPsoSP4组可有效改善小鼠银屑病样皮损，包括显著抑制小鼠体重减轻(第4~6天，P < 0.05，P < 0.01，P < 0.001)和皮损区红斑(第5~6天)、鳞屑(第4~6天)、皮肤增厚程度(第4~6天)及PASI评分(第3~6天)显著减少(P < 0.05，P < 0.001)。此外，与IMQ组相比，IMQ+rPsoSP4组皮损区组织病理损伤显著减轻，表皮厚度显著降低(P < 0.05)，浸润的炎性细胞数量极显著减少(P < 0.001)，皮损区IL-1β、IL-6和TNF-α mRNA水平极显著降低(P < 0.001，P < 0.000 1)，而IMQ+rPsoSP5组对小鼠银屑病样皮损无明显改善。综上，rPsoSP4可显著改善小鼠银屑病样皮损，具有显著的抗炎活性，而rPsoSP5的抗炎活性不明显。

关键词: 绵羊痒螨, rPsoSP4, rPsoSP5, 银屑病样皮肤炎症, 抗炎活性

Abstract:

Our team previously found that recombinant serine protease inhibitors 4 and 5 (rPsoSP4 and rPsoSP5) of Psoroptes ovis could inhibit the enzymatic activities of chymotrypsin, trypsin and elastase in vitro, thus we speculated that these two recombinant proteins might have anti-inflammation activities. To confirm the above speculation, we assessed whether rPsoSP4 and rPsoSP5 could improve psoriatic-like skin inflammation in mice. BALB/c mice were randomly divided into 5 groups (n=6), Vaseline or imiquimod cream (IMQ) was applied daily on the back skin continuously for 6 days to establish the control group and psoriasis model groups. During this period, the psoriasis model groups were intradermal injected with PBS (IMQ group) or pET32a(+)-vector protein (IMQ+pET32a(+) group) or rPsoSP4 (IMQ+rPsoSP4 group) or rPsoSP5(IMQ+rPsoSP5 group), then the changes of psoriasis-like skin lesions were observed every day, and mice were sacrificed on the 7^th day. Subsequently, the pathological changes and the transcription levels of IL-1β, IL-6, and TNF-α in the skin lesions among each group were compared. Results showed that IMQ induced psoriasis-like lesions were successfully established, and the mice appeared typical psoriasis-like lesions. Compared with the model group (IMQ group), IMQ+rPsoSP4 group can effectively attenuated psoriasis-like skin lesions, including significant inhibition of weight loss (from 4^th to 6^th day, P < 0.05, P < 0.01, P < 0.001) and reductions of skin erythema (5^th and 6^th day), skin scales (from 4^th to 6^th day), skin thickness (from 4^th to 6^th day) and PASI score (from 3^rd to 6^th day) in mice (P < 0.05, P < 0.001). Additionally, compared with the model group (IMQ group), IMQ+rPsoSP4 group showed that the pathological damage in the skin lesion was significantly reduced, accompanied by the significant decrease in epidermal thickness (P < 0.05) and the number of infiltrated inflammatory cells (P < 0.001). Moreover, there was a significantly decreased in mRNA levels of IL-1β, IL-6, and TNF-α in the skin lesion of IMQ+rPsoSP4 group in comparison with IMQ groups, while IMQ+rPsoSP5 group had no significant improvement on psoriatic lesions. In conclusion, rPsoSP4 could significantly improve psoriasis skin lesion in mice and has significant anti-inflammatory activity, while rPsoSP5 seemed to be no obvious effect.

Key words: Psoroptes ovis, rPsoSP4, rPsoSP5, psoriasis-like skin inflammation, anti-inflammatory activity

中图分类号:

S857.5

田焱, 杨富升, 李艳娥, 梁友萍, 樊洁, 吴芳燕, 古小彬. 绵羊痒螨重组丝氨酸蛋白酶抑制剂对小鼠银屑病样皮肤炎症的影响[J]. 畜牧兽医学报, 2024, 55(12): 5774-5783.

TIAN Yan, YANG Fusheng, LI Yan'e, LIANG Youping, FAN Jie, WU Fangyan, GU Xiaobin. Effect of Recombinant Serpins of Psoroptes ovis on Psoriasis-like Skin Inflammation in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5774-5783.

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