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  

Effect of Recombinant Serpins of Psoroptes ovis on Psoriasis-like Skin Inflammation in Mice

作　　者：田焱[1] 杨富升 李艳娥 梁友萍 樊洁 吴芳燕 古小彬[1] TIAN Yan;YANG Fusheng;LI Yan’e;LIANG Youping;FAN Jie;WU Fangyan;GU Xiaobin(College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川农业大学动物医学院,成都611130

出　　处：《畜牧兽医学报》2024年第12期5774-5783,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省科技计划重点研发项目(2019YFN0155);四川农业大学学科双支计划(2221993268)。

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,r Pso SP4和r Pso SP5)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r Pso SP4和r Pso SP5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r Pso SP4(IMQ+r Pso SP4组)或r Pso SP5(IMQ+r Pso SP5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+r Pso SP4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+r Pso SP4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-αmRNA水平极显著降低(P<0.001,P<0.0001),而IMQ+r Pso SP5组对小鼠银屑病样皮损无明显改善。综上,r Pso SP4可显著改善小鼠银屑病样皮损,具有显著的抗炎活性,而r Pso SP5的抗炎活性不明显。Our team previously found that recombinant serine protease inhibitors 4 and 5(r Pso SP4 and r Pso SP5)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 r Pso SP4 and r Pso SP5 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 r Pso SP4(IMQ+r Pso SP4 group)or r Pso SP5(IMQ+r Pso SP5 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-αin 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+r Pso SP4 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+r Pso SP4 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-αin the skin lesion of IMQ+r Pso SP4 group in comparison

关 键 词：绵羊痒螨 r Pso SP4 r Pso SP5 银屑病样皮肤炎症 抗炎活性 

分 类 号：S857.5[农业科学—临床兽医学]