目的观察成纤维胶原酶1(MMP1)和miR-222在增生性瘢痕(HS)成纤维细胞中的表达水平,探讨miR-222对HS发生、发展的调控机制。方法选取36例HS患者的HS组织,并各留取HS旁正常组织作为对照。分离培养出HS组织与正常组织的成纤维细胞;采用RT-PC...目的观察成纤维胶原酶1(MMP1)和miR-222在增生性瘢痕(HS)成纤维细胞中的表达水平,探讨miR-222对HS发生、发展的调控机制。方法选取36例HS患者的HS组织,并各留取HS旁正常组织作为对照。分离培养出HS组织与正常组织的成纤维细胞;采用RT-PCR法检测成纤维细胞MMP1 m RNA和miR-222表达水平;采用Western blot法检测成纤维细胞MMP1蛋白表达水平;采用MTT法检测成纤维细胞增殖情况;采用双荧光素酶报告实验检测miR-222是否可调控MMP1基因。结果与正常组织比较,HS组织成纤维细胞MMP1 m RNA、蛋白表达水平均下调(均P<0.05),miR-222表达水平上调(P<0.05)。与空白对照成纤维细胞比较,HS组织转染MMP1过表达质粒后的成纤维细胞MMP1 m RNA、蛋白表达水平均上调(均P<0.05),增殖速度明显减慢(P<0.05);转染antagomiR-222质粒后的成纤维细胞miR-222表达水平下调(P<0.05),MMP1 m RNA表达水平上调(P<0.05),增殖速度明显减慢(P<0.05)。双荧光素酶报告实验结果表明miR-222能与MMP1基因的3′-UTR区相结合,从而调控其表达。结论 miR-222在HS组织中存在过表达,miR-222或通过负调控其靶基因MMP1的表达而发挥其调控成纤维细胞增殖和凋亡的作用。展开更多
To establish a simple and reliable animal model of skin photo-damage, 20 mice were treated with 8-MOP and exposed to UVA （UVA 320-400 nm） for 24 h. After irradiation, the structure of the epidermis and dermis, colla...To establish a simple and reliable animal model of skin photo-damage, 20 mice were treated with 8-MOP and exposed to UVA （UVA 320-400 nm） for 24 h. After irradiation, the structure of the epidermis and dermis, collagen fibers, elastic fibers were observed by using HE staining and Weigert technique and compared with the normal controls. The acanthosis and epidemis proliferation with accompanying hyperkeratosis and parakeratosis were observed. Inflammatory infiltration was noted in the dermis. The elastic fibers became coarse, irregularly arranged and clustered, with their number increased. The collagen fibers showed obvious degeneration and some amorphous materials could also be observed. The blood vessels were irregularly dilated and vascular walls were thickened, with infiltration of inflammatory cells. It is concluded that murine photodamage model can be quickly, conveniently and reliably established by means of 8-MOP/UVA.展开更多
文摘目的观察成纤维胶原酶1(MMP1)和miR-222在增生性瘢痕(HS)成纤维细胞中的表达水平,探讨miR-222对HS发生、发展的调控机制。方法选取36例HS患者的HS组织,并各留取HS旁正常组织作为对照。分离培养出HS组织与正常组织的成纤维细胞;采用RT-PCR法检测成纤维细胞MMP1 m RNA和miR-222表达水平;采用Western blot法检测成纤维细胞MMP1蛋白表达水平;采用MTT法检测成纤维细胞增殖情况;采用双荧光素酶报告实验检测miR-222是否可调控MMP1基因。结果与正常组织比较,HS组织成纤维细胞MMP1 m RNA、蛋白表达水平均下调(均P<0.05),miR-222表达水平上调(P<0.05)。与空白对照成纤维细胞比较,HS组织转染MMP1过表达质粒后的成纤维细胞MMP1 m RNA、蛋白表达水平均上调(均P<0.05),增殖速度明显减慢(P<0.05);转染antagomiR-222质粒后的成纤维细胞miR-222表达水平下调(P<0.05),MMP1 m RNA表达水平上调(P<0.05),增殖速度明显减慢(P<0.05)。双荧光素酶报告实验结果表明miR-222能与MMP1基因的3′-UTR区相结合,从而调控其表达。结论 miR-222在HS组织中存在过表达,miR-222或通过负调控其靶基因MMP1的表达而发挥其调控成纤维细胞增殖和凋亡的作用。
基金a grant from Hubei Province Natural Science Foundation (No.302131041).
文摘To establish a simple and reliable animal model of skin photo-damage, 20 mice were treated with 8-MOP and exposed to UVA （UVA 320-400 nm） for 24 h. After irradiation, the structure of the epidermis and dermis, collagen fibers, elastic fibers were observed by using HE staining and Weigert technique and compared with the normal controls. The acanthosis and epidemis proliferation with accompanying hyperkeratosis and parakeratosis were observed. Inflammatory infiltration was noted in the dermis. The elastic fibers became coarse, irregularly arranged and clustered, with their number increased. The collagen fibers showed obvious degeneration and some amorphous materials could also be observed. The blood vessels were irregularly dilated and vascular walls were thickened, with infiltration of inflammatory cells. It is concluded that murine photodamage model can be quickly, conveniently and reliably established by means of 8-MOP/UVA.
