环丙沙星对博来霉素诱导小鼠真皮纤维化的抑制作用  

Inhibitory effect of ciprofloxacin on bleomycin-induced dermal fibrosis in mice

作　　者：陈艳[1] 雷铁池[1] 史赢[1] 徐世正[1] 

机构地区：[1]武汉大学人民医院皮肤科,430060

出　　处：《中华皮肤科杂志》2015年第8期559-563,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81371717）

摘　　要：目的：观察环丙沙星对博来霉素诱导实验性硬皮病小鼠模型真皮胶原合成及致纤维化基因表达的影响。方法用博来霉素连续4周皮下注射 BALB/c 小鼠背部建立实验性硬皮病小鼠模型，随后分为3组，分别给予1%环丙沙星乳膏（环丙沙星组）、2.5%积雪草苷乳膏（积雪草苷组）以及乳膏基质（模型组）连续外擦5周，另外，于小鼠背部皮下注射磷酸盐缓冲液后外擦乳膏基质设立空白对照（对照组）。连续治疗5周后取各组皮肤标本行 HE 染色、Masson 染色、Ⅰ型胶原蛋白以及基质金属蛋白酶1（MMP1）和金属蛋白酶组织抑制因子1（TIMP1）免疫组化分析；同时用半定量逆转录 PCR 技术测定结缔组织生长因子（CTGF）、转化生长因子1（TGFβ1）和 Smad3基因的表达水平，碱水解-分光光度法测定皮肤羟脯氨酸含量。各组间比较采用单因素方差分析，模型组和各处理组间两两比较采用 LSD 检验。结果博来霉素模型组小鼠注射区真皮厚度[（432.76±93.74）μm]较对照组[（301.69±79.47）μm]明显增厚（P 〈0.01），Masson 染色可见真皮胶原纤维束粗大致密，排列错乱，符合硬皮病真皮纤维化表现。环丙沙星和积雪草苷组小鼠皮肤胶原总含量、Ⅰ型胶原、TIMP1、MMP1染色强度比模型组明显减少（F 值分别为1628.54、33.29、84.82、224.81，均 P 〈0.01），但真皮厚度改变不明显（均P 〉0.05）。与模型组比较，积雪草苷不同程度下调 CTGF、TGFβ1和 Smad3基因表达水平（均 P 〈0.05），而环丙沙星仅抑制 TGFβ1（P 〈0.05）和 Smad3（P 〈0.05）的表达,对 CTGF 基因表达作用不明显（P 〉0.05）。结论环丙沙星可抑制 TGFβ1/Smad3通路，改变失衡的 MMP1和 TIMP1表达，显示抗真皮纤维化作用。Objective To investigate the effects of ciprofloxacin on dermal collagen synthesis and profibrotic gene expressions in an experimental mouse model of scleroderma induced by bleomycin. Methods Experimental mouse models of scleroderma were established by subcutaneous injection of bleomycin into the dorsal skin of 15 BALB/c mice for 4 consecutive weeks. Then, the mouse models were randomly and equally divided into 3 groups to be topically treated with 1% ciprofloxacin cream （ciprofloxacin group）, 2.5% asiaticoside cream （asiaticoside group）and cream vehicle （model group）respectively for 5 consecutive weeks. Five mice firstly injected with sterile phosphate buffered saline （PBS）for 4 weeks then topically treated with cream vehicle for 5 weeks served as the blank control group. After the 5-week topical treatment, all the mice were sacrificed, skin specimens were resected from the dorsal skin of them, and subjected to HE staining and Masson staining. Further more, an immunohistochemical assay was performed to measure the expressions of type I collagen （COL-1）, matrix metalloproteinase-1 （MMP1） and tissue inhibitor of matrix metalloproteinase-1 （TIMP1）, semi-quantitative reverse transcription PCR to quantify the expressions of connective tissue growth factor （CTGF）, transforming growth factor-β1 （TGFβ1）and Smad3 genes, and alkaline hydrolysis-spectrophotometry to determine the level of hydroxyproline in skin. Statistical analysis was carried out by one-way analysis of variance and the least significant difference（LSD）test with the SPSS 17.0 software. Results Compared with the blank control group, the model group showed increased dermal thickness at injection sites （432.76 ± 93.74 μm vs. 301.69 ± 79.47 μm, P 〈 0.01）. Masson staining revealed thick and dense collagen bundles in an irregular arrangement in the dermis in the model group, which was consistent with dermal fibrosis in scleroderma. The total content of collagen and staining intensity of COL-1, MMP1 and TIMP1 w

关 键 词：环丙沙星 硬皮病 局部性 纤维化 胶原 博来霉素 模型 动物 

分 类 号：R593.25[医药卫生—内科学]