吡非尼酮对硬皮病小鼠结缔组织生长因子和V型胶原含量的作用分析  

Effects of pirfenidone on connective tissue growth factor and the content of type V collagen in scleroderma mice

作　　者：覃玉花 唐开奖 佘柳君 宋星慧 以敏 蒋茵 QIN Yuhua;TANG Kaijiang;SHE Liujun;SONG Xinghui;YI Min;JIANG Yin(Department of Rheumatology and Immunology,Liuzhou Worker's Hospital,Guangxi,Liuzhou 545005,China;Department of Respiratory and Critical Care Medicine,Liuzhou Worker's Hospital,Guangxi,Liuzhou 545005,China;Department of Pathology,Liuzhou Worker's Hospital,Guangxi,Liuzhou 545005,China)

机构地区：[1]柳州市工人医院风湿免疫科,广西柳州545005 [2]柳州市工人医院呼吸与危重症医学科,广西柳州545005 [3]柳州市工人医院病理科,广西柳州545005

出　　处：《中国医药科学》2020年第11期1-5,共5页China Medicine And Pharmacy

基　　金：广西自然科学青年基金项目(2016GXNSFBA380136)。

摘　　要：目的探讨吡非尼酮(PFD)干预硬皮病小鼠对体内结缔组织生长因子(CTGF)和V型胶原(ColV)含量的影响。方法选择健康清洁级雌性BALB/c小鼠24只,随机分为四组,分别为正常对照组(6只),博来霉素(BLM)组(6只)、PFD干预组(6只)和BLM+PFD干预组(6只)。使用背部皮下注射BLM的方法建立硬皮病鼠模型,对照组注射磷酸盐缓冲液(PBS),1次/d,连续4周;PFD干预组采用灌胃的方法给药[300mg/(kg·d),1次/12h],从第11天到实验结束;BLM+PFD干预组先采用BLM皮下注射,在第11天开始予PFD进行干预,到第4周。采用样本碱水法检测皮肤羟脯氨酸(HYP)含量,HE和Masson染色法观察皮肤组织学改变,免疫组化染色法检测小鼠皮肤CTGF蛋白和ColV分布。结果BLM组小鼠皮肤HYP的含量明显高于BLM+PFD干预组,对照组和PFD干预组最低,差异有统计学意义(P<0.05)。HE和Masson染色显示,BLM组小鼠真皮层明显增厚,胶原纤维增粗、数量增多,血管增多、管腔狭窄、伴炎性细胞浸润;提示造模成功。免疫组化染色显示,BLM组真皮和基底细胞层CTGF蛋白均呈强阳性表达,ColV在真皮层、小血管及周围可聚集分布,BLM+PFD干预组较BLM组阳性表达率降低,差异有统计学意义(P<0.05)。结论硬皮病鼠皮肤中CTGF和ColV蛋白的表达增加,这可能是导致硬皮病发生发展的重要原因,给予PFD干预,均可降低两者的表达水平,这有可能是治疗硬皮病潜在的新靶点。Objective To explore the effects of pirfenidone(PFD)intervention on scleroderma mice on connective tissue growth factor(CTGF)and the content of collagen V(ColV).Methods 24 healthy and clean female BALB/c mice were randomly divided into 4 groups:normal control group(n=6),bleomycin(BLM)group(n=6),PFD intervention group(n=6)and BLM+PFD intervention group(n=6).A scleroderma mouse model was established by subcutaneous injection of BLM in the back,and the control group was injected with phosphate buffered saline(PBS)once a day for 4 weeks;the PFD intervention group was administered by gavage[300 mg/(kg·d),once/12h]from the 11th day to the end of the experiment;the BLM+PFD intervention group was first injected subcutaneously with BLM,and then the PFD intervention was started from the 11th day to the 4th week.The sample alkaline water method was applied to detect the content of skin hydroxyproline(HYP).HE and Masson staining methods were used to observe the skin histological changes.Immunohistochemical staining method was performed to detect the CTGF protein and ColV distribution in mouse skin.Results The content of HYP in the skin of the BLM group was significantly higher than that of the BLM+PFD intervention group,and the control group and the PFD intervention group were the lowest in the content of HYP in the skin,and the differences were statistically significant(P<0.05).HE and Masson staining showed that the dermal layer of the BLM group was significantly thickened,the collagen fibers were enlarged,the number was multiplied,the blood vessels were increased,the lumen was narrowed,and the inflammatory cells were infiltrated;suggesting that the modeling was successful.Immunohistochemical staining demonstrated that CTGF protein in the dermis and basal cell layer of the BLM group was strongly positively expressed,and ColV was aggregated and distributed in the dermis,small blood vessels and surroundings;the positive expression rate of the BLM+PFD intervention group was lower than that of the BLM group,and the difference

关 键 词：吡非尼酮 硬皮病 结缔组织生长因子 V型胶原 

分 类 号：R593.25[医药卫生—内科学]