虎杖苷对2型糖尿病小鼠溃疡创面愈合影响的实验研究  

Experimental study of polydatin on wound healing in type 2 diabetic of mice

作　　者：韩延忠 沈娟 吴立蓉 朱家勇 

机构地区：[1]广东药科大学基础学院药用生物活性物质研究所,广州510006 [2]广东药科大学中药学院,广州510006 [3]广东药科大学生命科学与生物制药学院,广州510006

出　　处：《中国临床药理学杂志》2017年第19期1934-1937,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81502520);广东药科大学"创新强校工程"青年创新人才类基金资助项目(2015KQNCX076)

摘　　要：目的研究虎杖苷对2型糖尿病小鼠溃疡创面愈合的作用效果。方法 C57BL/6J雄性小鼠持续喂以高脂饲料4周后,禁食12 h,腹腔注射链脲佐菌素40 mg·kg^(-1)体重,连续注射5 d,1周后,测量随机血糖值高于16.67mmo L·L^(-1)的小鼠为2型糖尿病小鼠。在小鼠背部正中两侧各剪一个0.8 cm的圆形创面深至筋膜,造成小鼠溃疡创面模型。按照血糖值随机将小鼠分为3组:高低2个剂量实验组、对照组,每组15只。造模当天创面局部给药且为实验第1天(实验周期共14 d),实验组分别局部给与400,100μg·m L^(-1)虎杖苷100μL,每天给药一次,连续给药5 d,第6~14天停止给药;对照组给予等量0.9%NaCl。计算实验第3,5,7,10,14天的创面愈合率,统计完全愈合时间,以HE染色观察实验第7,14天时组织形态学变化;以噻唑蓝(MTT)法与划痕实验测定虎杖苷组与磷酸盐缓冲液(PBS)对照组对成纤维细胞增殖趋化效果。以流式细胞仪测定虎杖苷组、白细胞介素-4组、虎杖苷+白细胞介素-4联用组、PBS对照组这4组的原代巨噬细胞极化为M2型巨噬细胞量。结果实验后第14天,高剂量实验组与对照组的创面愈合率分别是(80.72±5.33)%,(95.93±1.01)%,与对照组比较差异有统计学意义(P<0.001)。创面愈合时间:高剂量实验组与对照组分别为(16.6±1.1),(13.2±0.8)d,与对照组比较差异有统计学意义(P<0.001)。实验后第14天,实验组的胶原纤维排列紧密整齐已愈合,对照组未愈合。实验组的促成纤维细胞增殖最佳浓度为0.5μg·m L^(-1)。实验第1天,实验组与PBS对照组的迁移率分别是(90.20±3.90)%,(76.8±4.87)%,与PBS对照组比较差异有统计学意义(P<0.01),说明虎杖苷明显具有促成纤维细胞迁移作用。极化为M2型巨噬细胞百分比:实验组与PBS对照组分别是(3.7±0.36)%,(1.97±0.23)%,与PBS对照组比较差异有统计学意义(P<0.05)。结论虎杖苷具有促进2型糖尿病小鼠溃疡创面愈合的作用。Objective To explore the effect of polydatin on type 2diabetic of mice. Methods C57BL/6J male mice continued for 4 weeks after feeding with high fat diet,fasting 12 h per day,intraperitoneal injection of streptozotocin 40 mg·kg^-1 weight. A continuous injection of 5 days. One week later,measurements of random blood glucose values were higher than 16. 67 mmo L · L^-1 for type 2 diabetic mice.Disinfection of back shaving after anesthesia,a 0. 8 cm circular wound was cut on each side of the back of the mouse with a sterile surgical scissors, deep to fascia. The mice were randomly divided into experimental-H,experimental-L and control groups,15 rats in eachgroup. The wound was locally administered on that day and served as the first day of the experiment. The experimental period for 14 days. The local concentration of experimental-H and experimental-L were 400 and 100 μg · m L^-1 polydatin for 5 d,take the medicine once a day,and finish it for fifth days. Discontinue medication between 6^-14days; the control group was given the same dose of 0. 9% NaCl and sterile gauze. Wound healing rate and wound healing time were statistically analyzed each at 3,5,7,10 and 14 days. The 100% wound healing time were statistically analyzed. Morphological changes were observed by HE staining each at 7,14 days. Polydatin group and phosphate buffer（ PBS） control group on proliferation of fibroblast chemotaxis effect were detected by MTT method and scratch test. The percentage of polarized M2 macrophage in primary macrophages was detected by flow cytometry in the polydatin group,the interleukin（ IL）-4 group,the polydatin ＋ IL-4 group,the PBS control group. Results The wound healing rates of the experimental-H and the control were（ 95. 93 ± 1. 01） % and（ 80. 72 ± 5. 33） %,the wound healing rate of the experimental-H was significantly increased（ P〈0. 001）. Compared with the control,the wound healing time of the experimental-H was shortened,and shortened from（ 16. 6 ± 1. 1） d to（ 13. 2 ± 0. 8） d with

关 键 词：虎杖苷 2型糖尿病小鼠 溃疡创面 创面愈合率 迁移率 

分 类 号：R28[医药卫生—中药学]