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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 study the effect of polydatin on ulcer wound healing in type 2 diabetic mice. Methods C57BL / 6J male mice were fed with high-fat diet for 4 weeks. After fasting for 12 hours, rats were injected intraperitoneally with streptozotocin (40 mg · kg -1) and injected continuously for 5 days. After 1 week, Mice with a random blood glucose level above 16.67 mmo L · L -1 were type 2 diabetic mice. In the back of the mouse on both sides of the middle cut a 0.8 cm round wound deep fascia, resulting in mouse ulcer wound model. The mice were randomly divided into 3 groups according to the blood glucose level: 2 doses of experimental group and control group, 15 rats in each group. On the first day of the experiment (14 days), the wounds in the experimental group were administrated with 100μg of 400,100μg · m L -1 of polydatin locally once a day for 5 days d, stop administration on the 6th to the 14th day; the control group was given the same amount of 0.9% NaCl. The wound healing rate was calculated on the 3rd, 5th, 7th, 10th and 14th days. The complete healing time was calculated. The histological changes were observed on the 7th and the 14th day by HE staining. The MTT assay and scratch test Chemotaxis of fibroblasts to PD and PBS control groups. The macrophages of two groups, polydatin-4, polydatin-4 and interleukin-4, and PBS control were determined by flow cytometry . Results On the 14th day after the experiment, the wound healing rates of the experimental group and the control group were (80.72 ± 5.33)% and (95.93 ± 1.01)%, respectively, which were significantly different from those of the control group (P <0.001). The healing time of wounds was (16.6 ± 1.1) days and (13.2 ± 0.8) days in high-dose experimental group and control group, respectively. There was significant difference between the experimental group and the control group (P <0.001). On the 14th day after the experiment, collagen fibers in the experimental group arranged neatly and neatly, and the control group did not heal. The optimal concentration of fibroblast proliferation in experimental group was 0.5 μg · m L -1. On the first day of experiment, the migration rates of the experimental group and the PBS control group were (90.20 ± 3.90)% and (76.8 ± 4.87)%, respectively, which were significantly different from those of the PBS control group (P <0.01) Has contributed to the migration of fibroblasts. Polarization to type M2 macrophages: The experimental group and the PBS control group were (3.7 ± 0.36)% and (1.97 ± 0.23)%, respectively, which were significantly different from the PBS control group (P <0.05). Conclusion Polydatin can promote wound healing in type 2 diabetic mice.