薯蓣丸联合龙血竭胶囊对MKR转基因2型糖尿病小鼠创面愈合的影响

Effect of Shuyu Pill Combinined Longxuejie Capsules on Wound Healing in MKR Type 2 Diabetic Mice

目的通过薯蓣丸联合龙血竭胶囊对MKR转基因2型糖尿病小鼠创面进行干预,探讨薯蓣丸联合龙血竭胶囊对糖尿病创面愈合的影响。方法选取8周龄MKR小鼠80只,雌雄各半,按照小鼠的空腹血糖情况及体重随机分为5组:模型组、薯蓣丸+龙血竭组[薯蓣丸提取液(37.2 g/kg·d)]、薯蓣丸组、龙血竭组、二甲双胍组,每组16只小鼠。另设16只C57/BL6小鼠作为正常组。制造全层皮肤缺损的创面模型后分别给药14天,观察空腹血糖、创面愈合率、病理形态结构、晚期糖基化终末产物(AGE)及T细胞亚群(CD3^+、CD4^+、CD8^+、CD4^+/CD8^+)含量。结果与模型组比较,给药14天时薯蓣丸+龙血竭组、薯蓣丸组、二甲双胍组空腹血糖降低(P<0.01);给药3、7、10、14天时各给药组创面愈合率升高(P<0.01)。与二甲双胍组比较,给药3、7、10、14天时薯蓣丸+龙血竭组创面愈合率升高(P<0.01)。给药7天后,薯蓣丸+龙血竭组、薯蓣丸组、二甲双胍组、龙血竭组HE染色可见成纤维细胞,与模型组比较炎性细胞浸润减少;给药14天后,薯蓣丸+龙血竭组、薯蓣丸组、二甲双胍组、龙血竭组无明显炎性增生,炎症细胞较用药7天时明显减少。与模型组比较,给药7天时二甲双胍组、薯蓣丸+龙血竭组、龙血竭组、薯蓣丸组CD3^+、CD4^+百分比及CD4^+/CD8^+比值升高(P<0.05,P<0.01),CD8^+百分比降低(P<0.01);给药14天时,薯蓣丸+龙血竭组、龙血竭组及二甲双胍组CD4^+/CD8^+比值升高,薯蓣丸+龙血竭组、龙血竭组、薯蓣丸组、二甲双胍组CD3^+、CD4^+、CD8^+百分比降低(P<0.05,P<0.01)。结论薯蓣丸联合龙血竭胶囊能促进糖尿病创面愈合,其机制可能与降低血糖、AGE,提高机体细胞免疫以及改善炎症反应有关。

Objective To investigate the effect of Shuyu Pill and Longxuejie Capsules on wound healing in MKR type 2 diabetic mice.Methods Eighty MKR mice,including 40 males and 40 females,were recruited at 8 weeks of age.These mice were divided into 5 groups,i.e.the diabetic control group,Shuyu Pill+Longxuejie Capsules group,Shuyu Pill group,Longxuejie Capsules group,and Metformin group-by fasting blood sugar and weight,16 in each group.Another 16 C57/BL6 mice were recruited as the normal healing group.After building wound models of full-thickness skin loss,all mice were administrated for 14 days to observe their fasting blood sugar,wound healing rate,pathological structure,advanced glycation end(AGE)products,and T-cell subsets(CD3^+,CD4^+,CD8^+,CD4^+/CD8^+).Results Compared to the diabetic control group,the Shuyu Pill+Longxuejie Capsules group,Shuyu Pill group,and Metformin group exhibited a significant decrease in fasting blood sugar after 14 days of intervention(P<0.01);on the 3rd,7th,10th,and 14th days,the 4 groups on medication showed improvement in their wound healing rates(P<0.01).On the 3rd,7th,10th,and 14th days,the Shuyu Pill+Longxuejie Capsules group revealed a wound healing rate higher than the Metformin group(P<0.01).After 7 days of administration,hematoxylin-eosin stained fibroblasts were seen in the Shuyu Pill+Longxuejie Capsules group,Shuyu Pill group,Metformin group,and Longxuejie Capsules group,and these four groups had a smaller scale of inflammatory cell infiltration compared to the diabetic control group;when receiving medication for 14 days,no obvious inflammatory hyperplasia was detected in the Shuyu Pill+Longxuejie Capsules group,Shuyu Pill group,Metformin group,and Longxuejie Capsules group,and inflammatory cells were significantly reduced compared to the level on the 7th day of intervention.When the Metformin group,Shuyu Pill+Longxuejie Capsules group,Longxuejie Capsules group,and Shuyu Pill group had been given medication for 7 days,the CD3^+and CD4^+proportions and the CD4^+/CD8^+ratio of the Shuyu Pill group were higher than those of the diabetic model group(P<0.05,P<0.01),and yet,the Shuyu Pill group had a lower proportion of CD8^+(P<0.01);after 14 days of medication,the CD4^+/CD8^+ratios of the Shuyu Pill+Longxuejie Capsules group,Longxuejie Capsules group,Shuyu Pill group,and Metformin group were increased at varied degrees while their CD3^+,CD4^+,CD8^+proportions were reduced significantly(P<0.05,P<0.01).Conclusion As a combined medication,Shuyu Pill and Longxuejie Capsules effectively promote wound healing in mouse models of type 2 diabetes,and the underlying mechanism is probably associated with the decline and fall of blood sugar and AGEs and the improvement in cell immunity and inflammatory reaction.