改良保存自体血回输促进糖尿病小鼠创面愈合的机制:与MALAT1的关系

Mechanism of promoting wound healing by modified autologous blood transfusion in diabetic mice: relationship with MALAT1

目的评价改良保存的自体血回输促进糖尿病小鼠创面愈合的机制与转移相关肺腺癌转录因子1(MALAT1)的关系。方法SPF级ICR小鼠,体重21~25 g,取糖尿病建模成功的小鼠20只,采用随机数字表法分为2组(n=10):改良保存组(I组)和普通保存组(O组)。收集外周静脉血样,分别放入相应的保存液中保存7 d,随后测定其血小板聚集率、血糖、血清糖化血红蛋白(GHB)和磷酸二酯酶(DPG)浓度、WBC。制备小鼠创伤模型后即刻回输自体血。于自体血回输后7、10和14 d时计算创面愈合面积百分比,回输后14 d时分别采用Western blot、免疫组化和RT-qPCR法检测缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF-A)、基质金属蛋白酶-9(MMP-9)、β-肌动蛋白(FAP)、Ⅰ-型胶原(ColⅠ)、ColⅢ及其mRNA及MALAT1表达。结果与O组比较,I组自体血血糖、GHB、DPG、WBC降低,血小板聚集率升高,小鼠创面愈合面积百分比升高,ColⅠ和ColⅢ的阳性染色率升高,HIF-1α、VEGF-A、MMP-9、ColⅠ、ColⅢ、FAP及其mRNA和MALAT1表达上调(P<0.05)。结论改良保存的自体血回输促进糖尿病小鼠创面愈合的机制可能与上调MALAT1表达有关。

Objective To evaluate the relationship between the mechanism of promoting wound healing by modified autologous blood transfusion and metastasis-associated lung adenocarcinoma transcript 1(MALAT1)in diabetic mice.Methods Twenty SPF ICR mice,weighing 21-25 g,in which the diabetic model was successfully established,were divided into 2 groups(n=10 each)using a random number table method:modified preservation group(group I)and ordinary preservation group(group O).Peripheral venous blood samples were collected and stored in the corresponding preservation solution for 7 days.The platelet aggregation rate,blood glucose,serum glycosylated hemoglobin(GHB)and phosphodiesterase(DPG)concentrations and WBC were measured.Autologous blood was transfused back immediately after the wound model was established.The percentage of wound healing area was calculated at 7,10 and 14 days after autologous blood transfusion.The expression of hypoxia-inducible factor-1α,vascular endothelial growth factor,matrix metalloproteinase-9,β-actin,typeⅠcollagen(ColⅠ),ColⅢprotein and mRNA and MALAT1 was determined by Western blot,immunohistochemistry and quantitative real-time polymerase chain reaction respectively,at 14 days after transfusion.Results Compared with group O,the blood glucose,serum concentrations of GHB and DPG,and WBC were significantly decreased,platelet aggregation rate was increased,the percentage of wound healing area was increased,the positive staining rate of ColⅠand ColⅢwas increased,and the expression of hypoxia-inducible factor-1α,vascular endothelial growth factor,matrix metalloproteinase-9,ColⅠ,ColⅢandβ-actin protein and mRNA and MALAT1 was up-regulated in group I(P<0.05).Conclusion The mechanism by which modified autologous blood transfusion promotes wound healing may be related to up-regulating MALAT1 expression in diabetic mice.