葛花总黄酮对链脲佐菌素诱导糖尿病视网膜病变小鼠玻璃体血管内皮生长因子及高迁移率族蛋白B1信号通路的影响

Effects of Total Flavonoids Extracted from Puerariae Flos on Vitreous Vascular Endothelial Growth Factor and High Mobility Group Protein B1 Signaling Pathway in Streptozotocin-Induced Diabetic Retinopathy Mice

目的:探讨葛花总黄酮对链脲佐菌素诱导糖尿病视网膜病变小鼠玻璃体血管内皮生长因子(VEGF)及高迁移率族蛋白B1(HMGB1)信号通路的影响。方法:选取清洁级健康雄性C57BL/6小鼠90只,随机分为正常对照组15只及造模组75只。造模组小鼠采用6 mg/ml链脲佐菌素溶液0.1 ml/10 g腹腔注射建立糖尿病视网膜病变模型,正常对照组小鼠予以柠檬酸缓冲液(pH=4.2)0.1 ml/10 g腹腔注射。将造模成功的小鼠依据血糖水平均匀分为模型组、羟苯磺酸钙组和葛花总黄酮低、中及高剂量组,每组15只。模型组及正常对照组小鼠灌胃予以0.9%氯化钠溶液0.1 ml/10 g,1日1次;羟苯磺酸钙组小鼠灌胃予以0.1 g/ml羟苯磺酸钙溶液0.1 ml/10 g,1日1次;葛花总黄酮低、中及高剂量组小鼠分别灌胃予以5、10及20 mg/ml葛花总黄酮溶液0.1 ml/10 g,1日1次。各组小鼠均给药10周。检测并比较各组小鼠血糖水平,视网膜血管密度,玻璃体VEGF、核因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)及视网膜HMGB1蛋白表达水平。结果:与模型组比较,各组小鼠血糖水平、视网膜血管密度明显降低,玻璃体VEGF、NF-κB及TNF-α水平明显降低,视网膜HMGB1蛋白表达水平明显降低,且葛花总黄酮低剂量组>葛花总黄酮中剂量组>葛花总黄酮高剂量组和羟苯磺酸钙组,差异均有统计学意义(P<0.05)。结论:葛花总黄酮能降低糖尿病视网膜病变小鼠血糖水平,抑制视网膜血管异常增殖,降低玻璃体VEGF、NF-κB及TNF-α水平,呈剂量依赖性,其作用可能与抑制HMGB1信号通路活性有关。

OBJECTIVE:To probe into the effects of total flavonoids extracted from puerariae flos on vitreous vascular endothelial growth factor(VEGF)and high mobility group protein B1(HMGB1)signaling pathway in streptozotocin(STZ)-induced diabetic retinopathy mice.METHODS:Totally 90 clean healthy male C57BL/6 mice were selected to be randomly divided into the control group(n=15)and the model establishment group(n=75).The model establishment group was established diabetic retinopathy model by intraperitoneal injection of 6 mg/ml STZ solution 0.1 ml/10 g,the control group received intraperitoneal injection of citric acid buffer(pH=4.2)0.1 ml/10 g.The successful model of the mice were divided into the model group,calcium dobesilate group and total flavonoids extracted from puerariae flos low,medium and high-concentration group according to the blood glucose levels,with 11 mice in each group.The model group and control group were intragastrically given 0.9%sodium chloride solution 0.1 ml/10 g by gavage,qd;the calcium dobesilate group was given 0.1 g/ml calcium dobesilate solution 0.1 ml/10 g by gavage,qd;the total flavonoids extracted from puerariae flos low-concentration group,medium-concentration group,high-concentration group were given 5 mg/ml,10 mg/ml,20 mg/ml total flavonoids extracted from puerariae flos solution 0.1 ml/10 g by gavage,qd.All mice were treated for 10 weeks.The blood glucose levels,retinal vascular density,VEGF,nuclear factor-κB(NF-κB),tumor necrosis factor-α(TNF-α)levels in vitreous,retinal HMGB1 protein expression levels were detected and compared.RESULTS:Compared with the model group,the blood glucose levels and retinal vascular density of mice in each group reduced significantly,the levels of VEGF,NF-κB and TNF-αin the vitreous decreased significantly,and the expression levels of HMGB1 protein in the retina decreased significantly;and total flavonoids extracted from puerariae flos low-concentration group>total flavonoids extracted from puerariae flos medium-concentration group>total flavonoids extracted from puerariae flos high-concentration group and calcium dobesilate group,the differences were statistically significant(P<0.05).CONCLUSIONS:Total flavonoids extracted from puerariae flos can reduce blood glucose levels in diabetic retinopathy mice in a dose-dependent manner,inhibit abnormal proliferation of retinal blood vessels,decrease the levels of VEGF,NF-κB and TNF-α,the effect may be related to the inhibition of HMGB1 signaling pathway activity.