蛭草茶合剂对糖尿病小鼠视网膜Müller细胞的保护作用及炎症因子表达的影响

The protective effect of Zhicao Tea Mixture on Müller cells and the expression of inflammatory factors in mice with diabetic retinopathy

目的:观察蛭草茶合剂对糖尿病小鼠视网膜Müller细胞的保护作用及对炎症因子表达的影响。方法:健康清洁级雄性C57BL/6J小鼠75只,采用随机数字表法随机分为正常对照组、糖尿病组、低浓度蛭草茶合剂治疗组(低浓度治疗组)、中浓度蛭草茶合剂治疗组(中浓度治疗组)、高浓度蛭草茶合剂治疗组(高浓度治疗组),每组15只。糖尿病组、不同浓度治疗组小鼠按60 mg/kg剂量腹腔注射STZ诱导糖尿病动物模型。建模后4周,低浓度治疗组、中浓度治疗组、高浓度治疗组大鼠分别按30、60、120 ml/kg的浓度行蛭草茶合剂灌胃治疗。每两周测定各组小鼠体重及血糖。第8周后,采用Western blot检测各组小鼠视网膜Müller细胞中胶质纤维酸性蛋白(GFAP)表达;免疫荧光检测GFAP、谷氨酰胺合成酶(GS)表达;实时荧光定量PCR(RT-qPCR)、ELISA分别检测VEGF、TNF-α、IL-1β、IL-6 mRNA和蛋白表达。组间比较采用单因素方差分析。结果:糖尿病组小鼠体重较正常对照组降低,血糖升高;不同浓度治疗组小鼠血糖呈降低趋势。与正常对照组比较,糖尿病组小鼠视网膜Müller细胞GFAP蛋白表达升高(t=38.318,P<0.001),GS蛋白表达降低(t=29.737,P<0.001),差异有统计学意义;与糖尿病组比较,低浓度治疗组、中浓度治疗组、高浓度治疗组小鼠Müller细胞GFAP蛋白表达下降(t=13.677、19.387、16.305,P<0.05),GS蛋白表达升高(t=5.170、19.399、6.705,P<0.05),差异均有统计学意义。糖尿病组小鼠视网膜Müller细胞VEGF、TNF-α、IL-1β、IL-6 mRNA和蛋白表达均升高,不同浓度治疗组均降低。结论:蛭草茶合剂可降低糖尿病小鼠血糖,减轻糖尿病视网膜炎症反应。

Objective To observe the protective effect of Zhicao Tea Mixture on Müller cells and the expression of inflammatory factors in mice with diabetic retinopathy.Methods Seventy-five C57BL/6J mice were randomly divided into the normal control group,diabetes mellitus(DM)group,low concentrations group,medium concentrations group and high concentrations group,with 16 mice in each group.The diabetes model of mice in all groups except the normal control group were established by intraperitoneal injection of STZ(60 mg/kg).Four weeks after the successful modeling,the Zhicao Tea Mixture with low(30 ml/kg),medium(60 ml/kg)and high concentrations(120 ml/kg)were respectively administered by gavage.Weight and blood glucose of mice in each group were measured every two weeks.After 8 weeks,Western blot method was used to detect the mice retina Müller cells activation marker gelatinous fibrous acidic protein(GFAP).Immunofluorescence was performed to detect the expression GFAP and glutamine synthetase(GS).Real-time quantitative PCR(RT-qPCR)and ELISA were used to determine the mRNA and protein expression levels of mouse retinal VEGF,TNF-α,IL-1βand IL-6 respectively.Results The weight of mice in the DM group was lower than that of the normal control group,and the blood glucose was increased.Zhicao Tea Mixture had no effect on the weight of DM mice,but had a significant hypoglycemic effect.The GFAP expression(t=38.318,P<0.001)in the retina of mice in the DM group was increased and GS expression(t=29.737,P<0.001)was decreased compared with the control group.The GFAP expression(t=13.677,19.387,16.305;P<0.05)in the retina of mice in the low,medium and high concentrations group were decreased and GS expression(t=5.170,19.399,6.705;P<0.05)were increased compared with the DM group.The expressions of retinal inflammatory factors VEGF,TNF-α,IL-1βand IL-6 in DM group all increased,while the expressions of the above-mentioned inflammatory factors in the retina of mice decreased in the low,medium and high concentrations group.Conclusion Zhicao Tea Mixture can decrease the blood glucose of DM mice and reduces the diabetic retinal inflammatory response.