摘要
目的探讨密蒙花方防治糖尿病视网膜病变大鼠早期视网膜神经细胞损伤的作用机制,为临床实践提供实验依据。方法采用清洁级雄性Sprague-Dawley(SD)大鼠120只,其中正常组30只,其余大鼠采用链脲佐菌素诱导糖尿病大鼠动物模型,造模成功后随机分为模型组及密蒙花方组,密蒙花方组采用密蒙花方灌胃3个月,模型组用与密蒙花方组等容的蒸馏水灌胃。给药1个月后,采用TUNEL法检测大鼠视网膜神经节细胞(retinal ganglion cells,RGCs)凋亡情况;分别在给药1个月、2个月、3个月后采用高效液相色谱法检测各组大鼠视网膜中谷氨酸含量; PCR以及Western blot法检测视网膜中Sigma-1R mRNA和蛋白表达情况。结果给药1个月后,TUNEL法检测结果示,密蒙花方组RGCs凋亡数少于模型组。高效液相色谱法检测结果示,给药1个月、2个月、3个月后,模型组较正常组谷氨酸含量均增加(均为P <0. 01),密蒙花方组谷氨酸含量较模型组均降低(均为P <0. 01)。给药3个月后,正常组、模型组和密蒙花方组视网膜Sigma-1R mRNA相对表达量分别为1. 71±0. 11、0. 67±0. 09和1. 50±0. 14,Sigma-1R蛋白相对表达量分别为0. 39±0. 02、0. 16±0. 02和0. 30±0. 01;与正常组相比,模型组Sigma-1R mRNA及蛋白水平均降低(均为P <0. 01);与模型组相比,密蒙花方组Sigma-1R mRNA及蛋白水平均升高(均为P <0. 01);密蒙花方组与正常组之间差异均无统计学意义(均为P> 0. 05)。结论糖尿病视网膜病变早期即出现视网膜神经细胞的凋亡及损伤,密蒙花方可通过谷氨酸和Sigma-1R介导机制干预糖尿病大鼠早期视网膜神经细胞损伤,从而起到阻止糖尿病视网膜病变进一步发展的作用。
Objective To study the mechanism of the mimeng flower decoction to prevent and treat early diabetic retinopathy rats retinal nerve cell injury,to provide experimental basis for clinical practice.Methods Totally 120 clean level male Sprague-Dawley(SD)rats,in which 30 rats were normal group,and others were used streptozototin(STZ),After successful modeling,randomly divided into model group and mimeng flower group.In mimeng flower group,mimeng flower decoction were administered for 3 months.In model group,distilled water with the same capacity was administered.After one month,apoptosis of retinal ganglion cells in rats were detected by TUNEL.After one month,two months,three months,the content of glutamate in retinal tissue was determined by high performance liquid chromatography.The expression of Sigma-1R mRNA and protein in retina were detected by PCR and Western blot.Results STZ-induced diabetic rats with a stable and persistent hyperglycemic state showed typical symptoms of a little more than three.After one month,apoptosis of retinal ganglion cells in diabetic rats were detected by TUNEL method,and the numbers of apoptosis cells in miming flower group were less than that in model group.The results of glutamate contents in retinal tissue detected by high-performance liquid chromatography showed that the contents of glutamate increased in the model group compared with that in the normal group(P<0.01).After the drugs were used,the contents of glutamate in the mimeng flower group were reduced compared with the model group(P<0.01).The contents of glutamate in normal group reached the highest after two months,then decreased,while the contents of glutamate in model group and mimeng flower group increased gradually time dependent.The relative expressions of Sigma-1R mRNA in the normal group,the model group and the mimeng flower group respectively were 1.71±0.11,0.67±0.09 and 1.50±0.14.About the ratio of the retinal Sigma-1R protein expressed to the Sigma-1R/β-actin,the normal group,the model group and the mimeng flo
作者
严京
严华
王谦
葛东宇
吴正正
接传红
宫喜梅
宋小花
YAN Jing;YAN Hua;WANG Qian;GE Dong-Yu;WU Zheng-Zheng;JIE Chuan-Hong;GONG Xi-Mei;SONG Xiao-Hua(Eye Hospital of China Academy of Traditional Chinese Medical Science Beijing 100040,China;National Institutes for Food and Drug Control,Beijing 100050,China;Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《眼科新进展》
CAS
北大核心
2018年第12期1109-1113,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:81001540)~~
