白头翁皂苷B4对糖尿病大鼠视网膜凋亡相关蛋白表达的影响  

Effects of anemoside B4 on the expression of apoptosis-associated proteins in the retina of diabetic rats

作　　者：牛童童 黄江[1] 娄慧[1] 许琼明[2] 游雨虹 王雪[1] 张天琦 徐国旭[1] Niu Tongtong;Huang Jiang;Lou Hui;Xu Qiongming;You Yuhong;Wang Xue;Zhang Tianqi;Xu Guoxu(Department of Ophthalmology,the Second Affiliated Hospital of Soochow University,Suzhou 215000,China;College of Pharmaceutical Science,Soochow University,Suzhou 215123,China)

机构地区：[1]苏州大学附属第二医院,215000 [2]苏州大学药学院,215123

出　　处：《中华老年医学杂志》2020年第8期954-957,共4页Chinese Journal of Geriatrics

摘　　要：目的探讨白头翁皂苷B4对糖尿病大鼠视网膜细胞凋亡的作用。方法采用随机数字法将60只雄性SD大鼠随机分为正常对照组、糖尿病模型对照组及糖尿病+白头翁皂苷B4处理组,每组20只。模型对照组、糖尿病+白头翁皂苷B4处理组大鼠腹腔注射60 mg/kg链脲佐菌素(STZ)建立糖尿病模型。建模成功3 d后,糖尿病+白头翁皂苷B4处理组以5 mg/kg的剂量腹腔注射白头翁皂苷B4,2次/d,持续腹腔注射8周,正常对照组、模型对照组大鼠腹腔注射同等剂量生理盐水。于糖尿病+白头翁皂苷B4处理组腹腔注射8周后,获取视网膜组织,行视网膜石蜡切片苏木精-伊红染色(HE),观察视网膜组织结构及形态改变;采用蛋白质印迹法(Western blot)检测Bcl-2及Bax蛋白表达水平;行荧光定量PCR(Q-PCR)检测视网膜组织中Caspase-3 mRNA表达水平。结果 HE染色显示,正常对照组大鼠视网膜结构完整,形态清晰;模型对照组大鼠视网膜结构紊乱,各层变薄,细胞排列紊乱、稀疏;而白头翁皂苷B4治疗后的大鼠视网膜结构与形态好转;与正常对照组相比,模型对照组大鼠视网膜Bcl-2蛋白相对表达量减弱(t=57.81,P<0.01),Bax表达增强(t=10.47,P<0.01),Bcl-2/Bax比值下降(t=23.98,P<0.01),差异均具有统计学意义;与模型对照组比较,糖尿病+白头翁皂苷B4处理组大鼠视网膜Bcl-2蛋白相对表达量增强(t=41.07,P<0.01),Bax表达减弱(t=6.811,P<0.01),Bcl-2/Bax比值上升(t=14.70,P<0.01),差异均具有统计学意义;模型对照组大鼠视网膜组织Caspase-3 mRNA表达较正常对照组增高(t=7.916,P<0.01),而糖尿病+白头翁皂苷B4处理组大鼠视网膜组织中Caspase-3 mRNA表达较模型对照组降低(t=6.221,P<0.01),差异均有统计学意义。结论白头翁皂苷B对糖尿病大鼠视网膜细胞凋亡具有一定的抑制作用,其机制可能与增强Bcl-2表达及下调Bax、Caspase-3表达有关。Objective To investigate the effects of anemoside B4 on apoptosis of retinal cells in diabetic rats.Methods Sixty Sprague-Dawley rats were randomized into three groups:the normal control(control),diabetic rats(DM)and diabetic rats treated with Anemoside B4(B4)groups(n=20,each group).Rats in the DM and B4 groups were rendered diabetic with an intraperitoneal injection of streptozotocin(STZ,60 mg/kg).After 3 days of successful modeling,rats in the B4 group were intraperitoneally injected with anemoside B4(5 mg/kg),twice/day,for 8 weeks,while rats in the control and DM groups were injected with an equivalent volume of normal saline.After 8 weeks of anemoside B4 and normal saline-injection,rats were sacrificed and retinas were harvested for examination.Paraffin sections of retina were stained with the hematoxylin-eosin(H-E)method for morphological evaluation.Protein levels of Bax and Bcl-2 were detected by using Western blot.The expression of caspase-3 mRNA was detected with quantitative PCR.Results H-E staining results showed the control group had intact retinal structure and clear morphological features,whereas disordered retinal structure,thinner layers'and sparse and disorganized cells were seen in the DM group.However,retinal structure and morphology were improved after treatment with anemoside B4.Compared with the control group,the protein expression of Bcl-2 was lower(t=57.81,P<0.01),the protein expression of Bax was higher(r=10.47,P<0.01),and the Bcl-2/Bax ratio was lower(t=23.98,P<0.01)in the DM group.Compared with the DM group,the protein expression of Bcl-2 was higher(t=41.07,P<0.01),the protein expression of Bax was lower(i=6.811,P<0.01),and the Bcl-2/Bax ratio was higher(t=14.70,P<0.01)in the B4 group.Caspase-3 mRNA expression was higher in the DM group than in the control group(t=7.916,P<0.01)>but was lower in the B4 group compared with the DM group (t=6.221,P<0.01).Conclusions Anemoside B4 can inhibit the apoptosis of retinal cells by up-regulating Bcl-2 expression and down-regulating Bax and caspase-3

关 键 词：白头翁皂苷B4 糖尿 视网膜病变 凋亡 

分 类 号：R285.5[医药卫生—中药学]