[1]罗影,左中夫,张俏,等.葛花总黄酮对糖尿病视网膜病变大鼠视网膜Müller细胞的保护作用[J].眼科新进展,2020,40(11):1033-1037.[doi:10.13389/j.cnki.rao.2020.0231]
 LUO Ying,ZUO Zhongfu,ZHANG Qiao,et al.Protective effect of total flavonoids of Pueraria on retinal Müller cell in diabetic rats[J].Recent Advances in Ophthalmology,2020,40(11):1033-1037.[doi:10.13389/j.cnki.rao.2020.0231]
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葛花总黄酮对糖尿病视网膜病变大鼠视网膜Müller细胞的保护作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年11期
页码:
1033-1037
栏目:
实验研究
出版日期:
2020-11-05

文章信息/Info

Title:
Protective effect of total flavonoids of Pueraria on retinal Müller cell in diabetic rats
作者:
罗影左中夫张俏薛坤刘畅闵连秋
121001 辽宁省锦州市,锦州医科大学附属第一医院神经内科(罗影,张俏,薛坤,刘畅,闵连秋); 121001 辽宁省锦州市,锦州医科大学基础医学院解剖学教研室(左中夫)
Author(s):
LUO Ying1ZUO Zhongfu2ZHANG Qiao1XUE Kun1LIU Chang1MIN Lianqiu1
1.Department of Neurology,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China
2.Department of Anatomy,School of Basic Medicine,Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China
关键词:
糖尿病视网膜病变Müller细胞葛花总黄酮氧化应激Nrf2/HO-1信号通路
Keywords:
diabetic retinopathy Müller cell total flavonoids of Pueraria oxidative stress Nrf2/HO-1 signaling pathway
分类号:
R587
DOI:
10.13389/j.cnki.rao.2020.0231
文献标志码:
A
摘要:
目的 探讨葛花总黄酮(total flavonoids of pueraria,TFP)对糖尿病大鼠视网膜Müller细胞的保护作用。方法 取SPF级雄性SD大鼠60只,采用单次腹腔注射链脲佐菌素(55 mg·kg-1)制作大鼠糖尿病模型。将造模成功的糖尿病大鼠随机分成糖尿病组、TFP组及TFP+Brusatol组(Brusatol为核因子NF-E2相关因子信号通路抑制剂),每组15只。另取15只正常大鼠作为对照组。造模12周后,检测各组大鼠血糖浓度、体质量、视网膜谷氨酸含量、丙二醛(MDA)及超氧化物歧化酶(SOD)含量;采用免疫组织化学法检测视网膜神经胶质纤维酸性蛋白(GFAP)的表达,免疫荧光染色检测大鼠视网膜Nrf2表达,Western blot检测大鼠视网膜Nrf2、血红素氧合酶-1(HO-1)及谷氨酰胺合成酶(GS)蛋白表达。结果 造模12周后, 糖尿病组大鼠血糖浓度高于对照组,且均大于16.7 mmol·L-1;TFP组及TFP+Brusatol组血糖浓度均低于糖尿病组(均为P<0.05);糖尿病组、TFP组及TFP+Brusatol组大鼠体质量均低于对照组(均为P<0.05)。糖尿病组和TFP+Brusatol组大鼠视网膜谷氨酸含量高于对照组(均为P<0.05);TFP组谷氨酸含量低于糖尿病组(P<0.05)。糖尿病组和TFP+Brusatol组MDA含量均高于对照组,SOD活性均低于对照组(均为P<0.05)。TFP组MDA含量低于糖尿病组,SOD活性高于糖尿病组(P<0.05)。糖尿病组和TFP+Brusatol组大鼠视网膜GFAP蛋白表达阳性率均高于对照组(均为P<0.05);TFP组GFAP蛋白表达阳性率低于糖尿病组(P<0.05)。将对照组Nrf2蛋白免疫荧光强度设定为100.00%,糖尿病组Nrf2蛋白免疫荧光强度高于对照组(P<0.05)。TFP组Nrf2蛋白免疫荧光强度高于糖尿病组(P<0.05),而TFP+Brusatol组与对照组相比差异无统计学意义(P>0.05)。糖尿病组大鼠视网膜Nrf2蛋白表达高于对照组,糖尿病组和TFP+Brusatol组大鼠视网膜HO-1和GS蛋白表达均低于对照组(均为P<0.05)。TFP组大鼠视网膜Nrf2、HO-1及GS蛋白表达均高于糖尿病组(均为P<0.05),而TFP+Brusatol组与对照组大鼠视网膜Nrf2蛋白表达差异无统计学意义(P>0.05)。结论 TFP可通过降低血糖浓度,增强大鼠视网膜的抗氧化能力,进而保护糖尿病状态下受损的Müller细胞,其机制与激活Nrf2/HO-1信号通路有关。
Abstract:
Objective To investigate the effect and mechanism of total flavonoids of Pueraria (TFP) on Müller of retina in diabetes mellitus (DM) rats.Methods A total of 60 SPF male SD rats were collected, and streptozotocin (STZ) was injected intraperitoneally at a dose of 55 mg·kg-1 to induce diabetes model of rats. Diabetic rats were randomly divided into three groups: diabetic group (DM), TFP group and TFP+Brusatol group (Brusatol is NF-E2-related factor-2(Nrf2) signal pathway inhibitor), 15 rats in each group. Another 15 normal rats were taken as the control (CON) group. After 12 weeks, blood glucose, body weight, glutamate content, malondialdehyde (MDA) and superoxide dismutase (SOD) content were detected in retina, and the expression levels of glial fibrillary acidic protein (GFAP) and Nrf2 were detected by immunohistochemical staining. The expression levels of Nrf2, heme oxygenase-1(HO-1) and glutamine synthetase (GS) proteins were detected by Western blot.Results Twelve weeks after modeling, the blood glucose concentration of the diabetic group was higher than that of the control group, and both were greater than 16.7 mmol·L-1; the blood glucose con-centration of the TFP group and the TFP+Brusatol group were lower than the diabetic group (all P<0.05). The body masses of rats in the diabeticgroup, TFP group and TFP+Brusatol group were lower than those of the control group (all P<0.05). The glutamate content in the retina of rats in the diabetic group and the TFP+Brusatol group was higher than that in the control group (both P<0.05); the TFP group was lower than diabetic group (P<0.05). The MDA content of the diabetic group and the TFP+Brusatol group were higher than that of the control group, and the SOD activity was lower than that of the control group (all P<0.05). The MDA content of the TFP group was lower than that of the diabetic group, and the SOD activity was higher than that of the diabetic group (P<0.05).The positive rate of GFAP protein expression in the retina of rats in the diabetic group and TFP+Brusatol group was higher than that in the control group (both P<0.05); the TFP group was lower than diabetic group (P<0.05). The Nrf2 protein immunofluorescence intensity of the control group was set to 100.00%, and the Nrf2 protein immunofluorescence intensity of the diabetic group was higher than that of the control group (P<0.05).The immunofluorescence intensity of Nrf2 protein in the TFP group was higher than that in the diabetic group (P<0.05), while the difference between the TFP+Bru satol group and the control group was not statistically significant (P>0.05). The expression level of Nrf2 protein in the retina of the diabetic group was higher than that of the control group, and the expression levels of HO-1 and GS protein in the retina of the diabetic group and the TFP+Brusatol group were lower than those of the control group (all P<0.05). The expressions levels of Nrf2 protein, HO-1 protein, and GS protein in the retina of rats in the TFP group were higher than those in the diabetic group (all P<0.05), while the expression level of Nrf2 protein in the retina of rats in the TFP+Brusatol group and the control group was not significantly different (P>0.05).Conclusion TFP can protect Müller cells from damage by reducing blood glucose and enhancing the antioxidant capacity of retina, which may be related to the activation of Nrf2/HO-1 signaling pathway.

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备注/Memo

备注/Memo:
中国博士后科学基金资助(编号:2017M612870);辽宁省自然科学基金资助(编号:201602340)
更新日期/Last Update: 2020-11-05