[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]
点击复制

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
分享到：

《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 Ying1; ZUO Zhongfu2; ZHANG Qiao1; XUE Kun1; LIU Chang1; MIN 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.

参考文献/References:

［1］　POWERS M，GREVEN M，KLEINMAN R，NGUYEN Q D，DO D.Recent advances in the management and understanding of diabetic retinopathy［J］.F1000 Res，2017，6(1):2063-2071.
［2］　NASIRI A，ZIAMAJIDI N，ABBASALIPOURKABIR R，GOODARZI M T，SAIDIJAM M，BEHROUJ H，et al.Beneficial effect of aqueous garlic extract on inflammation and oxidative stress status in the kidneys of type 1 diabetic rats［J］.Indian J Clin Biochem，2016，32(3):329-336.
［3］　VOHRA R，ALDANA B I，SKYTT D M，FREUDE K，WAAGEPE-TERSEN H，BERGERSEN L H，et al.Essential roles of lactate in müller cell survival and function［J］.Mol Neurobiol，2018，55(1):1-14.
［4］　COUGHLIN B A，FEENSTRA D J，MOHR S.Müller cells and diabetic retinopathy［J］.Vision Res，2017，139(3) :93-100.
［5］　艾明，杨芳，孙明，李岱.葛花总黄酮对糖尿病小鼠视网膜MDA、SOD的影响［J］.临床眼科杂志，2012，20(4)：374-376.
AI M，YANG F，SUN M，LI D.The effect of total flavonoids from flos puerariae lobatae on MDA and SOD in the retina of diabetic mice［J］.J Clin Ophthalmol，2012，20(4):374-376.
［6］　李阔，王丽轩，姬春雪，张俊玲.葛花总黄酮对1型糖尿病小鼠脑内氧化应激及认知功能的影响［J］.广东医学，2016，37(5)：665-669.
LI K，WANG L X，JI C X，ZHANG J L.Effects of total flavonoids of Pueraria lobata on oxidative stress and cognitive function in brain of type 1 diabetic mice［J］.Guangdong Med，2016，37(5):665-669.
［7］　王昆，罗小娜，李时光，冯丽君，刘新生.Nrf2/HO-1信号通路在6-羟基多巴胺帕金森病模型大鼠神经损伤中的作用［J］.中国医药生物技术，2019，14(3)：237-244.
WANG K，LUO X N，LI S G，FENG L J，LIU X S.Role of Nrf2/HO-1 signaling pathway in 6-hydroxydopamine-induced nerve injury of Parkinson’s disease model rats［J］.Chin Med Biotechnol，2019，14(3):237-244.
［8］　MAHENDRAN G，THAMOTHARAN G，SENGOTTUVELU S,NARMATHA BAI V.RETRACTED: Anti-diabetic activity of Swertia corymbosa(Griseb.) Wight ex C.B.Clarke aerial parts extract in streptozotocin induced diabetic rats［J］.J Ethnopharmacol，2014，151(3):1175-1183.
［9］　ESPINO J，RODRGUEZ，ANA B.Melatonin and oxidative stress in the diabetic state: clinical implications and potential therapeutic applications［J］.Curr Med Chemist，2019，26(22):4178-4190.
［10］　ZHANG L W，XIA H M，HAN Q Q.Effects of antioxidant gene therapy on the development of diabetic retinopathy and the TFP+Brusatolabolic memory phenomenon［J］.Graefes Arch Clin Exp Ophthalmol，2015，253(2):249-259.
［11］　LUO W，HU L，LI W，XU G，XU L，ZHANG C，et al.Epo inhibits the fibrosis and migration of Müller glial cells induced by TGF-β and high glucose［J］.Graefes Arch Clin Exp Ophthalmol，2016，254(5):881-890.
［12］　RUIPING G，XINYI D，WENYI T，BOYA L，CHEN J，GEZHI X A.Synthesized glucocorticoid- induced leucine zipper peptide inhibits retinal müller cell gliosis［J］.Front Pharmacol，2018，9(4):331-340.
［13］　张俏，刘文强，左中夫，刘学政.褪黑素通过Müller细胞保护糖尿病视网膜神经节细胞的机制［J］.眼科新进展，2019，39(7)：611-614.
ZHANG Q，LIU W Q，ZUO Z F，LIU X Z.Mechanism of melatonin protecting diabetic retinal ganglion cells through Müller cells［J］.Rec Adv Ophthalmol，2019，39(7):611-614.

相似文献/References:

[1]杜玮 刘子扬 周艳艳 雒雷鸣.糖尿病视网膜病变与血清胆红素水平的关系[J].眼科新进展,2012,32(5):000.
[2]范松涛 卢建民.阿司匹林与糖尿病患者玻璃体出血以及玻璃体切割术疗效的相关性研究[J].眼科新进展,2012,32(11):000.
[3]李艳 李筱荣 袁佳琴 潘斌.糖尿病大鼠视网膜中VEGF、PEDF的表达与血-视网膜屏障损伤[J].眼科新进展,2013,33(1):000.
[4]李朝晖 崔治华 胡晓英 孟丽珠 张敬维.糖尿病视网膜病变激光面积与疗效的分析[J].眼科新进展,2013,33(2):000.
[5]冯冬梅 朱鸿 施彩虹.CXC趋化因子及其受体在糖尿病视网膜病变中的作用[J].眼科新进展,2013,33(6):000.
[6]牛淑玲.糖尿病视网膜病变患者HbAlc、FPG与血小板参数的变化及危险因素分析[J].眼科新进展,2013,33(7):000.
[7]毕春潮 王睿 王建洲 雷春灵 董晓娟 王小莉 薛晓辉.Ad-PEDF对糖尿病视网膜病变大鼠视网膜新生血管的抑制作用[J].眼科新进展,2013,33(8):000.
[8]杨萍 孙书明 李晓鹏.辛伐他汀对糖尿病视网膜病变和炎症因子的影响[J].眼科新进展,2013,33(8):000.
[9]罗文婷 孙大卫.血管黏附蛋白-1在眼科疾病中的研究进展[J].眼科新进展,2013,33(8):000.
[10]李小璐 马雅玲.糖尿病视网膜病变大鼠视网膜VEGF和PEDF的动态表达[J].眼科新进展,2013,33(9):000.
[11]梁汇珉,李赵伟,李铮,等.脑源性神经营养因子对糖尿病大鼠视网膜Müller细胞的保护作用[J].眼科新进展,2017,37(12):1110.[doi:10.13389/j.cnki.rao.2017.0280]
　LIANG Hui-Min,LI Zhao-Wei,LI Zheng,et al.Effect of brain-derived neurotrophic factor on retinal Müller cells in diabetic rats[J].Recent Advances in Ophthalmology,2017,37(11):1110.[doi:10.13389/j.cnki.rao.2017.0280]
[12]李赵伟,梁汇珉,李铮,等.罗格列酮对糖尿病视网膜Müller细胞的保护作用以及对Müller细胞胶质纤维酸性蛋白（GFAP）及炎症因子表达的影响[J].眼科新进展,2018,38(9):825.[doi:10.13389/j.cnki.rao.2018.0195]
　LI Zhao-Wei,LIANG Hui-Min,LI Zheng,et al.The protective effect of rosiglitazone on diabetic retinal Müller cells and its effects on the expression of glial fibrillary acidic protein (GFAP) and inflammatory factors in Müller cells[J].Recent Advances in Ophthalmology,2018,38(11):825.[doi:10.13389/j.cnki.rao.2018.0195]
[13]冯闯,左中夫,刘文强,等.抑制JAK/STAT信号通路对糖尿病早期大鼠视网膜Müller细胞的保护作用及对谷氨酸转运蛋白表达的影响[J].眼科新进展,2019,39(2):105.[doi:10.13389/j.cnki.rao.2019.0024]
　FENG Chuang,ZUO Zhong-Fu,LIU Wen-Qiang,et al.Inhibition of JAK/STAT signaling pathway protects retinal Müller cells and affects the expression of glutamate transporter in early diabetic rats[J].Recent Advances in Ophthalmology,2019,39(11):105.[doi:10.13389/j.cnki.rao.2019.0024]
[14]张俏,刘文强,左中夫,等.褪黑素通过Müller细胞保护糖尿病视网膜神经节细胞的机制[J].眼科新进展,2019,39(7):611.[doi:10.13389/j.cnki.rao.2019.0141]
　ZHANG Qiao,LIU Wen-Qiang,ZUO Zhong-Fu,et al.Protective effects of melatonin on diabetic retinal ganglion cell via Müller cell[J].Recent Advances in Ophthalmology,2019,39(11):611.[doi:10.13389/j.cnki.rao.2019.0141]
[15]于洋,刘学政.CDK抑制剂对糖尿病大鼠视网膜Müller细胞胶质增殖及新生血管形成的抑制作用[J].眼科新进展,2019,39(9):809.[doi:10.13389/j.cnki.rao.2019.0184]
　YU Yang,LIU Xue-Zheng.Inhibitory effects of CDK inhibitors on the glial proliferation of retinal Müller cells and retinal neovascularization in diabetic rats[J].Recent Advances in Ophthalmology,2019,39(11):809.[doi:10.13389/j.cnki.rao.2019.0184]
[16]吴传玲,刘安琪,左中夫,等.Raf-1激酶抑制蛋白(RKIP)对糖尿病大鼠视网膜神经损伤的保护作用[J].眼科新进展,2020,40(4):323.[doi:10.13389/j.cnki.rao.2020.0074]
　WU Chuanling,LIU Anqi,ZUO Zhongfu,et al.Protective effect of RKIP on retinal nerve injury in diabetic rats through p38-MAPK pathway[J].Recent Advances in Ophthalmology,2020,40(11):323.[doi:10.13389/j.cnki.rao.2020.0074]
[17]王坤,朱曼辉,陈莉莉,等.长链非编码RNA 母系表达基因3调控miR-34a对糖尿病视网膜病变Müller细胞活化及炎症因子分泌的影响[J].眼科新进展,2021,41(1):042.[doi:10.13389/j.cnki.rao.2021.0009]
　WANG Kun,ZHU Manhui,CHEN Lili,et al.Effect of lncRNA MEG3 regulating miR-34a on the activation of Müller cells and the secretion of inflammatory cytokines in diabetic retinopathy[J].Recent Advances in Ophthalmology,2021,41(11):042.[doi:10.13389/j.cnki.rao.2021.0009]
[18]罗影,张俏,单伟.神经胶质成熟因子-β对糖尿病大鼠视网膜Müller细胞活化的影响及可能机制[J].眼科新进展,2023,43(3):190.[doi:10.13389/j.cnki.rao.2023.0038]
　LUO Ying,ZHANG Qiao,SHAN Wei.Effect of glia maturation factor-β on the activation of retinal Müller cells in diabetic rats and its mechanism[J].Recent Advances in Ophthalmology,2023,43(11):190.[doi:10.13389/j.cnki.rao.2023.0038]

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