CAJ | 학술논문

目的探讨玻璃体内注射熊果酸(UA)对糖尿病视网膜病变(DR)小鼠视网膜损伤的保护作用及其机制.方法使用四氧嘧啶建立糖尿病小鼠模型.12周龄清洁级健康C57BL/6雄性小鼠60只,随机分为6组:正常对照组、糖尿病模型组、阳性对照组(曲安奈德)和UA干预(低、中、高剂量)组,每组10只小鼠.药物干预1周后处死小鼠,取眼球制作石蜡切片,采用组织病理学方法检测各组小鼠视网膜新生血管情况;实时PCR检测视网膜血管内皮生长因子(VEGF)、环氧化酶2(COX-2)、基质金属蛋白酶2(MMP-2) mRNA的表达水平;Western印迹法检测视网膜VEGF、COX-2、MMP-2蛋白的表达水平;试剂盒检测超氧化物歧化酶(SOD)活力、丙二醛含量.结果 HE染色和过碘酸雪夫(PAS)染色结果显示,正常对照组内界膜平整,血管内皮细胞排列整齐,几乎不见新生血管;糖尿病模型组可见大量新生血管;低剂量UA干预组与糖尿病模型组结果相似;高剂量UA干预组与阳性对照组结果相似,内界膜平整,基本不见新生血管.VEGF、COX-2、MMP-2 mRNA表达水平糖尿病模型组显著高于正常对照组和中、高剂量UA干预组(均P＜0.001);VEGF、COX-2、MMP-2蛋白表达水平糖尿病模型组显著高于正常对照组(P＜0.001)和低、中、高剂量UA干预组(均P＜0.05).糖尿病模型组视网膜组织丙二醛含量显著高于正常对照组(P =0.001)和中、高剂量UA干预组(P=0.026、0.007),SOD活性显著低于正常对照组(P ＜0.001)和低、中、高剂量UA干预组(P=0.040、0.021、0.003).结论熊果酸可能通过抑制DR小鼠视网膜VEGF、COX-2、MMP-2的表达抑制视网膜新生血管形成,并通过明显的抗氧化应激作用缓解DR.
목적 탐토파리체내주사웅과산(UA)대당뇨병시망막병변(DR)소서시망막손상적보호작용급기궤제.방법 사용사양밀정건립당뇨병소서모형.12주령청길급건강C57BL/6웅성소서60지,수궤분위6조:정상대조조、당뇨병모형조、양성대조조(곡안내덕)화UA간예(저、중、고제량)조,매조10지소서.약물간예1주후처사소서,취안구제작석사절편,채용조직병이학방법검측각조소서시망막신생혈관정황;실시PCR검측시망막혈관내피생장인자(VEGF)、배양화매2(COX-2)、기질금속단백매2(MMP-2) mRNA적표체수평;Western인적법검측시망막VEGF、COX-2、MMP-2단백적표체수평;시제합검측초양화물기화매(SOD)활력、병이철함량.결과 HE염색화과전산설부(PAS)염색결과현시,정상대조조내계막평정,혈관내피세포배렬정제,궤호불견신생혈관;당뇨병모형조가견대량신생혈관;저제량UA간예조여당뇨병모형조결과상사;고제량UA간예조여양성대조조결과상사,내계막평정,기본불견신생혈관.VEGF、COX-2、MMP-2 mRNA표체수평당뇨병모형조현저고우정상대조조화중、고제량UA간예조(균P＜0.001);VEGF、COX-2、MMP-2단백표체수평당뇨병모형조현저고우정상대조조(P＜0.001)화저、중、고제량UA간예조(균P＜0.05).당뇨병모형조시망막조직병이철함량현저고우정상대조조(P =0.001)화중、고제량UA간예조(P=0.026、0.007),SOD활성현저저우정상대조조(P ＜0.001)화저、중、고제량UA간예조(P=0.040、0.021、0.003).결론 웅과산가능통과억제DR소서시망막VEGF、COX-2、MMP-2적표체억제시망막신생혈관형성,병통과명현적항양화응격작용완해DR.
Objective To explore the protective effect and its mechanisms of intravitreal injection of ursolic acid (UA) on retinal injury in diabetic retinopathy (DR) mice.Methods DR model was set up by using alloxan.60 C57BL/6 male mice (clean animal,aged 12 weeks) were randomly and evenly divided into 6 groups:blank control group,DR model group,positive control group (triamcinolone acetonide),UA intervention groups (low,moderate and high dose).Mice were killed one week after drug intervention and the eyeballs were removed to prepare paraffin section.Retinal new blood vessel was examined by histopathological methods.The mRNA and protein expressions of vascular endothelial growth factor (VEGF),cyclooxygenase 2 (COX-2),and Matrix Metalloproteinase 2 (MMP-2) in retinal tissues were detected by real-time PCR and Western blotting,respectively.The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected by related kits.Results The HE and PAS staining showed uniform inner limiting membrane structure and orderly vascular endothelial ceils in blank control group,and neovascularization was seldom detected.In DR model group,numerous new blood vessels were formed beyond inner limiting membrane.UA intervention group with low dose had almost the same results with DR group,while the results of UA intervention group with high dose were similar with positive control group,in which the inner limiting membrane structure was uniform and new blood vessels were seldom seen.The mRNA expressions of VEGF,COX-2,and MMP-2 in DR model group were significantly higher than those of blank control group (P ＜0.001) and UA intervention group with moderate and high dose (P ＜0.001).The protein expressions of VEGF,COX-2 and MMP-2 in DR model group were significantly higher than those of blank control group (P ＜ 0.001) and UA intervention group with low,moderate and high dose (all P ＜ 0.05).The MDA content of retinal tissue in DR model group was significantly higher than those of blank control group (P =0.001) and UA intervention groups with moderate and high dose (P =0.026,0.007).The activity of SOD was significantly lower in DR model group than those of blank control group (P ＜ 0.001) and UA intervention groups with low,moderate and high dose (P =0.040,0.021,0.003).Conclusion UA may inhibit the formation of new blood vessels through reducing the expressions of VEGF,COX-2 and MMP-2 in retinal tissues,and promote a remission from DR by obvious resistance to oxidative stress.