中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
7期
613-616
,共4页
刘超%耿燕%张振华%顾艳枝
劉超%耿燕%張振華%顧豔枝
류초%경연%장진화%고염지
槲皮素%糖尿病%视网膜%血管生成素样蛋白2%血管生成素样蛋白2受体
槲皮素%糖尿病%視網膜%血管生成素樣蛋白2%血管生成素樣蛋白2受體
곡피소%당뇨병%시망막%혈관생성소양단백2%혈관생성소양단백2수체
Quercetin%Diabetes mellitus%Retina%Angiopoietin-like protein 2%Angiopoietin-like protein 2 receptor
背景 糖尿病视网膜病变(DR)的特征性病理改变为微血管病变,血管生成素样蛋白2(Ang-2)是与血管发生密切相关的新蛋白,槲皮素具有多种药理作用,可改善微循环、降低毛细血管通透性,但其对DR的作用机制尚不清楚.目的 探讨槲皮索脂质体对视网膜组织Ang-2及其受体(Tie2)表达的影响.方法 选取60只清洁级雄性成年Wistar大鼠以随机数字表法分为6个组,其中10只大鼠作为空白对照组,其他60只大鼠采用链脲佐菌素(STZ)35 mg/kg一次性腹腔内注射制作糖尿病模型.槲皮索分为50 mg/(kg·d)(低剂量)、150 mg/(kg·d)(中剂量)和250 mg/(kg·d)(高剂量),并用脂质体包裹,分别对模型鼠进行灌胃3 ～5 ml共12周,其他模型鼠分别用生理盐水和羟苯磺酸钙混悬液灌胃作为阴性对照和阳性对照.给药12周后处死各组大鼠并分离视网膜,用ELISA法检测视网膜组织中Ang-2的吸光度(A450)值,并用逆转录聚合酶链反应(RT-PCR)检测视网膜中Tie2 mRNA的A值(Tie2 mRNA/GAPDH mRNA).结果 ELISA检测表明,槲皮素中剂量组、高剂量组Ang-2的A450值分别为0.796±0.057和0.842±0.043,较生理盐水组的1.012±0.046明显下降,差异均有统计学意义(q=2.95、2.698,P＜0.05).RT-PCR检测表明,槲皮素中剂量组、高剂量组的Tie2 mRNA在视网膜中的表达量分别为0.712±0.092和0.821±0.087,均明显低于阴性对照组的1.182±0.098,差异均有统计学意义(q=3.497、2.852,P＜0.05).Ang-2蛋白及Tie2 mRNA表达量的下降均以中剂量组最为明显.结论 槲皮素脂质体对视网膜中Ang-2及其受体Tie2的表达有抑制作用,可对糖尿病大鼠的视网膜微循环异常有改善作用.
揹景 糖尿病視網膜病變(DR)的特徵性病理改變為微血管病變,血管生成素樣蛋白2(Ang-2)是與血管髮生密切相關的新蛋白,槲皮素具有多種藥理作用,可改善微循環、降低毛細血管通透性,但其對DR的作用機製尚不清楚.目的 探討槲皮索脂質體對視網膜組織Ang-2及其受體(Tie2)錶達的影響.方法 選取60隻清潔級雄性成年Wistar大鼠以隨機數字錶法分為6箇組,其中10隻大鼠作為空白對照組,其他60隻大鼠採用鏈脲佐菌素(STZ)35 mg/kg一次性腹腔內註射製作糖尿病模型.槲皮索分為50 mg/(kg·d)(低劑量)、150 mg/(kg·d)(中劑量)和250 mg/(kg·d)(高劑量),併用脂質體包裹,分彆對模型鼠進行灌胃3 ～5 ml共12週,其他模型鼠分彆用生理鹽水和羥苯磺痠鈣混懸液灌胃作為陰性對照和暘性對照.給藥12週後處死各組大鼠併分離視網膜,用ELISA法檢測視網膜組織中Ang-2的吸光度(A450)值,併用逆轉錄聚閤酶鏈反應(RT-PCR)檢測視網膜中Tie2 mRNA的A值(Tie2 mRNA/GAPDH mRNA).結果 ELISA檢測錶明,槲皮素中劑量組、高劑量組Ang-2的A450值分彆為0.796±0.057和0.842±0.043,較生理鹽水組的1.012±0.046明顯下降,差異均有統計學意義(q=2.95、2.698,P＜0.05).RT-PCR檢測錶明,槲皮素中劑量組、高劑量組的Tie2 mRNA在視網膜中的錶達量分彆為0.712±0.092和0.821±0.087,均明顯低于陰性對照組的1.182±0.098,差異均有統計學意義(q=3.497、2.852,P＜0.05).Ang-2蛋白及Tie2 mRNA錶達量的下降均以中劑量組最為明顯.結論 槲皮素脂質體對視網膜中Ang-2及其受體Tie2的錶達有抑製作用,可對糖尿病大鼠的視網膜微循環異常有改善作用.
배경 당뇨병시망막병변(DR)적특정성병리개변위미혈관병변,혈관생성소양단백2(Ang-2)시여혈관발생밀절상관적신단백,곡피소구유다충약리작용,가개선미순배、강저모세혈관통투성,단기대DR적작용궤제상불청초.목적 탐토곡피색지질체대시망막조직Ang-2급기수체(Tie2)표체적영향.방법 선취60지청길급웅성성년Wistar대서이수궤수자표법분위6개조,기중10지대서작위공백대조조,기타60지대서채용련뇨좌균소(STZ)35 mg/kg일차성복강내주사제작당뇨병모형.곡피색분위50 mg/(kg·d)(저제량)、150 mg/(kg·d)(중제량)화250 mg/(kg·d)(고제량),병용지질체포과,분별대모형서진행관위3 ～5 ml공12주,기타모형서분별용생리염수화간분광산개혼현액관위작위음성대조화양성대조.급약12주후처사각조대서병분리시망막,용ELISA법검측시망막조직중Ang-2적흡광도(A450)치,병용역전록취합매련반응(RT-PCR)검측시망막중Tie2 mRNA적A치(Tie2 mRNA/GAPDH mRNA).결과 ELISA검측표명,곡피소중제량조、고제량조Ang-2적A450치분별위0.796±0.057화0.842±0.043,교생리염수조적1.012±0.046명현하강,차이균유통계학의의(q=2.95、2.698,P＜0.05).RT-PCR검측표명,곡피소중제량조、고제량조적Tie2 mRNA재시망막중적표체량분별위0.712±0.092화0.821±0.087,균명현저우음성대조조적1.182±0.098,차이균유통계학의의(q=3.497、2.852,P＜0.05).Ang-2단백급Tie2 mRNA표체량적하강균이중제량조최위명현.결론 곡피소지질체대시망막중Ang-2급기수체Tie2적표체유억제작용,가대당뇨병대서적시망막미순배이상유개선작용.
Background The special pathological change of diabetic retinopathy(DR)is microvascular disorder.Angiopoietin-like protein 2(Ang-2)is a new protein associated with genesis of blood vessels.Quercetin has multiple pharmacological action,including improving the microcircularion and the permeability of blood capillary.However,the action mechanism of Ang-2 on DR was unclear.Objective The present study was to investigate the effects of quercetin on Ang-2 and its receptor Tie2 expression in retina with diabetes mellitus.Methods Sixty clean male Wistar rats were randomized into 7 groups and 10 rats for each group,and 10 rats served as blank control group.Streptozotocin of 35 mg/kg was intraperitoneally injected in 60 rats to establish the diabetic models.Quercetins encapsulated by liposome with the doses of 50,150 and 250 mg/(kg · d)(3-5 ml)were used to gavage in different groups of models for 12 weeks,and normal saline solution and calcium dobesilate were used at the same fashion as the negative control group and positive control group,respectively.Twelve weeks later,the animals were sacrificed and retinas were isolated.Expressions of Ang-2 protein and Tie2 mRNA in retinas were detected by ELISA and RT-PCR,respectively.The usage and rearing of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Sciences and Technology Commission.Results ELISA showed that the A450 of Ang-2 in 150 and 250 mg/(kg · d)quercetin groups was 0.796±0.057 and 0.842±0.043 respectively and was lower than that negative group(1.012±0.046),showing statistically significant differences(q =2.95,2.698,P＜0.05).RT-PCR assay showed that expression of Tie2 mRNA(Tie2 mRNA/GAPDH mRNA)in retinas was 0.712±0.092 and 0.821±0.087,presenting statistically significant differences in comparison with negative group(1.182±0.098)(q =3.497,2.852,P＜0.05).The expression levels of Ang-2 and Tie2 mRNA in retina were lowest in 150 mg/(kg · d)quercetin group.Conclusions Quercetin can improve the retinal microcirculation by downregulating the expressions of Ang-2 and its receptor in early period of diabetic rats.
