中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2014年
4期
443-446
,共4页
贺玲%郑晓龙%王晓%温中华%郭晶晶
賀玲%鄭曉龍%王曉%溫中華%郭晶晶
하령%정효룡%왕효%온중화%곽정정
熊果酸/药理学%缺血%视网膜疾病%血管内皮生长因子类/代谢%基质金属蛋白酶2/代谢%前列腺素内过氧化物合酶类/生物合成
熊果痠/藥理學%缺血%視網膜疾病%血管內皮生長因子類/代謝%基質金屬蛋白酶2/代謝%前列腺素內過氧化物閤酶類/生物閤成
웅과산/약이학%결혈%시망막질병%혈관내피생장인자류/대사%기질금속단백매2/대사%전렬선소내과양화물합매류/생물합성
URSOLIC ACID/pharmacology%Ischemia%Retinal diseases%Vascular endothelial growth factors/metabolism%Matrix metalloproteinase 2/metabolism%Prostaglandin-endoperoxide synthases/biosynthesis
目的：观察熊果酸对氧诱导小鼠视网膜组织中血管内皮细胞生长因子（VEGF）、环氧化酶-2（COX-2）及基质金属蛋白酶-2（MMP-2）表达的影响，从而探讨熊果酸（UA）的抗新生血管机制。方法将60只7d龄清洁级C57BL/6J小鼠，按随机数字表法分为6组：模型对照组（PBS液）、阳性对照组（曲安奈德）、UA干预组（低、中、高剂量）和空白对照组，每组10只小鼠（各20只眼）。空白对照组小鼠在空气中喂养，其它各组制备缺氧小鼠视网膜新生血管模型。模型成功后，各组立即给予相应的药物治疗：模型对照组小鼠玻璃体腔注射无菌PBS液3μl，UA干预组（低、中、高剂量）小鼠分别于玻璃体腔注射1.5、3.0、6.0μgUA各3μl，阳性对照组小鼠玻璃体腔注射曲安奈德（1ml∶40mg）3μl。17d龄时过量麻醉处死各组小鼠，摘除眼球，制作视网膜组织病理切片，利用免疫组化方法检测视网膜组织中VEGF、COX-2及MMP-2的阳性表达；制备视网膜组织匀浆液，应用Westernblot方法分别检测每组视网膜组织中VEGF、COX-2及MMP-2的阳性表达和蛋白测定；以及应用RT-PCR法分别检测每组视网膜组织中VEGF、COX-2及MMP-2的mRNA表达。结果不同组别视网膜组织中COX-2、VEGF和MMP-2阳性表达、蛋白测定和mRNA的表达比较，模型对照组与空白对照组比较，各细胞因子均明显升高（P＜0.05）；高剂量UA干预组与模型对照组比较明显降低（P＜0.05）；高剂量UA干预组与阳性对照组比较差异无统计学意义（P＞0.05）。高剂量UA干预组VEGF、COX-2及MMP-2的表达明显少于低剂量UA干预组（P＜0.05）。结论UA抑制小鼠视网膜缺血模型中VEGF、COX-2及MMP-2阳性表达、蛋白测定和mRNA的表达，从而对视网膜新生血管的形成有抑制作用，并与UA的剂量呈正相关。
目的：觀察熊果痠對氧誘導小鼠視網膜組織中血管內皮細胞生長因子（VEGF）、環氧化酶-2（COX-2）及基質金屬蛋白酶-2（MMP-2）錶達的影響，從而探討熊果痠（UA）的抗新生血管機製。方法將60隻7d齡清潔級C57BL/6J小鼠，按隨機數字錶法分為6組：模型對照組（PBS液）、暘性對照組（麯安奈德）、UA榦預組（低、中、高劑量）和空白對照組，每組10隻小鼠（各20隻眼）。空白對照組小鼠在空氣中餵養，其它各組製備缺氧小鼠視網膜新生血管模型。模型成功後，各組立即給予相應的藥物治療：模型對照組小鼠玻璃體腔註射無菌PBS液3μl，UA榦預組（低、中、高劑量）小鼠分彆于玻璃體腔註射1.5、3.0、6.0μgUA各3μl，暘性對照組小鼠玻璃體腔註射麯安奈德（1ml∶40mg）3μl。17d齡時過量痳醉處死各組小鼠，摘除眼毬，製作視網膜組織病理切片，利用免疫組化方法檢測視網膜組織中VEGF、COX-2及MMP-2的暘性錶達；製備視網膜組織勻漿液，應用Westernblot方法分彆檢測每組視網膜組織中VEGF、COX-2及MMP-2的暘性錶達和蛋白測定；以及應用RT-PCR法分彆檢測每組視網膜組織中VEGF、COX-2及MMP-2的mRNA錶達。結果不同組彆視網膜組織中COX-2、VEGF和MMP-2暘性錶達、蛋白測定和mRNA的錶達比較，模型對照組與空白對照組比較，各細胞因子均明顯升高（P＜0.05）；高劑量UA榦預組與模型對照組比較明顯降低（P＜0.05）；高劑量UA榦預組與暘性對照組比較差異無統計學意義（P＞0.05）。高劑量UA榦預組VEGF、COX-2及MMP-2的錶達明顯少于低劑量UA榦預組（P＜0.05）。結論UA抑製小鼠視網膜缺血模型中VEGF、COX-2及MMP-2暘性錶達、蛋白測定和mRNA的錶達，從而對視網膜新生血管的形成有抑製作用，併與UA的劑量呈正相關。
목적：관찰웅과산대양유도소서시망막조직중혈관내피세포생장인자（VEGF）、배양화매-2（COX-2）급기질금속단백매-2（MMP-2）표체적영향，종이탐토웅과산（UA）적항신생혈관궤제。방법장60지7d령청길급C57BL/6J소서，안수궤수자표법분위6조：모형대조조（PBS액）、양성대조조（곡안내덕）、UA간예조（저、중、고제량）화공백대조조，매조10지소서（각20지안）。공백대조조소서재공기중위양，기타각조제비결양소서시망막신생혈관모형。모형성공후，각조립즉급여상응적약물치료：모형대조조소서파리체강주사무균PBS액3μl，UA간예조（저、중、고제량）소서분별우파리체강주사1.5、3.0、6.0μgUA각3μl，양성대조조소서파리체강주사곡안내덕（1ml∶40mg）3μl。17d령시과량마취처사각조소서，적제안구，제작시망막조직병리절편，이용면역조화방법검측시망막조직중VEGF、COX-2급MMP-2적양성표체；제비시망막조직균장액，응용Westernblot방법분별검측매조시망막조직중VEGF、COX-2급MMP-2적양성표체화단백측정；이급응용RT-PCR법분별검측매조시망막조직중VEGF、COX-2급MMP-2적mRNA표체。결과불동조별시망막조직중COX-2、VEGF화MMP-2양성표체、단백측정화mRNA적표체비교，모형대조조여공백대조조비교，각세포인자균명현승고（P＜0.05）；고제량UA간예조여모형대조조비교명현강저（P＜0.05）；고제량UA간예조여양성대조조비교차이무통계학의의（P＞0.05）。고제량UA간예조VEGF、COX-2급MMP-2적표체명현소우저제량UA간예조（P＜0.05）。결론UA억제소서시망막결혈모형중VEGF、COX-2급MMP-2양성표체、단백측정화mRNA적표체，종이대시망막신생혈관적형성유억제작용，병여UA적제량정정상관。
Objective To investigate the influence of ursolic acid on vascular endothelial growth factor ( VEGF) , cycloxygen-ase-2 (COX-2), and matrix metalloproteinases-2 (MMP-2) expressed in the mouse retinal ischemic model , and to explore the mecha-nisms of anti-angiogenesis.Methods Sixty 7-day clean-class C57BL/6J mice were divided randomly into 6 groups [ n =10 mice (20 eyes) per group]:blank control, model control (PBS), positive control (triamcinolone), and ursolic acid (UA) intervention (low-dose, medium-dose, and high-dose).Mice in the blank control group were raised in air , and mice in other groups in(75%±2%)O2 high-oxygen environment for 5 consecutive days .Mice in the model control group and breastfeeding mice were put back in air environ-ment (21%O2 ) on the 12th day after the new-born mice to induce the generation of retinal neovascularization .When models were suc-cessful, the drug treatments were applied immediately to the corresponding groups , with injection of 3μl of sterile PBS in model control group, 3 μl of 1.5, 3.00 and 6.0 μg UA in UA intervention group, and 3 μl of triamcinolone (1 ml∶40 mg) in positive control group, respectively.All mice were killed after overdose anesthesia on the 17th day.Their eyeballs were made into samples and retinal tissue pathological sections with H-E dying method.The positive expressions of VEGF , COX-2, and MMP-2 were detected with immu-nohistochemical method .The fresh retinal tissue homogenate was prepared to detect the protein expressions of VEGF , COX-2, and MMP-2 in retinal tissue with western blot method ,and mRNA expressions of VEGF , COX-2, and MMP-2 were detected with real-time fluorescent quantitative polymerase chain reaction ( RT-PCR) .Results According to protein and mRNA expressions of VEGF , COX-2,and MMP-2 in retinal tissue among six groups , protein expressions of VEGF , COX-2, and MMP-2 in model group were significantly higher than those in blank group ( P <0.05 ) .Each protein expression in the high UA intervention group was significantly lower than that in the model group ( P <0.05 ) .Each protein expression in the high UA intervention group was not significantly different from that in the positive group ( P >0.05 ) .Each protein expression in the high UA intervention group was significantly lower than that in the low UA intervention group( P <0.05).Conclusions UA inhibited expressions of VEGF, COX-2, and MMP-2 in retinal ischemia model .UA also played an inhibitory role in the formation of neovascularization , and this role was positively correlated with UA dose .
