丹参酮IIA对BN大鼠脉络膜新生血管HIF-1α和VEGF表达的影响

Impacts of Tanshinone IIA on HIF-1αand VEGF expression of retinal choroidal neovascular in Brown Norway rats

目的观察中药单体丹参酮IIA对脉络膜新生血管(CNV)动物模型中HIF-1α和VEGF表达的影响,探讨丹参酮IIA对CNV的干预作用及其机制。方法将7~8周龄雄性BN大鼠60只随机分为空白对照组(A组)、模型对照组(B组)、雷珠单抗组(C组)、丹参酮IIA低剂量组(D组)、丹参酮IIA中剂量组(E组)、丹参酮IIA高剂量组(F组)各10只。给药后第7、14、21 d行右眼底照相、荧光素眼底血管造影(FFA)检查、脉络膜血管造影(ICGA)检查。RT-qPCR法检测HIF-1α和VEGF的表达情况。结果(1)CNV形成率:7 d时CNV形成率增高,14 d时CNV形成率达高峰值,21 d趋向稳定。各时间点各组CNV形成率均达到56.7%以上。(2)荧光素渗漏平均光密度:14 d时各组荧光素渗漏平均光密度比较(F=47.894 P=0.000);雷珠单抗组和丹参酮3个剂量组均较模型组降低,均有统计学意义(P<0.05);丹参酮3个剂量组间两两比较无统计学意义(P>0.05)。(3)VEGF表达:7 d时,除模型组外,雷珠单抗组和丹参酮各组均能对VEGF产生抑制作用(F=40.775 P=0.000),且雷珠单抗组抑制VEGF作用最强,与模型组(t=16.720,P=0.000)、丹参酮低剂量(t=3.440,P=0.026)、中剂量(t=23.518,P=0.025)、高剂量(t=5.621,P=0.005)比较,均有统计学意义;丹参酮各组间两两比较,差异无统计学意义(P>0.05),VEGF蛋白表达量并不随丹参酮浓度升高而呈现剂量依赖性。(4)HIF-1α表达:各组间HIF-1αmRNA比较(F7 d=19.544,P=0.000、F14 d=221.593、F21 d=67.754,均P=0.000);各组间HIF-1αmRNA相对表达量趋势相似。结论丹参酮IIA可降低CNV动物模型眼脉络膜视网膜中HIF-1α和VEGF的表达,抑制由半导体激光光凝而产生的实验性CNV。

OBJECTIVE To observe the effect of monomer from Tanshinone IIA on the expression of HIF-1α and VEGF in choroidal neovascular(CNV)of animal model,and to explore the intervention effect and mechanism of Tanshinone IIA on CNV.METHODS Sixty large male Brown Norway(BN)rats aged between 7~8 weeks were randomly and evenly divided into 6 groups:blank group(A),model group(B),Lucentis group(C),Tanshinone IIA low dose group(D),Tanshinone IIA medium dose group(E),Tanshinone IIA high dose group(F).At the 7th,14th,21st day after treatment,fundus photography,fluorescein fundus angiography(FFA)and indocyanine green angiography(ICGA)were performed for right eye of all subjects.Meanwhile,PCR was used to detect the HIF-1αand VEGF expression.RESULTS(1)The formation rate of CNV increased at the 7th day,reached a peak at 14th day,and tended to be stable at the 21st day.At each time point,CNV formation rate of each group was over 56.7%.(2)The mean optical density of fluorescein leakage in each group was compared at the 14th day(F=47.894,P=0.000).Results of three Tanshinone groups and Lucentis group were lower than that of model group,with statistical significance(P<0.05);There was no statistical significance among three Tanshinone groups(P>0.05).(3)At the 7th day,except for the model group,Lucentis group and Tanshinone groups all showed inhibiting effects on VEGF(F=40.775,P=0.000).Lucentis group showed the strongest effect,compared with the model group(t=16.720,P=0.000),low Tanshinone group(t=3.440,P=0.026),medium Tanshinone group(t=23.518,P=0.025)and high Tanshinone dose(t=5.621,P=0.005).Tanshinone inhibited VEGF regardless of its dose,but there was no significant difference between the two groups(P>0.05).In other words,VEGF protein expression did not show dose-dependent trend with the increase of Tanshinone concentration;(4)The expression of HIF-1α was compared among groups(F7 d=19.544,F14 d=221.593,F21 d=67.754,all P=0.000).The trend of HIF-1αmRNA expression was similar among the groups.CONCLUSIONS Tanshinone IIA reduced the expression of HIF-1αand VEGF in choroid retina of animal models of CNV,thus it inhibited the growth of experimental CNV generated by semiconductor laser photocoagulation in fundus.