姜黄素抑制BN大鼠脉络膜新生血管的机制研究  

Mechanism of curcumin inhibiting choroidal neovascularization in brown Norway rat

作　　者：陈水龄 亢泽峰[2] 褚文丽 郝雪莲 陶方方 张明明[2] 李书娇 Shui-Ling Chen;Ze-Feng Kang;Wen-Li Chu;Xue-Lian Hao;Fang-Fang Tao;Ming-Ming Zhang;Shu-Jiao Li(Department of Ophthalmology,Xiyuan Hospital of China Academy of Chinese Medical Science,Beijing 100091,China;Eye Hospital of China Academy of Chinese Medical Science,Beijing 100040,China)

机构地区：[1]中国中医科学院西苑医院眼科,北京市100091 [2]中国中医科学院眼科医院,北京市100040

出　　处：《国际眼科杂志》2023年第4期537-545,共9页International Eye Science

基　　金：国家自然科学基金资助项目(No.81574032)。

摘　　要：目的:探讨姜黄素抑制BN大鼠脉络膜新生血管(CNV)的机制。方法:激光光凝诱导BN大鼠36只建立CNV模型,随机分为6组,每组6只。正常组BN大鼠正常饲养,不做干预;实验组BN大鼠行532nm倍频激光光凝后,根据不同分组分别干预14d:模型组:生理盐水灌胃14d;雷珠单抗组:光凝后第2d玻璃体腔注射雷珠单抗注射液(10mg/mL,0.2mL/支)1次5μL。姜黄素组:低[100mg/(kg·d)]、中[200mg/(kg·d)]、高[400mg/(kg·d)]剂量的姜黄素混悬液分别灌胃14d。光凝14d后进行眼底照相、FFA与ICGA检查。制作BN大鼠CNV眼组织病理学标本,采用HE染色法观察不同组CNV中央厚度。制作BN大鼠CNV眼组织病理学标本,采用免疫组织化学法观察不同组CNV眼组织中AKT/p-AKT/HIF-1α/VEGF信号通路各因子蛋白的表达情况。采用RT-qPCR法检测CNV眼组织中AKT/HIF-1α/VEGF信号通路各因子mRNA的相对表达量。采用蛋白免疫印迹(Western blot)法检测CNV眼组织中AKT/p-AKT/HIF-1α/VEGF信号通路各因子蛋白的相对表达量。结果:模型组,雷珠单抗组,姜黄素低、中、高剂量组CNV生成率分别为78.18%、73.21%、77.19%、75.86%、74.55%,均大于70%;荧光素渗漏平均光密度值分别为182.12±6.59、119.22±8.03、166.45±8.33、164.34±5.69、149.22±6.45;雷珠单抗组较模型组显著降低(P<0.05);姜黄素低、中、高剂量组较雷珠单抗组显著升高(P<0.05),其中姜黄素高剂量组较模型组显著降低(P<0.05)。HE染色结果显示:正常组BN大鼠视网膜组织各层结构清晰、排列整齐。光凝后14d,雷珠单抗组CNV中央厚度较模型组显著减少(P<0.05);姜黄素高剂量组较模型组显著减少(P<0.05),较雷珠单抗组显著增加(P<0.05)。免疫组化结果显示:正常组BN大鼠视网膜组织结构中AKT、p-AKT、HIF-1α、VEGF因子表达呈阴性,未见棕黄色反应物。光凝后14d,模型组AKT、p-AKT、HIF-1α、VEGF因子的表达高于正常组(P<0.05);雷珠单抗组低于模型组(P<0.05);姜�English_abstract AIM:To investigate the mechanism of curcumin inhibiting the choroidal neovascularization(CNV)of brown Norway(BN)rats.METHODS:CNV model of 36 BN rats was established through laser photocoagulation induction,and they were divided into 6 groups with 6 rats in each group.Normal group was fed normally with no intervention,while 532nm laser photocoagulation was used to establish a experimental CNV model in BN rats.Rats after modeling were respectively intervened for 14d and divided into model group,ranibizumab group,curcumin low[100mg/(kg·d)],medium[200mg/(kg·d)],and high[400mg/(kg·d)]dose group.The model group was given intragastric administration of saline for 14d,ranibizumab(10mg/mL,0.2mL/dose)was injected at 2d after photocoagulation with 5μL once for rats in ranibizumab group,and different concentrations of curcumin were intragastrically administrated to the rats in low,medium and high groups for 14d.Fundus photography,fundus fluorescein angiography(FFA)and indocyanine green angiography(ICGA)examination were performed at 14d after photocoagulation.Ocular histopathological specimens of rats with CNV were made,and the central thickness of CNV were observed by HE staining.Ocular histopathological specimens were made,and the expressions of AKT/p-AKT/HIF-1α/VEGF signaling pathway-related proteins were observed by immunohistochemistry.The mRNA relative expressions of AKT/HIF-1α/VEGF factor in CNV tissues were detected by RT-qPCR,and the protein expressions of AKT/p-AKT/HIF-1α/VEGF factor in CNV tissues were detected by Western-blot.RESULTS:CNV generation rates in the model group,the ranibizumab group,and the low,medium and high-dose curcumin groups were 78.18%,73.21%,77.19%,75.86%,74.55%,respectively,which were higher than 70%.The average absorbance were 182.12±6.59,119.22±8.03,166.45±8.33,164.34±5.69,149.22±6.45,respectively;the ranibizumab group was significantly lower than the model group(P<0.05);the low-dose,medium-dose and high-dose groups were significantly higher than the ranibizumab

关 键 词：脉络膜新生血管 姜黄素 棕色挪威(BN)大鼠 血管内皮生长因子 缺氧诱导因子-1α(HIF-1α) 蛋白激酶B(AKT) 

分 类 号：R285[医药卫生—中药学]