加减驻景方对脉络膜新生血管动物模型缺氧信号因子表达的影响  

The effects of modified Zhujing formula on the expression of hypoxia signal factors in choroidal neovascularization animal model

作　　者：陶方方[1,2] 亢泽峰 陈水龄[1] 褚文丽 刘健 周志豪[3] TAO Fangfang;KANG Zefeng;CHEN Shuiling(Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)

机构地区：[1]中国中医科学院眼科医院,北京100040 [2]中国中医科学院西苑医院,北京100091 [3]北京市门头沟区中医院,北京102300

出　　处：《中国中医眼科杂志》2021年第5期309-315,共7页China Journal of Chinese Ophthalmology

基　　金：国家自然科学基金项目(81574032)。

摘　　要：目的观察加减驻景方对CNV动物模型缺氧信号因子表达的影响。方法氪激光诱导BN大鼠建立CNV动物模型,造模后大鼠被随机分为模型组、中药组、西药组和联合组。模型组大鼠用生理盐水灌胃,中药组用加减驻景方灌胃,西药组用玻璃体腔注射康柏西普,联合组用加减驻景方灌胃+玻璃体腔注射康柏西普。光凝后7 d、14 d、21 d处死大鼠取材,通过免疫组织化学法、实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白免疫印迹(Western blotting)技术检测CNV组织中AKT/mTOR/HIF-1α/VEGF信号通路mRNA和蛋白的相对表达量。结果 (1)免疫组化:模型组和各治疗组AKT、mTOR、HIF-1α、VEGF蛋白表达均呈阳性,随光凝时间延长,各治疗组目的蛋白表达量均低于模型组(P<0.05)。(2)RT-qPCR:模型组与各治疗组各目的基因mRNA相对表达量较空白组均显著增多(P<0.05)。除mTOR以外,各治疗组目的基因mRNA相对表达量均低于模型组(P<0.05)。各治疗组AKT、mTOR、VEGF mRNA相对表达量由高到低依次为中药组、西药组和联合组(P<0.05)。中药组HIF-1αmRNA相对表达量高于西药组和联合组(P<0.05),西药组和联合组之间比较无统计学意义(P>0.05)。(3)Western blotting:模型组与各治疗组目的蛋白相对表达量,较空白组均显著增多(P<0.05),除AKT蛋白以外,各治疗组目的蛋白相对表达量均低于模型组(P<0.05)。AKT和HIF-1α蛋白相对表达量中药组高于西药组和联合组(P<0.05)。mTOR与VEGF蛋白相对表达量由高到低依次为中药组、西药组和联合组,差异有统计学意义(P<0.05)。结论 AKT/mTOR/HIF-1α/VEGF信号通路的激活促进了CNV的生成,加减驻景方可通过抑制AKT/mTOR/HIF-1α/VEGF信号通路活性,发挥抑制CNV的作用,联合组作用最强。OBJECTIVE To observe the effects of modified Zhujing formula on the expression of hypoxia signal factors in choroidal neovascularization(CNV) animal model. METHODS The CNV models were induced by krypton laser on BN rats. After photocoagulation, the rats were randomly divided into model group, Chinese medicine group, western medicine group and combination group. The model group, the Chinese medicine group and the western medicine group were treated with normal saline, modified Zhujing formula and intravitreal injection of conbercept respectively, while the combination group were treated with both Chinese medicine and western medicine.Rats were killed at the 7th, 14th and 21st day after photocoagulation. The m RNA and protein expression of AKT/m TOR/HIF-1α/VEGF signal pathway in CNV tissues were detected by immunohistochemistry, RT-q PCR and Western blotting. RESULTS(1)The results of immunohistochemistryshowed that AKT, m TOR, HIF-1α and VEGF were all positive in the experimental groups. With the prolongation of photocoagulation time, the expression of target protein in each treatment group was lower than that in the model group(P<0.05).(2)RT-q PCR results showed that the relative m RNA expression of each target gene in the experimental groups were significantly higher than that in the blank group(P<0.05). Except m TOR, the relative expression of target genes m RNA in each treatment group were lower than those in the model group(P <0.05). The relative expression of AKT,m TOR and VEGF m RNA in Chinese medicine group, western medicine group and combination group showed downward trend(P <0.05). The relative expression of HIF-1α m RNA in Chinese medicine group was higher than those in western medicine group and combination group(P<0.05),but there was no significant difference between western medicine group and combination group(P>0.05). Western blotting results showed that the relative expression of each target protein in the experimental group were significantly higher than those in the blank group(P<0.05). Except

关 键 词：加减驻景方 脉络膜新生血管 雷帕霉素靶蛋白信号通路 瞳神络病 

分 类 号：R285.5[医药卫生—中药学]