摘要
目的观察加减驻景方对实验性CNV动物模型炎症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4组: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,西药组玻璃体腔注射康柏西普,联合组用加减驻景方灌胃和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、RT-qPCR和Western blotting技术检测CNV组织中TLR-4、NF-κB、IL-6 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TLR-4、NF-κB、IL-6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,治疗组中西药组表达量最高,中药组次之,联合组最低(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)。各治疗组目的基因mRNA相对表达量均低于模型组(P<0.05)。治疗组NF-κB 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)。TLR-4 mRNA相对表达量联合组最低(P<0.05),西药组与中药组比较无统计学意义(P>0.05)。IL-6 mRNA相对表达量中药组最低(P<0.05),西药组与联合组比较无统计学意义(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)。各治疗组目的蛋白相对表达量均低于模型组(P<0.05)。西药组TLR-4蛋白相对表达量高于中药组和联合组(P<0.05)。中药组NF-κB蛋白相对表达量高于西药组和联合组(P<0.05)。中药组IL-6蛋白相对表达量低于西药组与联合组(P<0.05)。均有统计学意义。结论TLR-4/NF-κB/IL-6信号通路的激活促进了CNV的生成,加减驻景方通过抑制TLR-4/NF-κB/IL-6信号通路活性,发挥抑制CNV的作用。
OBJECTIVE To observe the effects of modified Zhujing formula on the expression of inflammatory 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 concept respectively,while the combination group was treated with both Chinese Medicine and Western Medicine.Rats were killed at the 7th,14th and 21st day after photocoagulation.The mRNA and protein expression of TLR-4,NF-κB,IL-6 in CNV tissues were detected by immunohistochemistry,RT-qPCR and western blotting.RESULTS(1)Immunohistochemistry:The TLR-4,NF-κB and IL-6 were all positive in 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.In the treatment groups,the expression was the highest in the Western Medicine group,the second in the Chinese Medicine group,and the lowest in the combination group(P<0.05).(2)RT-qPCR:The relative mRNA expression of each target gene in the experimental groups was significantly higher than that in the blank group(P<0.05).The relative expression of target gene mRNA in each treatment group was lower than that in the model group(P<0.05).From high to low,the relative expression of NF-κB mRNA in the treatment group was Chinese Medicine group,Western Medicine group and combination group(P<0.05).The relative expression of TLR-4 mRNA in combination group was the lowest(P<0.05),and there was no significant difference between Western Medicine group and Chinese Medicine group(P>0.05).The relative expression of IL-6 mRNA was the lowest in Chinese Medicine group(P<0.05),but there was no significant difference between Western Medicine group and combination group(P>0.05)
作者
陶方方
亢泽峰
陈水龄
褚文丽
刘健
周志豪
TAO Fangfang;KANG Zefeng;CHEN Shuiling(Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)
出处
《中国中医眼科杂志》
2020年第12期842-848,共7页
China Journal of Chinese Ophthalmology
基金
国家自然科学基金项目(81574032)。
关键词
加减驻景方
脉络膜新生血管
核因子-ΚB信号通路
瞳神络病
modified Zhujing formula
choroidal neovascularization
nuclear factor-kappa B signaling pathway
supernatural pupil collateral disease
