摘要
目的观察枸杞加丹参对视网膜色素变性大鼠视网膜Bcl-2、Bid表达的影响。方法实验对象分五组。空白组:RCS(rdy+/+,p+/+)大鼠,8只,雌雄均4只,灌胃生理盐水;将RCS(rdy-/-,p-/-)雌性及雄性大鼠均随机分为4组,分别为:模型组、枸杞组、丹参组和枸杞加丹参组(杞参组),每组雌雄均4只,共8只,分别给予灌胃生理盐水、生理盐水、枸杞、丹参、枸杞加丹参。灌胃28天后,免疫组化、Real-time PCR、WB检测各组RCS大鼠视网膜Bcl-2、Bid的相对表达量。结果①免疫组化检测显示:RCS(rdy-/-,p-/-)杞参组大鼠视网膜上Bcl-2蛋白阳性表达较RCS(rdy-/-,p-/-)大鼠模型组明显增多;RCS(rdy-/-,p-/-)杞参组大鼠视网膜上Bid蛋白阳性表达较RCS(rdy-/-,p-/-)大鼠模型组明显减少,但较RCS(rdy+/+,p+/+)大鼠空白组多。②RT-qPCR检测显示:RCS(rdy-/-,p-/-)杞参组Bcl-2 mRNA相对表达量明显高于RCS(rdy-/-,p-/-)模型组,差异有统计学意义(P<0.05),且接近RCS(rdy+/+,p+/+)大鼠空白对照组。RCS(rdy-/-,p-/-)杞参组Bid mRNA相对表达量明显低于RCS(rdy-/-,p-/-)模型组,差异有统计学意义(P<0.05),且接近RCS(rdy+/+,p+/+)大鼠空白对照组。③WB检测显示:RCS(rdy-/-,p-/-)杞参组Bcl-2蛋白相对表达量明显高于RCS(rdy-/-,p-/-)模型组,差异有统计学意义(P<0.05),且接近RCS(rdy+/+,p+/+)大鼠空白对照组。RCS(rdy-/-,p-/-)杞参组Bid蛋白相对表达量明显低于RCS(rdy-/-,p-/-)模型组,差异有统计学意义(P<0.05),且接近RCS(rdy+/+,p+/+)大鼠空白对照组。结论视网膜色素变性经典动物模型RCS(rdy-/-,p-/-)大鼠视网膜组织中Bcl-2表达下降、Bid表达上升;枸杞加丹参灌胃促进RCS(rdy-/-,p-/-)大鼠视网膜抗凋亡因子Bcl-2表达上调,同时,下调促凋亡因子Bid表达,在抗线粒体凋亡途径上减轻视网膜色素变性大鼠的视网膜细胞凋亡,从而保护视功能。
Objective To observe the effects of Radix Salviae Miltiorrhizae on the expression of Bcl-2 and Bid in the retina of rats with retinitis pigmentosa were observed.Methods The experimental subjects were divided into five groups.Blank group:RCS(rdy+/+,p+/+)rats,8 rats,4 males and females,(n=8),normal saline;RCS(rdy-/-,p-/-)females and Male rats were randomly divided into 4 groups,namely:model group,sputum group,Danshen group and sputum plus salvia miltiorrhiza group(Cangshen group),4 males and 4 s in each group,8 in each group,respectively given saline,saline,sputum,salvia,sputum and salvia miltiorrhiza.After 28 days of intragastric administration,the relative expression of Bcl-2 and Bid in the retina of RCS rats was detected by immunohistochemistry,Real-time PCR and WB.Results①Immunohistochemistry showed that the positive expression of Bcl-2 protein in the retina of RCS(rdy-/-,p-/-)ginseng group was higher than that of RCS(rdy-/-,p-/-)rat model.The group significantly increased;the positive expression of Bid protein in the retina of RCS(rdy-/-,p-/-)sputum group was significantly lower than that of the RCS(rdy-/-,p-/-)rat model group,but compared with RCS(rdy+/+,p+/+).There were many rats in the blank group.②RT-qPCR showed that the relative expression of Bcl-2 mRNA in RCS(rdy-/-,p-/-)ginseng group was significantly higher than that in RCS(rdy-/-,p-/-)model group.It was statistically significant(P<0.05)and was close to the RCS(rdy+/+,p+/+)rat blank control group.The relative expression of Bid mRNA in RCS(rdy-/-,p-/-)ginseng group was significantly lower than that in RCS(rdy-/-,p-/-)model group,the difference was statistically significant(P<0.05),and Close to the RCS(rdy+/+,p+/+)rat blank control group.③WB assay showed that the relative expression of Bcl-2 protein in RCS(rdy-/-,p-/-)ginseng group was significantly higher than that in RCS(rdy-/-,p-/-)model group.The significance of the study(P<0.05),and close to the RCS(rdy+/+,p+/+)rat blank control group.The relative expression of Bid protein in RCS(rdy-/-,p-/-)ginseng group was significantly lower than that in RCS(rdy-/-,p-/-)model group,the difference was statistically significant(P<0.05),and Close to the RCS(rdy+/+,p+/+)rat blank control group.Conclusion In the classical animal model of retinitis pigmentosa,RCS(rdy-/-,p-/-)rats showed decreased expression of Bcl-2 and increased Bid expression;sputum plus salvia miltiorrhiza was promoted by RCS(rdy-/-,p-/-).The expression of anti-apoptotic factor Bcl-2 was up-regulated in rat retina,and the expression of pro-apoptotic factor Bid was down-regulated,and the apoptosis of retinal cells in rats with retinitis pigmentosa was alleviated in the antimitochondrial apoptotic pathway,thereby protecting visual function.
作者
彭俊
王英
蒋鹏飞
刘家琪
潘坤
周亚莎
徐剑
彭清华
Peng Jun;Wang Ying;Jiang Pengfei;Liu Jiaqi;Pan Kun;Zhou Yasha;Xu Jian;Peng Qinghua(First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410007,China;Hunan University of Traditional Chinese Medicine,Changsha 410208,China;Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Eye,Ear,Nose and Throat Diseases in Hunan Province,Changsha 410208,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第4期1166-1175,共10页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81804150):补虚活血中药对虚中夹瘀证RP模型大鼠感光细胞凋亡相关基因表达的干预作用及机制,负责人:彭俊
湖南省教育厅创新平台开放基金项目(16K065):补虚活血法对RP虚中夹瘀证模型大鼠Rho、XBP1、Caspase12表达的研究,负责人:彭清华
湖南中医药大学中医诊断学国家重点学科开放基金优先资助项目(No.2015ZYZD02):基于Rho、XBPI表达探讨补虚活血法对虚中夹瘀证RP模型大鼠的干预研究,负责人:徐剑、彭清华
湖南省科技厅中医药防治五官科疾病湖南省重点实验室建设项目(2017TP1018):中医药防治五官科疾病湖南省重点实验室,负责人:彭清华
长沙市科技局长沙市科技计划项目(kc1704005)
长沙市科技计划项目(kc1704005):中医药防治五官科疾病湖南省重点实验室,负责人:彭清华
作者简介
通讯作者:彭清华,二级教授,主任医师,博士研究生导师,主要研究方向:青光眼、眼底病;通讯作者:徐剑,医学博士,主任医师,主要研究方向:眼底病,青光眼。
