四君子汤调控线粒体凋亡保护脾气虚模型大鼠视网膜脱离复位后感光细胞的作用  

Effect of Sijunzi Decoction Protecting Rats’ Photoreceptor Cells of Reattached Retina with Pi-Qi Deficiency Syndrome by Regulating Mitochondrial Apoptosis Pathway

作　　者：任秀瑜 康紫薇 张妍春 李京珂 薛大喜 崔阳阳 REN Xiu-yu;KANG Zi-wei;ZHANG Yan-chun;LI Jing-ke;XUE Da-xi;CUI Yang-yang(Ophthalmology Department,Shaanxi Eye Hospital,Xi'an People's Hospital,Xi'an Fourth(Hospital),Affiliated People's Hospital of Northwest University,Xi'an,710004;Xi'an Ocular Fundus Disease Research Institute,Xi'an,710004;Department of Chinese Medicine,First Clinical Medical College,Shaanxi University of Traditional Chinese Medicine,Shaanxi,712000)

机构地区：[1]西安市人民医院(西安市第四医院)陕西省眼科医院,西北大学附属人民医院眼科,西安710004 [2]西安市眼底病研究所,西安710004 [3]陕西中医药大学第一临床医学院中医系,陕西712000

出　　处：《中国中西医结合杂志》2023年第2期219-226,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：陕西省重点研发计划项目(No.2021SF-162);西安市科技计划重大研究项目[No.201805104YX12SF38(3)];白求恩-朗沐科研发展专项基金项目(No.BJ2020IIT001)。

摘　　要：目的观察四君子汤是否通过线粒体凋亡信号通路发挥对脾气虚型大鼠视网膜脱离(RD)复位后感光细胞的保护作用。方法成年SD大鼠52只随机分为对照组(A组)、RD自动复位组(B组)、脾气虚RD自动复位组(C组)、四君子汤组(D组)。C、D组使用破气耗气加饥饱失常法制备脾气虚证动物模型,实验开始6周后B、C、D组使用视网膜下注射0.1%透明质酸钠的方法制备RD自动复位模型,D组随后灌服四君子汤(20 g/kg)至实验结束。分别于实验开始后8、9周随机分批处死动物取材。记录大鼠进食量、体重及精神状态;比色法测定红细胞膜钠钾ATP酶(Na+-K+-ATPase)活性;JC-1荧光探针法检测视网膜组织线粒体膜电位(ΔΨm)水平;Western Blot检测各组视网膜组织B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)及视网膜细胞核细胞凋亡诱导因子(AIF)表达量变化;免疫荧光法检测AIF、细胞色素C(Cyt C)在视网膜组织中的分布。结果与A、B组比较,C、D组大鼠在实验开始后1周出现便溏、精神萎靡、毛色枯黄等脾气虚症状,6周红细胞膜Na+-K+-ATPase活性明显降低(P<0.05),C组上述症状持续至实验结束,D组于9周时上述症状好转,红细胞膜Na+-K+-ATPase活性较C组升高(P<0.05)。与A组比较,B组8、9周视网膜组织ΔΨm降低(P<0.01);与B组比较,C、D组8周时ΔΨm降低(P<0.01);随着实验时间延长,至9周时D组ΔΨm较C组显著升高(P<0.05)。与A组比较,8、9周时C组视网膜组织Bax升高(P<0.05),D组则较C组显著降低(P<0.05),同时9周时C组Bax水平亦高于B组(P<0.01)。8周时C组Bcl-2/Bax较A组降低(P<0.05),9周时较A、B、D组降低(P<0.01)。8周时,与B组比较,C、D组细胞核AIF蛋白表达量升高(P<0.01);至9周时,C组细胞核AIF蛋白表达量仍持续升高,且高于D组(P<0.05)。免疫荧光结果显示B、C、D组AIF及Cyt C较A组着色弥散,其中B组AIF特异性表达于少量外核层细胞核;C组AIF及Cyt C在外核层特异性着色Objective To observe whether Sijunzi Decoction(SJZD)protects rats’ photoreceptor cells of reattached retina with Pi-Qi deficiency syndrome by regulating mitochondrial apoptosis pathway.Methods Fifty-two adult SD rats were randomly divided into 4 groups,the control group(Group A),retinal spontaneous reattached group(Group B),retinal spontaneous reattached with Pi-Qi deficiency syndrome group(Group C)and SJZD treated group(Group D). The rats in in Group C and Group D were established with PiQi deficiency syndrome by herbs inducing exhaustion,dissipating stagnant qi and irregular diet. After 6 weeks,retinal spontaneous reattached model was prepared by sub-retinal injection of 0.1% sodium hyaluronate in Group B,C and D,then Group D was given SJZD(20 g/kg) until the end of the experiment. Animals were sacrificed randomly at 8th and 9th week respectively. Food intake,body weight,and mental status of rats were recorded.Na+-K+-ATPase of erythrocyte membrane was detected by colorimetric method,and mitochondrial transmembrane potential(ΔΨm)of retinal tissue was detected by JC-1 fluorescent probe. The retinal expressions of BCL2-associated X(Bax),B-cell lymphoma-2(Bcl-2)and apoptosis inducing factor(AIF)were measured by Western Blot. The AIF and Cytochrome C(Cyt C)were detected by immunofuorescence staining. Results Rats in Group C and D showed symptoms of Pi-Qi deficiency syndrome,such as loose stool,listlessness,dull fur at 1th week,and the Na+-K+-ATPase of erythrocyte membrane was significantly lower than Group A and B at 6weeks(P<0.05). In Group C,the above symptoms lasted until the end of the experiment. At 9 weeks,compared with Group C,the above symptoms in Group D were improved and the Na+-K+-ATPase of erythrocyte membrane was significantly increased(P<0.05). At 8th and 9th week,compared with Group A,ΔΨm in Group B reduced significantly(P<0.01). Compared with Group B,the ΔΨm decreased at 8th week in Group C and D(P<0.01),with the extension of experiment time,the ΔΨm in Group D was significantly higher than

关 键 词：四君子汤 脾气虚证 视网膜脱离复位 线粒体膜透化 感光细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]