中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
1期
13-17
,共5页
王毅%马玉兰%李娟%曾庆华%李罗翔
王毅%馬玉蘭%李娟%曾慶華%李囉翔
왕의%마옥란%리연%증경화%리라상
血脂异常%ApoE基因缺失小鼠%视网膜/外核层、视网膜色素上皮、Bruch膜%年龄相关性黄斑变性%传统中医药/健脾祛瘀
血脂異常%ApoE基因缺失小鼠%視網膜/外覈層、視網膜色素上皮、Bruch膜%年齡相關性黃斑變性%傳統中醫藥/健脾祛瘀
혈지이상%ApoE기인결실소서%시망막/외핵층、시망막색소상피、Bruch막%년령상관성황반변성%전통중의약/건비거어
Dyslipidemia%ApoE-/-mouse%Retina/outer nuclear layer,retinal pigment epithelium,Bruch membrane%Age-related macular degeneration%Traditional Chinese medicine/tonifying spleen and promoting blood circulation
背景 血脂异常是年龄相关性黄斑变性(AMD)的发病因素之一,AMD的治疗方法一直是研究的热点. 目的 观察健脾祛瘀中药制剂对血脂异常ApoE基因缺失(ApoE-/-)小鼠视网膜和Bruch膜的治疗作用.方法 采用随机数字表法根据小鼠体质量分层随机分组.将36只2月龄ApoE-/-小鼠分为普通饮食组、高脂模型组及健脾祛瘀治疗组,每组12只.高脂模型组及健脾祛瘀治疗组小鼠均用高脂饲料喂养4个月,健脾祛瘀治疗组小鼠于6月龄开始给予健脾祛瘀制剂灌胃,每次0.3ml,每日2次,持续1个月；普通饮食组和高脂模型组小鼠同时间点开始用等量生理盐水灌胃,从6月龄开始3个组均给予普通饲料喂养.于小鼠7月龄时抽取股静脉血检测各组小鼠血浆总胆固醇(TC)、低密度脂蛋白(LDL)和血浆甘油三脂(TG)浓度,抽血后以过量麻醉法处死,制备视网膜组织切片并行Masson染色,光学显微镜和透射电子显微镜下观察视网膜和Bruch膜的组织形态学改变.采用Mias 2000型图像分析系统半定量检测各组小鼠视网膜外核层细胞、视网膜色素上皮(RPE)层细胞数及Bruch膜厚度.结果 7月龄健脾祛瘀治疗组小鼠TC、LDL和TG浓度分别为(6.47±0.49)、(1.46±0.10)、(0.62±0.21) mmol/L,比同龄高脂模型组小鼠的(10.53±0.30)、(1.90±0.13) mmol/L和(1.15±0.29) mmol/L以及普通饮食组小鼠的(9.63±0.18) mmol/L、(1.12±0.15)、(0.88±0.21) mmol/L均明显降低,差异均有统计学意义(P＜0.05～0.01).7月龄高脂模型组小鼠及同龄普通饮食组小鼠外核层和RPE层细胞排列紊乱,细胞萎缩明显,Bruch膜纤维分叉,高脂模型组部分小鼠可见玻璃膜疣；而同龄健脾祛瘀治疗组小鼠外核层和RPE层细胞排列较整齐,细胞萎缩减轻,Bruch膜纤维未见分叉,未见玻璃膜疣.7月龄健脾祛瘀治疗组小鼠外核层和RPE层细胞数分别为(23 124.00±755.18)个及(10.75±0.59)个,比同龄高脂模型组小鼠的(19 107.00±1436.82)个和(8.55±1.11)个以及普通饮食组的(21 663.00±1073.27)个和(9.75±0.58)个明显增多,差异均有统计学意义(P＜0.05).7月龄健脾祛瘀治疗组小鼠Bruch膜厚度为(0.89±0.05) μm,明显比高脂模型组小鼠的(1.02±0.13)μm薄,差异有统计学意义(P＜0.01).7月龄健脾祛瘀治疗组小鼠RPE、Bruch膜组织的超微结构形态明显改善. 结论 健脾祛瘀中药制剂可降低血脂异常ApoE-/-小鼠的高脂血症,改善视网膜外核层、RPE层、Bruch膜的组织结构形态.
揹景 血脂異常是年齡相關性黃斑變性(AMD)的髮病因素之一,AMD的治療方法一直是研究的熱點. 目的 觀察健脾祛瘀中藥製劑對血脂異常ApoE基因缺失(ApoE-/-)小鼠視網膜和Bruch膜的治療作用.方法 採用隨機數字錶法根據小鼠體質量分層隨機分組.將36隻2月齡ApoE-/-小鼠分為普通飲食組、高脂模型組及健脾祛瘀治療組,每組12隻.高脂模型組及健脾祛瘀治療組小鼠均用高脂飼料餵養4箇月,健脾祛瘀治療組小鼠于6月齡開始給予健脾祛瘀製劑灌胃,每次0.3ml,每日2次,持續1箇月；普通飲食組和高脂模型組小鼠同時間點開始用等量生理鹽水灌胃,從6月齡開始3箇組均給予普通飼料餵養.于小鼠7月齡時抽取股靜脈血檢測各組小鼠血漿總膽固醇(TC)、低密度脂蛋白(LDL)和血漿甘油三脂(TG)濃度,抽血後以過量痳醉法處死,製備視網膜組織切片併行Masson染色,光學顯微鏡和透射電子顯微鏡下觀察視網膜和Bruch膜的組織形態學改變.採用Mias 2000型圖像分析繫統半定量檢測各組小鼠視網膜外覈層細胞、視網膜色素上皮(RPE)層細胞數及Bruch膜厚度.結果 7月齡健脾祛瘀治療組小鼠TC、LDL和TG濃度分彆為(6.47±0.49)、(1.46±0.10)、(0.62±0.21) mmol/L,比同齡高脂模型組小鼠的(10.53±0.30)、(1.90±0.13) mmol/L和(1.15±0.29) mmol/L以及普通飲食組小鼠的(9.63±0.18) mmol/L、(1.12±0.15)、(0.88±0.21) mmol/L均明顯降低,差異均有統計學意義(P＜0.05～0.01).7月齡高脂模型組小鼠及同齡普通飲食組小鼠外覈層和RPE層細胞排列紊亂,細胞萎縮明顯,Bruch膜纖維分扠,高脂模型組部分小鼠可見玻璃膜疣；而同齡健脾祛瘀治療組小鼠外覈層和RPE層細胞排列較整齊,細胞萎縮減輕,Bruch膜纖維未見分扠,未見玻璃膜疣.7月齡健脾祛瘀治療組小鼠外覈層和RPE層細胞數分彆為(23 124.00±755.18)箇及(10.75±0.59)箇,比同齡高脂模型組小鼠的(19 107.00±1436.82)箇和(8.55±1.11)箇以及普通飲食組的(21 663.00±1073.27)箇和(9.75±0.58)箇明顯增多,差異均有統計學意義(P＜0.05).7月齡健脾祛瘀治療組小鼠Bruch膜厚度為(0.89±0.05) μm,明顯比高脂模型組小鼠的(1.02±0.13)μm薄,差異有統計學意義(P＜0.01).7月齡健脾祛瘀治療組小鼠RPE、Bruch膜組織的超微結構形態明顯改善. 結論 健脾祛瘀中藥製劑可降低血脂異常ApoE-/-小鼠的高脂血癥,改善視網膜外覈層、RPE層、Bruch膜的組織結構形態.
배경 혈지이상시년령상관성황반변성(AMD)적발병인소지일,AMD적치료방법일직시연구적열점. 목적 관찰건비거어중약제제대혈지이상ApoE기인결실(ApoE-/-)소서시망막화Bruch막적치료작용.방법 채용수궤수자표법근거소서체질량분층수궤분조.장36지2월령ApoE-/-소서분위보통음식조、고지모형조급건비거어치료조,매조12지.고지모형조급건비거어치료조소서균용고지사료위양4개월,건비거어치료조소서우6월령개시급여건비거어제제관위,매차0.3ml,매일2차,지속1개월；보통음식조화고지모형조소서동시간점개시용등량생리염수관위,종6월령개시3개조균급여보통사료위양.우소서7월령시추취고정맥혈검측각조소서혈장총담고순(TC)、저밀도지단백(LDL)화혈장감유삼지(TG)농도,추혈후이과량마취법처사,제비시망막조직절편병행Masson염색,광학현미경화투사전자현미경하관찰시망막화Bruch막적조직형태학개변.채용Mias 2000형도상분석계통반정량검측각조소서시망막외핵층세포、시망막색소상피(RPE)층세포수급Bruch막후도.결과 7월령건비거어치료조소서TC、LDL화TG농도분별위(6.47±0.49)、(1.46±0.10)、(0.62±0.21) mmol/L,비동령고지모형조소서적(10.53±0.30)、(1.90±0.13) mmol/L화(1.15±0.29) mmol/L이급보통음식조소서적(9.63±0.18) mmol/L、(1.12±0.15)、(0.88±0.21) mmol/L균명현강저,차이균유통계학의의(P＜0.05～0.01).7월령고지모형조소서급동령보통음식조소서외핵층화RPE층세포배렬문란,세포위축명현,Bruch막섬유분차,고지모형조부분소서가견파리막우；이동령건비거어치료조소서외핵층화RPE층세포배렬교정제,세포위축감경,Bruch막섬유미견분차,미견파리막우.7월령건비거어치료조소서외핵층화RPE층세포수분별위(23 124.00±755.18)개급(10.75±0.59)개,비동령고지모형조소서적(19 107.00±1436.82)개화(8.55±1.11)개이급보통음식조적(21 663.00±1073.27)개화(9.75±0.58)개명현증다,차이균유통계학의의(P＜0.05).7월령건비거어치료조소서Bruch막후도위(0.89±0.05) μm,명현비고지모형조소서적(1.02±0.13)μm박,차이유통계학의의(P＜0.01).7월령건비거어치료조소서RPE、Bruch막조직적초미결구형태명현개선. 결론 건비거어중약제제가강저혈지이상ApoE-/-소서적고지혈증,개선시망막외핵층、RPE층、Bruch막적조직결구형태.
Background Dyslipidemia is one of the major causes of age-related macular degeneration (AMD).At present,the study of the preventive and treating methods of A MD is still a hot spot.Objective The purpose of this study was to observe the effect of tonifying the spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Thirty-six ApoE-/-mice aged 2 months were randomly divided into the normal diet group,high fat diet group and medicine group.A diet with a higher content of fat was given for 5 consecutive months to the mice of the high fat diet group and medicine group,and in the last month,a concoction that tonifies the spleen and promotes blood circulation was gavagely administered in the medicine group,and an equivalent volumes of normal saline solution was administered in the same way in the normal diet group and high fat diet group.Total plasma cholesterol (TC),low density lipoprotein (LDL)and triglyceride (TG) were detected by (ELISA? Name of assay?) using the 7170 Hitachi automatic biochemical analyzer,and morphological changes of the retina and Bruch membrane were examined by light microscopy and transmission electron microscopy.The number of outer nuclear layer (ONL) cells,retinal pigment epithelial (RPE) cells and the thickness of the Bruch membrane were examined by semi-quantitative histopathology with the Mias 2000 Imaging Analyzer System.Results The concentrations of TC,LDL and TG were (6.47 ±0.49) mmol/L,(1.46 ±0.10)mmol/L and (0.62 ±0.21) mmol/L,respectively,in 7-month-old mice of the medicine group,showing a significant reduction in comparison with (10.53 ±0.30) mmol/L,(1.90±0.13) mmol/L,(1.15±0.29) mmol/L of the high fat diet group,and (9.63 ± 0.18) mmol/L,(1.12 ± 0.15) mmol/L,(0.88 ± 0.21) mmol/L in the normal diet group (P＜0.05-0.01).The disorder and atrophy of ONL and RPE cells,divergence of fiber of the Bruch membranes were found in both the high fat diet group and normal diet control group under the light microscope,and drusen formed in some of the mice in the high fat diet group.However,ONL and RPE were well organized in the medicine group.The cell numbers in the ONL and RPE layer in the 7-month-old mice were (23 124.00±755.18) and (10.75±0.59),respectively,in the medicine group,(19 107.00 ± 1436.82) and (8.55 ± 1.11),respectively,in the high fat diet group,(21 663.00± 1073.27) and (9.75 ±0.58),respectively,in the normal diet group,with significant differences among them (P＜0.05-0.001).Thickness of the Bruch membrane in the medicine group extensively reduced in high fat diet group and normal diet control group (P＜0.01).The ultrastructures of the RPE and Bruch membrane were much more improved in the mdedicine group.Conclusions Tonifying the spleen and promoting blood circulation can attenuate hyperlipemia in ApoE-/-mouse;furthermore,it lessens the pathological abnormalities in the ONL,RPE and Bruch membrane.
