中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2011年
6期
521-526
,共6页
胡艳滨%张静楷%孙智勇%袁志刚%刘玉梅%毛春洁%颜华
鬍豔濱%張靜楷%孫智勇%袁誌剛%劉玉梅%毛春潔%顏華
호염빈%장정해%손지용%원지강%류옥매%모춘길%안화
糖尿病视网膜病变%微循环%纤维化%细胞凋亡%结缔组织生长因子
糖尿病視網膜病變%微循環%纖維化%細胞凋亡%結締組織生長因子
당뇨병시망막병변%미순배%섬유화%세포조망%결체조직생장인자
Diabetic retinopathy%Microcirculation%Fibrosis%Apoptosis%Connective tissue growth factor
目的 研究糖尿病大鼠视网膜凋亡细胞和结缔组织生长因子(CTGF)表达情况及其在微循环改变中的作用.方法 实验研究.55只成年雄性Wistar大鼠,以随机数字表法,随机分为正常对照(CON)组(10只鼠)和糖尿病(DM)组(45只鼠).根据病程不同,将存活的30只DM组大鼠再分为2个月组(DM2)、4个月组(DM4)及6个月组(DM6),每组各10只鼠.用末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测视网膜凋亡细胞,用免疫组织化学法测定CTGF表达水平,以视网膜消化铺片过碘酸雪夫(PAS)染色法观察视网膜血管改变情况.观察并比较不同病程的DM大鼠视网膜凋亡细胞及CTGF表达情况、视网膜微血管形态变化及周细胞数量变化.对各组大鼠视网膜的细胞凋亡指数、CTGF表达阳性率及血管周细胞数量进行比较采用LSD-t检验法,两变量间的相关性分析采用线性相关法.结果 CON组大鼠视网膜细胞凋亡和CTGF表达均阴性.DM2组、DM4组、DM6组大鼠视网膜细胞凋亡指数分别为0.05±0.01、0.25±0.03及0.52±0.02;组间两两比较,差异均有统计学意义(DM2与DM4组比较t=21.432,DM2与DM6组比较t=50.843,DM4与DM6组比较t=29.410;P<0.05),表明随DM病程延长,视网膜凋亡指数逐渐增加.DM2组、DM4组、DM6组大鼠视网膜CTGF表达阳性率分别为(22.79±2.99)%、(41.73±2.59)%、(55.27±2.68)%,组间两两比较,差异均有统计学意义(DM2与DM4组比较t=15.345,DM2与DM6组比较t=26.316,DM4与DM6组比较t=10.971;P<0.05),表明随DM病程延长,视网膜CTGF表达水平逐渐增高.CON及DM2组大鼠视网膜血管走形良好,DM4组大鼠视网膜部分血管渐变僵硬、狭窄;DM6组大鼠视网膜血管主干僵硬,部分微血管狭窄明显.与CON组视网膜血管周细胞数量对比,DM2组未见明显变化;随病程延长,DM4组和DM6组大鼠视网膜血管周细胞数量较CON组逐渐减少,差异均有统计学意义(t=3.367,6.667;P<0.05).DM大鼠视网膜细胞凋亡指数与CTGF阳性表达水平呈显著正相关(r=0.958,P<0.05),凋亡指数和CTGF阳性表达水平与视网膜血管周细胞数量均呈显著负相关(r=-0.540,-0.595;P<0.05).结论 在DM大鼠视网膜组织中,凋亡细胞和致纤维化因子CTGF的产生均早于微循环血管走形及周细胞的改变.(中华眼科杂志,2011,47:521-526)
目的 研究糖尿病大鼠視網膜凋亡細胞和結締組織生長因子(CTGF)錶達情況及其在微循環改變中的作用.方法 實驗研究.55隻成年雄性Wistar大鼠,以隨機數字錶法,隨機分為正常對照(CON)組(10隻鼠)和糖尿病(DM)組(45隻鼠).根據病程不同,將存活的30隻DM組大鼠再分為2箇月組(DM2)、4箇月組(DM4)及6箇月組(DM6),每組各10隻鼠.用末耑脫氧覈糖覈痠轉移酶介導的dUTP缺口末耑標記(TUNEL)法檢測視網膜凋亡細胞,用免疫組織化學法測定CTGF錶達水平,以視網膜消化鋪片過碘痠雪伕(PAS)染色法觀察視網膜血管改變情況.觀察併比較不同病程的DM大鼠視網膜凋亡細胞及CTGF錶達情況、視網膜微血管形態變化及週細胞數量變化.對各組大鼠視網膜的細胞凋亡指數、CTGF錶達暘性率及血管週細胞數量進行比較採用LSD-t檢驗法,兩變量間的相關性分析採用線性相關法.結果 CON組大鼠視網膜細胞凋亡和CTGF錶達均陰性.DM2組、DM4組、DM6組大鼠視網膜細胞凋亡指數分彆為0.05±0.01、0.25±0.03及0.52±0.02;組間兩兩比較,差異均有統計學意義(DM2與DM4組比較t=21.432,DM2與DM6組比較t=50.843,DM4與DM6組比較t=29.410;P<0.05),錶明隨DM病程延長,視網膜凋亡指數逐漸增加.DM2組、DM4組、DM6組大鼠視網膜CTGF錶達暘性率分彆為(22.79±2.99)%、(41.73±2.59)%、(55.27±2.68)%,組間兩兩比較,差異均有統計學意義(DM2與DM4組比較t=15.345,DM2與DM6組比較t=26.316,DM4與DM6組比較t=10.971;P<0.05),錶明隨DM病程延長,視網膜CTGF錶達水平逐漸增高.CON及DM2組大鼠視網膜血管走形良好,DM4組大鼠視網膜部分血管漸變僵硬、狹窄;DM6組大鼠視網膜血管主榦僵硬,部分微血管狹窄明顯.與CON組視網膜血管週細胞數量對比,DM2組未見明顯變化;隨病程延長,DM4組和DM6組大鼠視網膜血管週細胞數量較CON組逐漸減少,差異均有統計學意義(t=3.367,6.667;P<0.05).DM大鼠視網膜細胞凋亡指數與CTGF暘性錶達水平呈顯著正相關(r=0.958,P<0.05),凋亡指數和CTGF暘性錶達水平與視網膜血管週細胞數量均呈顯著負相關(r=-0.540,-0.595;P<0.05).結論 在DM大鼠視網膜組織中,凋亡細胞和緻纖維化因子CTGF的產生均早于微循環血管走形及週細胞的改變.(中華眼科雜誌,2011,47:521-526)
목적 연구당뇨병대서시망막조망세포화결체조직생장인자(CTGF)표체정황급기재미순배개변중적작용.방법 실험연구.55지성년웅성Wistar대서,이수궤수자표법,수궤분위정상대조(CON)조(10지서)화당뇨병(DM)조(45지서).근거병정불동,장존활적30지DM조대서재분위2개월조(DM2)、4개월조(DM4)급6개월조(DM6),매조각10지서.용말단탈양핵당핵산전이매개도적dUTP결구말단표기(TUNEL)법검측시망막조망세포,용면역조직화학법측정CTGF표체수평,이시망막소화포편과전산설부(PAS)염색법관찰시망막혈관개변정황.관찰병비교불동병정적DM대서시망막조망세포급CTGF표체정황、시망막미혈관형태변화급주세포수량변화.대각조대서시망막적세포조망지수、CTGF표체양성솔급혈관주세포수량진행비교채용LSD-t검험법,량변량간적상관성분석채용선성상관법.결과 CON조대서시망막세포조망화CTGF표체균음성.DM2조、DM4조、DM6조대서시망막세포조망지수분별위0.05±0.01、0.25±0.03급0.52±0.02;조간량량비교,차이균유통계학의의(DM2여DM4조비교t=21.432,DM2여DM6조비교t=50.843,DM4여DM6조비교t=29.410;P<0.05),표명수DM병정연장,시망막조망지수축점증가.DM2조、DM4조、DM6조대서시망막CTGF표체양성솔분별위(22.79±2.99)%、(41.73±2.59)%、(55.27±2.68)%,조간량량비교,차이균유통계학의의(DM2여DM4조비교t=15.345,DM2여DM6조비교t=26.316,DM4여DM6조비교t=10.971;P<0.05),표명수DM병정연장,시망막CTGF표체수평축점증고.CON급DM2조대서시망막혈관주형량호,DM4조대서시망막부분혈관점변강경、협착;DM6조대서시망막혈관주간강경,부분미혈관협착명현.여CON조시망막혈관주세포수량대비,DM2조미견명현변화;수병정연장,DM4조화DM6조대서시망막혈관주세포수량교CON조축점감소,차이균유통계학의의(t=3.367,6.667;P<0.05).DM대서시망막세포조망지수여CTGF양성표체수평정현저정상관(r=0.958,P<0.05),조망지수화CTGF양성표체수평여시망막혈관주세포수량균정현저부상관(r=-0.540,-0.595;P<0.05).결론 재DM대서시망막조직중,조망세포화치섬유화인자CTGF적산생균조우미순배혈관주형급주세포적개변.(중화안과잡지,2011,47:521-526)
Objective To study the cell apoptosis and the expression of connective tissue growth factor (CTGF) in the retina of diabetic rats and to explore their contributions to the changes of microcirculation. Methods It was a experiment study. Fifty-five adult male Wistar rats were divided into two groups, normal control group (CON,10 rats) and diabetes mellitus group (DM, 45 rats). The 30 surviving rats in the DM group were further divided into 3 groups based on the time of observation, 2 month (DM2), 4 month (DM4) and 6 month (DM6) groups, with 10 rats in each group. Cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). Expression of CTGF was determined by immunohistochemical study. Retinal vessels were observed by retinal digest stretched periodic acid Schiff (PAS) staining. Results Immunohistochemical staining and TUNEL staining revealed negative results in normal control group. Retinal cell apoptosis index increased gradually from DM2 to DM6, the differences between any two groups were statistically significant (t2-4,2-6,4-6=21.432, 50.843, 29.410;P<0.05). Expression of CTGF in the retina increased from DM2-DM6, the differences between any two groups were statistically significant (t2-4,2-6,4-6=15.345, 26.316, 10.971;P<0.05). PAS staining of retinal blood vessels obtained negative results in the CON and DM2 groups. Part of retinal capillaries were slightly stiff and narrow in DM4 group. Retinal capillaries in DM6 group were trunk stiff and were narrowed obviously. The number of pericytes was reduced in DM4, and progressed following the course of diabetes. The number of pericytes in the DM2 group did not different from that in the CON group (t=0.875,P=0.387). The number of pericytes in the DM4 and DM6 group were significantly decreased as compared to the CON group (t=3.367,6.667;P<0.05). Retinal cellular apoptosis index had a significant positive correlation to the expression of CTGF (r=0.958,P<0.05). Number of pericytes was significantly correlated (negative correlation) with retinal cellular apoptosis index and the expression of CTGF (r=-0.540, -0.595; P<0.05). Conclusions The appearances of cellular apoptosis and fibrosing factor CTGF in the retina of diabetic rats occurred earlier than the changes of microcirculation and the number of capillary pericytes.
