姜黄素治疗大鼠角膜碱烧伤的实验研究

Study of Curcumin in Treating Corneal Alkaline Burn of Rats

目的观察姜黄素对大鼠角膜碱烧伤新生血管(CNV)的抑制作用和对CD11b及角膜上皮凋亡细胞表达的影响,初步探讨其治疗角膜碱烧伤的作用机制。方法选用雄性SD大鼠24只,随机分为4组(每组6只):A组为空白组,B组为模型组,C组为姜黄素组,D组为二甲基亚砜(DMSO)组。除空白组外,采用氢氧化钠眼内烧伤造模,并予相应干预,第3、7、10、14天裂隙灯下观察角膜CNV形态并眼前节照相记录CNV长度,计算CNV面积,进行统计学分析;通过H-E染色法观察角膜病理形态学变化;免疫组化法观察CD11b表达;TUNEL染色法检测角膜上皮细胞凋亡表达。结果裂隙灯下观察B组和D组CNV生长旺盛,管腔粗大,C组CNV稀疏管腔细小;3 d时三组CNV面积分析差异无统计学意义(P>0.05),7、10、14 d时CNV面积比较C组CNV面积显著降低(P<0.05);H-E染色B组和D组角膜基质层内大量的CNV管腔和血细胞,C组基质层偶见CNV,管腔细小;A组CD11b、角膜上皮凋亡细胞在角膜仅少量表达,B组和D组CD11b在基质层新生血管内壁上高表达,角膜上皮层凋亡细胞高表达,C组CD11b低表达,角膜上皮层凋亡细胞低表达。结论姜黄素能有效的抑制大鼠角膜碱烧伤CNV治疗角膜碱烧伤,通过下调CD11b抗炎和下调角膜上皮凋亡细胞抗凋亡发挥作用。

Objective To study the inhibition of curcumin on corneal neovascularization（CNV） in the corneal alkaline burned rats, and the effect on CD11 b and corneal epithelium apoptosis. Then investigate its mechanism on treatment of corneal alkaline burn. Methods 24 male SD rats were randomly divided into 4 groups（6 rats in each group）： Group A was the blank control group, Group B was the model group, Group C was the curcumin group, and Group D was the DMSO group. Except for the blank group, the other groups were modeled by sodium hydroxide to induce the burn in eyes and with the corresponding intervention. On the 3th day, 7th day, 10 th day and 14 th day, CNV was observed and taken photos with slit-lamp microscope. The CNV area was calculated accordingly, and all data were analyzed statistically. The pathological change was observed by H-E staining; the expression of CD11 b in corneal was detected with immunohistochemical method; the corneal epithelial cell apoptosis was detected by TUNEL staining. Results The CNV of Group B and Group D was vigorous, its lumen was wider, CNV of Group C was scattered and small; The CNV area of three groups at 3th day was not statistically different（P〉0.05）; The CNV area of Group C was decreased significantly at 7th,10 th, 14 th day（P〈0.05）. After HE staining, there were amount of lumen of CNV and haemocytes in the stromal layer, CNV of group C was few and with small lumen. CD11 b and TUNEL cells of Group A were lowly expressed in epithelium of cornea; CD11 b of Group B and D in the CNV wall and corneal epithelium apoptosis were highly expressed, while that in Group C were with low expression. In B group, they were obviously expressed and had statistically significant conversely.Compared with B group, they were lower in C group. The expression level between D group and B group was no significant different. Conclusion Curcumin could inhibit CNV effectively in the corneal alkaline burned rats to anti-inflammatory and anti-apoptosis effects through down regulating CD11 b and corneal epithelium apoptosis.