补肾养血明目方对实验性视网膜变性的干预作用

Effect of Bushen Yangxue Mingmu formula on experimental retinal degeneration

目的观察补肾养血明目方对实验性视网膜变性的干预作用,为临床应用提供客观依据。方法将碘酸钠腹腔注射诱导的视网膜变性C57BL/6N小鼠按随机数字表法随机分为3组:蒸馏水组,补肾养血明目方组和正常对照组,前两组自造模后第1天分别用蒸馏水、补肾养血明目方煎剂灌胃治疗,正常组小鼠不予造模,常规饲养。分别于治疗后7 d、14 d时采用视网膜电生理检查技术、病理学及TUNEL方法观察各组视网膜的生理功能和病变程度。结果治疗后7 d,3组小鼠的暗适应视网膜电图(d-ERG)、振荡电位(OPs)和明适应视网膜电图(l-ERG)的振幅相比较,差异有统计学意义(P<0.05)。病理切片显示蒸馏水组及补肾养血明目方组的视网膜色素上皮(RPE)细胞数量均减少,外核层出现排列紊乱和波浪状改变,但补肾养血明目方组的病变程度较蒸馏水组轻,两组病变范围得分分别为3.18±0.603和3.64±0.924,组间差异无统计学意义(P>0.05)。TUNEL法检测补肾养血明目方组和蒸馏水组RPE细胞的凋亡率分别为36.2%和60.7%,差异有统计学意义(P<0.05)。治疗后14 d,3组小鼠视网膜电图d-ERG、OPs、l-ERG间的振幅差异有统计学意义(P<0.05)。HE染色结果显示2组造模小鼠的视网膜外核层细胞数目也减少,但病变的程度和范围较治疗后7 d时轻,补肾养血明目方组和蒸馏水组病变范围得分分别为2.09±0.539和2.73±0.647,2组相比差异有统计学意义(P<0.05)。TUNEL检测显示补肾养血明目方组RPE凋亡细胞数少于蒸馏水组,且外核层未见细胞凋亡现象,补肾养血明目方组和蒸馏水组凋亡率分别为28.8%和51.5%,差异有统计学意义(P<0.05)。结论补肾养血明目方对实验性视网膜变性有干预作用,可以减轻视网膜病变程度和减少视网膜色素上皮细胞和感光细胞的凋亡,其作用机制有待进一步探讨。

OBJECTIVE To observe the intervention effect of Bushen Yangxue Mingmu formula on experi- mental retinal degeneration, thus to provide evidence for further clinical application. METHODS C57BL/6N mice were induced retinitis degeneration with sodium iodate and divided randomly into two groups： distilled water group, Bushen Yangxue Mingmu group. The two groups of mice were gavaged with distilled water, Bushen Yangxue Mingmu formula respectively from day 1 after sodium iodate treatment. Ten normal mice were used as the normal control without any treatment. On day 7, 14 after treatment, retinal electrophysiology examination, histopathology study and TUNEL test were performed to evaluate the retinopathy. RESULTS On day 7 after treatment, analysed the amplitude of dark adapted electroretinogram （d-ERG）, oscillatory potentials （OPs） and light adapted electroretinogram （1- ERG） in the distilled water group and Bushen Yangxue Mingmu group, and there was no significant difference （P〉 0.05 ）. The retinal pigment cells in distilled water group and Bushen Yangxue Mingmu group were decreased, and the outer nuclear layers appeared disorder and wavy change, but there was less retinopathy in Bushen Yangxue Mingmu group than distilled water group. The lesion grades were 3.18±0.603 and 3.64±0.924, and the difference among the two groups was significant （P〈0.01）, The apoptosis rates determined by TUNEL were 36.2% and 60.7% in the two groups, and the difference among them was significant （P〈0.05）. On day 14, there was significant difference betweenthe two groups in amplitude of retinal d-ERG, OPs, 1- ERG （P〈0.05）.Retinal outer nuclear layer cells in each group was also decreased, but the degree and scope of lesions than the day 7.The lesion grades were 2.09±0.539 and 2.73±0.647 respectively in Bushen Yangxue Mingmu group and distilled water group, there was significant difference among the groups （P〈0.05）. In TUNEL test, it was shown that there were less apoptosis aells in the treament groups, and no apoptosis was found in the outer nuclear layer which was found in the distilled water group. The apoptosis rated in the two was 28.8% and 51.5%, which Bushen Yangxue Mingmu group was significantly less than that of distilled water group （P〈0.05）. CONCLUSIONS Bushen Yangxue Mingmu formula had intervention effect on the experimental retinal degeneration, and could allevi- ate the severity of retinopathy, decrease apoptosis of retinal pigment epithelial cells and photoreceptor cells, though its mechanism needs further study.