青光安Ⅱ号方及其有效组分对DBA/2J小鼠视网膜Ras、MEK及ERK蛋白表达的影响

Effect of Qingguang'an Ⅱ Decoction and its effective components on the expression of Ras,MEK and ERK proteins in retina of DBA/2J mice

目的观察青光安Ⅱ号方及其有效组分对DBA/2J小鼠视网膜细胞中Ras、MEK与ERK蛋白表达的影响,探讨其对视神经保护的作用机制。方法将8只雌性C57BL/6J小鼠作为正常对照组(A组),采用随机数字表法将48只雌性DBA/2J小鼠分为6组:模型组(B组)、阳性对照组(C组)、青光安Ⅱ号方汤剂组(D组)、青光安Ⅱ号方有效组分低剂量组(E组)、青光安Ⅱ号方有效组分中剂量组(F组)、青光安Ⅱ号方有效组分高剂量组(G组),每组8只。A、B组给予12.91 mL/(kg·d)蒸馏水;C组给予0.31 g/(kg·d)益脉康分散片;D组给予9.67 g/(kg·d)青光安Ⅱ号方汤剂;E、F、G组分别给予0.85、1.70、3.40 g/(kg·d)青光安Ⅱ号方有效组分。每4周进行眼压测量以及裂隙灯检查;灌胃4周后取视网膜组织行Western blot检测各组小鼠视网膜Ras、MEK与ERK蛋白的表达水平。结果C57BL/6J小鼠不同时相点的眼压不随年龄增长而变化(P>0.05);DBA/2J小鼠眼压在第18~38周高于C57BL/6J小鼠(P<0.05)。裂隙灯检查C57BL/6J小鼠眼前段均未见明显异常;DBA/2J小鼠随周龄增长逐渐出现角膜钙化、虹膜基质萎缩、瞳孔后粘连等眼前节病变。与A组比较,B组Ras、MEK、ERK蛋白表达水平均降低(P<0.05);与B组比较,C、D、E、F、G组Ras蛋白表达水平均升高(P<0.05),G组MEK、ERK蛋白表达水平均升高(P<0.05);与C组比较,G组Ras蛋白表达水平升高(P<0.05);与D组比较,G组MEK蛋白表达水平升高(P<0.05);与E、F组比较,G组Ras蛋白表达水平升高(P<0.05)。结论青光安Ⅱ号方及其有效组分低、中、高剂量对DBA/2J小鼠视网膜Ras、MEK、ERK蛋白可起到不同程度的上调作用,以青光安Ⅱ号方有效组分高剂量组效果最佳。

Objective To observe the effect of Qingguang'an Ⅱ Decoction and its effective components on the expression of Ras,MEK and ERK proteins in retina of DBA/2J mice,and to explore its mechanism of action on optic nerve protection.Methods 8 female C57BL/6J mice were divided into normal control group(group A).A total of 48 female DBA/2J mice were divided into 6 groups by random number table method:model group(group B),positive control group(group C),Qingguang'an Ⅱ Decoction group(group D),Qingguang'an Ⅱ Decoction effective component low-dose group(group E),Qingguang'anⅡDecoction effective component medium-dose group(group F),Qingguang'an Ⅱ Decoction effective component high-dose group(group G),with 8 mice in each group.Groups A and B were given 12.91 mL/(kg·d)distilled water;group C was given 0.31 g/(kg·d)Yimaikang dispersible tablets;group D was given 9.67 g/(kg·d)Qingguang'anⅡDecoction;groups E,F and G were given 0.85,1.70,3.40 g/(kg·d)Qingguang'an Ⅱ Decoction effective component,respectively.Intraocular pressure measurement and slit lamp examination were performed every 4 weeks.After 4 weeks of intragastric administration,retinal tissues were collected for Western blot to detect the relative expression of Ras,MEK and ERK proteins in the retina of mice in each group.Results The intraocular pressure of C57BL/6J mice at different time points did not change with age(P>0.05).The intraocular pressure of DBA/2J mice was higher than C57BL/6J mice at 18-38 weeks(P<0.05).Slit lamp examination showed no abnormal anterior segment of C57BL/6J mice;DBA/2J mice gradually developed corneal calcification,iris matrix atrophy,retropupil adhesion and other anterior segment lesions with age.Compared with group A,the protein expression levels of Ras,MEK and ERK in group B were decreased(P<0.05);compared with group B,the expression of Ras protein in groups C,D,E,F and G increased(P<0.05),and the protein expression levels of MEK and ERK in group G increased(P<0.05);compared with group C,the expression level of Ras protein in group G was increased(P<0.05);compared with group D,the expression of MEK protein in group G increased(P<0.05);compared with groups E and F,the expression of Ras protein in group G increased(P<0.05).Conclusion Qingguang'an Ⅱ Decoction and its effective component at low,medium and high doses could up-regulate Ras,MEK and ERK proteins in retina of DBA/2J mice to different degrees.The effect of the high-dose group of Qingguang'an Ⅱ Decoction is the best.