青光安Ⅱ号方对DBA/2J小鼠视网膜中Rho/ROCK相关因子的影响

Effect of Qingguangan Ⅱ on Rho/ROCK associated factors in the retina of DBA/2J mice

目的:观察青光安Ⅱ号方对DBA/2J小鼠视网膜中RhoA mRNA、ROCK mRNA、Caspase-3mRNA及Bcl-2 mRNA转录的影响。方法:使用48只DBA/2J小鼠随机平均分成模型组、青光安Ⅱ号汤剂组、青光安Ⅱ号有效组分低、中、高浓度组及阳性对照组(益脉康分散片组)6组,同时使用8只C57BL/6J小鼠作为空白组,DBA/2J小鼠喂养到38周后形成青光眼模型,干预4周后,采用实时荧光定量PCR(Quantitative Real-time,PCR)检测DBA/2J小鼠视网膜中RhoA mRNA、ROCK mRNA、Caspase-3 mRNA及Bcl-2 mRNA的转录情况。结果:干预4周后,在RhoA mRNA、ROCK mRNA和Caspase-3 mRNA的转录中,与空白组相比,其它6组的相对表达量均升高,模型组、益脉康分散片组及青光安Ⅱ号方有效组分低浓度组有统计学差异(P<0.05);与模型组相比较,其它6组的相对表达量均低于模型组,其中RhoA mRNA转录中的青光安Ⅱ号方汤剂组和青光安Ⅱ号方有效组分高浓度组有统计学差异(P<0.05);ROCK mRNA和Caspase-3 mRNA的转录中,模型组与青光安Ⅱ号方有效组分中、高浓度组统计学有差异(P<0.05);在Bcl-2 mRNA转录中,与空白组比较,其它6组相对表达量均下降,模型组、青光安Ⅱ号方汤剂组和青光安Ⅱ号方有效组分低浓度组统计学有差异(P<0.05);青光安Ⅱ号方有效组分中、高浓度组的Bcl-2 mRNA的相对表达量均高于模型组,差异有统计学意义(P<0.05)。结论:高浓度青光安Ⅱ号方可能是通过抑制Rho/ROCK信号通路而减弱了视网膜神经节细胞(RGCs)中Caspase-3的转录,激活了Bcl-2的表达,从而减弱了RGCs的凋亡。

Objective:The effect of Qingguangan Ⅱ on the transcription of RhoA mRNA,ROCK mRNA,Caspase-3mRNA and Bcl-2 mRNA in the retina of DBA/2J mice was observed. Methods:Forty-eight DBA/2J mice were randomly divided into six groups:model groups,Qingguangan Ⅱ decoction group,low concentration,medium concentration and high concentration group of Qingguangan Ⅱ effective ingredient and positive control group(Yimaikang tablet group),and eight C57BL/6 mice were used as blank group,DBA/2J mice were fed until 38 weeks before forming a glaucoma model,The transcription of RhoA mRNA,ROCK mRNA,Caspase-3 mRNA and Bcl-2 mRNA in the retinal of DBA/2J mice was detected using real-time fluorescence quantitative PCR(Quantitative Real-time PCR)after 4 weeks of intervention. Results:Four weeks after the intervention,In the transcription of the RhoA mRNA,ROCK mRNA and the Caspase-3 mRNA,Compared to the blank groups,Relative expression was increased in the other 6 groups,There are statistical differences in the model group,Yimaikang tablet group and low concentration group(P<0.05);In comparison to the model groups,The other 6 groups were lower than the model group,Among them,there are statistical differences between the effective groups of Qingguangan Ⅱ decoction and high concentration group of Qingguangan Ⅱ effective ingredient in RhoA mRNA transcription(P<0.05);In the transcription of the ROCK mRNA and the Caspase-3 mRNA,Statistics have differences between the model group and the effective component of the medium and high concentration group(P<0.05);In the Bcl-2 mRNA transcription,Compare them to blank groups,Relexpression expression decreased in the other 6 groups,Statistics have differences between model group,Qingguangan Ⅱ decoction group and low concentration groups(P<0.05);The relative expression of Bcl-2 mRNA in high concentration group of effective component is higher than that of the model group,There are differences in statistics(P<0.05). Conclusion:The high concentration of QingguanganⅡ prescription probably attenuated Caspase-3 transcription in retinal ganglion cells by inhibiting the Rho/ROCK signaling pathway and activated Bcl-2 expression by inhibiting ROCK signaling,which attenuated apoptosis in retinal ganglion cells.