电针对负透镜诱导型近视豚鼠视网膜中MMP-3和TIMP-3及Col3α1表达的影响

Effects of electroacupuncture on the expression of matrix metalloproteinase-3,tissue inhibitor of metalloproteinase-3 and collagen typeⅢalpha 1 in the retina of guinea pigs with negative lens-induced myopia

目的:探讨电针对负透镜诱导型近视豚鼠视网膜中基质金属蛋白酶(MMP)-3、金属蛋白酶组织抑制剂(TIMP)-3和III型胶原α1(Col3α1)表达的影响。方法:将80只豚鼠随机分为正常对照组、负透镜诱导型近视组、电针干预组和假穴组,每组20只。正常对照组不做任何干预,负透镜诱导型近视组、电针干预组和假穴组,右眼均配戴-6.0 D透镜,左眼不戴镜。戴镜同时电针干预组在合谷穴与太阳穴给予电针刺激,假穴组豚鼠在假穴位进行干预。造模前,造模2、4 wk检影验光检测屈光度,A超检测眼轴长度,HE染色观察视网膜组织结构变化,定量聚合酶链反应(Q-PCR)和蛋白免疫印迹(WB)检测视网膜中MMP-3、TIMP-3、Col3α1 mRNA和蛋白表达的情况。结果:造模2、4 wk,负透镜诱导型近视组与正常对照组相比眼轴长度均明显增加(均P<0.05),屈光度均明显降低(均P<0.05);与负透镜诱导型近视组相比,电针干预组干预后眼轴长度均减少(均P<0.05),屈光度均增加(均P<0.05)。HE染色显示,正常对照组豚鼠视网膜组织各层分界明显,排列规则;负透镜诱导型近视组视网膜厚度、内外核层厚度及细胞数量减少,排列不规则;电针干预组视网膜整体结构较为完善,排列较规则,组织各层形态结构未出现明显异常。Q-PCR和WB检测结果显示,负透镜诱导型近视组视网膜中MMP-3、TIMP-3和Col3α1 mRNA及蛋白表达均比正常对照组明显升高(均P<0.05);而电针干预组干预后视网膜中MMP-3、TIMP-3和Col3α1mRNA及蛋白表达均较负透镜诱导型近视组明显降低(均P<0.05)。结论:电针能够延缓负透镜诱导型近视豚鼠眼轴增长,下调负透镜诱导型近视豚鼠视网膜中的MMP-3、TIMP-3及Col3α1 mRNA及蛋白表达。

AIM:To investigate the effect of electroacupuncture on the expression of matrix metalloproteinase-3(MMP-3),tissue inhibitor of metalloproteinase-3(TIMP-3)and collagen type III alpha 1(Col3α1)in the retina of guinea pigs with lens-induced myopia.METHODS:A total of 80 guinea pigs were randomly divided into the normal control(NC)group,negative lens-induced myopia(LIM)group,electro-acupuncture(EA)group,and sham group,with 20 guinea pigs in each group.The NC group did not receive any intervention,the LIM,EA and the sham groups were all wearing-6.0 D lens in the right eye and no lens in the left eye.The EA group was given electroacupuncture stimulation at Hegu Point and temple,and guinea pigs in the sham group were given intervention at false points.The refraction was measured by optometry,the axial length was measured by A-ultrasound,and the changes in retinal tissue structure were observed by HE staining.Quantitative polymerase chain reaction(Q-PCR)and western blot(WB)were used to detect the expression of MMP-3,TIMP-3,and Col3α1 at mRNA and protein levels before modeling and at 2 and 4 wk after modeling.RESULTS:The axial length of the negative LIM group was significantly increased compared with the NC group at 2 and 4 wk after modeling(all P<0.05),and the diopter was significantly decreased(all P<0.05);the axial length of the eye in the EA group was decreased compared with the negative LIM group(all P<0.05),and the diopter was increased(all P<0.05).HE staining showed that the boundaries of retinal tissue in the NC group were obvious and arranged regularly.The retinal thickness,the thickness of the inner and outer nuclear layers,and the number of cells were reduced and irregularly arranged in the negative LIM group.The overall structure of the retina in the group EA was relatively perfect,the arrangement was more regular,and the morphological structure of each layer of the tissue did not appear obvious abnormalities.Q-PCR and WB detection results showed that the mRNA and protein relative expression levels of MMP-3,TIMP-3,and Col3α1 in the negative LIM group were significantly higher than those in the NC group(all P<0.05);the expression levels of the EA group was significantly decreased after intervention compared with the negative LIM group(all P<0.05).CONCLUSION:Electroacupuncture can delay axial growth in negative LIM guinea pigs,and downregulate negative LIM induced expression of MMP-3,TIMP-3,and Col3α1 in the retina.