“益阳坚阴”复方中药对形觉剥夺性近视豚鼠眼眼轴轴、MMP-2及COL-I表达的时效研究

Time-dependent study of the effects of the compound Chinese medicine "Yiyang Jianyin" on form deprivation myopia in guinea pigs,as well as MMP-2 and COL-I ex-pression

目的 观察“益阳坚阴”复方中药对形觉剥夺性近视(FDM)豚鼠的眼轴、MMP-2及COL-I表达的时效研究。方法 选取3周龄雄性花豚鼠72只,随机分为空白对照组(A组)、模型组(B组)、中药干预组(C组);B组及C组豚鼠均缝合右眼;A组和B组每日予0.9%氯化钠注射液10 mL/kg灌胃,C组每日予10 mL/kg的“益阳坚阴”复方中药灌胃。连续干预2周及4周后取材,所有豚鼠干预前后行A超测量眼轴,干预后取材行Western-blot法检测豚鼠视网膜组织中基质金属蛋白酶-2(MMP-2)及巩膜组织中巩膜I型胶原(COL-I)的含量,对数据进行统计分析。结果 (1)眼轴:①干预2周后,3组干预前后比较,眼轴均有增长(Z_(A)=-5.504,Z_(B)=-7.491,Z_(C)=-7.382,均P=0.000)。3组间眼轴比较,差异有统计学意义(H=20.439,P=0.000)。与A组比较,B组豚鼠眼轴明显增长(Z=-3.941,P=0.000);而C组眼轴无明显增长(P>0.05)。与B组比较,C组眼轴较短(Z=-3.392,P=0.001)。②干预4周后,3组干预前后比较,眼轴均增长(Z_(A)=-6.819,Z_(B)=-8.466,Z_(C)=-8.853,均P=0.000)。3组间眼轴比较,差异有统计学意义(H=17.997,P=0.000)。与A组比较,B组及C组豚鼠干预后眼轴均有增长(Z_(B)=-3.449,P=0.001;Z_(C)=-3.881,P=0.000);与B组比较,C组眼轴差异无统计学意义(P>0.05)。(2)MMP-2、COL-I表达水平:①干预2周后,3组间MMP-2、COL-I蛋白表达水平比较,差异具有统计学意义(F_(MMP-2)=69.529,P=0.000;F_(COL-I)=23.753,P=0.001)。与A组比较,B、C组的MMP-2蛋白表达水平升高(t_(B)=-10.353,P=0.000;t_(c)=-3.733,P=0.020),B组的COL-I蛋白表达水平降低(t=8.155,P=0.001),而C组的COL-I蛋白表达水平比较差异无统计学意义(P>0.05);与B组比较,C组的MMP-2蛋白表达水平降低(t=7.860,P=0.001)、COL-I蛋白表达水平升高(t=-4.670,P=0.010),差异均有统计学意义。②干预4周后,3组间MMP-2、COL-I蛋白表达水平比较,差异具有统计学意义(F_(MMP-2)=143.191,P=0.000;F_(COL-I)=19.765,P=0.002)。与A组比较,B组及C组的MMP-2蛋白表达水平均升高(t_(B)=-19.549,P=0.000;t_(C)=-7.315,P=0.002),COL-I蛋白表达水平均降低(t_(B)=4.837,P=0.008;t_(C)=5.234,P=0.006);与B组比较,C组的MMP-2蛋白表达水平降低(t_(C)=8.483,P=0.001),差异有统计学意义;而COL-I蛋白表达水平,差异无统计学意义(P>0.05)。结论 “益阳坚阴”复方中药在干预2周后可以抑制FDM豚鼠眼轴的发展,推测可能与下调MMP-2的表达,减少COL-I胶原降解有关。

OBJECTIVE To observe the time-dependent effects of the compound Chinese medicine"Yiyang Jianyin"on induced form deprivation myopia (FDM) in guinea pigs,including changes in eye axial length,MMP-2 expression,and COL-I expression.METHODS Seventy-two male guinea pigs at the age of three weeks were randomly divided into three groups:The blank control group (Group A),the model group (Group B),and the intervention group with the compound Chinese medicine (Group C).Guinea pigs in Groups B and C had their right eyes sutured.Group A and Group B received daily intragastric administration of 10 mL/kg of 0.9%sodium chloride injection,while Group C received daily intragastric administration of 10 mL/kg of the compound Chinese medicine"Yiyang Jianyin".After two weeks and four weeks of continuous intervention,eye axial length was measured using A-scan before and after the intervention.Western blot analysis was conducted on the retinal tissue to measure the content of matrix metalloproteinase-2 (MMP-2),and the content of type I collagen (COL-I) was measured in the scleral tissue.Statistical analysis was performed on the data.RESULTS (1) Eye axial length:①After two weeks of intervention,all three groups showed an increase in eye axial length compared to before the intervention (Z_A=-5.504,Z_B=-7.491,Z_C=-7.382,all P=0.000).There was a statistically significant difference in eye axial length among the three groups (H=20.439,P=0.000).Compared to Group A,the eye axial length of guinea pigs in Group B significantly increased (Z=-3.941,P=0.000),while there was no significant increase in eye axial length in Group C (P>0.05).Compared to Group B,the eye axial length in Group C was shorter (Z=-3.392,P=0.001).②After four weeks of intervention,all three groups showed an increase in eye axial length compared to before the intervention (Z_A=-6.819,Z_B=-8.466,Z_C=-8.853,all P=0.000).There was a statistically significant difference in eye axial length among the three groups (H=17.997,P=0.000).Compared to Group A,both Group B and Group C showed an increase in eye axial length after intervention (Z_B=-3.449,P=0.001;Z_C=-3.881,P=0.000).There was no statistically significant difference in eye axial length between Group B and Group C (P>0.05).(2) MMP-2 and COL-I expression levels:①After two weeks of intervention,there were statistically significant differences in MMP-2 and COL-I protein expression levels among the three groups (F_(MMP-2)=69.529,P=0.000;F_(COL-I)=23.753,P=0.001).Compared to Group A,both Group B and Group C showed an increase in MMP-2 protein expression levels (t_B=-10.353,P=0.000;t_c=-3.733,P=0.020),while Group B showed a decrease in COL-I protein expression levels (t=8.155,P=0.001),and there was no statistically significant difference in COL-I protein expression levels in Group C (P>0.05).Compared to Group B,Group C showed a decrease in MMP-2 protein expression levels (t=7.860,P=0.001) and an increase in COL-I protein expression levels (t=-4.670,P=0.010),with statistically significant differences.②After four weeks of intervention,there were statistically significant differences in MMP-2 and COL-I protein expression levels among the three groups(F_(MMP-2)=143.191,P=0.000;F_(COL-I)=19.765,P=0.002).Compared to Group A,both Group B and Group C showed an increase in MMP-2 protein expression levels (t_B=-19.549,P=0.000;t_C=-7.315,P=0.002) and a decrease in COL-I protein expression levels (t_B=4.837,P=0.008;t_C=5.234,P=0.006).Compared to Group B,Group C showed a decrease in MMP-2 protein expression levels (t_C=8.483,P=0.001),with statistically significant differences.However,there was no statistically significant difference in COL-I protein expression levels (P>0.05).CONCLUSIONS The compound Chinese medicine"Yiyang Jianyin"can inhibit the development of FDM-induced axial elongation in guinea pigs after two weeks of intervention,which may be related to the downregulation of MMP-2 expression and the reduction of COL-I collagen degradation.