木犀草素与青蒿琥酯联合用药防治实验性外伤性增生性玻璃体视网膜病变

Prevention and treatment of experimental traumatic proliferative vitreoretinopathy with luteolin and artesunate

目的探讨木犀草素(LU)与青蒿琥酯(ART)联合用药在外伤性增生性玻璃体视网膜病变(TPVR)中的作用。方法选取16只月龄、体重相近的健康成年雄性青紫蓝兔,实验眼(右眼)玻璃体内注射富含血小板血浆0.3 mL制作TPVR模型。将实验兔随机分成4组,每组4只,对照组玻璃体内注射0.2 mL生理盐水;单用药物组分别在玻璃体内注射0.2 mL的LU(LU组)及0.2 mL的ART(ART组),联合用药组(联合组)玻璃体内注射LU及ART各0.1 mL。注药后第4周通过眼部B超及眼底照相检查观察玻璃体混浊情况及视网膜改变情况并划分相对应的增生性玻璃体视网膜病变(PVR)等级;取眼球组织行Western blot实验检测各组α平滑肌肌动蛋白(α-SMA)表达水平,制作眼球组织切片行HE染色观察结构改变。PVR分级等级资料比较采用秩和检验;多组间比较采用单因素方差分析,组间差异比较使用Bonferroni检验。结果眼部B超及眼底照相检查观察结果提示,ART组、LU组及联合组兔玻璃体改变均优于对照组。ART组、LU组、联合组PVR分级均优于对照组,差异均有统计学意义(均为P<0.05)。Western blot检测结果显示,联合组兔玻璃体内α-SMA表达水平最低;联合组、ART组、LU组α-SMA表达水平均低于对照组,差异均有统计学意义(均为P<0.05);其中,联合组α-SMA与LU组比较,差异有统计学意义(P<0.05),联合组α-SMA与ART组比较,差异无统计学意义(P>0.05)。HE染色结果显示,对照组视网膜脱离、水肿最严重,联合组兔视网膜浅脱离,结构大体平整,损伤轻于ART组及LU组。结论玻璃体内注射LU、ART的联合用药可有效抑制实验动物模型TPVR的发展,优于LU、ART的单用治疗效果。未来应进一步研究联合用药的具体协同机制,为治疗TPVR的研究打下基础。

Objective To investigate the effect of luteolin(LU)and artesunate(ART)in the treatment of traumatic proliferative vitreoretinopathy(TPVR).Methods TPVR models were established by injecting 0.3 mL of platelet-rich plasma into the vitreous cavity of the experimental eyes(right eyes)of 16 healthy adult blue rabbits of a similar weight and age.The experimental rabbits were randomly divided into 4 groups,with 4 rabbits in each group.Those in the control group were intravitreally injected with 0.2 mL of normal saline;those in the LU group were intravitreally injected with 0.2 mL of LU;those in the ART group were intravitreally injected with 0.2 mL of ART;and those in the combination group were intravitreally injected with 0.1 mL of LU and ART,respectively.At the 4th week after injection,the vitreous opacity and retinal changes were recorded with the aid of an ophthalmic B-scan ultrasound device and a fundus camera,and proliferative vitreoretinopathy(PVR)grades were determined.Eyeball tissues were removed for Western blotting to measure the expression level of α smooth muscle actin(α-SMA).Eyeball slices were stained with hematoxylin&eosin(HE)to survey structural changes.PVR grading data were compared using the rank sum test.Multi-group comparisons were conducted using the one-way analysis of variance.Differences among groups were analyzed using the Bonferroni test.Results The observations under the ophthalmic B-scan ultrasound device and fundus camera showed that changes in the vitreous cavity in the ART,LU,and combination groups were better than those in the control group.The PVR grading in the ART,LU,and combination groups was superior to that in the control group,and the differences were statistically significant(all P<0.05).Western blot results showed that the α-SMA level in the ART,LU,and combination groups was lower than that in the control group(all P<0.05)and the lowest in the combination group,and the difference between the combination group and the LU group was statistically significant(P<0.05)but not significant between the combination group and the ART group(P>0.05).The HE staining results showed that the retinal detachment and edema in the control group were the most serious,while the retinal detachment in the combination group was shallow,the structure was generally flat,and the damage was milder than that in the ART and LU groups.Conclusion Intravitreal injection of both LU and ART can effectively inhibit the development of TPVR in experimental animal models,which is superior to the efficacy of LU and ART alone.Further research needs to be conducted on the synergistic mechanisms of the drug combination to lay the foundation for the treatment of TPVR.