Yu Juan, Luo Haomin, Li Ning, Duan Xuanchu
Invest Ophthalmol Vis Sci. 2015 Sep;56(10):6019-28. doi: 10.1167/iovs.15-16558.
We investigated the efficacy of miR-29b in inhibiting scar formation in rabbits who undergo glaucoma filtering surgery (GFS).
Trabeculectomy was performed on 60 rabbits diagnosed with glaucoma. The rabbits were divided into 5 groups: a blank group, single surgery group, positive control group that was treated with intraoperative mitomycin C (MMC), negative control group that was treated twice with empty vector postoperatively, and experimental group that was treated twice with Lentivirus-mediated miR-29b after being subjected to trabeculectomy. The operated eyes were tracked and followed up from postoperative days 1 to 28 (D1-D28). After the surgery, real-time PCR and Western blot analysis were performed on D28.
At 1 week after undergoing GFS, the IOP was significantly lower in the eyes having filtering blebs. No statistically significant difference was found in the four treatment groups. After 21 days, the filtering bleb function score of the experimental group was the highest; however, their IOP was the lowest. On postoperative D28, the mean number of fibroblasts in the experimental group was significantly the lowest. The experimental group had the least collagen content according to Sircol assay. In the experimental group, the level of Col1A1 expression also was reduced in the sclera and conjunctival areas.
A subconjunctival injection of lentivirus-mediated miR-29b lowers postoperative IOP and sustains the function of filtering bleb. It inhibits the proliferation of fibroblasts and reduces collagen deposition by repressing the PI3K/Akt/Sp1 pathway in rabbits subjected to GFS.
我们研究了miR-29b对接受青光眼滤过手术(GFS)的兔子瘢痕形成的抑制作用。
对60只诊断为青光眼的兔子进行小梁切除术。将兔子分为5组:空白组、单纯手术组、术中使用丝裂霉素C(MMC)治疗的阳性对照组、术后用空载体处理两次的阴性对照组,以及小梁切除术后用慢病毒介导的miR-29b处理两次的实验组。对手术眼从术后第1天至第28天(D1-D28)进行跟踪随访。术后第28天进行实时PCR和蛋白质印迹分析。
在接受GFS后1周,有滤过泡的眼睛眼压显著降低。四个治疗组之间未发现统计学上的显著差异。21天后,实验组的滤过泡功能评分最高;然而,其眼压最低。术后第28天,实验组的成纤维细胞平均数量显著最低。根据Sircol测定法,实验组的胶原蛋白含量最少。在实验组中,巩膜和结膜区域的Col1A1表达水平也降低。
结膜下注射慢病毒介导的miR-29b可降低术后眼压并维持滤过泡功能。它通过抑制GFS兔子的PI3K/Akt/Sp1途径来抑制成纤维细胞增殖并减少胶原蛋白沉积。