PTEN基因对兔晶状体上皮细胞增殖抑制作用的实验研究

发布时间：2019-01-12 08:27

【摘要】：目的研究白内障术后晶状体上皮细胞增殖过程中是否存在PTEN基因对PI-3k/PKB信号转导途径的抑制作用,为后发性白内障的防治提供实验依据。方法将健康白色家兔42只(84眼)随机分入实验组(36只)和对照组(6只)。实验组兔行双眼透明晶状体皮质吸出术,对照组不做处理。在术后1 d、3 d、1周、2周、1个月和2个月各处死随机选择的6只实验组兔。应用免疫组织化学、原位杂交和RT-PCR方法检测赤道部晶状体上皮细胞中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及PTEN mRNA、PKB mRNA、Ser473-R的表达。结果对照组赤道部LEC核可见PCNA弱阳性表达,实验组术后1d时吸光度(A)值迅速升高为0.86±0.02,与对照组相比,差异有显著统计学意义(P0.001);术后1~7 d PCNA的表达维持在高水平状态;2周后,随着时间的延长,PCNA表达逐渐减弱;术后1个月、2个月时大致恢复至术前状态。对照组兔晶状体上皮细胞胞浆中有PTEN mRNA的表达,吸光度值(A)为0.54±0.03。实验组术后1 d PTEN mRNA相对含量明显下降为0.20±0.02(P0.001),术后1周时略有恢复0.25±0.04(P0.001),术后1个月时恢复术前水平0.53±0.04(P0.001)。PKB mRNA的表达恒定,对照组与实验组各时间点差异均无统计学意义(均为P0.05)。对照组晶状体上皮细胞中Ser473-R的表达较少(0.15±0.01),实验组术后1 d时Ser473-R的吸光度(A)值迅速升高为0.54±0.03,两组差异有显著统计学意义(P0.001);术后1~3 d Ser473-R的表达维持在高水平状态;1周时,Ser473-R表达开始减弱;术后1个月、2个月时大致恢复术前状态。PTEN mRNA与PCNA的表达呈负相关(r=-0.954,P0.001);PKB mRNA相对含量与PCNA表达呈负相关(r=-0.319,P=0.039);Ser473-R与PCNA的表达呈显著正相关(r=0.955,P0.001);Ser473-R与PTEN mRNA的表达呈显著负相关(r=-0.969,P0.001)。结论 PTEN mRNA在正常兔晶状体上皮细胞中有表达,在白内障术后,它可能通过抑制PI-3k信号转导途径中的磷酸化过程而抑制晶状体上皮细胞的增殖。
[Abstract]:Objective to investigate the inhibitory effect of PTEN gene on PI-3k/PKB signal transduction pathway during lens epithelial cell proliferation after cataract surgery, and to provide experimental evidence for the prevention and treatment of post-cataract. Methods 42 healthy white rabbits (84 eyes) were randomly divided into experimental group (36) and control group (6). The experimental group received binocular clear lens cortex aspiration, while the control group did not. Six experimental rabbits were randomly selected for 1 week, 2 weeks, 1 month and 2 months after operation. The expression of proliferating cell nuclear antigen (proliferating cell nuclear antigen,PCNA) and PTEN mRNA,PKB mRNA,Ser473-R in equatorial lens epithelial cells were detected by immunohistochemistry, in situ hybridization and RT-PCR. Results there was a weak positive expression of PCNA in the LEC nucleus of the equatorial part of the control group. The absorbance (A) value of the experimental group increased rapidly to 0.86 卤0.02 on the 1st day after operation, which was significantly different from that of the control group (P0.001). The expression of PCNA was maintained at a high level at 1 and 7 days after operation, and gradually decreased with the prolongation of time 2 weeks later, and returned to the preoperative state at 1 month and 2 months after operation. The expression of PTEN mRNA was found in the cytoplasm of rabbit lens epithelial cells in the control group, and the absorbance value (A) was 0.54 卤0.03. In the experimental group, the relative content at 1 d PTEN mRNA decreased significantly to 0.20 卤0.02 (P0.001), and recovered slightly to 0.25 卤0.04 (P0.001) at 1 week after operation. The expression of P0. 001). PKB mRNA was constant at 1 month after operation. There was no significant difference between the control group and the experimental group at each time point (P0.05). The expression of Ser473-R in lens epithelial cells of the control group was less (0.15 卤0.01). The absorbance (A) value of Ser473-R in the experimental group increased rapidly to 0.54 卤0.03 on the 1st day after operation, and there was a significant difference between the two groups (P0.001). The expression of Ser473-R was maintained at a high level on the 1st and 3rd day after operation, the expression of Ser473-R began to decrease at 1 week, and the expression of. PTEN mRNA was negatively correlated with the expression of PCNA at 1 month and 2 months after operation (r = 0.954, P 0.001). The relative content of PKB mRNA was negatively correlated with the expression of PCNA (r-0.319, P0.039), the expression of Ser473-R was positively correlated with the expression of PCNA (r = 0.955, P 0.001), and the expression of Ser473-R and PTEN mRNA was negatively correlated with the expression of PTEN mRNA (r-0.969, P0.001). Conclusion PTEN mRNA is expressed in normal rabbit lens epithelial cells. It may inhibit the proliferation of lens epithelial cells by inhibiting the phosphorylation of PI-3k signal transduction pathway after cataract surgery.
【作者单位】：中国医科大学附属第一医院眼科;
【基金】：辽宁省自然科学基金资助(编号:201102259)~~
【分类号】：R776.1

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