电压门控性氯通道3在高眼压模型大鼠小梁网中的表达

目的: 检测电压门控性氯离子通道3(ClC-3)在大鼠小梁网中的表达情况,探讨其在青光眼发病机制中的可能作用。方法: 30只健康雄性大鼠。左眼作为正常对照,右眼前房内注射玻璃酸钠,每周1次,连续10周,制备高眼压模型,每次全麻后玻璃酸钠注射前采用Tono-Pen XL眼压计测量双眼眼压并记录。实验分为正常对照组、玻璃酸钠注射3周组和10周组。分别取3组大鼠的小梁网组织,采用RT-PCR法检测ClC-3mRNA表达水平,免疫组织化学法检测ClC-3蛋白的表达。结果: 玻璃酸钠注射后1、3和10周,大鼠右眼眼压均较注射前增高(P<0.01),注射后1周眼压值较注射前轻微增高,注射后3周眼压较注射前明显增高,但眼压值未达到高眼压成模标准,注射后10周眼压值较注射前明显增高,达到高眼压成模标准。RT-PCR法检测,与正常对照组比较,玻璃酸钠注射3周组大鼠小梁网组织中ClC-3 mRNA表达水平明显增高(t=7.88,P<0.05);玻璃酸钠注射10周组大鼠小梁网组织中ClC-3 mRNA表达水平明显降低(t=15.93,P<0.05)。免疫组织化学检测,3组大鼠小梁网组织中ClC-3蛋白表达均为阳性,玻璃酸钠注射3周组大鼠小梁网组织中ClC-3蛋白表达强度较正常对照组大鼠增强,玻璃酸钠注射10周组大鼠小梁网组织中ClC-3蛋白表达强度较正常对照组大鼠减弱。结论: 大鼠小梁组织中存在ClC-3的表达,ClC-3可能与小梁网的病理调节有关联,进而参与青光眼的发病。

Expression of voltage-gated chloride channel-3 in trabecular meshwork of rats

Objective To detect the expressions of voltage-gated chloride channel 3(ClC-3) in the trabecular meshwork of the rats, and to investigate its possible role in the pathogenesis of glaucoma. Methods The anterior chamber of right eyes of 30 healthy male rats were injected with sodium hyaluronate once a week for ten weeks to set up high intraocular pressure models;the left eyes were used as the blank control.The eye IOP were measured and recorded by Tono-Pen XL after anesthesia and before the injection of sodium hyaluronate.Then the rats were divided into normal control group, sodium hyaluronate injection 3 weeks group and sodium hyaluronate injection 10 weeks group.The trabecular meshwork tissue of the rats in the three groups were taken to detect the ClC-3 gene mRNA levels by RT-PCR method, and to detect the expression levels of ClC-3 protein by immunohistochemical method. Results The IOP of right eyes at 1 week, 3 weeks and 10 weeks after injection of sodium hyaluronate were higher than before injection(P<0.01).The IOP of right eyes at 1 week after injection was increased slightly, and was significantly increased at 3 weeks after injection, but did not reach to the high intraocular pressure model standard.The IOP of right eyes at 10 week after injection was significantly higher than before injection and reached to the high intraocular pressure model standard.The RT-PCR results showed that the ClC-3 gene mRNA expression level in the trabecular meshwork of the rats in sodium hyaluronate injection 3 weeks group was increased compared with normal control group(t=7.88, P<0.05);the ClC-3 gene mRNA expression level in the trabecular meshwork of the rats in sodium hyaluronate injection 10 weeks group was reduced compared with normal control group(t=15.93, P<0.05).The immunohistochemistry results showed that the ClC-3 protein expressions in the trabecular meshwork tissue of the rats in three groups were all positive;the ClC-3 protein expression of the rats in sodium hyaluronate injection 3 weeks group was increased compared with normal control group, and the ClC-3 protein expression of the rats in sodium hyaluronate injection 10 weeks group was reduced compared with normal control group. Conclusion ClC-3 can express in the trabecular tissue of the rats, and ClC-3 may be associated with the pathological regulation of trabecular meshwork, then paticipates in the pathogenesis of glaucoma.