激肽原1作为增生性玻璃体视网膜病变血清分子标志物的验证

Verification of kininogen 1 as a serum molecular marker of proliferative vitreoretinopathy

目的验证激肽原1是否可以作为增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)的血清分子标志物。方法收集PVR患者A、B、C、D级玻璃体(n=8)和相应的PVR患者术前和术后6个月血清样本(n=20),正常捐献眼玻璃体样本(n=8)和健康体检者血清(n=20)做正常对照,PVR术后不同状态的血清样本,包括环扎术后未愈者(n=8)和硅油眼(n=8)。对玻璃体和相应的血清样本分别进行激肽原1的蛋白质印迹法(Western blotting)分析和酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)。结果Western blotting分析显示激肽原1在PVR患者玻璃体和血清中均可检测到,而在正常捐献眼和正常人的血清中未检测到,且在PVR C、D级中明显高于PVR B级。激肽原1可以在所有的PVR患者玻璃体和血清样本中检测到(24/24,100%)。ELISA显示严重PVR(C、D级合并)患者激肽原1浓度玻璃体中为(281.0±63.0)μg·L-1,血清中为(499.8±153.8)μg·L-1,明显高于轻度PVR(A、B级合并)患者[玻璃体:(237.5±32.1)μg·L-1,血清:(402.8±85.5)μg·L-1],二者差异有统计学意义(P<0.05)。PVR患者术后6个月血清中激肽原1明显下降至(81.9±18.6)μg.L-1(P<0.05),仍高于正常对照组(57.9±8.9)μg.L-1(P<0.05)。但环扎术后未愈者血清中激肽原1浓度(116.8±45.1)μg·L-1明显高于正常对照组和硅油眼(51.6±14.1)μg·L-1(P<0.01),亦高于玻璃体切割术后6个月PVR患者(P<0.05)。结论激肽原1是PVR患者玻璃体和血清的特异蛋白质,且与PVR的严重程度和预后评估相关,显示了作为PVR血清分子标志物的可能性。

Objective To detect kininogen 1 as the serum molecular marker of proliferative vitreoretinopathy （PVR） or not. Metbods The vitreous （ n = 8 ） of PVR at grade A,B, C,D and the corresponding serum samples preoperatively and at 6 months postoperatively（ n = 20 ） were collected. The donor vitreous （ n = 8 ） and healthy serum samples （ n = 20） were treated as normal controls. After PVR, different serum samples included irrecoverable eyes after cerclage（ n = 8 ） and silicon oil eyes（ n = 8 ）. All the vitreous and corresponding serum samples were underwent Western blotting analysis and enzyme linked immunosorbent assay（ELISA）. Results The Western blotting analysis outcomes presented that kininogen 1 was only detected in the vitreous and serum samples of PVR, but not in the donor vitreous and normal serum samples. Furthermore, it expressed higher in samples of PVR at C grade and PVR at D grade than those of PVR at B grade. All vitreous and serum samples of PVR patients were detected with kininogen 1 （24/24,100%）. The ELISA outcomes demonstrated that concentrations of kininogen 1 in vitreous（281.0 ± 63.0） μg·L^-1 and in serum samples（499.8 ± 153.8）μg·L^-1 of severe PVR（ C grade combined with D grade） were significantly higher than in vitreous （237. 5 ± 32.1 ） μg·L^-1 and in serum （ 402.8 ± 85.5 ）μg·L^-1 of light PVR （ A grade combined with B grade） （ P 〈 0.05 ）. And the concentration of ldninogen 1 in serum samples significantly decreased at 5 months postoperatively（81.9 ± 18.6）μg·L^-1,bdt was still higher than that in normal serum samples （ 57.9 ± 8.9 ） μg·L^-1（ both P 〈 0.05 ）. The concentration of kininogen 1 in serum of irrecoverable eyes after cerclage （ 116. 8 ± 45. 1 ） μg·L^-1 was significantly higher than that of normal controls, higher than that of silicone oil group（51.6 ± 14. 1 ） μg·L^-1 （ both P 〈 0.01 ）, and also higher than that of PVR patients at 6 months after vitrectomy （P 〈 0.05 ）. Conelusions Kininogen 1 is the specific protein in vitreous and serum of the PVR patients, and it correlates with the severity of PVR and prognostic evaluation. Therefore, kininogen 1 can be the serum biomarker of PVR.