联合抗原修复法在免疫组化法检测家兔颈动脉粥样硬化陈旧性蜡块组织MMP-9及TIMP-1表达中的应用

Application of joint antigen retrieval method in detecting MMP-9 and TIMP-1 expressions of rabbit carotid atherosclerosis old wax tissue by immunohistochemical staining

目的探讨联合抗原修复法对免疫组织化学(免疫组化)法检测家兔颈动脉粥样硬化陈旧性蜡块组织基质金属蛋白酶-9(MMP-9)、组织金属蛋白酶抑制物-1(TIMP-1)表达的影响。方法对家兔颈动脉粥样硬化陈旧性蜡块组织连续切片198张,分别采用胰蛋白酶、加热、联合抗原修复法,检测MMP-9、TIMP-1的免疫组化染色效果。结果 0.1%胰蛋白酶(pH 7.6)37℃孵育20 min为胰蛋白酶抗原修复法的最佳条件,0.01 mol/L乙二胺四乙酸缓冲液(pH 8.0)进行微波抗原修复为加热抗原修复法的最佳条件,胰蛋白酶与微波联合进行的抗原修复法明显优于单用胰蛋白酶抗原修复法、加热抗原修复法,差异均有统计学意义(P<0.05)。结论联合抗原修复法更适于免疫组化法检测家兔颈动脉粥样硬化陈旧性蜡块中MMP-9、TIMP-1的表达。

Objective To investigate the influence of the joint antigen retrieval method for detecting MMP-9 and TIMP-1expression of rabbit carotid atherosclerosis old wax tissue by immunohistochemical staining. Methods The serial tissue sections of rabbit carotid atherosclerosis old wax were respectively examined the immunohistochemical staining effects of MMP-9 and TIMP-1 by the pancreatic enzyme,heating and joint antigen retrieval methods. Results The best condition of pancreatic enzyme antigen retrieval method was incubated for 20 min with 0.1% trypsin（p H 7.6） at 37 ℃. The best condition of heating antigen retrieval method was 0.01 mol/L EDTA buffer（p H 8.0） for conducting microwave antigen repair. The antigen retrieval method of pancreatic enzymes associated with microwave was superior to the single use of trypsin retrieval or heating retrieval,the difference was statistically significant（P〈0.05）. Conclusion The joint antigen retrieval method is more suitable to detect the MMP-9 and TIMP-1 expressions of rabbit carotid atherosclerosis old wax by immunohistochemical staining method.