电泳法检测肝和血清中醇脱氢酶同工酶

Detection of alcohol dehydrogenase isoenzymes in liver and serum by electrophoresis in liver disease

目的:建立琼脂糖凝胶电泳法检测人血清醇脱氢酶(alcoholdehydrogenase isoenzymes ADH EC 1.1.1.1)同工酶。方法:采用自制琼脂糖凝胶板,摸索实验条件,在pH8.6,74mmol/L巴比妥电泳缓冲系统中进行20mA、20min电泳,可清晰地分离出ADH同工酶三条区带。结果:检测了67例患者血清ADH总活性0.013-0.021Kat/L,同工酶ADHⅠ占ADH总活性0.0l-0.30,ADHⅡ占ADH总活性0.12-0.31,ADHⅢ占ADH总活性0.39-0.80。检测了10例人健康肝组织匀浆的ADH总活性为0.136-0.196Kat/L,同工酶ADHⅠ占总活性的0.07-0.25,同工酶ADHⅡ占总活性的0.19-0.27和同工酶ADHⅢ占总活性0.56-0.73。结论:正常人血清中ADH酶活性很低,为0-1.8×10^(-3)Kat/L,其同工酶不易被检测出。而肝脏组织中含有大量ADH酶活性,当肝脏受到严重损伤时,ADH即从肝细胞内逸出,进入血液,使血清中ADH酶活性明显升高,同时通过琼脂糖电泳对其同工酶的检测可测出三条区带,帮助临床进一步了解肝细胞损伤程度,对患者预后和病程的监测具有一定的临床意义。

AIM: To establish an agaroe gel electrophoretic method for detecting alcohol dehydrogenase isoenzymes in human serum or liver tissue. METHODS: The samples of human liver tissue or serum were electrophoresed with 74 mmol/L diethylbarbital buffer (pH 8.6)of 10 g/L agarose gel.Electrophoresis can separate the Alcohol Dehydrogenase(ADH)isoenzymes to three bands clearly at 20 mA for 20 min in serum. RESULTS: The total ADH activity in serum of sixty-seven patients with liver diseases was ranged from 0.013 Kat/L to 0.021 Kat/L.ADH Ⅰ isoenzyme was ranged from 0.01 to 0.30 of the total activities,ADH Ⅲ isoenzyme was from 0.12 to 0.31.ADH Ⅲ was from 0.39 to 0.80.Total ADH activity in liver tissues of ten healthy subjects was ranged from 0.136 Kat/L to 0.196 Kat/L.ADH Ⅰ isoenzyme was from 0.07 to 0.25 of the total activities.ADH Ⅲ isoenzyme was ranged from 0.19 to 0.27.ADH Ⅲ was from 0.56 to 0.73. CONCLUSION: ADH activity is high is normal human liver. Three ADH isoenzymes can be separated with agarose gel electrophoresis.But serum ADH activity is low. High activities were obtained in serum from persons suffering from serious liver diseases.