摘要
对云南树鼩(Tupaia belangeri chinensis)心肌、肺、颌下腺、脾脏、肾脏、肾上腺、骨骼肌和大脑8种组织乳酸脱氢酶(LDH)同工酶进行琼脂糖凝胶电泳分析。结果表明,8种组织均呈现为LDH-M型和LDH-H型两种亚基组合而成的5种不同分子形式同工酶。采用分光光度比色定量法测得5种同工酶组分的相对酶活性。对其H亚基和M亚基百分率以及H/M亚基比率进行了统计分析。并就不同酶谱特征与基因表达状况进行了分析讨论。
Lactate dehydrogenase (EC 1.1.1.27) is an oxidoreductase which catalyzes the resversion conversion of phruvate to lactate. The five isoenzymes of lactate dehydrogenase are tetramers composed of four polypeptide chains of two types subunits A and B(or M and H) .The five isoenzymes have been demonstrated in all mammalian tissues,cells and serum. Tree shrew is a specific kind of animal between insectivora and primate. It is one of the most suitable research materials for experimental medicine, experimental pathology and evolutional biology.
The present paper deals with the analysis by agarose gel electrophoresis of
lactate dehydrogenase isoenzymes of eight different tissues of tree shrew
(Tupaia belangeri chinansis) . The eight tissues are, heart, lung, submaxil- lary gland, spleen, kidney, adrenal gland, skeletal muscle and brain. The electrophoresis was performed on 0.5% agarose gel thin layer in a pH 8.6 buffer containing 0.06 mol/L sodium diethylbarbiturate, 0.2 N hydrochloric acid. The isoenzymic activity was visualized with a staining solution consisting of sodium DL-lactate, nicotinamide adenine dinucleotide (NAD), N-methylphenazinium methylsulphate(PMS) and nitro-tetrazolium blue chloride (NBT). The electropherograms of LDH isoenzymes was scanned record at Xs = 570 nm and Xr = 500 nm, scanning speed for 20mm/min. Five different molecular forms of isoenzymes and alterations of isoenzymic patterns were obserbed in the eight tissues of tree shrew. The iyoenzymic phenotypes of five molecular forms are LCH1, LDH2, LDH3, LDH4, LDH5 in heart, submaxi-llary gland and brain, only the phenotype of skeletal muscle were LDH5, LDH4, LDH3, LDH2, LDH1. The LDH4 to LDH1 isoenzymes migrated towards the anode and LDH5 isoenzyme toward the cathode in the pH 8.6 buffer. LDH, is the fastest and LDH5 the slowest in the electrophoresis. The mean values of the five components of LDH isoenzymes in eight itssues were measured. The percentage of H subunit was calculated with the formula H% = LDH1% + 3/4 LDH2% +1/2 LDH3% + 1/4 LDH4%.TheH/M subunits ratio was 2.3, 0.9, 1.5, 1.0, 1.6, 0.94, 0.55 and 1.8 in above mentioned eight tissues, respectively.
关键词
树QU
乳酸脱氢酶
同功酶
Tree shrew, Lactate dehydrogenase isoenzyme, Agarose gelelectrophoresis
