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目的探讨血清苹果酸脱氢酶(MDH)和嘌呤核苷磷酸化酶(PNP)对肝损伤早期诊断的应用价值。方法 C57小鼠随机分为对照组、低剂量组、中剂量组和高剂量组,各剂量组分别灌胃给予对乙酰氨基酚(APAP)200、350和500 mg/kg,对照组给予生理盐水。于给药后不同时间点(3、6、12和24 h)采集血清,用全自动生化分析仪测定血清谷丙转氨酶(ALT)的水平,紫外分光光度计测定血清MDH的水平,ELISA测定血清PNP的水平。解剖后取肝脏计算肝指数,并进行HE染色,观察肝组织病理形态学变化。结果与对照组同时间点相比,给APAP后3 h,各剂量组MDH均显著升高(P<0.05或P<0.01),PNP在中剂量组及高剂量组显著升高(P<0.05或P<0.01),ALT则无明显变化;给APAP后6 h,各剂量组ALT、MDH和PNP均明显升高(P<0.05或P<0.01);给APAP后12 h,中剂量组及高剂量组ALT和MDH显著升高(P<0.01),PNP在各剂量组均显著升高(P<0.01);给APAP后24 h,各指标水平均有所下降,和对照组同时间点相比,ALT、MDH和PNP仅在高剂量组有显著性差异(P<0.01)。与对照组相比,各时间点高剂量组肝指数均显著升高(P<0.05或P<0.01),显微镜下肝组织病理切片可见明显的肝细胞脂肪变性,高剂量组出现水肿。结论 MDH和PNP在肝损伤早期ALT没有发生明显变化时就显著升高,具有较好的灵敏性,且变化趋势与ALT相同,与病理结果一致,可以作为APAP致肝损伤早期的生物学标志。
Objective To investigate the value of serum malate dehydrogenase (MDH) and purine nucleoside phosphorylase (PNP) in the early diagnosis of liver injury. Methods C57 mice were randomly divided into control group, low dose group, middle dose group and high dose group. Each dose group was given orally paracetamol (APAP) at 200, 350 and 500 mg / kg respectively. The control group was given normal saline . Serum was collected at different time points after administration (3, 6, 12 and 24 h), serum alanine aminotransferase (ALT) level was measured by automatic biochemical analyzer, serum MDH level was measured by UV spectrophotometer, PNP level. After liver dissection, the liver index was calculated and HE staining was performed to observe the pathological changes of liver tissue. Results Compared with the control group at the same time point, the MDH of each dose group was significantly increased 3 h after APAP (P <0.05 or P <0.01), while the PNP was significantly increased in the medium and high dose groups (P <0.05 (P <0.05 or P <0.01), while there was no significant difference in ALT. At 6 h after APAP, ALT, MDH and PNP in each dose group were significantly increased (P <0.05 or P <0.01) The levels of ALT and MDH in the high dose group were significantly increased (P <0.01), and the levels of PNP in each dose group were significantly increased (P <0.01). After 24 h of APAP, the levels of each index decreased, ALT, MDH and PNP were significantly different only in the high-dose group (P <0.01). Compared with the control group, the liver index in the high-dose group was significantly increased at each time point (P <0.05 or P <0.01). The hepatic steatosis was obvious under the microscope and the edema occurred in the high-dose group. Conclusions MDH and PNP are significantly elevated in the early stage of liver injury and have good sensitivity. ALT is the same with ALT. It is consistent with pathological findings and can be used as a biomarker of APAP-induced early liver injury.