摘要
目的探讨外源性腐胺对大鼠肝功能和肝细胞凋亡的影响。方法将90只健康清洁级SD大鼠按随机数字表法分为3组：对照组10只（腹腔注射生理盐水2mL）、低剂量腐胺组40只、高剂量腐胺组40只。后2组大鼠分别按25、50μg／g剂量，腹腔注射2．5、5．0g／L腐胺溶液约2mL。对照组于注射后24h，另2组分别于注射后24、48、72、96h，每组取10只大鼠处死，心脏采咀测定血清中ALT、AST含量；摘取肝脏大体观察并行liE染色观察组织形态，原位末端标记法检测肝细胞凋亡情况并计算细胞凋亡指数（AI）。对数据行3组间及组内各时相点单因素方差分析或Welch检验，两两比较行LSD或DunnettT3检验；另行低、高剂量腐胺组两因素析因设计方差分析。结果（1）3组大鼠肝脏大体观察无明显异常。对照组大鼠肝组织形态正常；低、高剂量腐胺组大鼠肝组织偶见轻微水肿，未见坏死细胞。（2）低剂量腐胺组大鼠注射后24、48、96h的ALT含量与注射后72、96h的AST含量分别为（38±10）、（45±6）、（34±4）、（207±18）、（196±19）U／L，高剂量腐胺组大鼠注射后72、96h的ALT含量与注射后24、72、96h的AST含量分别为（58±6）、（48±5）、（213±43）、（209±40）、（230±29）U／L，均分别明显高于对照组注射后24h的（29±5）、（163±42）U／L（P值均小于0．01）。低、高剂量腐胺组大鼠注射后48、72、96hALT含量以及注射后96hAST含量组间比较，差异均有统计学意义（P值均小于0。05）。与组内注射后24h比较，低剂量腐胺组大鼠注射后48h与高剂量腐胺组大鼠注射后96hALT含量显著增高（P值均小于0．05），低剂量腐胺组大鼠注射后48、72、96h与高剂量腐胺组大鼠注射后48hAST含量显著增高或降低（P值均小于0．05）。低、高剂量腐胺组析图分析显示，不同腐胺浓度对ALT含量的影响差异无统计学意义（F=0．0l，P=0．974），但对AST含量的影响存在明显差异（F=12．21，P=0．001）。（3）低剂量腐胺组大鼠注射后24、48、72h的AI值分别为（5．69±0．38）％、（13．80±1．66）％、（11．56±1．74）％，高剂量腐胺组大鼠注射后72、96h的AI值分刖为（10．29±1．43）％、（15．29±1．41）％，均明显高于对照组注射后24h的（3．50-4-0．30）％（P值均小于O．01）。低、高剂量腐胺组大鼠注射后24、48、96hAI值组间比较，差异均有统计学意义（P值均小于0．05）。与组内注射后24h比较，低、高剂量腐胺组大鼠注射后48、72、96hAI值显著增高或降低（P值均小于0．05）。低、高剂量腐胺组析因分析显示，不同腐胺浓度及作用时间对AI值的影响均存在明显差异（F值分别为22．95、130．44，P值均小于0．01）。结论按25、50pμg／g剂量腹腔注射外源性腐胺可导致大鼠一定程度的肝功能损害，诱导肝细胞异常凋亡。腐胺剂量越高，作用时间越长，造成的损害和凋亡程度越明显。
Objective To explore the influence of exogenous putrescine on the function of liver and apoptosis of liver cells in rats.Methods Ninety healthy clean SD rats were divided into control group (C,n =10,intraperitoneally injected with 2 mL normal saline),low dosage putrescine group (LP,n =40),and high dosage putrescine group (HP,n =40) according to the random number table.Rats in the latter two groups were intraperitoneally injected with approximately 2 mL putrescine (2.5 or 5.0 g/L) with the dosage of 25 or 50 μg/g.Ten rats from group C at post injection hour (PIH) 24 and 10 rats from each of the latter two groups at PIH 24,48,72,96 were sacrificed.Heart blood was obtained for determination of serum contents of ALT and AST.Liver was harvested for gross observation and histomorphological observation with HE staining.Apoptosis was shown with in situ end labeling,and apoptosis index (AI) was calculated.Data among the three groups and those at different time points within one group were processed with one-way analysis of variance or Welch test; LSD or Dunnett's T3 test was used for paired comparison; factorial design analysis of variance of two factors was applied for data between group LP and group HP.Results (1) No obvious abnormality was observed at gross observation of liver of rats in each group.Liver tissue of rats in group C was normal.Light edema was observed occasionally in liver of rats in groups LP and HP,but necrotic cells were not seen.(2) Content of ALT at PIH 24,48,96 and content of AST at PIH 72 and 96in group LP were respectively (38 ± 10),(45 ±6),(34 ±4),(207 ± 18),(196 ± 19) U/L,and content of ALT at PIH 72 and 96 and content of AST at PIH 24,72,96 in group HP were respectively (38 ±6),(48 ±5),(213 ±43),(209 ±40),(230 ±29) U/L.They were significantly higher than those of rats in group C [(29 ± 5),(163 ± 42) U/L,with P values all below 0.01].There were statistically significant differences between group LP and group HP in the content of ALT at PIH 48,72,96 and content of AST at PIH 96 (with P values all below 0.05).Compared with that at PIH 24 of each group,content of ALT of rats in group LP at PIH 48 and that of rats in group HP at PIH 96,as well as content of AST of rats in group LP at PIH 48,72,96 and that of rats in group HP at PIH 48 were significantly increased or decreased (with P values all below 0.05).Factorial analysis showed that the differences due to different concentration of putrescine on content of AST were statistically significant (F =12.21,P =0.001),but not on content of ALT (F =0.01,P =0.974) between group LP and group HP.(3) AI values of rats in group LPat PIH24,48,72 were respectively (5.69 ±0.38)％,(13.80 ± 1.66)％,(11.56 ± 1.74)％,and AI values of rats in group HP at PIH 72 and 96 were respectively (10.29 ± 1.43) ％,(15.29 ± 1.41) ％.They were all obviously higher than AI value of control group at PIH 24 [(3.50 ± 0.30) ％,with P values all below 0.01].There were statistically significant differences between group LP and group HP in AI value at PIH 24,48,96(with P values all below 0.05).Compared with that at PIH 24 of each group,AI value of rats in groups LP and HP at PIH 48,72,96 were significantly increased or decreased (with P values all below 0.05).Factorial analysis showed that the differences in the influence of concentration of putrescine and stimulation time on AI value were statistically significant (with F values respectively 22.95 and 130.44,P values all below 0.01).Conclusions Intraperitoneal injection of exogenous putrescine in the dosage of 25 or 50 μg/g could lead to certain degree of functional damage of liver and apoptosis of liver cells of rat.The higher the dosage and the longer the stimulation time,the more obvious the damage and apoptosis would be.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2014年第1期46-50,共5页
Chinese Journal of Burns
