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[外源性腐胺对大鼠肝脏功能及肝细胞凋亡的影响]

[Influence of exogenous putrescine on the function of liver and apoptosis of liver cells in rats].

作者信息

Zhou Yueping, Rong Xinzhou, Fan Guicheng, Liu Sirong, Wei Yaming

机构信息

Department of Burns, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou 510180, China.

Department of Burns, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou 510180, China. Email:

出版信息

Zhonghua Shao Shang Za Zhi. 2014 Feb;30(1):46-50.

Abstract

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 96 in 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 LP at PIH 24, 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.

摘要

目的

探讨外源性腐胺对大鼠肝脏功能及肝细胞凋亡的影响。

方法

将90只健康清洁级SD大鼠按随机数字表法分为对照组(C组,n = 10,腹腔注射2 mL生理盐水)、低剂量腐胺组(LP组,n = 40)和高剂量腐胺组(HP组,n = 40)。后两组大鼠分别按25或50 μg/g的剂量腹腔注射约2 mL腐胺(2.5或5.0 g/L)。于注射后24小时处死C组10只大鼠,后两组分别于注射后24、48、72、96小时各处死10只大鼠。采集心脏血液测定血清谷丙转氨酶(ALT)和谷草转氨酶(AST)含量。取肝脏进行大体观察及HE染色组织形态学观察。采用原位末端标记法显示凋亡情况并计算凋亡指数(AI)。三组间及组内不同时间点的数据采用单因素方差分析或Welch检验;配对比较采用LSD或Dunnett's T3检验;LP组和HP组之间的数据采用两因素析因设计方差分析。

结果

(1)各组大鼠肝脏大体观察均无明显异常。C组大鼠肝脏组织正常。LP组和HP组大鼠肝脏偶见轻度水肿,但未见坏死细胞。(2)LP组注射后24、48、96小时的ALT含量及72、96小时的AST含量分别为(38±10)、(45±6)、(34±4)、(207±18)、(196±19)U/L,HP组注射后72、96小时的ALT含量及24、72、96小时的AST含量分别为(38±6)、(48±5)、(213±43)、(209±40)、(230±29)U/L。均显著高于C组大鼠[(29±5)、(163±42)U/L,P值均<0.01]。LP组与HP组在注射后48、72、96小时的ALT含量及96小时的AST含量差异有统计学意义(P值均<0.05)。与各组注射后24小时比较,LP组注射后48小时大鼠的ALT含量、HP组注射后96小时大鼠的ALT含量,以及LP组注射后48、72、96小时大鼠的AST含量和HP组注射后48小时大鼠的AST含量均显著升高或降低(P值均<0.05)。析因分析显示,LP组和HP组之间,不同浓度腐胺对AST含量的影响差异有统计学意义(F = 12.21,P = 0.001),但对ALT含量的影响差异无统计学意义(F = 0.01,P = 0.974)。(3)LP组注射后24、48、72小时大鼠的AI值分别为(5.69±0.38)%、(13.80±1.66)%、(11.56±1.74)%,HP组注射后72、96小时大鼠的AI值分别为(10.29±1.43)%、(15.29±1.41)%。均明显高于对照组注射后24小时的AI值[(3.50±0.30)%,P值均<0.01]。LP组与HP组在注射后24、48、96小时的AI值差异有统计学意义(P值均<0.05)。与各组注射后小时比较,LP组和HP组注射后48、72、96小时大鼠的AI值均显著升高或降低(P值均<0.05)。析因分析显示,腐胺浓度和刺激时间对AI值影响的差异均有统计学意义(F值分别为22.95和130.44,P值均<0.01)。

结论

腹腔注射25或50 μg/g剂量外源性腐胺可导致大鼠肝脏出现一定程度的功能损害及肝细胞凋亡。剂量越高、刺激时间越长,损害及凋亡越明显。

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