磷酸肌酸对离体大鼠肝脏冷保存的保护作用

曹经琳; 王淼; 王洋; 任贵军; 史艳敏; 窦剑

doi:10.3969/j.issn.1674-7445.2015.03.007

磷酸肌酸对离体大鼠肝脏冷保存的保护作用

doi: 10.3969/j.issn.1674-7445.2015.03.007

石家庄，河北医科大学第三医院肝胆外科(曹经琳、王洋、任贵军、史艳敏、窦剑)；河北邢台市人民医院小儿外科(王淼)

基金项目:

河北省省级重大医学科研课题 zd2013031

详细信息

通讯作者:
窦剑, Email:2423978344@qq.com

中图分类号: R617
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出版历程
- 收稿日期: 2015-01-26
- 网络出版日期: 2021-01-19
- 刊出日期: 2015-05-15

Protective effect of creatine phosphate on isolated rat liver against cold preservation

Department of Hepatobiliary Surgery, the Third Hospital of Hebei Medical University, Shijiazhuang 050051, China

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Corresponding author: Dou Jian, Email:2423978344@qq.com

摘要

摘要: 目的探讨磷酸肌酸(CP)对大鼠离体肝脏冷保存的保护作用。方法建立大鼠肝脏单纯冷保存离体灌注模型, 对照组予单纯威斯康星大学保存液(UW液)灌注肝脏, 低剂量组以UW液为基液加入1 g/100 ml CP灌注肝脏, 中剂量组以UW液为基液加入2 g/100 ml CP灌注肝脏; 高剂量组以UW液为基液加入3 g/100 ml CP灌注肝脏。各组大鼠肝脏分别于4℃相应灌注液中冷保存后0、6、12、18、24 h共5个时间点, 分别检测肝下下腔静脉内保存液的丙氨酸转氨酶(ALT)、乳酸脱氢酶(LDH)含量, 检测肝脏组织丙二醛(MDA)含量、髓过氧化物酶(MPO)活性, 观察肝脏组织肝细胞的凋亡指数(AI)和肝脏组织核因子-κB阳性表达率, 光学显微镜下观察肝脏组织的病理学变化。结果低、中、高剂量组大鼠肝脏在冷保存12 h后, ALT及LDH含量均低于对照组(均为P < 0.05);冷保存18 h后低、中、高剂量组大鼠肝脏组织的MDA、MPO含量均低于对照组(均为P < 0.05);在冷保存12 h及18 h时, 低、中、高剂量组大鼠肝脏的肝细胞AI及核因子-κB阳性表达率均低于对照组(均为P < 0.05);冷保存24 h后, 高剂量组保存液的ALT、MDA含量均明显高于对照组及低、中剂量组(均为P < 0.05)。病理检查结果显示, 高、中、低剂量组大鼠肝脏的损伤明显轻于对照组, 各剂量组之间比较无明显差别。结论在UW液中加入CP对大鼠离体肝脏冷保存有较好的保护作用, 优于单纯应用UW液保存。
- 磷酸肌酸 /
- 肝移植 /
- 器官保存 /
- 冷保存损伤 /
- 保护
Abstract: Objective To discuss the protective effect of creatine phosphate(CP) on isolated rat liver against cold preservation. Methods Isolated perfused rat liver model under simple cold preservation was established. The liver of the control group was perfused with pure University of Wisconsin(UW) solution. With UW solution as the base fluid, the liver of the low-dose group was perfused with 1 g/100 ml CP in UW solution; the liver of the middle-dose group was perfused with 2 g/100 ml CP in UW solution; the liver of the high-dose group was perfused with 3 g/100 ml CP in UW solution. The livers of each group were cold preserved in the corresponding perfusion fluid at 4℃. The content of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in preservation solution in infrahepatic vena cava were determined. The content of malondialdehyde (MDA) and activity of myeloperoxidase (MPO) in liver tissues were detected. The apoptosis index (AI) of liver cells in liver tissues and positive expression rate of NF-κB in liver tissues were observed. Pathologic changes of liver tissues were observed under optical microscope. Results At 12 h after the cold preservation, the content of ALT and LDH in the rat livers of low-, middle-and high-dose groups were lower than those of the control group (all in P < 0.05). At 18 h after the cold preservation, the content of MDA and MPO in the liver tissues of low-, middle-and high-dose groups were lower than those of the control group (all in P < 0.05). At 12 h and 18 h after the cold preservation, AI and positive expression rate of NF-κB in liver cells in the rat livers of low-, middle-and high-dose groups were lower than those of the control group (all in P < 0.05). At 24 h after the cold preservation, the content of ALT and MDA in preservation solution of the high-dose group was obviously higher than that of the control group as well as the low-and middle-dose groups (all in P < 0.05). The results of pathological examination indicated that the injuries to the livers of the high-, middle-and low-dose groups were obviously lighter than that of the control group. There was no obvious difference among each dose group. Conclusions CP in UW solution may well protect the isolated rat liver against cold preservation, which is better than pure UW solution.
- Creatine phosphate /
- Liver transplantation /
- Organ preservation /
- Cold preservation injury /
- Protection

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表 1 各组大鼠肝脏冷保存后保存液的肝功能指标变化

Table 1. Change of liver function indicators of preservation solution after cold storage of rats' liver in each group(x±s)

组别	n	ALT(U/L)					LDH(IU/L)
组别	n	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h
对照组	10	46.5±1.5	53.1±2.7	63.7±3.2	64.5±1.3	73.8±2.0^b	1 641±102	1 916±94	2 272±103	2 549±108	2 809±99
低剂量组	10	45.7±2.3	51.6±1.5	58.5±1.9^a	64.9±3.6	71.3±2.3^b	1 653±115	1 890±84	2 165±86^a	2 458±147	2 735±96
中剂量组	10	45.3±2.1	51.7±1.7	58.5±1.8^a	65.5±2.2	71.7±1.8^b	1 591±109	1 884±103	2 172±88^a	2 434±131	2 767±165
高剂量组	10	46.1±1.6	52.5±1.6	58.9±3.3^a	66.2±4.4	75.7±5.1	1 664±87	1 997±104	2 174±60^a	2 501±700	2 779±62
注：与对照组比较，^aP＜0.05；与高剂量组比较，^bP＜0.05

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表 2 各组大鼠肝脏冷保存后肝脏组织的丙二醛、髓过氧化物酶含量变化

Table 2. Change of MDA and MPO contents of liver tissue after cold storage of rats' liver in each group (x±s)

组别	n	MDA(nmol/mg protein)					MPO(U/g)
组别	n	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h
对照组	10	2.83±0.17	3.52±0.22	4.13±0.25	5.11±0.21	5.87±0.19^b	0.78±0.14	1.19±0.11	1.70±0.19	2.10±0.09	2.53±0.13
低剂量组	10	2.77±0.15	3.42±0.44	4.12±0.21	4.71±0.26^a	5.67±0.17^b	0.74±0.11	1.22±0.10	1.66±0.09	1.97±0.08^a	2.48±0.02
中剂量组	10	2.82±0.15	3.44±0.13	4.24±0.15	4.82±0.25^a	5.57±0.23^b	0.80±0.13	1.22±0.08	1.60±0.18	1.95±0.13^a	2.58±0.14
高剂量组	10	2.67±0.50	3.32±0.24	4.23±0.10	4.97±0.30^a	6.23±0.64	0.73±0.14	1.16±0.15	1.55±0.21	1.94±0.15^a	2.48±0.11
注：与对照组比较，^aP＜0.05；与高剂量组比较，^bP＜0.05

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表 3 各组大鼠肝脏冷保存后肝脏组织的细胞凋亡指数变化

Table 3. Change of apoptosis index of liver tissue after cold storage of rats' liver in each group(x±s)

组别	n	凋亡指数(%)
组别	n	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h
对照组	10	10.4±3.5	17.3±4.7	27.3±3.9	32.2±3.9	35.7±2.5
低剂量组	10	9.5±2.7	13.8±2.6	18.2±2.9^a	26.0±4.4^a	34.8±3.3
中剂量组	10	9.0±3.6	13.0±4.1	19.7±3.9^a	28.0±4.1^a	35.2±1.5
高剂量组	10	9.2±2.9	14.2±1.8	19.6±3.3^a	29.4±3.2^a	36.9±2.9
注：与对照组比较，^aP＜0.05

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表 4 各组大鼠肝脏冷保存后肝脏组织核因子-κB阳性表达率变化

Table 4. Change of NF-κB positive expression rate of liver tissue after cold storage of rats' liver in each group (x±s)

组别	n	核因子-κB阳性表达率(%)
组别	n	保存0 h	保存6 h	保存12 h	保存18 h	保存24 h
对照组	10	22.7±2.4	30.3±3.0	39.3±4.1	53.7±3.1	64.7±3.9
低剂量组	10	19.7±4.5	28.9±2.9	30.9±2.0^a	48.9±2.8^a	62.3±3.8
中剂量组	10	19.6±3.5	26.7±3.7	35.3±3.2^a	49.3±2.0^a	60.2±3.7
高剂量组	10	18.6±4.6	29.2±5.8	34.4±2.7^a	47.6±5.8^a	60.9±7.6
注：与对照组比较，^aP＜0.05

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