胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体、白细胞介素－6的表达及肝功能恢复的影响

doi:10.3877/cma.j.issn.1673-9450.2018.06.010

中华损伤与修复杂志(电子版) ›› 2018, Vol. 13 ›› Issue (06) : 450 -454. doi: 10.3877/cma.j.issn.1673-9450.2018.06.010

所属专题：文献；

论著

胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体、白细胞介素－6的表达及肝功能恢复的影响

杜汉朋¹^,^†(

), 陈伟², 孔凡东¹, 吴铭¹

^1. 511400　广州，南方医科大学附属何贤纪念医院普通外科
^2. 510080　广州，中山大学附属第一医院胆胰外科

收稿日期:2018-10-12 出版日期:2018-12-01
通信作者: 杜汉朋
基金资助:
广东省科技计划项目(2017ZC0432)

Effect of bile acid on expression of farnesoid X receptor, interleukin-6 and recovery of liver function after hepatectomy in rats with cirrhosis of liver

Hanpeng Du¹^,^†(), Wei Chen², Fandong Kong¹, Ming Wu¹

^1. Department of General Surgery, Hexian Memorial Hospital, Affiliated to Southern Medical University, Guangzhou 511400, China
^2. Department of Hepatobiliary Surgery, First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, China

Received:2018-10-12 Published:2018-12-01
Corresponding author: Hanpeng Du
About author:
Corresponding author: Du hanpeng, Email: 13450201177@qq.com

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引用本文：

杜汉朋, 陈伟, 孔凡东, 吴铭. 胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体、白细胞介素－6的表达及肝功能恢复的影响[J/OL]. 中华损伤与修复杂志(电子版), 2018, 13(06): 450-454.

Hanpeng Du, Wei Chen, Fandong Kong, Ming Wu. Effect of bile acid on expression of farnesoid X receptor, interleukin-6 and recovery of liver function after hepatectomy in rats with cirrhosis of liver[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2018, 13(06): 450-454.

目的

探讨胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体(FXR)、白细胞介素-6(IL-6)的表达和肝功能恢复的影响作用。

方法

将24只SD大鼠采用0.03%硫代乙酰胺溶液喂养方法制备肝硬化模型，行肝切除术后采用简单随机法分为对照组和实验组，每组各12只。其中对照组用普通标准饲料喂养，而实验组用含0.2%胆汁酸的饲料喂养。1周后抽取下腔静脉血液行肝功能检查，随后处死实验动物，迅速切取残肝组织备用。分别采用蛋白质印迹法和全自动化学发光分析等方法检测FXR、IL-6的表达及肝功能恢复情况。数据比较采用t检验。

结果

实验组中天冬氨酸基转移酶(AST)、丙氨酸转氨酶(ALT)和体内胆汁酸水平分别为(156.4±32.5) U/L、(203.3±38.6) U/L、(6.68±3.90) μmol/L，均低于对照组(215.6±42.3) U/L、(273.8±42.1) U/L、(6.51±4.30) μmol/L，差异均有统计学意义(t＝4.432、2.566、2.114，P＝0.004、0.025、0.035)；实验组中FXR的表达量为0.45±0.16，而对照组的表达量为0.28±0.08，差异有统计学意义(t＝4.47，P＝0.003)；实验组中肝脏再生指标肝细胞有丝分裂指数(MI)、肝细胞增殖细胞核抗原(PCNA)标记指数和细胞核DNA含量分别为(2.89±0.67)%，(4.76±0.89)%和(8.82±2.12)%，均高于对照组(1.12±0.34)%，(2.46±0.68)%和(3.98±1.16)%，差异均有统计学意义(t＝4.688、5.069、6.858，P＝0.003、0.006、0.005)；而实验组中IL-6的水平为(1.34±0.56) ng/mL，对照组为(1.68±0.55) ng/mL，实验组明显低于对照组，差异有统计学意义(t＝2.29，P＝0.02)。

结论

胆汁酸可以通过启动FXR、减少IL-6炎性介质、促进大鼠肝功能恢复和肝脏再生。

关键词: 肝硬化, 肝切除术, 肝再生, 胆汁酸

Objective

To evaluate the effect of bile acid on the expression of farnesoid X receptor, interleukin-6 and recovery of liver function after hepatectomy in rats with cirrhosis of liver.

Methods

The liver cirrhosis model was prepared by feeding 24 SD rats with 0.03% thioacetamide solution. After the hepatectomy, the rats were divided into control group and experimental group by simple random method, 12 rats in each group. The control group was fed with normal standard feed, while the experimental group was fed with a diet containing 0.2% bile acid. One week later, the inferior vena cava blood was taken for liver function test, and then the experimental animals were sacrificed, and the residual liver tissue was quickly taken out for use. The expression of FXR and IL-6 and the recovery of liver function were detected by Western blotting and automatic chemiluminescence analysis. Data comparison was performed using the t test.

Results

Aspartate aminotransferase(AST), alanine transaminase(ALT) and in vivo bile acid levels in the experimental group were (156.4±32.5) U/L, (203.3±38.6) U/L, (6.68±3.90) μmol/L, which was lower than the control group (215.6±42.3) U/L, (273.8±42.1) U/L, (6.51±4.30) μmol/L, the difference was statistically significant (t=4.432, 2.566, 2.114, P=0.004, 0.025, 0.035); The hepatic mitotic index(MI), hepatocyte proliferating cell nuclear antigen(PCNA) labeling index and nuclear DNA content in the experimental group were (2.89±0.67)%, (4.76±0.89)% and (8.82±2.12)%, which were higher than the control group (1.12±0.34)%, (2.46±0.68)% and (3.98±1.16)%, the difference was statistically significant (t=4.688, 5.069, 6.858, P=0.003, 0.006, 0.005). The level of IL-6 in the experimental group was (1.34±0.56) ng/mL, and that in the control group was (1.68±0.55) ng/mL. The experimental group was significantly lower than the control group, and the difference was statistically significant (t=2.29, P=0.02).

Conclusion

Bile acid can promote FXR, reduce IL-6 inflammatory mediators, promote liver function recovery and liver regeneration in rats.

Key words: Liver cirrhosis, Hepatectomy, Liver regeneration, Bile acid

表1 探针和引物序列

扩增基因		序列
核受体X		?
?	探针	5′-Fam-TGGGAAATGTGCCTGAATGT-tamra-3′
?	上游引物	5′-TGCGGTTGAAGCTATGTTCCT-3′
?	下游引物	5′-TGAGTCATTTTCAGTTCCCCAAT-3′
GAPDH		?
?	探针	5′-Fam-TCCATCCTGGCCCCTCGCTGTGCA-tamra-3′
?	上游引物	5′-CGAATTTGCCTACAGCAACAG-3′
?	下游引物	5′-GACTGAGTGTGGCAGGGACTC-3′

表1 探针和引物序列

扩增基因		序列
核受体X		?
?	探针	5′-Fam-TGGGAAATGTGCCTGAATGT-tamra-3′
?	上游引物	5′-TGCGGTTGAAGCTATGTTCCT-3′
?	下游引物	5′-TGAGTCATTTTCAGTTCCCCAAT-3′
GAPDH		?
?	探针	5′-Fam-TCCATCCTGGCCCCTCGCTGTGCA-tamra-3′
?	上游引物	5′-CGAATTTGCCTACAGCAACAG-3′
?	下游引物	5′-GACTGAGTGTGGCAGGGACTC-3′

表2 2组肝硬化大鼠肝切除术后肝功能水平比较(±s)

组别	鼠数(只)	AST(U/L)	ALT(U/L)	白蛋白(g/L)	体内胆汁酸(μmol/L)
实验组	12	156.4±32.5	203.3±38.6	22.5±3.9	6.68±3.90
对照组	12	215.6±42.3	273.8±42.1	30.6±5.6	6.51±4.30
t值	?	4.432	2.566	3.449	2.114
P值	?	0.004	0.025	0.008	0.035

表2 2组肝硬化大鼠肝切除术后肝功能水平比较(±s)

组别	鼠数(只)	AST(U/L)	ALT(U/L)	白蛋白(g/L)	体内胆汁酸(μmol/L)
实验组	12	156.4±32.5	203.3±38.6	22.5±3.9	6.68±3.90
对照组	12	215.6±42.3	273.8±42.1	30.6±5.6	6.51±4.30
t值	?	4.432	2.566	3.449	2.114
P值	?	0.004	0.025	0.008	0.035

图1 2组肝硬化大鼠肝切除术后FXR表达水平比较。FXR为法尼酯X受体；GAPDH为甘油醛－3－磷酸脱氢酶

表3 2组肝硬化大鼠肝切除术后肝脏再生指标的比较(±s)

组别	鼠数(只)	MI(%)	PCNA标记指数(%)	细胞核DNA含量(%)
实验组	12	2.89±0.67	4.76±0.89	8.82±2.12
对照组	12	1.12±0.34	2.46±0.68	3.98±1.16
t值	?	4.688	5.069	6.858
P值	?	0.003	0.006	0.005

表3 2组肝硬化大鼠肝切除术后肝脏再生指标的比较(±s)

组别	鼠数(只)	MI(%)	PCNA标记指数(%)	细胞核DNA含量(%)
实验组	12	2.89±0.67	4.76±0.89	8.82±2.12
对照组	12	1.12±0.34	2.46±0.68	3.98±1.16
t值	?	4.688	5.069	6.858
P值	?	0.003	0.006	0.005

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