曲美他嗪对胆管缺血再灌注损伤大鼠模型的影响及其机制

张锦锐; 吴昊; 白易; 张雅敏

doi:10.3969/j.issn.1001-5256.2022.11.021

曲美他嗪对胆管缺血再灌注损伤大鼠模型的影响及其机制

DOI: 10.3969/j.issn.1001-5256.2022.11.021

张锦锐^{1, 2},
吴昊^{1, 2},
白易³,
张雅敏^3, , ,

1.
天津医科大学一中心临床学院，天津 300070
2.
卫生部危重病急救医学重点实验室，天津 300192
3.
天津市第一中心医院肝胆外科，天津 300192

基金项目:

天津市卫生健康科技重点项目 (TJWJ2021ZD002)

伦理学声明：本研究方案于2016年3月4日经由天津市第一中心医院实验动物伦理委员会审批，批号：2016-03-A1，符合实验室动物管理与使用准则。

利益冲突声明：本研究不存在研究者、伦理委员会成员以及与公开研究成果有关的利益冲突。

作者贡献声明：张锦锐负责课题设计，实施实验，数据分析及撰写论文；吴昊参与实验，修改论文；白易负责实验指导，修改论文；张雅敏负责指导撰写文章并最后定稿。

详细信息

通信作者:
张雅敏，13802122219@163.com

计量
- 文章访问数: 466
- HTML全文浏览量: 137
- PDF下载量: 41
- 被引次数: 0
出版历程
- 收稿日期: 2022-04-18
- 录用日期: 2022-05-20
- 出版日期: 2022-11-20

Effect of trimetazidine on a rat model of bile duct ischemia-reperfusion injury and its mechanism

Jinrui ZHANG^{1, 2},
Hao WU^{1, 2},
Yi BAI³,
Yamin ZHANG^{3
, ,
,}

1.
The First Central Clinical College of Tianjin Medical University, Tianjin 300070, China
2.
Ministry of Health Key Laboratory of Critical Illness and Emergency Medicine, Tianjin 300192, China
3.
Department of Hepatobiliary Surgery, Tianjin First Central Hospital, Tianjin 300192, China

Research funding:

Tianjin Health Science and Technology Project (TJWJ2021ZD002)

More Information

Corresponding author: ZHANG Yamin, 13802122219@163.com(ORCID: 0000-0003-3764-7281)

摘要

摘要: 目的探究曲美他嗪对大鼠缺血性胆管损伤的作用及其相关机制。方法雄性SD大鼠40只，随机分为假手术组(Sham组，n=10)、胆管缺血再灌注损伤组(BIRI组，n=10)、曲美他嗪预处理6 h组(TMZ-6h组，n=10)以及曲美他嗪预处理3 d组(TMZ-3d组，n=10)。缺血时间30 min，再灌注24 h后取材；HE染色病理切片观察各组胆管损伤情况，检测血清ALT、ALP、DBil水平，胆管组织内超氧化物歧化酶(SOD)酶活力和丙二醛(MDA)水平，Western Blot检测氧化应激和凋亡相关信号分子，如核因子红系2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、抗凋亡蛋白B细胞淋巴瘤-2(Bcl-2)和半胱天冬酶-3(Caspase-3)。计量资料多组间比较采用单因素方差分析，进一步两两比较采用Sidak t检验。结果 HE染色示BIRI组大鼠胆管上皮细胞部分连续性中断、坏死脱落，TMZ-6 h组胆管组织无明显变化，TMZ-3 d组胆管上皮细胞边缘较清晰、少量坏死脱落细胞。与Sham组相比，BIRI组ALT、ALP、DBil、MDA水平均显著升高(P值均＜0.05)，TMZ-3 d组ALT、ALP、DBil、MDA水平与BIRI组相比均显著降低(P值均＜0.05)，TMZ-6 h组与BIRI组相比无明显变化(P值均＞0.05)。与Sham组相比，BIRI组SOD酶活力显著下降(P＜0.05)，TMZ-3 d组与BIRI组相比显著增高(P＜0.05)，TMZ-6 h组与BIRI组相比无明显变化(P＞0.05)。与Sham组相比，BIRI组Nrf2、HO-1、Caspase-3表达水平均显著升高(P值均＜0.05)，Bcl-2表达水平显著降低(P＜0.05)；TMZ-3 d组Nrf2、HO-1、Bcl-2表达水平与BIRI组相比显著增加(P值均＜0.05)，Caspase-3表达水平显著降低(P＜0.05)；TMZ-6 h组Nrf2、HO-1、Bcl-2、Caspase-3与BIRI组相比无明显变化(P值均＞0.05)。结论曲美他嗪可降低大鼠胆管缺血再灌注损伤，其机制可能与抑制胆管细胞内氧化应激水平降低细胞凋亡有关。
- 再灌注损伤 /
- 胆管 /
- 曲美他嗪 /
- 大鼠, Sprague-Dawley
Abstract: Objective To investigate the effect of trimetazidine on ischemic bile duct injury in rats and related mechanism. Methods A total of 40 male Sprague-Dawley rats were randomly divided into sham-operation group (Sham group with 10 rats), bile duct ischemia-reperfusion injury group (BIRI group with 10 rats), 6-hour trimetazidine pretreatment group (TMZ-6h group with 10 rats), and 3-day trimetazidine pretreatment group (TMZ-3d group with 10 rats). The ischemia time was 30 minutes, and samples were collected after 24 hours of reperfusion. HE staining was used to observe bile duct injury, and the serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), and direct bilirubin (DBil) were measured, as well as the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in bile duct tissue; Western Blot was used to measure the levels of the signal molecules related to oxidative stress and apoptosis, such as nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), B-cell lymphoma-2 (Bcl-2), and caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Sidak t-test was used for further comparison between two groups. Results HE staining showed continuous interruption, necrosis, and exfoliation of bile duct epithelial cells in the BIRI group and no significant change in bile duct tissue in the TMZ-6h group, and in the TMZ-3d group, the bile duct epithelial cells had clear boundaries with a small amount of necrotic and exfoliated cells. Compared with the Sham group, the BIRI group had significant increases in the levels of ALT, ALP, DBil, and MDA (all P < 0.05); compared with the BIRI group, the TMZ-3d group had significant reductions in the levels of ALT, ALP, DBil, and MDA (all P < 0.05); there were no significant differences between the TMZ-6h group and the BIRI group (all P > 0.05). Compared with the Sham group, the BIRI group had a significant reduction in SOD activity (P < 0.05), the TMZ-3d group had a significant increase compared with the BIRI group (P < 0.05), and there was no significant difference between the TMZ-6h group and the BIRI group (P > 0.05). Compared with the Sham group, the BIRI group had significant increases in the expression levels of Nrf2, HO-1, and caspase-3 (all P < 0.05) and a significant reduction in the expression level of Bcl-2 (P < 0.05); compared with the BIRI group, the TMZ-3d group had significant increases in the expression levels of Nrf2, HO-1, and Bcl-2 (P < 0.05) and a significant reduction in the expression level of caspase-3 (P < 0.05); there were no significant differences in Nrf2, HO-1, Bcl-2, and caspase-3 between the TMZ-6h group and the BIRI group (all P > 0.05). Conclusion Trimetazidine can reduce bile duct ischemia-reperfusion injury in rats, possibly by inhibiting the level of oxidative stress in bile duct cells and reducing cell apoptosis.
- Reperfusion Injury /
- Bile Ducts /
- Trimetazidine /
- Rats, Sprague-Dawley

HTML全文

图 1 各组大鼠肝外胆管组织病理学变化(HE染色，×200)

注：a，Sham组；b，BIRI组；c，TMZ-6h组；d，TMZ-3d组。

Figure 1. Histopathological changes of extrahepatic bile duct of rats in each group (HE, ×200)

下载: 全尺寸图片幻灯片

图 2 各组大鼠胆管组织氧化应激与凋亡相关蛋白表达水平

Figure 2. Expression levels of oxidative stress and apoptosis related proteins in bile duct tissue of rats in each group

下载: 全尺寸图片幻灯片

表 1 各组大鼠血生化指标和胆管组织氧化应激指标比较结果

Table 1. Comparison of blood biochemical indexes and bile duct tissue oxidative stress indexes in each group

指标	Sham组	BIRI组	TMZ-6h组	TMZ-3d组	F值	P值
ALT(U/L)	35.20±4.32	431.20±22.84¹⁾	400.60±22.13	286.60±28.38²⁾	352.7	＜0.001
ALP(U/L)	81.80±7.85	452.40±31.09¹⁾	459.40±44.67	288.40±32.34²⁾	155.2	＜0.001
DBil(μmol/L)	4.06±0.73	57.21±5.26¹⁾	55.46±6.17	9.52±3.17²⁾	216.0	＜0.001
SOD(U/mg)	315.00±8.51	95.60±7.79¹⁾	92.20±10.23	187.40±11.67²⁾	469.2	＜0.001
MDA(nmol/mg)	1.96±0.41	6.50±0.58¹⁾	6.36±0.85	4.18±0.49²⁾	50.1	＜0.001
注：与Sham组比较，1)P＜0.05；与BIRI组比较，2)P＜0.05。ALT、ALP、DBil水平检测每组样本量为10，SOD、MDA水平检测每组样本量为5。

下载: 导出CSV

参考文献(15)

[1]	van LEEUWEN OB, van REEVEN M, van der HELM D, et al. Donor hepatectomy time influences ischemia-reperfusion injury of the biliary tree in donation after circulatory death liver transplantation[J]. Surgery, 2020, 168(1): 160-166. DOI: 10.1016/j.surg.2020.02.005.
[2]	ZHANG YC, LIU W, FU BS, et al. Therapeutic potentials of umbilical cord-derived mesenchymal stromal cells for ischemic-type biliary lesions following liver transplantation[J]. Cytotherapy, 2017, 19(2): 194-199. DOI: 10.1016/j.jcyt.2016.11.005.
[3]	van RIJN R, van LEEUWEN OB, MATTON APM, et al. Hypothermic oxygenated machine perfusion reduces bile duct reperfusion injury after transplantation of donation after circulatory death livers[J]. Liver Transpl, 2018, 24(5): 655-664. DOI: 10.1002/lt.25023.
[4]	TSIOUFIS K, ANDRIKOPOULOS G, MANOLIS A. Trimetazidine and cardioprotection: facts and perspectives[J]. Angiology, 2015, 66(3): 204-210. DOI: 10.1177/0003319714530040.
[5]	SETTAF A, MORIN D, LAMCHOURI F, et al. Trimetazidine ameliorates the hepatic injury associated with ischaemia-reperfusion in rats[J]. Pharmacol Res, 1999, 39(3): 211-216. DOI: 10.1006/phrs.1998.0427.
[6]	KAMADA N, CALNE RY. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage[J]. Transplantation, 1979, 28(1): 47-50.
[7]	ZHAO HF, ZHANG GW, ZHOU J, et al. Biliary tract injury caused by different relative warm ischemia time in liver transplantation in rats[J]. Hepatobiliary Pancreat Dis Int, 2009, 8(3): 247-254.
[8]	ZHANG GD, LIU BX, BAO MS. Ischemia-reperfusion injury of intrahepatic bile duct epithelium in rats and the protective effect of Salvia Miltiorrhiza[J]. Chin J Hepatobiliary Surg, 2005, 11(3): 193-194. DOI: 10.3760/cma.j.issn.1007-8118.2005.03.020. 张国栋, 刘冰新, 鲍民生. 大鼠肝内胆管上皮缺血再灌注损伤及丹参的保护作用[J]. 中华肝胆外科杂志, 2005, 11(3): 193-194. DOI: 10.3760/cma.j.issn.1007-8118.2005.03.020.
[9]	MARZILLI M, VINEREANU D, LOPASCHUK G, et al. Trimetazidine in cardiovascular medicine[J]. Int J Cardiol, 2019, 293: 39-44. DOI: 10.1016/j.ijcard.2019.05.063.
[10]	GUPTA K, PANDEY S, BAGANG N, et al. Trimetazidine an emerging paradigm in renal therapeutics: Preclinical and clinical insights[J]. Eur J Pharmacol, 2021, 913: 174624. DOI: 10.1016/j.ejphar.2021.174624.
[11]	MAHFOUDH BOUSSAID A, SELMI R, BEJAOUI M, et al. Effectiveness of a single versus repeated administration of trimetazidine in the protection against warm ischemia/reperfusion injury of rat liver[J]. Turk J Med Sci, 2016, 46(4): 1258-1264. DOI: 10.3906/sag-1505-102.
[12]	ZAOUALI MA, BONCOMPAGNI E, REITER RJ, et al. AMPK involvement in endoplasmic reticulum stress and autophagy modulation after fatty liver graft preservation: a role for melatonin and trimetazidine cocktail[J]. J Pineal Res, 2013, 55(1): 65-78. DOI: 10.1111/jpi.12051.
[13]	LI Z, ZENG YH, WANG K, et al. Research advances in the prevention and treatment of hepatic ischemia-reperfusion injury with traditional Chinese medicine components[J]. J Clin Hepatol, 2022, 38(2): 471-476. DOI: 10.3969/j.issn.1001-5256.2022.02.044. 李振, 曾意豪, 王科, 等. 中医药成分防治肝缺血再灌注损伤的研究进展[J]. 临床肝胆病杂志, 2022, 38(2): 471-476. DOI: 10.3969/j.issn.1001-5256.2022.02.044.
[14]	TONELLI C, CHIO ⅡC, TUVESON DA. Transcriptional regulation by Nrf2[J]. Antioxid Redox Signal, 2018, 29(17): 1727-1745. DOI: 10.1089/ars.2017.7342.
[15]	XUE R, QIU J, WEI S, et al. Lycopene alleviates hepatic ischemia reperfusion injury via the Nrf2/HO-1 pathway mediated NLRP3 inflammasome inhibition in Kupffer cells[J]. Ann Transl Med, 2021, 9(8): 631. DOI: 10.21037/atm-20-7084.