苦参碱对大鼠肝缺血再灌注损伤的抑制作用及机制
| 作者: |
1,2白宁,
2王栋,
2欧阳锡武,
2周乐杜,
2王志明
1 中南大学湘雅医院急诊科，湖南 长沙 410008 2 中南大学湘雅医院普通外科，湖南 长沙 410008 |
| 通讯: |
王志明
Email: wzmxycsu@hotmail.com |
| DOI: | 10.3978/.2018.01.013 |
摘要
目的：探讨苦参碱对大鼠肝缺血再灌注损伤的抑制作用及机制。
方法：大鼠肝缺血再灌注损伤模型采用阻断肝血流60 min再灌注120 min诱导；将40只SD大鼠随机均分为假手术组、肝缺血再灌注损伤模型组（模型组）、低剂量苦参碱（25 mg/kg）预处理+肝缺血再灌注损伤模型组（低剂量苦参碱组）、高剂量苦参碱（50 mg/kg）预处理+肝缺血再灌注损伤模型组（高剂量苦参碱组），低、高剂量苦参碱组在肝缺血前30 min，经门静脉主干注入各自剂量的苦参碱溶液，假手术组与模型组大鼠则以相同的方式注入等体积的生理盐水；120 min再灌注结束后，收集各组大鼠血标本行血清转氨酶、炎症因子水平检测，收集肝组织标本行组织病理学、肝细胞凋亡检测，以及TRAIL、BAX、激活型caspase-3（cleaved caspase-3）蛋白表达检测。
结果：组织病理学检测结果显示，除假手术组外，其余各组肝组织均有肝损伤表现，但损伤程度轻重不一（模型组>低剂量苦参碱组>高剂量苦参碱组）。与假手术组比较，其余各组血清转氨酶、炎症因子水平均明显升高，肝组织中肝细胞凋亡明显增加，TRAIL、BAX、cleaved caspase-3蛋白表达明显上调（均P<0.05），但以上指标的变化程度在低、高剂量苦参碱组均明显小于模型组，且在高剂量苦参碱组更为明显（均P<0.05）。
结论：苦参碱对大鼠肝缺血再灌注损伤具有抑制作用，机制可能与其抑制TRAIL的表达，减少BAX与caspase-3的活化，从而抑制肝细胞凋亡有关。
关键词：
肝；再灌注损伤；苦参碱；TNF相关凋亡诱导配体；细胞凋亡
方法：大鼠肝缺血再灌注损伤模型采用阻断肝血流60 min再灌注120 min诱导；将40只SD大鼠随机均分为假手术组、肝缺血再灌注损伤模型组（模型组）、低剂量苦参碱（25 mg/kg）预处理+肝缺血再灌注损伤模型组（低剂量苦参碱组）、高剂量苦参碱（50 mg/kg）预处理+肝缺血再灌注损伤模型组（高剂量苦参碱组），低、高剂量苦参碱组在肝缺血前30 min，经门静脉主干注入各自剂量的苦参碱溶液，假手术组与模型组大鼠则以相同的方式注入等体积的生理盐水；120 min再灌注结束后，收集各组大鼠血标本行血清转氨酶、炎症因子水平检测，收集肝组织标本行组织病理学、肝细胞凋亡检测，以及TRAIL、BAX、激活型caspase-3（cleaved caspase-3）蛋白表达检测。
结果：组织病理学检测结果显示，除假手术组外，其余各组肝组织均有肝损伤表现，但损伤程度轻重不一（模型组>低剂量苦参碱组>高剂量苦参碱组）。与假手术组比较，其余各组血清转氨酶、炎症因子水平均明显升高，肝组织中肝细胞凋亡明显增加，TRAIL、BAX、cleaved caspase-3蛋白表达明显上调（均P<0.05），但以上指标的变化程度在低、高剂量苦参碱组均明显小于模型组，且在高剂量苦参碱组更为明显（均P<0.05）。
结论：苦参碱对大鼠肝缺血再灌注损伤具有抑制作用，机制可能与其抑制TRAIL的表达，减少BAX与caspase-3的活化，从而抑制肝细胞凋亡有关。
Inhibitory effect of matrine against hepatic ischemia-reperfusion injury in rats and its mechanism
CorrespondingAuthor:WANG Zhiming Email: wzmxycsu@hotmail.com
Abstract
Objective: To investigate the inhibitory effect of matrine against hepatic ischemia-reperfusion injury (HIRI) in rats and the mechanism.
Methods: HIRI in rats was induced by 60 min hepatic ischemia followed by 120 min reperfusion. Forty SD rats were equally randomized into sham operation group, HIRI model group (model group), low dose matrine (25 mg/kg) pretreatment plus HIRI model group (low dose matrine group) and high dose matrine (50 mg/kg) pretreatment plus HIRI model group (high dose matrine group). Rats in low and high dose matrine groups were injected with matrine of respective dose via the main trunk of the portal vein 30 min before hepatic ischemia, while those in sham operation group and model group received the same volume of normal saline by the same fashion. After the 120 min reperfusion, blood samples were drawn for measuring the serum levels of transaminases and inflammatory factors, and liver tissue samples were harvested for histopathological examination and analysis of hepatic cell apoptosis as well as determination of the protein expressions of TRAIL, BAX and activated caspase-3 (cleaved caspase-3).
Results: The results of histopathological examination showed that there were liver injuries in all groups except in sham operation group, but the injuries varied in degree from severe to mild (model group>low dose matrine group>high dose matrine group). Compared with sham operation group, the serum levels of transaminases and inflammatory factors were significantly increased, the liver cell apoptosis was increased, and the protein expressions of TRAIL, BAX and cleaved caspase-3 were all up-regulated in the remaining groups (all P<0.05), but the changing amplitudes in above parameters in low and high dose group matrine groups were significantly milder than those in model group, and were more evident in high dose matrine group (all P<0.05).
Conclusion: Matrine has inhibitory effect against HIRI in rats, and the mechanism may probably be associated with its inhibiting TRAIL expression and then reducing BAX and caspase-3 activation, and thereby suppressing liver cell apoptosis.
Keywords：
Liver; Reperfusion Injury
MATRINE; TNF-Related Apoptosis-Inducing Ligand; Apoptosis
Methods: HIRI in rats was induced by 60 min hepatic ischemia followed by 120 min reperfusion. Forty SD rats were equally randomized into sham operation group, HIRI model group (model group), low dose matrine (25 mg/kg) pretreatment plus HIRI model group (low dose matrine group) and high dose matrine (50 mg/kg) pretreatment plus HIRI model group (high dose matrine group). Rats in low and high dose matrine groups were injected with matrine of respective dose via the main trunk of the portal vein 30 min before hepatic ischemia, while those in sham operation group and model group received the same volume of normal saline by the same fashion. After the 120 min reperfusion, blood samples were drawn for measuring the serum levels of transaminases and inflammatory factors, and liver tissue samples were harvested for histopathological examination and analysis of hepatic cell apoptosis as well as determination of the protein expressions of TRAIL, BAX and activated caspase-3 (cleaved caspase-3).
Results: The results of histopathological examination showed that there were liver injuries in all groups except in sham operation group, but the injuries varied in degree from severe to mild (model group>low dose matrine group>high dose matrine group). Compared with sham operation group, the serum levels of transaminases and inflammatory factors were significantly increased, the liver cell apoptosis was increased, and the protein expressions of TRAIL, BAX and cleaved caspase-3 were all up-regulated in the remaining groups (all P<0.05), but the changing amplitudes in above parameters in low and high dose group matrine groups were significantly milder than those in model group, and were more evident in high dose matrine group (all P<0.05).
Conclusion: Matrine has inhibitory effect against HIRI in rats, and the mechanism may probably be associated with its inhibiting TRAIL expression and then reducing BAX and caspase-3 activation, and thereby suppressing liver cell apoptosis.