印尼姜黄对刀豆球蛋白A所致小鼠肝损伤的保护作用及其影响TGF-β1/Smad通路的机制探讨  

Protective Effect of Curcuma zanthorrhiza Rhizome on Hepatitis Induced by Concanavalin A in Mice and Effect on TGF-β_1/ Smad Signaling Pathway

作　　者：汪蕾[1] Nuriza Rahmadini 高天慧[1] 余佳丽[1] 甘彦雄 廖婉[1] 傅超美[1] 

机构地区：[1]成都中医药大学药学院,中药材标准化教育部重点实验室,四川省中药资源系统研究与开发利用重点实验室--省部共建国家重点实验室培育基地,成都611137

出　　处：《中国实验方剂学杂志》2017年第7期127-133,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年项目(81403103);四川省科技支撑计划项目(2014SZ0156)

摘　　要：目的:研究印尼姜黄对刀豆蛋白A(Con A)诱导的小鼠肝损伤的保护作用,并探究其可能机制。方法:SPF级小鼠随机分为正常组、模型组、联苯双酯阳性药组及印尼姜黄高、中、低剂量组。灌胃给药7 d,末次给药后2 h除正常组外,其余各组小鼠均一次性尾静脉注射Con A 30 mg·kg^(-1)诱导小鼠肝损伤,禁食不禁水8 h后称量体重,摘眼球取血并脱颈椎处死动物,计算小鼠的肝脏、脾脏指数,检测血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),乳酸盐脱氢酶(LDH)活性及肝组织匀浆中总超氧化物歧化酶(T-SOD),丙二醛(MDA),一氧化氮(NO)的含量,苏木素-伊红(HE)观察肝组织病理学变化,采用实时荧光定量PCR(Real-time PCR)和蛋白质免疫印迹法(Western blot)检测肝脏转化生长因子-β_1(TGF-β_1),Smad3,Smad7的mRNA和蛋白表达。结果:与正常组比较,模型组肝脏、脾脏指数、血清中ALT,AST,LDH及肝匀浆MDA,NO的水平显著升高,肝匀浆T-SOD水平显著降低,肝脏组织中TGF-β1,Smad3的mRNA及蛋白表达显著升高,Smad7的mRNA和蛋白表达显著降低(P<0.01),模型组的肝脏病变较为明显;与模型组比较,印尼姜黄与联苯双酯均可明显降低肝炎小鼠的肝脏、脾脏指数,降低血清中ALT,AST,LDH及肝匀浆MDA,NO的水平,肝匀浆T-SOD水平明显升高;印尼姜黄高剂量组肝脏病理改变明显减轻,印尼姜黄高剂量组的小鼠肝脏组织中TGF-β_1,Smad3的mRNA及蛋白表达下降,Smad7的mRNA和蛋白表达升高(P<0.05,P<0.01)。结论:印尼姜黄对Con A诱导的小鼠肝损伤具有显著保护作用,通过多位点影响肝组织TGF-β_1/Smad通路可能是其阻断肝损伤进程的机制。Objective： To study the protective effects of Curcuma zanthorrhiza rhizome on liver injury induced by concanavalin A （Con A） in mice, and investigate its possible mechanism. Method： SPF mice were randomly divided into normal group, model group, biphenyl group, C. zanthorrhiza rhizome high dose group, middle dose group and low dose group. All the mice except thoae in normal group received Con A （30 mg·kg^-1） by injection via tail vein to induce mice liver injury. The drugs were given for 7 days by gavage administration, and the mice in normal group received the equal volume of normal saline. After fasting （but water was given） for 8 h, their body weight was measured; blood was taken from eyeball, and the mice were killed by taking off the cervical vertebrae. Then their liver index, spleen index, contents of alanine aminotransferase （ALT）, aspartate amino transferase （AST）, and lactate dehydrogenase （LDH） in serum and total superoxide dismutase （T-SOD）, malondialdehyde （MDA）, nitric oxide （NO） in liver homogenates were measured. Hepatic tissue was observed by hematoxylin-eosin staining. Furthermore, real-time fluorescence quantitative PCR （Real-time PCR） and Western blot assay were used to detect mRNA and protein expression levels of liver tramsforming gravth factor-β1 （TGF- β1）, Smad3 and Smad7. Result： As compared with the normal group, the liver index, spleen index, levels of ALT, AST and LDH in serum as well as MDA and NO levels in liver homogenates were significantly increased in model group; T-SOD level in liver homogenates was significantly reduced; mRNA and protein expression levels of liver TGF-β1, Smad3 were significantly increased, while mRNA and protein expression levels of Smad3 were significantly reduced （P 〈 0.01 ） , with more obvious liver lesions in model group. As compared with the model group, both C. zanthorrhiza rhizome and biphenyl significantly reduced the liver index and spleen index; decreased the levels of ALT, AST, LDH in serum a

关 键 词：印尼姜黄 刀豆球蛋白A 肝损伤 TGF-β1/Smad通路 

分 类 号：R285.5[医药卫生—中药学]