【作者】 阮文福（NGUYEN VAN PHUC）；

【导师】 黄仁彬；

【作者基本信息】 广西医科大学 ， 药理学， 2014， 博士

【摘要】 目的：探讨玉郎伞多糖对双氯芬酸钠(Diclofenac sodium，DICF)、尼美舒利(Nimesulide，NIM)、布洛芬(Ibuprofen，IBU)致小鼠肝损伤的保护作用及其机制研究。方法：1.实验动物分组、给药及模型建立把昆明种小鼠随机分为正常对照组(NC)，损伤模型组，YLSPS高剂量组(YLSPSH，600mg/kg)，YLSPS中剂量组(YLSPSM，300mg/kg)，YLSPS低剂量组(YLSPSL，150mg/kg)和联苯双酯组(200mg/kg，DDB)。以联苯双酯为阳性对照药和YLSPS高、中、低剂量组治疗，每日灌胃给药一次。除正常组给相等容量生理盐水外，其余所有小鼠给予相应的致肝损伤药物，每日一次，连续10天(尼美舒利)，14天(双氯芬酸钠、布洛芬)。实验结束前动物禁食20h，取血及脏器进行检测肝功能指标。2.计算肝脏重量指数，脾脏重量指数3. HE染色观察各组小鼠肝细胞损伤程度4.在DICF模型中，测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总胆红素（Total bilirubin, TBIL)的含量及血清中肿瘤坏死因子-α（TNF-α)、白介素-1β (IL-1β)及肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)和过氣化氢酶(CAT)的水平。实时荧光定量PCR方法检测肝组织Bax、Bc1-2基因的mRNA表达。5.在NIM模型中，测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总抗氧化能力(Tota1antioxide capacity，T-AOC)，血清肿瘤坏死因子-α（TNF-α)、白介素-6(IL-6)及肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)含量。Westem-Blot检测Bax、Bc1-2蛋白表达。6.在IBU模型中，测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总抗氧化能力（Tota1antioxide capacity，T-AOC）及肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)含量。Westem-Blot检测和免疫组化技术检测各组小鼠肝组织Bax、Bc1-2蛋白的表达。结果：1.在DICF模型中，与模型组比较，YLSPS各剂量组用能降低肝脏指数和脾脏指数；可降低血清ALT、AST、ALP、TBIL水平和肝组织MDA含量；降低血清中TNF-α与IL-1β；提高肝组织SOD、CAT、GSH-Px活性（P<0.05）；病理切片结果表明YLSPS各剂量组能够不同程度地减轻肝细胞的损伤程度。YLSPS能降低Bax mRNA的表达；增加肝组Bcl-2mRNA的表达(P<0.05)。2.在NIM模型中，与模型组比较，YLSPS各剂量组能降低肝脏指数和脾脏指数；可降低血清ALT、AST、ALP和肝组织MDA含量；降低血清TNF-α与IL-6；提高血清T-AOC水平和肝组织SOD、GSX-Px活性（P<0.05）；病理切片结果表明YLSPS各组能够不同程度地减轻肝细胞的损伤程度。YLSPS能降低Bax蛋白表达；增加肝组织Bcl-2蛋白表达(P<0.05)。3.在IBU模型中，与模型组比较，YLSPS各剂量组能降低肝脏指数和脾脏指数；可降低血清ALT、AST、ALP和肝组织MDA含量；提高血清T-AOC水平和肝组织SOD、GSX-Px活性（P<0.05）；病理切片结果表明YLSPS各组能够不同程度地减轻肝细胞的损伤程度。YLSPS能降低Bax蛋白表达；增加肝组织Bcl-2蛋白表达(P<0.05)。结论：YLSPS对双氯芬酸钠、尼美舒利和布洛芬所致小鼠肝损伤具有保护作用，其机制可能与其抗氧自由基、抑制脂质过氧化、抑制细胞凋亡作用有关。更多还原

【Abstract】 Objective： To observe the protective effect of Yulangsan Polysaecharide(YLSPS) on diclofenac sodium，nimesulide and ibuprofen-induced liver injuryin Kunming mice.Method：1.Animal and treatmentKunming mice were randomly divided into NC group，liver injured modelgroup with， high-，middle-，low-YLSPS group (600，300，150mg/kg) anddimethyl diphenyl bicarboxylate (200mg/kg DDB) group. The treatment groupswere orally administered once per day，whereas the normal and model groupswere orally administered with saline. Except normal mice, all the other micewere treated with drug induced liver injury for10days (nimesulide),14days(diclofenac, ibuprofen). At the end of the experimental period the mice were sacrificed and blood, liver samples were obtained.2.The indexs of the liver and spleen were observed3.Hematoxylineosin (HE) stain was used to examine the degree of hepaticinjury.4.Diclofenac sodium model：Estimation of ALT，AST and ALP, the content ofTBIL, TNF-α and IL-1β in serum and the activities of SOD, GSH–Px and CAT，the content of MDA in liver tissue. The levels of expression of Bax and Bcl-2mRNA in liver were examined by Real-time quantitative-PCR.5. Nimesulide model：Estimation of ALT，AST, ALP and T-AOC，the contentof TNF-α，IL-6in serum；the activities of SOD，GSH–Px and the content ofMDA in liver tissue. The expressions of Bax and Bcl-2in liver were detected byWestern Blot.6.Ibuprofen model：Estimation of of ALT，AST and ALP，the content ofT-AOC in serum；the activities of SOD，GSH-Px and the content of MDA inliver tissue were investigated. The expressions of Bax and Bcl-2in liver weredetected by immunohistochemical and Western Blot.Results：1.Diclofenac model： Compared with model group, YLSPS could obviouslyreduce the weight of the liver and spleen. The activities of ALT，AST andALP，the content of TBIL，MDA，TNF-α and IL-1β and the expression ofBax could be decreased. The activities of SOD，GSH-Px，CAT and theexpression of Bcl-2could be increased (P<0.05), and the degree of hepaticinjury could be lessened.2.Nimesulide model：Compared with model group， YLSPS could obviouslyreduce the activities of ALT，AST and ALP，the content of MDA，TNF-α， IL-6the index of liver, spleen and the expression of Bax. The activities ofSOD，GSH-Px，T-AOC and the expression of Bcl-2could be increased(P<0.05)；the degree of hepatic injury could be lessened.3.Ibuprofen model：Compared with model group，YLSPS could be decreasedthe index of liver and spleen，the activities of ALT，AST and ALP，the contentof MDA and the expression of Bax. The activities of SOD，GSH-Px，T-AOCand the expression of Bcl-2could be increased (P<0.05); the degree of hepaticinjury could be lessened.Conclusion：YLSPS has protective effect on diclofenac sodium, nimesulide andibuprofen-induced liver injury in mice. The mechanism may be related toattenuating free radical and inhibiting the effect on lipid peroxidation andapoptosis.更多还原