本文關(guān)鍵詞：姜黃素對慢性酒精肝損傷保護作用的研究，由筆耕文化傳播整理發(fā)布。

        第一部分姜黃素對乙醇誘導(dǎo)的大鼠原代肝細(xì)胞氧化損傷的保護作用目的：以Sparague-Dawley大鼠（簡稱SD大鼠）原代肝細(xì)胞為研究對象，建立酒精肝損傷模型，探討酒精對肝細(xì)胞的氧化損傷以及姜黃素對其的保護作用，并探討姜黃素對肝細(xì)胞HO-1活性表達的誘導(dǎo)。方法：1、分離并培養(yǎng)SD大鼠原代肝細(xì)胞，用不同濃度乙醇（0~200mmol/L）對大鼠原代肝細(xì)胞染毒8h，或者100mmol/L無水乙醇染毒不同時間（0~24h），檢測細(xì)胞的LDH釋放水平、AST水平以及氧化/抗氧化系統(tǒng)水平（MDA水平、GSH含量和SOD活性）。2、100mmol/L無水乙醇染毒細(xì)胞前1h用不同劑量姜黃素（0~50μmol/L）預(yù)處理，或者乙醇染毒前不同時間（0~5h）用15μmol/L姜黃素預(yù)處理，檢測細(xì)胞LDH、AST、MDA水平，GSH含量和SOD活性。3、梯度離心法分離提取各組肝細(xì)胞微粒體，檢測HO-1酶的活性。結(jié)果：1、隨著乙醇濃度的增加和作用時間的延長，大鼠原代肝細(xì)胞LDH釋放水平和AST水平顯著增高，MDA水平升高，GSH含量和SOD活性下降，，在濃度為100mmol/L和時間為8h時影響程度明顯，P值均<0.05。2、與乙醇對照組相比，提前用姜黃素干預(yù)可有效抑制LDH的釋放、抑制AST和MDA水平的升高，提高GSH含量以及SOD活性，預(yù)作用時間為1h、劑量為15μmol/L時差異有統(tǒng)計學(xué)意義（P <0.05）。3、姜黃素能上調(diào)HO-1蛋白表達，提高肝細(xì)胞內(nèi)HO-1酶的活性，以15μmol/L的劑量預(yù)作用1h時效果最明顯（P <0.05）。結(jié)論：乙醇誘導(dǎo)的大鼠原代肝細(xì)胞氧化損傷程度與給予乙醇的濃度及其作用時間呈正相關(guān)，姜黃素能夠有效降低乙醇誘導(dǎo)的肝細(xì)胞氧化損傷，并能誘導(dǎo)HO-1發(fā)揮抗氧化作用。第二部分姜黃素對BALB/c小鼠慢性酒精性肝損傷的干預(yù)作用目的：以BALB/c小鼠為研究對象，建立慢性酒精性肝損傷模型，并探討姜黃素對酒精誘導(dǎo)的肝臟氧化損傷的保護作用。方法：1、將實驗小鼠隨機分為4組（A：正常對照組，B：姜黃素組，C：酒精組，D：酒精+姜黃素組），C、D兩組梯度酒精暴露6周（前四周灌胃量為2.4g/kg/day，后兩周增加到4g/kg/day）建立慢性酒精性肝損傷模型，B、D兩組給予75mg/kg/d的姜黃素行灌胃干預(yù)。2、實驗結(jié)束后收集小鼠血液及肝組織標(biāo)本，制作肝組織病理切片，檢測血清轉(zhuǎn)氨酶（AST、ALT）水平和脂質(zhì)代謝（TC、TG、HDL-C）水平。3、檢測肝組織ROS含量和氧化/抗氧化系統(tǒng)水平（T-AOC、GSH、GPx、GST、MDA）。結(jié)果：1、正常對照組和姜黃素組小鼠體重增長明顯大于酒精灌胃的兩組（P值均<0.05），但姜黃素的補充并未明顯改善由酒精灌胃所造成的體重增長緩慢這一情況。2、與正常對照組相比，酒精灌胃組肝組織切片可見明顯的脂肪變性、血清轉(zhuǎn)氨酶AST、ALT水平提高（P <0.05），以及血清TC、TG含量升高（P <0.05）、HDL-C含量降低（P>0.05）。給予姜黃素干預(yù)后，可見肝臟脂肪變性有明顯減輕，AST、ALT水平下降，血脂紊亂恢復(fù)正常。3、與正常對照組相比，酒精灌胃組小鼠肝組織ROS含量顯著上升，T-AOC、GSH、GPx以及GST水平都顯著降低，MDA水平明顯升高，P值均<0.05。而酒精+姜黃素組小鼠肝組織ROS含量以及各項抗氧化、脂質(zhì)過氧化指標(biāo)均恢復(fù)正常。結(jié)論：姜黃素能減輕酒精誘導(dǎo)的肝臟脂肪變性、抑制血清轉(zhuǎn)氨酶的釋放、改善脂質(zhì)代謝紊亂，并且抑制ROS在肝臟中的蓄積、減緩肝組織的氧化損傷，從而對酒精誘導(dǎo)的肝損傷起保護作用。

    Part1Protective Effects of Curcumin on Oxidative DamageInduced by Ethanol in Rat Primary HepatocytesObjective: To establish alcoholic liver injury model in Sparague-Dawley rat primaryhepatocytes, and study the protective effects of curcumin on oxidative damage induced byethanol.Methods:1. Rat primary hepatocytes were treated with ethanol of different dosage (0-200mmol/L) for8h, or100mmol/L in different time (0-24h) to detect cell viability (LDH,AST levels) and oxidative/antioxidative system (MDA, GSH and SOD levels).2. On the basis of this, the hepatocytes were pre-treated with curcumin of differentdoses (0-50μmol/L) for1h, or15μmol/L in different time (0-5h) before ethanol exposure(100mmol/L,8h).Then detect LDH, AST, MDA, GSH and SOD levels.3. Extract the hepatocellular microsomes by gradient centrifugation and detect theHO-1enzyme activity.Results:1. There is remarkable increase of LDH and AST levels along with the increase ofethanol dosage and the extend of the time, as well as MDA formation in hepatocytes, and adecrease of GSH and SOD in ethanol group, it is significant difference at the dosage of100mmol/L or the time of8h (P <0.05).2. Compared with the alcohol group, pre-treatment of curcumin could significantlyprohibit the release of AST and LDH, the rise of MDA level and the decline of SODactivities and GSH content of hepatocytes exposed to ethanol (P <0.05).3. Compared with the control group, pre-treatment of curcumin could improve theprotein level and enzyme activity of HO-1at the dose of15μmol/L and time for1h.Conclusions: Ethanol-induced oxidative damage may occurred in rat primary hepatocytesat dose-and time-dependent manner. Curcumin could prevent its damage, as well asup-regulate HO-1protein expression and improve its enzyme activity. Part2Effects of Curcumin on Chronic Alcoholic Liver Injuryin BALB/c MiceObjective: To establish chronic alcoholic liver injury model in BALB/c mice, andinvestigate the protective effects of curcumin on oxidative damage.Methods:1. BALB/c mice were randomly divided into4groups (A: normal control group,B: curcumin group, C: ethanol group and D: ethanol+curcumin group), the chronicalcoholic liver injury model was established with gradient dosages of ethanol exposure by6weeks (from2.4g/kg/day to4g/kg/day). Group B and D were administered with curcuminof75mg/kg/day.2. Blood samples and liver tissues were collected at the end of the experiment, thehistopathological changes of the liver were observed, the enzyme activity of ALT, AST inserum and plasma levels of TC, TG, HDL-C were determined.3. The content of ROS in liver tissues was determined, as well as the level of T-AOC,GSH, GPx, GST and MDA.Results:1. The body weight gain in group A and B were significant higher than the othertwo groups (P <0.05), but no significant differences were found among group C and D (P>0.05).2. Compared with the control group, ethanol exposure significantly elevated the levelof serum AST, ALT, TC and TG (P>0.05), decline the level of HDL-C (P>0.05), andlipid droplet accumulation were observed. These were attenuated by curcuminsupplementation.3. Compared with the control group, ethanol exposure resulted in liver ROS generation,T-AOC, GSH, GPx and GST depletions and MDA elevention (P <0.05), which weresignificantly reversed by curcumin supplementation.Conclusion: Curcumin supplementation could alleviate pathological changes in the liverand hepatic enzymes release, attenuate the lipid disorder, improve antioxidant enzymeactivity and protect liver from ethanol-induced oxidative stress damage.

姜黃素對慢性酒精肝損傷保護作用的研究

縮略詞5-7摘要7-9Abstract9-10前言11-14第一部分 姜黃素對乙醇誘導(dǎo)的大鼠原代肝細(xì)胞氧化損傷的保護作用14-29    1 實驗材料14-16    2 實驗方法16-20    3 實驗結(jié)果20-26    4 討論26-29第二部分 姜黃素對 BALB/c 小鼠慢性酒精性肝損傷的保護作用29-43    1 實驗材料29    2 試驗方法29-33    3 實驗結(jié)果33-41    4 討論41-43小結(jié)43-46參考文獻46-51綜述51-65    參考文獻60-65附錄65-67致謝67

  本文關(guān)鍵詞：姜黃素對慢性酒精肝損傷保護作用的研究，由筆耕文化傳播整理發(fā)布。