中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
2015年
6期
591-595
,共5页
郑永平%肖亮生%李庆南%胡建凡%郑灿滨%林奕芬
鄭永平%肖亮生%李慶南%鬍建凡%鄭燦濱%林奕芬
정영평%초량생%리경남%호건범%정찬빈%림혁분
叶下红%肝脂肪变性%固醇调节元件结合蛋白1%磷酸化细胞外调节蛋白激酶1/2%Toll样受体4%高迁移率族蛋白B1%超氧化物歧化酶%丙二醛
葉下紅%肝脂肪變性%固醇調節元件結閤蛋白1%燐痠化細胞外調節蛋白激酶1/2%Toll樣受體4%高遷移率族蛋白B1%超氧化物歧化酶%丙二醛
협하홍%간지방변성%고순조절원건결합단백1%린산화세포외조절단백격매1/2%Toll양수체4%고천이솔족단백B1%초양화물기화매%병이철
Emilia sonchifolia%Hepatic steatosis%Sterol-regulatory element binding protein 1%Phosphorylated extracellular signal regulated protein kinase 1/2%Toll like receptor 4%High mobility group box-1 protein%Superoxide dismutase%Malonaldehyde
目的：探讨叶下红对实验性肝脂肪变大鼠的保护作用及其分子机制。方法将70只SD大鼠按随机数字表法分为正常对照组、模型组、高或低剂量叶下红组及高剂量叶下红+磷酸化细胞外调节蛋白激酶1/2（pERK1/2）抑制剂（PD98059）组（PD组）。正常对照组予以普通饲料；其余各组予以高脂低蛋白饲料联合30%四氯化碳（CCl4）花生油2 mL/kg每3 d皮下注射1次，连续3周制备动物模型。制模后叶下红组以高、低剂量叶下红提取物（5.0 g/kg和2.5 g/kg）灌胃，每日1次；PD组每日1次灌胃5.0 g/kg叶下红提取物后加用0.3 mg/kg PD98059每周1次尾静脉注射。3周后均改为普通饲料，第5周末取肝组织进行病理学观察；用全自动生化分析仪检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆固醇（TC）、甘油三酯（TG）水平，采用免疫组化染色、蛋白质免疫印迹试验（Western Blot）及流式细胞术检测固醇调节元件结合蛋白1（SREBP-1）、pERK1/2、Toll样受体4（TLR4）、高迁移率族蛋白B1（HMGB1）阳性细胞计数及蛋白表达；用羟胺法及硫代巴比妥酸（TBA）法检测肝组织超氧化物歧化酶（SOD）、丙二醛（MDA）水平。结果高、低剂量叶下红组及PD组肝组织小叶炎症较模型组减轻（分：1.50±0.53、1.80±0.43、1.20±0.42比2.30±0.48），ALT、AST、TC、TG、SREBP-1、MDA均较模型组减少，以高剂量叶下红组最为明显〔ALT（U/L）为51.91±6.95比66.50±12.15，AST （U/L）为125.70±5.62比147.10±10.52，TC（mmol/L）为1.79±1.04比2.81±1.08，TG（mmol/L）为0.87±0.55比1.17±0.67，SREBP-1为（30.60±5.56）%比（53.10±5.02）%，MDA（nmol/mg）为5.20±0.87比10.61±5.45， P＜0.05或P＜0.01〕；pERK1/2、TLR4、HMGB1相对表达量与模型组比较差异无统计学意义〔pERK1/2为（43.77±4.93）%比（46.83±5.27）%，TLR4（相对荧光强度）为69.12±24.64比69.08±24.32，HMGB1（相对荧光强度）为22.93±14.88比33.17±13.29，均P＞0.05〕；而PD组上述各指标值与高剂量叶下红组接近，说明细胞外调节蛋白激酶（MEK）抑制剂对叶下红作用无影响。结论叶下红能减轻实验性肝脂肪变肝损伤程度，明显减轻肝小叶炎症，其机制可能与减轻氧化应激反应有关，SREBP-1可能参与其作用的发挥。
目的：探討葉下紅對實驗性肝脂肪變大鼠的保護作用及其分子機製。方法將70隻SD大鼠按隨機數字錶法分為正常對照組、模型組、高或低劑量葉下紅組及高劑量葉下紅+燐痠化細胞外調節蛋白激酶1/2（pERK1/2）抑製劑（PD98059）組（PD組）。正常對照組予以普通飼料；其餘各組予以高脂低蛋白飼料聯閤30%四氯化碳（CCl4）花生油2 mL/kg每3 d皮下註射1次，連續3週製備動物模型。製模後葉下紅組以高、低劑量葉下紅提取物（5.0 g/kg和2.5 g/kg）灌胃，每日1次；PD組每日1次灌胃5.0 g/kg葉下紅提取物後加用0.3 mg/kg PD98059每週1次尾靜脈註射。3週後均改為普通飼料，第5週末取肝組織進行病理學觀察；用全自動生化分析儀檢測血清丙氨痠轉氨酶（ALT）、天鼕氨痠轉氨酶（AST）、總膽固醇（TC）、甘油三酯（TG）水平，採用免疫組化染色、蛋白質免疫印跡試驗（Western Blot）及流式細胞術檢測固醇調節元件結閤蛋白1（SREBP-1）、pERK1/2、Toll樣受體4（TLR4）、高遷移率族蛋白B1（HMGB1）暘性細胞計數及蛋白錶達；用羥胺法及硫代巴比妥痠（TBA）法檢測肝組織超氧化物歧化酶（SOD）、丙二醛（MDA）水平。結果高、低劑量葉下紅組及PD組肝組織小葉炎癥較模型組減輕（分：1.50±0.53、1.80±0.43、1.20±0.42比2.30±0.48），ALT、AST、TC、TG、SREBP-1、MDA均較模型組減少，以高劑量葉下紅組最為明顯〔ALT（U/L）為51.91±6.95比66.50±12.15，AST （U/L）為125.70±5.62比147.10±10.52，TC（mmol/L）為1.79±1.04比2.81±1.08，TG（mmol/L）為0.87±0.55比1.17±0.67，SREBP-1為（30.60±5.56）%比（53.10±5.02）%，MDA（nmol/mg）為5.20±0.87比10.61±5.45， P＜0.05或P＜0.01〕；pERK1/2、TLR4、HMGB1相對錶達量與模型組比較差異無統計學意義〔pERK1/2為（43.77±4.93）%比（46.83±5.27）%，TLR4（相對熒光彊度）為69.12±24.64比69.08±24.32，HMGB1（相對熒光彊度）為22.93±14.88比33.17±13.29，均P＞0.05〕；而PD組上述各指標值與高劑量葉下紅組接近，說明細胞外調節蛋白激酶（MEK）抑製劑對葉下紅作用無影響。結論葉下紅能減輕實驗性肝脂肪變肝損傷程度，明顯減輕肝小葉炎癥，其機製可能與減輕氧化應激反應有關，SREBP-1可能參與其作用的髮揮。
목적：탐토협하홍대실험성간지방변대서적보호작용급기분자궤제。방법장70지SD대서안수궤수자표법분위정상대조조、모형조、고혹저제량협하홍조급고제량협하홍+린산화세포외조절단백격매1/2（pERK1/2）억제제（PD98059）조（PD조）。정상대조조여이보통사료；기여각조여이고지저단백사료연합30%사록화탄（CCl4）화생유2 mL/kg매3 d피하주사1차，련속3주제비동물모형。제모후협하홍조이고、저제량협하홍제취물（5.0 g/kg화2.5 g/kg）관위，매일1차；PD조매일1차관위5.0 g/kg협하홍제취물후가용0.3 mg/kg PD98059매주1차미정맥주사。3주후균개위보통사료，제5주말취간조직진행병이학관찰；용전자동생화분석의검측혈청병안산전안매（ALT）、천동안산전안매（AST）、총담고순（TC）、감유삼지（TG）수평，채용면역조화염색、단백질면역인적시험（Western Blot）급류식세포술검측고순조절원건결합단백1（SREBP-1）、pERK1/2、Toll양수체4（TLR4）、고천이솔족단백B1（HMGB1）양성세포계수급단백표체；용간알법급류대파비타산（TBA）법검측간조직초양화물기화매（SOD）、병이철（MDA）수평。결과고、저제량협하홍조급PD조간조직소협염증교모형조감경（분：1.50±0.53、1.80±0.43、1.20±0.42비2.30±0.48），ALT、AST、TC、TG、SREBP-1、MDA균교모형조감소，이고제량협하홍조최위명현〔ALT（U/L）위51.91±6.95비66.50±12.15，AST （U/L）위125.70±5.62비147.10±10.52，TC（mmol/L）위1.79±1.04비2.81±1.08，TG（mmol/L）위0.87±0.55비1.17±0.67，SREBP-1위（30.60±5.56）%비（53.10±5.02）%，MDA（nmol/mg）위5.20±0.87비10.61±5.45， P＜0.05혹P＜0.01〕；pERK1/2、TLR4、HMGB1상대표체량여모형조비교차이무통계학의의〔pERK1/2위（43.77±4.93）%비（46.83±5.27）%，TLR4（상대형광강도）위69.12±24.64비69.08±24.32，HMGB1（상대형광강도）위22.93±14.88비33.17±13.29，균P＞0.05〕；이PD조상술각지표치여고제량협하홍조접근，설명세포외조절단백격매（MEK）억제제대협하홍작용무영향。결론협하홍능감경실험성간지방변간손상정도，명현감경간소협염증，기궤제가능여감경양화응격반응유관，SREBP-1가능삼여기작용적발휘。
Objective To investigate the preventive effects of emilia sonchifolia on experimental hepatic steatosis in rats and its molecular mechanism.Methods Seventy Sprague-Dawley (SD) rats were randomly divided into five groups: normal control, model, high dose emilia sonchifolia, low dose emilia sonchifolia groups and high dose emilia sonchifolia + phosphorylated extracellular signal regulated protein kinase 1/2 (pERK1/2) inhibitor (PD98059) group (PD group). In normal control group, the rats were fed with normal diet, and in the other four groups, the rats were fed with high fat and low protein diet combined with 30% carbon tetrachloride (CCl4) peanut oil 2 mL/kg subcutaneous injection, once every 3 days for consecutive 3 weeks to establish animal models with hepatic steatosis. In emilia sonchifolia high and low dose groups, 5.0 g/kg and 2.5 g/kg doses of emilia sonchifolia were given respectively by gavage, once a day. In PD group, after administration of emilia sonchifolia high dose by gavage once a day, additionally PD98059 0.3 mg/kg was injected through a tail vein, once a week. After 3 weeks, all rats were switched to normal diet and treatment continued as before. At the end of the 5th week, liver tissues were taken for pathological analyses. The serum levels of alanine transaminase (ALT), aspartate transaminase (AST), total cholesterol (TC), and triglyceride (TG) were determinated by automatic biochenical analyzer. The positive cell count and protein expressions of sterol-regulatory element binding protein 1 (SREBP-1), pERK1/2, toll like receptor 4 (TLR4) and high mobility group box-1 protein (HMGB1) were tested by immunohistochemistry, Western Blot and flow cytometry. The levels of superoxide dismutase (SOD) and malonaldehyde (MDA) in liver cell homogenate were detected by hydroxylamine and TBA method.Results Compared with the model group, the lobular inflammation in high and low dose emilia sonchifolia groups and PD group was attenuated (1.50±0.53, 1.80±0.43, 1.20±0.42 vs. 2.30±0.48), and ALT, AST, TC, TG, SREBP-1, and MDA were significantly decreased, the decrease in high dose emilia sonchifolia group being the most significant [ALT (U/L): 51.91±6.95 vs. 66.50±12.15, AST (U/L): 125.70±5.62 vs. 147.10±10.52, TC (mmol/L): 1.79±1.04 vs. 2.81±1.08, TG (mmol/L): 0.87±0.55 vs. 1.17±0.67, SREBP-1: (30.60±5.56)% vs. (53.10±5.02)%, MDA (nmol/mg): 5.20±0.87 vs. 10.61±5.45,P < 0.05 orP < 0.01]; the relative expression levels of pERK1/2, TLR4, and HMGB1 showed no statistically significant differences between each treated group and the model group [pERK1/2: (43.77±4.93)% vs. (46.83±5.27)%, TLR4 (rmfi): 69.12±24.64 vs. 69.08±24.32, HMGB1 (rmfi): 22.93±14.88 vs. 33.17±13.29, allP > 0.05]. While the above index values in PD group were close to those in high dose emilia sonchifolia group, showing that PD98059 had no impact on emilia sonchifolia's action.Conclusions Emilia sonchifolia can alleviate hepatic injury and attenuate lobular inflammation in rat experimental hepatic steatosis. Its mechanism is possibly related to the reduction of oxidative stress reaction, and SREBP-1 may be as a mediator involved in the action.
