中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2013年
2期
116-120
,共5页
炎症%胆固醇%脂肪肝%肝硬化
炎癥%膽固醇%脂肪肝%肝硬化
염증%담고순%지방간%간경화
Inflammation%Cholesterol%Fatty Liver%,Liver cirrhosis
目的 观察炎症对小鼠肝脏内胆固醇蓄积、内质网应激及纤维化的影响.方法 将8周龄雄性C57BL/6J小鼠随机分为对照组(n=6)和炎症组(n=6),均以高脂饮食,炎症组小鼠隔天皮下注射0.5ml 10％酪蛋白以建立慢性炎症模型,对照组注射等量等渗盐水,14周处死.收集血清和肝组织样本,测定血清及肝组织中炎症因子及脂质水平;油红O、天狼猩红和Masson三色染色观察肝脂质沉积、形态改变以及纤维化程度；荧光实时定量PCR检测肝组织单核细胞趋化蛋白1、肿瘤坏死因子(TNF)α 、胆固醇调节元件结合蛋白(SREBP)2、低密度脂蛋白受体、3-羟-3-甲基戊二酰辅酶A还原酶、活化转录因子6、葡萄糖调节蛋白(GRP)78、骨形成蛋白(BMP)7、转化生长因子(TGF)β、层黏连蛋白、Ⅰ/Ⅳ型胶原的mRNA水平.对数据进行两样本均数t检验或近似t检验.结果 血清中,炎症组白细胞介素(IL)6的水平[(32.41±7.42)pg/ml]高于对照组[(12.55±4.75)pg/ml],t=5.522,P＜ 0.01;总胆固醇水平为(10.62±0.48)mmol/L,低于对照组的(14.82±1.56)mmol/L,t=6.303,P＜0.01.肝组织中,炎症组TNFα的mRNA相对表达水平为2.12±0.72,高于对照组的1.05±0.35,t=3.132,P＜0.01;总胆固醇水平为(23.21±8.75)μg/mg,高于对照组的(12.10±2.57)μg/mg,t=2.984,P＜0.05.油红O染色表明,炎症组肝脏脂滴沉积异常增多；天狼猩红和masson三色染色可见炎症组有更为明显的肝纤维化改变.荧光定量PCR检测表明炎症组SREBP-2、GRP78的mRNA相对表达水平分别为2.63±0.13、2.21±0.99,高于对照组的1.01±0.19、1.07±0.47,t值分别为15.641和2.031,P值均＜0.05.TGF β、Ⅰ型胶原的mRNA相对表达水平分别为1.38±0.28、1.71±0.51,高于对照组的1.01±0.14、1.02±0.27,t值分别为3.032和3.152,P值均＜0.05.BMP-7的mRNA相对表达水平为0.55±0.25,低于对照组的1.01±0.15,t=3.765,P＜0.01.结论 慢性炎症可以明显促进小鼠肝脏胆固醇沉积而加重肝细胞损害,刺激内质网应激,从而增加促纤维化因子表达、抑制抗纤维化因子表达,促进肝纤维化发生.
目的 觀察炎癥對小鼠肝髒內膽固醇蓄積、內質網應激及纖維化的影響.方法 將8週齡雄性C57BL/6J小鼠隨機分為對照組(n=6)和炎癥組(n=6),均以高脂飲食,炎癥組小鼠隔天皮下註射0.5ml 10％酪蛋白以建立慢性炎癥模型,對照組註射等量等滲鹽水,14週處死.收集血清和肝組織樣本,測定血清及肝組織中炎癥因子及脂質水平;油紅O、天狼猩紅和Masson三色染色觀察肝脂質沉積、形態改變以及纖維化程度；熒光實時定量PCR檢測肝組織單覈細胞趨化蛋白1、腫瘤壞死因子(TNF)α 、膽固醇調節元件結閤蛋白(SREBP)2、低密度脂蛋白受體、3-羥-3-甲基戊二酰輔酶A還原酶、活化轉錄因子6、葡萄糖調節蛋白(GRP)78、骨形成蛋白(BMP)7、轉化生長因子(TGF)β、層黏連蛋白、Ⅰ/Ⅳ型膠原的mRNA水平.對數據進行兩樣本均數t檢驗或近似t檢驗.結果 血清中,炎癥組白細胞介素(IL)6的水平[(32.41±7.42)pg/ml]高于對照組[(12.55±4.75)pg/ml],t=5.522,P＜ 0.01;總膽固醇水平為(10.62±0.48)mmol/L,低于對照組的(14.82±1.56)mmol/L,t=6.303,P＜0.01.肝組織中,炎癥組TNFα的mRNA相對錶達水平為2.12±0.72,高于對照組的1.05±0.35,t=3.132,P＜0.01;總膽固醇水平為(23.21±8.75)μg/mg,高于對照組的(12.10±2.57)μg/mg,t=2.984,P＜0.05.油紅O染色錶明,炎癥組肝髒脂滴沉積異常增多；天狼猩紅和masson三色染色可見炎癥組有更為明顯的肝纖維化改變.熒光定量PCR檢測錶明炎癥組SREBP-2、GRP78的mRNA相對錶達水平分彆為2.63±0.13、2.21±0.99,高于對照組的1.01±0.19、1.07±0.47,t值分彆為15.641和2.031,P值均＜0.05.TGF β、Ⅰ型膠原的mRNA相對錶達水平分彆為1.38±0.28、1.71±0.51,高于對照組的1.01±0.14、1.02±0.27,t值分彆為3.032和3.152,P值均＜0.05.BMP-7的mRNA相對錶達水平為0.55±0.25,低于對照組的1.01±0.15,t=3.765,P＜0.01.結論 慢性炎癥可以明顯促進小鼠肝髒膽固醇沉積而加重肝細胞損害,刺激內質網應激,從而增加促纖維化因子錶達、抑製抗纖維化因子錶達,促進肝纖維化髮生.
목적 관찰염증대소서간장내담고순축적、내질망응격급섬유화적영향.방법 장8주령웅성C57BL/6J소서수궤분위대조조(n=6)화염증조(n=6),균이고지음식,염증조소서격천피하주사0.5ml 10％락단백이건립만성염증모형,대조조주사등량등삼염수,14주처사.수집혈청화간조직양본,측정혈청급간조직중염증인자급지질수평;유홍O、천랑성홍화Masson삼색염색관찰간지질침적、형태개변이급섬유화정도；형광실시정량PCR검측간조직단핵세포추화단백1、종류배사인자(TNF)α 、담고순조절원건결합단백(SREBP)2、저밀도지단백수체、3-간-3-갑기무이선보매A환원매、활화전록인자6、포도당조절단백(GRP)78、골형성단백(BMP)7、전화생장인자(TGF)β、층점련단백、Ⅰ/Ⅳ형효원적mRNA수평.대수거진행량양본균수t검험혹근사t검험.결과 혈청중,염증조백세포개소(IL)6적수평[(32.41±7.42)pg/ml]고우대조조[(12.55±4.75)pg/ml],t=5.522,P＜ 0.01;총담고순수평위(10.62±0.48)mmol/L,저우대조조적(14.82±1.56)mmol/L,t=6.303,P＜0.01.간조직중,염증조TNFα적mRNA상대표체수평위2.12±0.72,고우대조조적1.05±0.35,t=3.132,P＜0.01;총담고순수평위(23.21±8.75)μg/mg,고우대조조적(12.10±2.57)μg/mg,t=2.984,P＜0.05.유홍O염색표명,염증조간장지적침적이상증다；천랑성홍화masson삼색염색가견염증조유경위명현적간섬유화개변.형광정량PCR검측표명염증조SREBP-2、GRP78적mRNA상대표체수평분별위2.63±0.13、2.21±0.99,고우대조조적1.01±0.19、1.07±0.47,t치분별위15.641화2.031,P치균＜0.05.TGF β、Ⅰ형효원적mRNA상대표체수평분별위1.38±0.28、1.71±0.51,고우대조조적1.01±0.14、1.02±0.27,t치분별위3.032화3.152,P치균＜0.05.BMP-7적mRNA상대표체수평위0.55±0.25,저우대조조적1.01±0.15,t=3.765,P＜0.01.결론 만성염증가이명현촉진소서간장담고순침적이가중간세포손해,자격내질망응격,종이증가촉섬유화인자표체、억제항섬유화인자표체,촉진간섬유화발생.
Objective To investigate whether inflammatory stress exacerbates hepatic cholesterol accumulation and liver fibrosis using a C57BL/6J mouse model of chronic inflammation.Methods Twelve male C57BL/6J mice were given a high-fat diet(15.0％ fat,1.25％ cholesterol,0.5％ cholic acid)and randomly assigned to the normal control group(n =6;subcutaneously injected with 0.5 mL of isotonic saline,every other day for 14 weeks)or the chronic inflammation model group(n =6;subcutaneously injected with of 0.5 mL of 10％ casein,every other day for 14 weeks).At the end of week 14,the animals were sacrificed and blood was collected from the left ventricle for serological analysis of inflammatory markers and lipid profile,including serum amyloid A(SAA),interleulin-6(IL-6),total cholesterol(TC)and free cholesterol(FC),low-density lipoprotein(LDL),and high-density lipoprotein(HDL)).Extracted liver tissues were divided for use in histological analysis(lipid accumulation and fibrosis evaluated by Oil Red O,Sirius red and Masson's trichrome staining)and quantitative fluorescence real-time PCR(to measure b-actin normalized expression of TNF-α MCP 1,SREBP-2,LDLr,HMGCoA-r,ATF-6,GRP78,BMP-7,TGF-β,and collagens type Ⅰ and type Ⅳ).Comparisons between groups were made by the two-samples t-test or Satterthwaite t-approximation test,collagen type Ⅰ and type Ⅳ.Results Compared to the normal control group,the inflammation model group showed elevated serum IL-6(12.55 ± 4.75 vs.32.41 ± 7.42 pg/mL,P ＜ 0.01),reduced serum TC(14.82 ± 1.56 vs.10.62 ± 0.48 mmol/L,P ＜ 0.01),up-regulated hepatic TNF-α mRNA expression(1.05 ± 0.35 vs.2.12 ± 0.72,P ＜ 0.01),and elevated hepatic TC(12.10 ± 2.57 vs.23.21 ± 8.75mmol/L,P ＜ 0.05).In addition,the inflammation group showed abnormal lipid deposition,and increased and thickened reticular fibers.The livers of the inflammation group also showed up-regulated mRNA expression of SREBP-2(normal control:1.01 ± 0.19 vs.2.63 ± 0.13,P ＜ 0.05),GRP78(1.07 ± 0.47 vs.2.21 ± 0.99,P ＜ 0.05),TGF-β(1.01 ± 0.14 vs.1.38 ± 0.28,P ＜ 0.05),and collagen type Ⅰ(1.02 ± 0.27 vs.1.71 ± 0.51,P ＜ 0.05)and down-regulation of BMP-7(1.01 ±0.15 vs.0.55 ±0.25,P＜ 0.01).Conclusion Activation of the inflammatory system exacerbates hepatic cholesterol accumulation and hepatic fibrosis in C57BL/6J mice.
