中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2013年
2期
143-149
,共7页
张步春%李宪凯%车文良%李伟明%侯磊%魏毅东%徐亚伟
張步春%李憲凱%車文良%李偉明%侯磊%魏毅東%徐亞偉
장보춘%리헌개%차문량%리위명%후뢰%위의동%서아위
动脉粥样硬化%过氧化物酶体增殖物激活受体%糖尿病,2型%受体,LDL%小鼠,基因敲除
動脈粥樣硬化%過氧化物酶體增殖物激活受體%糖尿病,2型%受體,LDL%小鼠,基因敲除
동맥죽양경화%과양화물매체증식물격활수체%당뇨병,2형%수체,LDL%소서,기인고제
Atherosclerosis%Peroxisome proliferator-activated receptors%Diabetes mellitus,type 2%Receptors,LDL%Mice,knockout
目的 探讨过氧化物酶体增殖物激活受体(PPAR)α/γ双激动剂对低密度脂蛋白受体基因敲除(LDLr-/-)糖尿病小鼠动脉粥样硬化斑块稳定性的影响.方法 取4周龄雌性LDLr-/-小鼠45只,高糖高脂饮食喂养4周后采用随机数表法随机分为3组(每组15只),即对照组、2型糖尿病模型组(DM组)和PPARα/γ双激动剂替赛格列他(tesaglitazar)糖尿病治疗组(治疗组).对照组仅给予高糖高脂饮食喂养.DM组给予高糖高脂饮食联合小剂量链脲佐菌素诱导.治疗组饲料中混合20μg/kg tesaglitazar喂养.随机分组后继续高糖高脂饮食喂养6周.药物干预6周后处死小鼠,分别于药物干预前和处死前测定小鼠体质量、血清总胆固醇(TC)、甘油三酯(TG)和血糖(Glu)水平.免疫印迹和免疫组织化学染色检测头臂干动脉斑块炎症因子细胞间黏附分子1(ICAM-1)、血管内皮黏附分子1(VCAM-1)、单核细胞趋化蛋白-1(MCP-1)表达水平.苏木素伊红、油红O、天狼猩红染色观察头臂干动脉斑块,免疫组织化学染色检测头臂干动脉斑块平滑肌肌动蛋白(α-SMA)和巨噬细胞表面分子-3(Mac-3)表达.结果 DM组小鼠血清TC、TG和Glu水平均显著高于对照组,分别为(32.34±3.26) mmol/L比(16.17±1.91) mmol/L、(3.57 ±0.99) mmol/L比(2.21 ±0.11) mmol/L和(15.21±4.67) mmol/L比(6.89±0.83) mmol/L(P均＜0.01).DM组小鼠头臂干动脉斑块炎症因子ICAM-1、VCAM-1和MCP-1的蛋白表达水平均显著高于对照组,分别为2.31 ±0.35比1.34 ±0.21,1.65±0.14比0.82±0.26和2.27±0.16比1.56±0.23(P均＜0.01).DM组小鼠血管斑块面积显著大于对照组[(4.597±1.260)×103 μm2比(0.075 ±0.030)×103 μm2],脂质、α-SMA、Mac-3和胶原含量均显著高于对照组,分别为(47.23±2.64)％比(9.67±1.75)％、(5.54±1.17)％比(2.13±0.41)％、(19.15 ±3.51)％比(1.72±0.16)％和(4.27±0.74)％比(0.43±0.09)％(P均＜0.01).治疗组小鼠血清TC、TG和Glu水平均显著低于DM组(P均＜0.01),分别为(30.47 ±3.18)mmol/L、(3.14 ±0.71) mmol/L和(7.92±1.28) mmol/L.治疗组小鼠头臂干斑块炎症因子ICAM-1、VCAM-1和MCP-1蛋白表达水平均低于DM组(P＜0.05或P＜0.01),分别为1.84±0.22、1.27±0.11和1.83±0.24.治疗组小鼠血管斑块面积[(1.283 ±0.410)×103 μm2]小于DM组(P＜0.01),脂质含量和Mac-3表达水平亦低于DM组(P均＜0.01),分别为(23.52±1.39)％和(12.84±3.22)％.但治疗组小鼠头臂干动脉斑块内胶原含量[(6.32±1.15)％]和α-SMA含量[(9.46±1.47)％]均高于DM组(P均＜0.01).治疗组小鼠主动脉脂质沉积和Mac-3表达含量则低于DM组小鼠(P＜0.01).结论 tesaglitazar可以减轻2型糖尿病LDLr-/-小鼠血管斑块脂质沉积和炎症反应,增加斑块内胶原和α-SMA含量,利于斑块的稳定.
目的 探討過氧化物酶體增殖物激活受體(PPAR)α/γ雙激動劑對低密度脂蛋白受體基因敲除(LDLr-/-)糖尿病小鼠動脈粥樣硬化斑塊穩定性的影響.方法 取4週齡雌性LDLr-/-小鼠45隻,高糖高脂飲食餵養4週後採用隨機數錶法隨機分為3組(每組15隻),即對照組、2型糖尿病模型組(DM組)和PPARα/γ雙激動劑替賽格列他(tesaglitazar)糖尿病治療組(治療組).對照組僅給予高糖高脂飲食餵養.DM組給予高糖高脂飲食聯閤小劑量鏈脲佐菌素誘導.治療組飼料中混閤20μg/kg tesaglitazar餵養.隨機分組後繼續高糖高脂飲食餵養6週.藥物榦預6週後處死小鼠,分彆于藥物榦預前和處死前測定小鼠體質量、血清總膽固醇(TC)、甘油三酯(TG)和血糖(Glu)水平.免疫印跡和免疫組織化學染色檢測頭臂榦動脈斑塊炎癥因子細胞間黏附分子1(ICAM-1)、血管內皮黏附分子1(VCAM-1)、單覈細胞趨化蛋白-1(MCP-1)錶達水平.囌木素伊紅、油紅O、天狼猩紅染色觀察頭臂榦動脈斑塊,免疫組織化學染色檢測頭臂榦動脈斑塊平滑肌肌動蛋白(α-SMA)和巨噬細胞錶麵分子-3(Mac-3)錶達.結果 DM組小鼠血清TC、TG和Glu水平均顯著高于對照組,分彆為(32.34±3.26) mmol/L比(16.17±1.91) mmol/L、(3.57 ±0.99) mmol/L比(2.21 ±0.11) mmol/L和(15.21±4.67) mmol/L比(6.89±0.83) mmol/L(P均＜0.01).DM組小鼠頭臂榦動脈斑塊炎癥因子ICAM-1、VCAM-1和MCP-1的蛋白錶達水平均顯著高于對照組,分彆為2.31 ±0.35比1.34 ±0.21,1.65±0.14比0.82±0.26和2.27±0.16比1.56±0.23(P均＜0.01).DM組小鼠血管斑塊麵積顯著大于對照組[(4.597±1.260)×103 μm2比(0.075 ±0.030)×103 μm2],脂質、α-SMA、Mac-3和膠原含量均顯著高于對照組,分彆為(47.23±2.64)％比(9.67±1.75)％、(5.54±1.17)％比(2.13±0.41)％、(19.15 ±3.51)％比(1.72±0.16)％和(4.27±0.74)％比(0.43±0.09)％(P均＜0.01).治療組小鼠血清TC、TG和Glu水平均顯著低于DM組(P均＜0.01),分彆為(30.47 ±3.18)mmol/L、(3.14 ±0.71) mmol/L和(7.92±1.28) mmol/L.治療組小鼠頭臂榦斑塊炎癥因子ICAM-1、VCAM-1和MCP-1蛋白錶達水平均低于DM組(P＜0.05或P＜0.01),分彆為1.84±0.22、1.27±0.11和1.83±0.24.治療組小鼠血管斑塊麵積[(1.283 ±0.410)×103 μm2]小于DM組(P＜0.01),脂質含量和Mac-3錶達水平亦低于DM組(P均＜0.01),分彆為(23.52±1.39)％和(12.84±3.22)％.但治療組小鼠頭臂榦動脈斑塊內膠原含量[(6.32±1.15)％]和α-SMA含量[(9.46±1.47)％]均高于DM組(P均＜0.01).治療組小鼠主動脈脂質沉積和Mac-3錶達含量則低于DM組小鼠(P＜0.01).結論 tesaglitazar可以減輕2型糖尿病LDLr-/-小鼠血管斑塊脂質沉積和炎癥反應,增加斑塊內膠原和α-SMA含量,利于斑塊的穩定.
목적 탐토과양화물매체증식물격활수체(PPAR)α/γ쌍격동제대저밀도지단백수체기인고제(LDLr-/-)당뇨병소서동맥죽양경화반괴은정성적영향.방법 취4주령자성LDLr-/-소서45지,고당고지음식위양4주후채용수궤수표법수궤분위3조(매조15지),즉대조조、2형당뇨병모형조(DM조)화PPARα/γ쌍격동제체새격렬타(tesaglitazar)당뇨병치료조(치료조).대조조부급여고당고지음식위양.DM조급여고당고지음식연합소제량련뇨좌균소유도.치료조사료중혼합20μg/kg tesaglitazar위양.수궤분조후계속고당고지음식위양6주.약물간예6주후처사소서,분별우약물간예전화처사전측정소서체질량、혈청총담고순(TC)、감유삼지(TG)화혈당(Glu)수평.면역인적화면역조직화학염색검측두비간동맥반괴염증인자세포간점부분자1(ICAM-1)、혈관내피점부분자1(VCAM-1)、단핵세포추화단백-1(MCP-1)표체수평.소목소이홍、유홍O、천랑성홍염색관찰두비간동맥반괴,면역조직화학염색검측두비간동맥반괴평활기기동단백(α-SMA)화거서세포표면분자-3(Mac-3)표체.결과 DM조소서혈청TC、TG화Glu수평균현저고우대조조,분별위(32.34±3.26) mmol/L비(16.17±1.91) mmol/L、(3.57 ±0.99) mmol/L비(2.21 ±0.11) mmol/L화(15.21±4.67) mmol/L비(6.89±0.83) mmol/L(P균＜0.01).DM조소서두비간동맥반괴염증인자ICAM-1、VCAM-1화MCP-1적단백표체수평균현저고우대조조,분별위2.31 ±0.35비1.34 ±0.21,1.65±0.14비0.82±0.26화2.27±0.16비1.56±0.23(P균＜0.01).DM조소서혈관반괴면적현저대우대조조[(4.597±1.260)×103 μm2비(0.075 ±0.030)×103 μm2],지질、α-SMA、Mac-3화효원함량균현저고우대조조,분별위(47.23±2.64)％비(9.67±1.75)％、(5.54±1.17)％비(2.13±0.41)％、(19.15 ±3.51)％비(1.72±0.16)％화(4.27±0.74)％비(0.43±0.09)％(P균＜0.01).치료조소서혈청TC、TG화Glu수평균현저저우DM조(P균＜0.01),분별위(30.47 ±3.18)mmol/L、(3.14 ±0.71) mmol/L화(7.92±1.28) mmol/L.치료조소서두비간반괴염증인자ICAM-1、VCAM-1화MCP-1단백표체수평균저우DM조(P＜0.05혹P＜0.01),분별위1.84±0.22、1.27±0.11화1.83±0.24.치료조소서혈관반괴면적[(1.283 ±0.410)×103 μm2]소우DM조(P＜0.01),지질함량화Mac-3표체수평역저우DM조(P균＜0.01),분별위(23.52±1.39)％화(12.84±3.22)％.단치료조소서두비간동맥반괴내효원함량[(6.32±1.15)％]화α-SMA함량[(9.46±1.47)％]균고우DM조(P균＜0.01).치료조소서주동맥지질침적화Mac-3표체함량칙저우DM조소서(P＜0.01).결론 tesaglitazar가이감경2형당뇨병LDLr-/-소서혈관반괴지질침적화염증반응,증가반괴내효원화α-SMA함량,리우반괴적은정.
Objective To investigate the effects of peroxisome proliferator-activated receptor (PPAR) α/γ agonist on atherosclerotic plaque stabilization in diabetic LDL receptor knockout(LDLr-/-)mice.Methods Female 4-week-old LDLr-/-mice fed with high-glucose and high-fat diet for 4 weeks were randomly divided into three groups (n =15 each): control group (only fed with high-glucose and high-fat diet),diabetic group [induced by high-glucose and high-fat diet combined with a low-dose of streptozotocin (STZ)] without tesaglitazar and with tesaglitazar (20 μg/kg oral treatment).Mter 6 weeks,the mice were sacrificed,body weight,fasting blood glucose (Glu),total cholesterol (TC),triglyceride (TG) levels were measured.The expression of ICAM-1,VCAM-1,MCP-1 in the brachiocephalic atheroselerotic lesions were determined by Western blot and immunohistochemistry,respectively.Brachiocephalic artery was prepared for morphologic study (HE,oil red O,Sirius red staining) and immunohistochemical analysis (macrophage surface molecule-3,α-smooth muscle actin),respectively.Results Serum TC [(32.34 ± 3.26) mmol/Lvs.(16.17 ±1.91) mmol/L],TG[(3.57 ±0.99) mmol/L vs.(2.21 ±0.11) mmol/L] and Glu[(15.21 ±4.67) mmol/L vs.(6.89 ± 0.83)mmol/L] levels were significantly higher in diabetic group than in the control group (all P ＜ 0.01).The expression of ICAM-1 (2.31 ± 0.35 vs.1.34 ± 0.21),VCAM-1 (1.65 ±0.14 vs.0.82 ±0.26),MCP-1 (2.27 ± 0.16 vs.1.56 ± 0.23) were significantly upregulated in diabetic group compared with control group (all P ＜ 0.01).Brachiocephalic atheroselerotic plaque area [(4.597 ±1.260) ×103 μm2 vs.(0.075 ±0.030) ×103 μm2],lipid deposition [(47.23 ±2.64)％ vs.(9.67 ±1.75)％],Mac-3 positive area [(19.15 ± 3.51)％ vs.(1.72 ± 0.16)％],α-smooth muscle actin [(5.54 ± 1.17) ％ vs.(2.13 ± 0.41) ％] and collagen content [(4.27 ± 0.74) ％ vs.(0.43 ± 0.09) ％]were all significantly larger/higher in diabetic LDLr-/-mice than in the control group (all P ＜ 0.01).While tesaglitazar treatment significantly reduced serum TC [(30.47 ± 3.18) mmol/L],TG [(3.14 ± 0.71)mmol/L] and Glu [(7.92 ± 1.28) mmol/L] levels (all P ＜ 0.01).Similarly,the expression of ICAM-1 [(1.84 ± 0.22)],VCAM-1 [(1.27 ± 0.11)],MCP-1 [(1.83 ± 0.24)],brachiocephalic atheroselerotic lesion area [(1.283 ± 0.410) × 103 μm2],lipid deposition [(23.52 ± 1.39) ％] were also significantly reduced by tesaglitazar (all P ＜ 0.05).Moreover,tesaglitazar increased o-smooth muscle actin [(9.46 ±1.47) ％] and collagen content [(6.32 ± 1.15) ％] in diabetic LDLr-/-mice (all P ＜ 0.05).In addition,lipid deposition and Mac-3 positive areas [(10.67 ± 0.88)％ vs.(15.83 ± 1.01)％] in the aortic root were also reduced in tesaglitazar treated diabetic LDLr-/-mice (P ＜ 0.01).Conclusions Tesaglitazar has antiinflammatory effects in the diabetic LDLr-/-mice.Tesaglitazar could reduce lipid deposition,increase collagen and α-SMA content in the brachiocephalic atheroselerotic lesions,thus,stabilize atherosclerotic plaque in this model.
