黄连素抑制FSP27基因表达改善2型糖尿病地鼠内脏白色脂肪组织胰岛素抵抗的研究

摘要：目的 研究黄连素（BBR）对2型糖尿病（T2DM）中国地鼠内脏白色脂肪组织（VWAT）中脂肪特异蛋白27（FSP27）和PR结构域蛋白16 （PRDM16）信号通路基因mRNA表达的影响并探讨相关机制。方法 以高脂饮食诱导肥胖胰岛素抵抗（OIR）地鼠模型,然后给予小剂量链脲菌素建立T2DM地鼠模型,对照组喂以普通饲料。造模完成后随机分成对照组、OIR组、肥胖T2DM组和T2DM BBR组。BBR治疗9周后,应用实时定量PCR方法检测各组地鼠VWAT中FSP27和PRDM16信号通路及其靶基因的mRNA表达改变。结果 与对照组相比,OIR组和肥胖T2DM组地鼠VWAT中PRDM16、CtBP-1、CtBP-2、C/EBPβ、PPARγ、PGC1α、PGC-1β及棕脂组织特异基因UCP-1、Cidea、Elovl3、PPARα及Acox、Cpt1和Acadm的mRNA表达降低,而FSP27和白脂组织特异基因Resistin、MEST和Serpina3k的mRNA表达增加。BBR治疗降低肥胖T2DM组地鼠VWAT中FSP27的表达而增强PRDM16信号通路效应,诱导棕脂组织特异基因mRNA的表达,诱导VWAT棕色化基因表型,改善脂诱性胰岛素抵抗。结论 BBR降低FSP27表达而增加PRDM16的表达与其诱导VWAT棕色化的分子机制相关,有助于增强产热耗能, 改善VWAT的异常脂代谢,改善FIVWATIR,恢复VWAT的功能。     

RIP140/PGC-1α在AngⅡ调节心肌能量代谢中的作用研究

目的探讨转录辅助因子受体相互作用蛋白140(receptor interacting protein 140,RIP140)与过氧化物酶体增殖物受体γ共激活因子1α(peroxisome proliferator-activated receptor gamma coactivator-1α,PGC-1α)在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)调节心肌细胞能量代谢中的作用。方法借助腺病毒载体系统诱导RIP140和PGC-1α基因过表达;利用荧光检测系统测定乳鼠心肌细胞线粒体ATP的含量;实时荧光定量PCR和Western blot的方法检测RIP140和PGC-1α的表达情况。结果乳鼠心肌细胞给予100 nmol·L-1AngⅡ刺激36 h后,心肌细胞线粒体ATP的含量降低(P<0.01),同时伴随RIP140 mRNA与蛋白水平的升高,而PGC-1αmRNA与蛋白水平下调。AngⅡ诱导的ATP含量减少在过表达RIP140组中进一步下降,而在过表达PGC-1α组中有所减轻。结论AngⅡ诱导的ATP含量减少与RIP140表达上调和PGC-1α表达下调有关。     

腺病毒介导心脏特异性过表达RIP140对大鼠心脏功能及炎症通路的影响

目的探讨心脏组织特异性过表达受体相互作用蛋白140(receptor interacting protein 140,RIP140)对心脏功能及炎症通路的影响。方法大鼠心脏组织多点注射携带RIP140基因的腺病毒载体,使心肌组织过表达RIP140;免疫荧光验证RIP140蛋白在心脏组织中的过表达情况;超声心动图与血流动力学参数评估心脏功能;ELISA分析TNF-α、IL-2、IL-1β等炎症因子水平;Western blot分析p65、IκB-α的蛋白水平。结果腺病毒载体介导的外源基因RIP140可成功过表达于心脏组织,诱导心室扩张、左室射血分数下降、心功能受损,促进心肌组织TNF-α、IL-2、IL-1β炎症因子释放,p65蛋白入核、胞质IκB-α蛋白降解。结论腺病毒介导RIP140基因在心脏组织中的特异性过表达损伤心脏功能,激活NF-κB/p65炎症通路,促进炎症因子释放。     

软脂酸抑制C2C12肌细胞PGC-1α的表达

目的探讨不同脂肪酸类型及浓度对C2C12肌细胞PGC-1α表达的影响。方法分别用0．25mmol／L、0．5mmo]／L和0．75mmol／L三个浓度梯度的软脂酸（C16：0）、硬脂酸（C18：0）、棕榈酸（C16：1）、油酸（C18：1）以及亚油酸（C18：2）孵育C2C12肌细胞。采用实时荧光定量PCR检测细胞中PGC-1α的基因表达。结果在0．25mmol／L浓度时,各组肌细胞PGC-1α mRNA表达较对照组差异无统计学意义（P〉0．05）;在0．5mmoL／L和O．75mmol／L浓度时,软脂酸组肌细胞PGC-1α mRNA表达较对照组降低（P〈0．05）;其余各组表达无统计学差异（P〉0．05）。PGC-1α mRNA表达在在培养6h、12h和24h降低,其中24h降低最明显（P〈0．01）。结论软脂酸孵育的C2C12肌细胞PGC-1α的mRNA表达下降,并且有浓度和时间依赖性。而单不饱和或多不饱和脂肪酸培养的C2C12肌细胞对PGC-1α表达无明显影响。     

川续断皂苷Ⅵ对小鼠骨髓基质细胞ST-2 成脂分化的影响及其机制研究

目的观察川续断皂苷Ⅵ（Asperosaponin Ⅵ,ASAⅥ）对脂肪细胞分化的影响及Wnt 通路的调节。方法以小鼠骨髓基质细胞系ST-2 为研究对象,将其分为对照组、成脂分化组以及4 个不同剂量的ASAⅥ组。其中对照组加入溶媒,成脂分化组加入成脂诱导分化试剂,ASAⅥ组除加入成脂诱导分化试剂外,使用不同浓度（10-7、10-6、10-5、 10-4 mol/L）的ASAⅥ干预细胞。各组细胞处理5 d 后,行油红O 染色,观察脂滴形成情况并计算成脂率进行定量分析;荧光定量PCR（FQ-PCR）检测成脂相关基因PPARγ、FABP4 和Wnt/β-连环素（β-catenin）通路蛋白β-catenin 的 mRNA 表达水平;Wnt 通路抑制剂DKK1 干预诱导成脂分化的ST-2 细胞5 d,FQ-PCR 检测ASAⅥ所调节的 PPARγ、FABP4 和β-catenin mRNA 表达水平。结果与成脂分化组相比,10-5 mol/L 和10-4 mol/L ASAⅥ组中分化的脂肪细胞显著减少,10-5、10-4 mol/L ASAⅥ明显抑制PPARγ、FABP4 的mRNA 表达,但上调β-catenin mRNA 表达。 DKK1 能够逆转ASAⅥ对ST-2 细胞成脂分化的抑制作用,促进PPARγ、FABP4 的mRNA 表达,抑制β-catenin 的 mRNA 表达。结论ASAⅥ能显著抑制ST-2 细胞的成脂分化,这一作用可能是通过Wnt/β-catenin 通路的激活所介导的。     

RIP140与TNF-α对心肌细胞能量代谢的调控作用

目的观察RIP140与TNF-α对心肌细胞能量代谢的影响。方法体外培养H9c2心肌细胞,分为空载病毒组、过表达RIP140组、空载病毒+TNF-α组、过表达RIP140+TNF-α组,荧光定量PCR检测PPAR-α、PPAR-β/δ、PDK4的mRNA表达水平。使用过表达RIP140的腺病毒感染H9c2细胞,Western blot分析细胞核p65蛋白水平、胞质IκB-α蛋白水平;荧光定量PCR检测TNF-α、IL-1β、IL-2的mRNA表达水平; TNF-α刺激心肌细胞,荧光定量PCR检测RIP140的mRNA表达水平,Western blot分析RIP140蛋白表达水平。结果与过表达RIP140组比较,过表达RIP140+TNF-α刺激组PPAR-β/δ和PDK4的mRNA表达下降。过表达RIP140的心肌细胞核内p65蛋白水平升高,胞质IκB-α蛋白水平下降,TNF-α、IL-1β、IL-2的mRNA表达升高; TNF-α刺激心肌细胞,使RIP140的mRNA和蛋白表达水平升高。结论RIP140与TNF-α可相互作用,介导心肌细胞炎症反应和能量代谢紊乱。     

健脾消脂方异病同治2型糖尿病合并脂代谢紊乱的网络药理学机制分析

目的 探讨健脾消脂方异病同治2型糖尿病（T2DM）合并脂代谢紊乱潜在作用机制。方法 通过中药系统药理学分析平台（TCMSP）获得健脾消脂方中主要的活性成分及其潜在靶蛋白。利用GeneCards、OMIM和TTD数据库筛选出与T2DM、脂代谢紊乱相关的疾病靶点,并通过Cytoscape3.7.0软件构建“成分-靶点-基因-疾病”网络图;将T2DM、脂代谢紊乱的共同靶点导入STRING数据库构建蛋白质互作（PPI）网络图,并将PPI分析数据导入Cytoscape3.7.0软件中,筛选出健脾消脂方异病同治T2DM合并脂代谢紊乱的核心靶点,并构建核心靶点网络图。在DAVID数据库中进行GO富集分析及KEGG通路富集分析。结果 检索到健脾消脂方71种药物活性成分,262个有效靶点,2型糖尿病相关靶点2 078个、脂代谢紊乱相关靶点1 467个,药物与疾病的共同靶点133个。GO与KEGG分析表明健脾消脂方治疗T2DM合并脂代谢紊乱主要涉及晚期糖基化终末产物-糖基化终末产物受体（AGE-RAGE）信号通路、肿瘤坏死因子（TNF）信号通路、磷脂酰肌醇-3激酶/蛋白激酶B(PI3K-AKT)信号通路、...     

洋甘菊活性组分对LPS诱导人支气管上皮细胞的保护作用及机制研究

目的 基于驱动蛋白家族 3A 基因（Kif3a）和 Sonic hedgehog（SHH）信号通路探讨洋甘菊活性组分对脂多糖（LPS）诱导的人支气管上皮细胞（16HBE）的保护作用及机制。方法 用LPS诱导16HBE建立哮喘炎症细胞模型。CCK-8法检测不同浓度洋甘菊活性组分对LPS诱导16HBE的增殖的抑制作用，筛选出最佳洋甘菊活性组分给药浓度。将细胞分为空白对照组、LPS模型组、阳性药物组（1 µmol•L^-1地塞米松干预2 h）、洋甘菊活性组分组（200 µg•mL^-1，48 h）。Annexin V/PI结合流式细胞术检测各组的凋亡率。实时荧光PCR法和Western blot法检测Kif3a、SHH、Ptch1和Gli1的mRNA和蛋白表达变化。结果 LPS干预后，可显著降低16HBE活力（P < 0.05），并诱导其细胞凋亡（P < 0.01）。洋甘菊活性组分在浓度200 µg•mL^-1处理48 h时，可以显著逆转LPS对细胞的损伤并促进增殖（P < 0.01），减弱LPS诱导的细胞凋亡（P< 0.01）。与空白对照组比较，LPS诱导的16HBE内Kif3a在mRNA水平和蛋白表达水平显著降低（P < 0.05），SHH、Ptch1、Gli1在mRNA水平和蛋白表达水平显著升高（P < 0.05）；与LPS模型组相比较，洋甘菊活性组分处理后可显著逆转mRNA水平和蛋白表达（P< 0.05），增加Kif3a的mRNA水平和蛋白表达水平，并降低SHH、Ptch1、Gli1的mRNA水平和蛋白表达（P< 0.05）。结论 LPS诱导16HBE损伤，下调Kif3a水平从而上调SHH、Ptch1、Gli1水平，调控SHH信号通路处于异常激活状态。洋甘菊活性组分可显著逆转LPS诱导的细胞炎症损伤，上调Kif3a水平，下调SHH、Ptch1、Gli1水平，通过Kif3a调控SHH信号通路，对LPS诱导的16HBE发挥保护作用，改善哮喘气道炎症。     

溴氰菊酯对大鼠大脑组织CuZn-SOD、GR、GCS亚单位和Nrf2基因表达的影响

目的 研究溴氰菊酯（DM）染毒对大鼠大脑皮层和海马组织铜、锌超氧化物歧化酶（Cu、Zn-SOD）、谷胱苷肽还原酶（GR）和γ-谷氨酰半胱氨酸合成酶（GCS）轻亚单位（GCS1）mRNA表达的影响，以及DM对GCS重亚单位（GCSh）基因和转录因子NF-E2相关因子2（Nrf2）基因mRNA和蛋白表达的影响，并探讨DM对这些酶活力影响的机制。     

褪黑素受体激动剂Neu-p11改善T2DM大鼠的胰岛素抵抗作用研究

目的研究褪黑素受体激动剂Neu-p11改善2型糖尿病(type 2 diabetes mellitus,T2DM)大鼠的胰岛素抵(insulin resistance,IR)作用,并预测其可能机制。方法采用长期高脂肪饲养加小剂量链脲佐菌素(STZ,30 mg·kg^-1,ip)诱导雌性大鼠2型糖尿病动物模型,灌胃给药4周后处死,取肝脏组织。采用RT-PCR方法测定大鼠Sirt1、Nrf-1/2、ERRα等相关因子的mRNA表达量;Western Blot法测定PGC-1α、Mfn2的相对表达。测定大鼠INS,计算HOME-IR与ISI。测定大鼠肝脏ATP含量及ATP酶活力。结果与正常组相比,T2DM大鼠发生IR,PGC-1α、Mfn2蛋白表达上调,Sirt1等mRNA表达显著降低,ERRαmRNA表达升高,ATP含量显著减少,ATP酶活力减弱。与模型组相比,Neu-p11低剂量组降低T2DM大鼠INS;改善T2DM大鼠IR;上调大鼠肝脏蛋白和mRNA表达,下调ERRαmRNA的表达,增加大鼠肝脏ATP含量与ATP酶活力。结论褪黑素受体激动剂Neu-p11可能以调节T2DM大鼠肝脏线粒体平衡相关因子,维持线粒体功能正常为机制,改善IR。     

脂肪干细胞来源外泌体携带miR-27抑制白色脂肪的棕色化

目的 探讨脂肪干细胞（ASC）来源的外泌体在白色脂肪棕色化中的作用。方法 分离培养并鉴定小鼠ASC细胞;慢病毒转染构建过表达 miR-27的 ASC细胞;超高速离心提取正常及过表达 miR-27的 ASC细胞分泌的外泌体,实时荧光定量核酸扩增（qRT-PCR）分析外泌体中 miR-27的表达量;高脂喂食构建肥胖小鼠模型,分别注射过表达 miR-27外泌体（过表达组）、正常 ASC外泌体（正常表达组）及生理盐水（对照组）,在寒冷暴露条件下监测体质量变化;剥离小鼠附睾皮下脂肪组织和肩胛骨脂肪组织,比较组织大小,蛋白免疫印迹法（Western blot）检测解偶联蛋白 1（UCP1）、过氧化物酶体增殖剂激活受体 γ（PPARγ）、含 PR结构域的锌指蛋白 16（PRDM16）、过氧化物酶体增殖子激活受体 γ共激活因子 1α（PGC-1α）的表达。结果 成功提取 ASC及其外泌体,并成功转染 miR-27;ASC及其外泌体中的 miR-27显著高于白色脂肪组织（P＜0.05）。与对照组相比,过表达组及正常表达组的小鼠体质量下降幅度小,随时间的增加效果更加明显（P＜0.05）。过表达组及正常表达组的小鼠的附睾皮下脂肪组织较对照组的小鼠要稍大;而肩胛骨脂肪组织的大小无明显变化。正常表达组与过表达组的 UCP1、PPARγ、PRDM16及 PGC-1α的表达水平较对照组下降（P＜0.05）。结论 脂肪干细胞来源外泌体能够抑制白色脂肪细胞的棕色化。     

ERK信号转导途径在小檗碱抑制脂多糖诱导的COX一2表达中的作用

目的探讨小檗碱是否抑制脂多糖诱导的COX-2mRNA及蛋白表达,以及小檗碱是否通过ERK信号转导途径抑制cOx-2的表达。方法取健康志愿者外周血,分离及培养单核细胞,分为五组,分别为对照组;脂多糖（Lipopolysaccharide,LPS）组;LPS＋小檗碱25μmol／L组;LPS＋小檗碱50μmol/L组;LPS＋小檗碱100μmoL／L组。分别在培养后30min,6h,12h,24h提取细胞,行RT—PCR法测定COX-2mRNA水平,行westemblot法测定ERK、P—ERK及COX-2蛋白水平。同时加入选择性ERK抑制剂,分别测定C0X-2mRNA及蛋白水平。结果与对照组相比,LPS组COX-2mRNA及蛋白表达明显增强（P〈0．01）。与LPS组相比,小檗碱组COX-2mRNA及蛋白表达明显抑制（P〈0．05）,且随着浓度增加,抑制作用更明显,在给药后12h,小檗碱对COX-2抑制作用最强。与LPS组相比,小檗碱组ERK活性水平有明显统计学差异（P〈0．05）。加入ERK抑制剂之后,COX-2mRNA及蛋白水平降低明显（P〈0．05）。结论小檗碱能抑制人外周血单核细胞cOx-2mRNA及蛋白水平,其作用程度呈浓度依赖性,小檗碱对ERK活性蛋白表达有明显抑制作用。小檗碱可能通过ERK信号转导途径抑制人外周血单核细胞C0x_2mRNA及蛋白表达。     

小檗碱对p38MAPK通路及COX-2mRNA表达的作用

目的研究中药小檗碱是否通过p38MAPK信号转导途径抑制人外周血单核细胞COX-2mRNA及蛋白表达。方法取人外周静脉血分离及培养单核细胞,分为对照组、脂多糖(LPS)组、LPS+小檗碱25μmol/L组、LPS+小檗碱50μmol/L组、LPS+小檗碱100μmol/L组。分别在培养后0.5、6、12、24h提取细胞,行RT-PCR法测定COX-2mRNA水平,行Westernblot法测定p38MAPK、p-p38MAPK及COX-2蛋白水平。同时加入选择性p38MAPK抑制剂,分别测定COX-2mRNA及蛋白水平。结果与对照组相比,LPS组COX-2mRNA及蛋白表达明显增强(P<0.01)。与LPS组相比,小檗碱组COX-2mRNA及蛋白表达明显抑制(P<0.05),且随着浓度增加,抑制作用更明显,在给药后12h,小檗碱对COX-2抑制作用最强,但是与LPS组相比,小檗碱组p38MAPK活性水平无明显统计学差异(P>0.05)。加入p38MAPK抑制剂之后,COX-2mRNA及蛋白水平降低明显(P<0.05)。结论小檗碱能抑制人外周血单核细胞COX-2mRNA及蛋白水平,并呈浓度依赖性,p38MAPK与人外周血COX-2表达有关,而小檗碱对p38MAPK活性蛋白表达无明显抑制作用。     

A novel berberine‐metformin hybrid compound exerts therapeutic effects on obese type 2 diabetic rats

In this study, we investigated the biological activities of a novel berberine‐metformin hybrid compound (BMH473) as an anti‐diabetic agent. BMH473 exhibited significant anti‐hyperglycaemic and anti‐hyperlipidaemic effects on T2DM rats. In white adipose tissue, BMH473 reduced the perirenal and epididymal adipose tissue mass and modulated the lesions in perirenal adipose tissue, by inhibiting the protein expressions of PPAR‐?, C/EBP‐α and SREBP‐1c as well as the mRNA expressions of lipogenic genes. Moreover, BMH473 downregulated the levels of pro‐inflammatory cytokines in perirenal adipose tissue through the suppression of p‐NF‐κB. In liver, BMH473 reduced liver ectopic fat accumulation, by regulating the protein expression levels of SREBP‐1c and PPAR‐α as well as the mRNA expression levels of lipogenic genes. In addition, BMH473 inhibited hepatic gluconeogenesis by promoting the phosphorylation levels of AMPK α and ACC, and down‐regulating the mRNA expression levels of FBPase, G6Pase and PEPCK. Furthermore, BMH473 exhibited significant inhibitory effects on lipogenesis and lipid accumulation in 3T3‐L1 adipocytes by modulating the protein expression levels of PPAR‐?, C/EBP‐α and SREBP‐1 c as well as the mRNA expression levels of lipogenic genes. In conclusion, our results suggest that the newly synthesized BMH473 is beneficial for maintaining glucose and lipid homeostasis in type 2 diabetic rats, and exhibits better anti‐hyperlipidaemic effects compared to metformin and berberine.     

Berberine inhibits the interleukin-1 beta-induced inflammatory response via MAPK downregulation in rat articular chondrocytes

Osteoarthritis (OA) is one of the most chronic degenerative arthritic diseases, which gradually results in chondrocyte changes, articular cartilage degeneration, subchondral bone sclerosis, joint pain, swelling, and dysfunction. Berberine (BBR) has various confirmed biological activities, such as anti-inflammatory and antioxidant activities. However, the effect of BBR on the production of inflammation-associated proteins, including inducible nitric oxide synthase (iNOS), cyclooxygenase (Cox)-2, metalloproteinases (MMPs), Collagen II, TNF-α, and IL-6 via the MAPK (mitogen-activated protein kinases) pathway in IL-1β-stimulated rat chondrocytes, has not yet been studied. Thus, the purpose of this study was to evaluate whether BBR would decrease the production of inflammation-associated proteins through the MAPK signal pathway. Rat chondrocytes were cultured and pretreated with BBR at different concentrations (0, 25, 50, and 100 μM) and then stimulated with or without IL-1β (10 ng/mL). The mRNA expression of iNOS, COX-2, MMP-3, MMP-13, TNF-α, and IL-6 was measured by real-time polymerase chain reaction (RT-PCR), and the protein expression of iNOS, COX-2, Collagen II, MMP-3,MMP-13, and MAPKs were measured by Western blotting. The results showed that the expression of iNOS, COX-2, MMP-3, MMP-13, TNF-α, and IL-6 increased in the IL-1β-treated group and BBR showed an ability to inhibit the elevated expression under the pretreatment. Furthermore, the IL-1β-induced downregulation of Collagen II could be ameliorated by BBR. Moreover, the expression of MAPKs was significantly decreased by BBR. These results demonstrated that BBR had the anti-catabolic and anti-inflammation abilities that were through the MAPKs in IL-1β-induced rat chondrocytes. These findings may provide a novel therapeutic choice for treatment of OA using BBR.     

Berberine inhibits IFN-γ signaling pathway in DSS-induced ulcerative colitis

AimsThe potential signaling pathways and core genes in ulcerative colitis (UC) were investigated in this study. Furthermore, potential mechanisms of BBR in treating UC were also explored.MethodsExpression profiling by array of UC patients were obtained from Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were determined with the differential analysis. The biological functions of DEGs were analyzed through the Database for Annotation, Visualization and Integrated Discovery (DAVID). The Gene Set Enrichment Analysis (GSEA) was applied to analyze the expression differences between two different phenotype sample sets. Dextran sulfate sodium (DSS) was applied to establish UC model of mice and lipopolysaccharide (LPS) was utilized to induce inflammatory damage of NCM460 cells. Therapeutic effects of berberine (BBR) on disease performance, pathologic changes and serum supernatant indices were analyzed in vivo. To further investigate the potential mechanisms of BBR in treating UC, the expression of genes and proteins in vivo and in vitro were examined by RT-qPCR, immunohistochemical staining and western blotting.ResultsImmune-inflammatory genes were identified and up-regulated significantly in UC patients. In addition, IFN-γ signaling pathway and its core genes were significantly up-regulated in the phenotype of UC. All disease performance and the pathologic changes of UC in mice were evidently ameliorated by BBR treatment. The pro-inflammatory cytokines of serum, including CXCL9, CXCL1, IL-17 and TNF-α, in UC mice were significantly reduced by treatment of BBR. In terms of mechanisms of BBR in treating UC, the pro-inflammatory and immune-related genes, encoding IFN-γ, IRF8, NF-κB and TNF-α decreased significantly in UC mice followed by BBR treatment. Meanwhile, the expression of IFN-γ and its initiated targets, including IRF8, Ifit1, Ifit3, IRF1, were suppressed significantly by BBR treatment in vivo. The blocking of IFN-γ in vitro led to the silence of IFN-γ signaling pathway after exposure to BBR. Furthermore, the blocking of IFN-γ in vitro led to the silence of IFN-γ signaling pathway after exposure to BBR.ConclusionBBR holds anti-inflammatory activity and can treat UC effectively. The anti-inflammatory property of BBR is tightly related to the suppression of IFN-γ signaling pathway, which is crucial in immune-inflammatory responses of the colon mucosa.     

辛伐他汀对K562细胞Ras-MAPK信号通路的体内外研究

目的通过体内、外实验研究辛伐他汀对K562细胞Ras-MAPK信号通路的影响,探讨辛伐他汀诱导K562细胞凋亡的可能机制。方法体外培养CML细胞株K562细胞,做以下实验:①用流式细胞术(FCM)检测辛伐他汀处理K562细胞后其凋亡率的变化,②18只Balb/c-nu/nu裸小鼠皮下接种K562细胞,构建裸鼠的K562细胞移植瘤模型,③TUNEL法检测辛伐他汀诱导裸鼠体内K562细胞早期凋亡的变化,④RT-PCR检测裸鼠体内外K562细胞N-ras、c-Raf-1、ERK1mRNA水平的变化。结果辛伐他汀在体外能明显诱导K562细胞凋亡,Ras-MAPK通路大多数基因出现差异表达。不同剂量的辛伐他汀能够诱导裸鼠体内K562细胞凋亡,并随剂量的增加凋亡率逐渐增高(P<0.01);辛伐他汀能够引起N-ras、c-Raf-1、ERK1mRNA水平的明显差异表达(P<0.01)。结论辛伐他汀在体内外均能够引起参与K562细胞凋亡的Ras分子和Ras分子下游分子mRNA水平的变化,从一定角度说明辛伐他汀能依赖Ras-MAPK信号通路诱导K562细胞凋亡。