QUANTITATIVE STUDY ON APOPTOSIS INDUCED BY ELECTROMAGNETIC PULSES IN NIH-3T3 FIBROBLASTS

Aim: To observe the apoptotic changes following exposure to EMP and to explore the possible injury mechanism in NIH - 3T3 fibroblasts. Methods: Following NIH - 3T3 cells were exposed to EMP,the proliferation and viability of NIH - 3T3 fibroblasts were estimated by hemacytometer and MTT Measurements. Apoptotic rate was measured by flow cytometer. The imnmohistochemical SP method was used to determine bcl- 2, p53. The positively stained cells were analyzed with CMIAS- Ⅱ image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. Results: The proliferation and viability of NIH- 3T3 cells were markedly inhibited and significant apoptosis was induced following exposure to EMP.EMP could increase the number of non- adherent cells, the highest ratio (33.9%) of non- adherent cells also occurred at 6h. The As70 value of MTT decreased immediately at 1 h, 6h following the cells were exposed as compared with the control (P ＜ 0.05). The highest ratio of apoptosis was 15.07% at 6h, then decreased gradually. Down - regulation of bcl - 2 and up - regulation of p53 were induced by EMP ( P ＜ 0.05). Conclusions: EMP could promote apoptosis of NIH - 3T3 fibroblasts. EMP could also down - regulate bcl - 2 level and up - regulate p53 level in NIH - 3T3 fibroblasts. Bcl - 2 and p53 gene may take part in the process of apoptosis.

华泽兰根部乙酸乙酯萃取物基于NF-κB及NLR-P3炎症小体信号通路对NIH-3T3小鼠胚胎成纤维细胞功能的影响

目的探讨华泽兰乙酸乙酯萃取物对小鼠胚胎成纤维细胞(NIH-3T3)功能的影响。方法从炎症因子一氧化氮(NO)及氧化应激重要因素活性氧(ROS)介入,通过核因子κB(NF-κB)及核苷酸结合寡聚化结构域样受体蛋白3(NLR-P3)炎症小体等信号通路角度阐明其作用机制。体外培养NIH-3T3细胞,分为空白对照组(Control)、不同浓度华泽兰根部乙酸乙酯萃取物组,检测不同浓度下华泽兰根部乙酸乙酯萃取物对NIH-3T3细胞Toll增殖活性(MTT法)、细胞迁移(细胞划痕实验)、分泌NO及胞内ROS生成的影响,Western blot法检测其对Toll样受体4(TLR-4)、磷脂酰肌醇3激酶(PI3K)、NLR-P3、NF-κB、髓样分化因子(Myd88)、核因子抑制蛋白α(IκBα)、白细胞介素-18(IL-18)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、凋亡相关斑点样蛋白(ASC)、环氧化酶-2(COX-2)蛋白表达的影响。结果细胞增殖与细胞划痕实验表明,华泽兰根部乙酸乙酯萃取物对于NIH-3T3细胞的增殖与划痕愈合影响具有双向性。与对照组相比在低浓度下(<6.25μg·mL^(-1)),萃取物有促进NIH-3T3细胞增殖与迁移的作用,而高浓度(>25μg·mL^(-1))反而表现出对NIH-3T3细胞的增殖与迁移抑制作用,且不同阶段呈现出相应的剂量依赖性,与对照组相比有统计学意义(P<0.05)。在萃取物浓度为6.25μg·mL^(-1)时,NO生成量相较于低浓度组差异有显著统计学意义(P<0.01),而高浓度组(>12.5μg·mL^(-1))相较对照组差异无统计学意义(P>0.05)。细胞内ROS生成实验表明,经华泽兰根部乙酸乙酯萃取物处理后ROS生成量显著高于对照组(P<0.05,P<0.01)。Western blot实验表明,与对照组相比,萃取物作用浓度为6.25μg·mL^(-1)时,细胞内NF-κB、TLR-4、Myd88、NLR-P3、ASC表达显著升高(P<0.01),IκBα表达升高(P<0.05),IL-18表达显著降低(P<0.01),Caspase-3表达降低(P<0.05)。结论华泽兰乙酸乙酯萃取物对小鼠胚胎成纤维细胞(NIH-3T3)增殖的影响具有双向性,即低浓度促进细胞增殖而高浓度抑制细胞增殖。其作用机制可能与NF-κB及NLR-P3炎症小体信号通路和细胞凋亡有关。

槲皮素对小鼠体内和体外NIH-3T3细胞的抗氧化作用及其机理

目的比较槲皮素对小鼠体内和体外H2O2处理前/后的NIH-3T3细胞抗氧化效应的差异并探讨其作用机理。方法将NIH-3T3细胞分为4组,槲皮素前保护组(Qb)、槲皮素后保护组(Qa)、H2O2组(H2O2)和对照组(C)。用试剂盒检测细胞内T-AOC、SOD、GSH-Px、GSH、MDA、NOS和NO2-/NO3-的水平;MTT和TUNEL法检测细胞凋亡率和生存率;免疫印迹和免疫组化技术检测细胞的cyclin D1、PTEN、NF-κB、HSP-70、BCl-2、BAX、及caspase-3的表达。另将20只Wistar大鼠等分为对照组和实验组,检测槲皮素灌胃前、后1、2及24 h血浆内T-AOC、SOD、GSH-Px、GSH、MDA、NOS和NO2-/NO3-的水平。结果H2O2组细胞凋亡率增高,cyclin D1、PTEN及BCl-2的表达下调;BAX、HSP-70、NF-κB及caspase-3的表达上调;T-AOC,SOD,GSH-Px及GSH水平降低,NOS、NO2-/NO3-及MDA增高。动物灌胃后1 h、2 h血浆中T-AOC、SOD、GSH-Px、GSH,NOS及NO2-/NO3-的水平增高,MDA降低;24 h后基本恢复。结论槲皮素在体内外皆可通过调节抗氧化酶体系发挥抗氧化作用,其效应依据H2O2处理与否和处理前/后等细胞的异质性而呈现一定差异。

In vitro responses of human pulp cells and 3T3 mouse fibroblasts to six contemporary dental restoratives

In vitro responses of human primary pulp cells (HPCs) and 3T3 mouse fibroblasts to six contempo-rary commercial dental restoratives were evaluated using the WST-1 assay. The results show that Fuji II is not cytotoxic to both cells. Fuji II LC is not cyto-toxic to HPCs but cytotoxic to 3T3 cells, indicating that 3T3 cells are more vulnerable to 2-hydroxyethyl methacrylate (HEMA) than HPCs. Vitremer is very cytotoxic probably due to having diphenyliodonium chloride and HEMA in it. Z100 is very cytotoxic probably due to having triethylene glycol dimethacry-late (TEGDMA) in it. P60 is cytotoxic but less cyto-toxic than Z100 probably due to no TEGDMA in it. Durelon is the most cytotoxic among the six materials studied probably due to the high cytotoxicity of zinc ions. Additionally, the cytotoxcity of the tested mate-rials was found to be dose-dependent.

sIL-13Rα2抑制IL-13介导的NIH-3T3胶原I的合成及血吸虫病肝组织中sIL-13Rα2/IL-13的表达(英文)

目的研究在成纤维细胞NIH-3T3中IL-13可溶性受体sIL-13Rα2对IL-13的抑制作用,为进一步研究sIL-13Rα2对日本血吸虫感染小鼠体内肝纤维化的治疗作用奠定基础。方法用ELISA和RT-PCR检测感染日本血吸虫的BALB/c小鼠0、6、8、10和12w不同感染时期肝脏组织IL-13和sIL-13Rα2表达和转录水平。构建sIL-13Rα2表达质粒转染成纤维细胞NIH-3T3,用IL-13(50ng/mL)刺激转染后成纤维细胞NIH-3T3,用RT-PCR和Western blotting分别检测该细胞分泌的Ⅰ型胶原。结果感染后小鼠肝脏肉芽肿组织中IL-13和sIL-13Rα2的蛋白表达水平随感染时间的延长而逐渐增高,第8wIL-13水平达到高峰(16.1586pg/mL),随后逐渐降低但仍高于正常水平(3.4146pg/mLP=0.017);第10wsIL-13Rα2的分泌达到高峰(4827.426pg/mL),以后逐渐减低,但仍高于正常水平(4057.112pg/mLP=0.021)。IL-13和sIL-13Rα2的mRNA转录趋势和ELISA检测结果相符合,均随感染时间的延长而增高,分别在第8w和第10w达到最高峰,随后逐渐降低但仍高于正常组小鼠(P=0.033;P=0.025)。实验组(sIL-13Rα2=2mg/mL)Ⅰ型胶原mRNA转录水平较正常对照组减低8.83%(P=0.012);蛋白水平较对照组减低7.41%(P=0.031)。结论sIL-13Rα2在NIH-3T3细胞中对IL-13有抑制作用,提示sIL-13Rα2在治疗血吸虫病肝纤维化中具有潜在价值。

miR-433在NIH-3T3细胞纤维化和矽尘诱导的小鼠肺纤维化中的作用

目的:本研究旨在探究微小RNA-433(miR-433)在纤维化中的作用及机制。方法:采用TargetScan预测miR-433的潜在靶基因。检测转化生长因子β1(TGF-β1)处理小鼠胚胎成纤维细胞(NIH-3T3细胞)24h后miR-433表达的变化。检测miR-433 mimic对TGF-β1处理的NIH-3T3细胞中p-SMAD2、纤维连接蛋白(FN)、α-平滑肌肌动蛋白(α-SMA)和结缔组织生长因子(CTGF)表达水平的影响。采用CCK-8法和流式细胞术检测转染miR-433 mimic对TGF-β1诱导的NIH-3T3细胞生长和S期阻滞的影响。构建矽尘诱导的小鼠肺纤维化模型并使用agomiR-433进行干预,HE染色和Masson染色观察miR-433对小鼠肺纤维化的影响,采用免疫组化检测小鼠肺组织中α-SMA的表达。结果:miR-433能特异性结合SMAD2的3’-UTR,并抑制其蛋白和mRNA的表达。TGF-β1能下调NIH-3T3细胞中miR-433的表达水平,上调p-SMAD2蛋白的水平,同时也增强FN、α-SMA和CTGF蛋白和mRNA的表达,并增强细胞活力,诱导S期细胞数量增加;而miR-433 mimic能逆转TGF-β1对NIH-3T3细胞活力和S期阻滞的影响。在矽尘诱导的小鼠肺纤维化模型中,agomiR-433能抑制肺纤维化进程,减少小鼠肺组织中α-SMA的表达。结论:miR-433可能通过靶向调控SMAD2干预TGF-β1/SMAD2信号通路,从而参与调节纤维化的病理过程。

小鼠gdnf基因在真核细胞NIH-3T3中的表达

为了得到超量表达胶质细胞源神经营养因子(glial cell line-derived neuotrophic factor,GDNF)的NIH-3T3细胞株,用于制作精原干细胞(spermatogonial stem cells,SSCs)培养的饲养层,通过RT-PCR方法成功地从幼年小鼠睾丸中克隆了gdnf基因,构建了真核表达载体pcDNA3.1-gdnf,并用其转染NIH-3T3细胞.对筛选出的阳性细胞克隆进行的免疫荧光染色、RT-PCR和Western blotting的结果表明,获得了超量表达gdnf基因的NIH-3T3细胞株,这为精原干细胞的培养奠定了基础.

CREG knockdown promotes NIH3T3 Fibroblasts Apoptosis via activating Cathepsin

Background The CREG is an important lysosomal protein involved in a variety of cellular functions including promoting cell differentiation,sustaining mature homeostasis, and antagonizing apoptosis.Deficiency of CREG in cell and tissue results in a pathologic apoptosis.The present study aimed to elucidate the mechanism of CREG regulation apoptosis.Methods We firstly generated stable NIH3T3 fibroblasts by transfection of pDS_shCREGs vectors.Furthermore, PI-Annexin V and TUNEL staining were used to identify that CREG knockdown promoted the cell apoptosis in NIH3T3 fibroblasts.Western blotting and immunofluorescence staining was used to identify the expression and localization of M6P/ IGF2R and cathepsin L in cytoplasm.Results pDS_shCREGs vector transfection produced an approximately 80%decrease in CREG levels both in the lysate and in the media.The expression and localization of M6P/IGF2R and cathepsin L in cytoplasma changed obviously associated with down-regulated of CREG.In addition,the retention and secretion of cathepsin L enhanced significantly.Using the specific inhibitor or siRNA to block cathepsin L activation attenuated the apoptosis mediated by CREG downregulation.Conclusions Our findings indicated that inhibition of CREG expression in NIH3T3 fibroblasts leads to impaired cathepsin L sorting function mediated by M6P/IGF2R and subsequently promotes pathological cell apoptosis.

唾液对体外培养NIH-3T3及MEC-1细胞粘附的影响

目的：探讨人唾液对正常及肿瘤细胞在体外粘附有无不同影响。方法：接种ＮＩＨ—３Ｔ３及ＭＥＣ－１细胞在预先涂有或未涂唾液的２４孔培养板上，用细胞计数法分别在接种后２、４、６和８ｈ计数贴壁细胞。结果：ＮＩＨ－３Ｔ３细胞及ＭＥＣ－１细胞在涂有唾液的塑料底物上在２、４、６、和８ｈ其贴壁抑制率分别为３５％、４１％、２６％、３４％和１０％、９％、８％、４％。结论：人全唾液能抑制体外培养的ＮＩＨ－３Ｔ３细胞的粘附，但对ＭＥＣ－１细胞的影响不大。

金钱鱼凋亡相关基因SaAR对NIH-3T3细胞凋亡的影响

目的观察金钱鱼凋亡相关基因SaAR(Scatophagusargusapoptosis related gene)对成纤维细胞增殖的影响。方法体外培养NIH-3T3细胞,应用脂质体将不同浓度的pcDNA3-SaAR真核表达质粒分别转染细胞并设立对照组进行比较,应用流式细胞术测定细胞凋亡率。结果SaAR基因可显著诱导NIH-3T3细胞凋亡,并呈现剂量依赖性(P<0.05)。结论SaAR基因对体外培养的血管外膜成纤维细胞凋亡有一定的诱导作用。

槲皮素抗NIH-3T3细胞凋亡的作用

目的:探讨槲皮素对过氧化氢(H2O2)诱导NIH-3T3细胞凋亡的抑制作用。方法:体外培养小鼠成纤维细胞,取对数生长期的成纤维细胞分成四个实验组。对照组(C):仅加含有10%胎牛血清的DMEM培养基。槲皮素前保护组(Qb):先用含有50μmol/L的槲皮素培养基作用24 h,再换含有0.5 mmol/L H2O2作用细胞30 m in。槲皮素后保护组(Qa):先用0.5 mmol/L H2O2作用细胞30 m in,再换含有50μmol/L的槲皮素培养基作用24 h。H2O2实验组(H2):先用含有0.5 mmol/L H2O2作用细胞30 m in,再换仅含有10%胎牛血清的DMEM培养基作用24 h。取培养细胞提取DNA和制备1×107/m l细胞悬液的滴片,应用TUNEL和Ladder电泳检测细胞凋亡率;应用细胞免疫组化技术检测Bc l-2、Bax、Caspase-3蛋白质的表达。结果:由TUNEL和Ladder实验得出,Qb组的细胞凋亡率明显低于H2组和Qa组。由细胞免疫组化技术检测分析得出,Qb组与Qa、H2组相比,Bc l-2的表达增高,Bax、Caspase-3的表达减低。结论:槲皮素可能是通过上调Bc l-2的表达,下调Bax、Caspase-3的表达,对H2O2诱导NIH-3T3细胞的凋亡起到抑制作用。

逆转录病毒载体介导的LacZ基因在NIH-3T3细胞中的稳定表达(英文)

精原干细胞(SSCs)介导的转基因技术很可能成为制作转基因动物及治疗雄性不育的一条新途径。为了研究逆转录病毒载体介导法转染体外培养SSCs的可行性,用脂质体介导法将携带LacZ基因的重组逆转录病毒载体pLNCL导入包装细胞PA317,用含G418的培养液筛选得到5株稳定转染的产毒细胞。收集这些克隆的产毒上清,过滤后进行倍比稀释,用NIH-3T3细胞通过X-gal染色测定其浓缩前病毒滴度。结果显示,PA3173培养上清中病毒的浓缩前滴度最高,达1.1×103CFU/mL。再将筛选到的稳定转染的NIH-3T3细胞培养至单层,进行X-gal染色检测β-半乳糖苷酶的表达。结果显示,大多数稳定转染的NIH-3T3细胞均为X-gal+,表明这些细胞成功表达了目的基因LacZ。本研究结果为后期工作中用该载体感染体外培养SSCs奠定了基础。

对苯二甲酸对NIH-3T3细胞间隙连接通讯功能的影响

目的 研究对苯二甲酸 (TPA)对NIH 3T3细胞间隙连接通讯 (GJIC)功能的影响。方法 采用噻唑蓝(MTT)比色法及划痕标记示踪技术 (scrape loadinganddyetransfer,SLDT) ,以 0、 12 5、 2 5 0、 5 0 0、 10 0 0、 2 0 0 0 μg/ml的剂量染毒细胞 ,同时设立溶剂对照及阳性对照 ,观察TPA对GJIC的影响。结果 TPA对细胞GJIC功能抑制随着染毒剂量增大和染毒时间延长而逐渐增强 ,呈现剂量 效应和时间 效应关系 ,在 2 5 0 μg/ml或更高剂量染毒 1h或更长时间时 ,细胞GJIC功能被明显抑制 ,在染毒 8h时各剂量组细胞GJIC功能抑制效应均最显著 ;去除TPA后 ,GJIC功能有恢复倾向 ,但在 12h内仍不能恢复至正常水平 ;加入哺乳动物肝脏微粒体酶 (S9)能加重TPA对细胞GJIC功能抑制作用。结论 TPA原形及其代谢产物对NIH

Effects of salvianolic acid-A on NIH/3T3 fibroblast proliferation,collagen synthesis and gene expression

AIM To investigate the mechanisms ofsalvianolic acid A（SA-A）against liver fibrosisin vitro.METHODS NIH/3T3 fibroblasts were culturedroutinely,and incubated with 10-4mol/L-10-7mol/L SA-A for 22 h.The cell viability wasassayed by[3H]proline incorporation,cellproliferation by[3H]TdR incorporation,cellcollagen synthetic rate was measured with[3H]proline impulse and collagenase digestionmethod.The total RNA was prepared from thecontrol cells and the drug treated cellsrespectively,and α（1）I pro-collagen mRNAexpression was semi-quantitatively analyzedwith RT-PCR.RESULTS 10-4mol/L SA-A decreased cellviability and exerted some cytotoxiciy,while10-5mol/L-10-7mol/L SA-A did not affect cellviability,but inhibited cell proliferationsignificantly,and 10-6mol/L SA-A had the besteffect on cell viability among theseconcentrations of drugs.10-5mol/L-10-6mol/LSA-A inhibited intracellular collagen syntheticrate,but no significant influence on extracellularcollagen secretion.Both 10-5mol/L and10-6mol/L SA-A could decrease α（1）I pro-collagen mRNA expression remarkably.CONCLUSION SA-A had potent action againstliver fibrosis.It inhibited NIH/3T3 fibroblastproliferation,intracellular collagen syntheticrate and type I pro-collagen gene expression,which may be one of the main mechanisms of thedrug.

Cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells

Today it is generally accepted that most bonding agents are cytotoxic. In this study the relative cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells were investigated. Materials and Methods. Near-confluent mouse 3T3 fibroblast cells were exposed to Dulbecco Modified Eagle’s Medium containing extractions from the seven different bonding agents. The cell survival rate was then determined using the standard MTT assay. Results. The cell survival rate ranking is: iBond (94%) < Gbond (78%) < Xeno V (71%) < Adper Easy Bond (63%) < Xeno V+ (61%) < Adper Scotchbond SE (33%) < XP Bond (32%). Part A of Adper Scotchbond SE had a survival rate of 35% and part B 38%. These two parts did not differ significantly. Adper Scotchbond SE and XP Bond do not differ significantly. While Xeno V+, Xeno V and Adper Easy Bond do not differ. (p < 5%;Tukey-Kramer Multiple-Comparison Test). Conclusion. All of the tested adhesive bonding agents were cytotoxic with survival rate of 3T3 cells between 94% to 31%. Of the 7 bonding agents tested iBond was found to be only slightly toxic and by far the least toxic. The two bonding agents (XP Bond and Adper Scotchbond SE) containing UDMA plus TEGDMA plus HEMA plus camphorquinone were found to be the most toxic.

Reactive oxygen species and senescence modulatory effects of rice bran extract on 4T1 and NIH-3T3 cells co-treatment with doxorubicin

Objective:To determine the effect of rice bran extract(RBE)in combination with doxorubicin on 4 T1 triple-negative breast cancer cells and NIH-3 T3 cells.Methods:RBE was obtained by maceration with n-hexane.The phytochemical profile of RBE was observed using highperformance liquid chromatography.Cytotoxic effect of RBE was evaluated through MTT assay.In addition,flow cytometry was used for cell cycle and apoptosis analysis.Cellular senescence was observed using SA-β-Gal assay and intracellular reactive oxygen species(ROS)levels were evaluated using DCFDA staining.The pro-oxidant property of RBE was also evaluated through 1-chloro-2,4-dinitrobenzene spectrophotometry and molecular docking.Results:RBE was obtained with a yield of 18.42%w/w and contained tocotrienols as the major compound.RBE exerted no cytotoxic effect on 4 T1 and NIH-3 T3 cells.However,RBE in combination with doxorubicin decreased 4 T1 cell viability synergistically(combination index<0.9)and induced apoptosis and senescence on 4 T1 cells.RBE significantly decreased senescence in doxorubicin-treated NIH-3 T3 cells.Additionally,RBE did not increase ROS levels in doxorubicin-treated 4 T1 cells.Meanwhile,the combination of RBE and doxorubicin reduced ROS levels in NIH-3 T3 cells.RBE significantly reduced glutathione-S-transferase activity and alpha-tocotrienol interacted with glutathione-Stransferase in the glutathione binding site.Conclusions:Rice bran may be used as a co-chemotherapeutic agent to improve the therapeutic effectiveness of doxorubicin while protecting against the cellular senescence effects of doxorubicin on healthy cells.

3,4-Oxo-isopropylidene-shikimic acid promotes adiopkine expression during murine 3T3-L1 fibroblast differentiation into adipocytes

Objective:3,4-Oxo-isopropylidene-shikimic acid(ISA),a derivative of shikimic acid,has exhibited ameliorative effect on cognitive impairment in experimental animal models of dementia.This study investigated the effect of ISA on lipid accumulation and adipokine secretion during differentiation of 3T3-L1 fibroblasts to adipocytes.Methods:3T3-L1 cells were cultured and treated with ISA(50e800 mM)from days 3e8.Lipid accumulation and triglyceride content were measured.Gene expression of adipokines(adiponectin,leptin,and resistin),CCAAT/enhancer binding protein(C/EBP)b,C/EBP a and peroxisome proliferator-activated receptor g(PPAR g)and PPAR target genes,including adipocyte fatty acid binding protein(aP2)and fatty acid synthase(FAS)were investigated.Results:ISA promoted 3T3-L1 fibroblast differentiation to adipocytes and increased triglyceride content by 26%.On mechanistic levels,ISA increased expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS.Moreover,ISA stimulated expressions of adipokines secreted by adipocytes,including adiponectin,leptin,and resistin.Conclusions:These findings demonstrated that ISA promoted adipogenesis by up-regulating expressions of C/EBP b,PPAR g,C/EBP a,aP2 and FAS,and also stimulated adipokines during adipocyte differentiation.Further study should clarify the relationship between stimulation of adipokines and cognitive enhancing effect of ISA.

Human ovarian neoplasm cell CD147 stimulates production and activation of matrix metalloproteinases in co-cultures with mouse fibroblasts

Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases(MMPs). Methods: The expression of CD147 on different human ovarian neoplasm cell lines was studied by western blotting. Co-culture was carried out to investigate the stimulative effect of the positive expression CD147 cell HO-8910 on the production of MMPs of fibroblast cell in vitro. Zymography and immune blotting were used to study the production and activity of positive MMPs, at the time, to explore the relation between CD147 and MMPs. Results: CD147 was positively presented in 2 ovarian neoplasm cell lines(HO-8910,3-AO), but in SKOV3, TC-1,NIN3T3 cell was negative. MMP-2 and MMP-9 were detected by HO-8910 cell line, mouse fibroblast cell and co-culture cells; but the expression in co-culture cell is obviously higher than individual cultures of each type alone.CD147 stimulated MMPs in dose-dependent manner. Conclusion: CD147 causes increased production and activation of MMP-2, MMP-9.CD147 is probably a indirect marker of some ovarian cancer cells with invasion and metastasis.

钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响

目的:观察钩藤散对NIH-3T3细胞衰老模型增殖与凋亡的影响。方法:NIH-3T3细胞接种在含10%胎牛血清的DMEM培养至20代,然后分为年轻、空白、模型3个组,传至30代时,用H2 O2处理,β-半乳糖苷酶染色试剂盒染色,观察衰老细胞形态、衰老指征,确认衰老模型是否成功建立;用钩藤散(17,8.5,4.25,2.15 g.kg-1)ig SD大鼠,每天2次,连续给药3 d,于末次ig 1 h后麻醉、腹主动脉采血,制备含药血清;用钩藤散含药血清处理衰老模型,观察衰老细胞形态、衰老指征、增殖与凋亡等指标。结果:模型组细胞较对照组形态有显著改变,细胞衰老比例明显增加(P<0.05),因此H2O2氧化应激方法可成功建立细胞衰老模型;钩藤散含药血清能明显减少细胞形态的改变、改善衰老指征、促进细胞增殖(P<0.01)、降低细胞凋亡(P<0.01),尤其10倍剂量组最为明显。结论:钩藤散能有效促进细胞增殖,降低细胞凋亡,从而防止衰老。