高糖对人类肾小球系膜细胞PKC及MMPs/TIMPs的影响

目的：研究体外高糖环境下人类肾小球系膜细胞(NHMC)中PKC激活或使用PKC抑制剂干预后 MMP2,9/TIMP1,2的表达,探讨PKC与MMPs/TIMPs信号转导在糖尿病肾病发生发展中的作用。方法：将NHMC分4组： N组（对照组,5 mmol/L葡萄糖）;H组（高糖组,30 mmol/L葡萄糖）;P组（抑制剂组,30 mmol/L葡萄糖+10-5mol/L白屈菜红碱）;M组(甘露醇组,5 mmol/L葡萄糖+25 mmol/L甘露醇)。分别于上述4种不同成分培养基中进行细胞培养,第24,48,72 h用MTT法测定NHMC增殖;并于培养后第24,48 h收集细胞,抽提mRNA及蛋白质,用ELISA方法测定胞膜、胞核蛋白质PKC活性。用RT-PCR和Western 印迹方法检测MMP2,9, TIMP1,2 mRNA和蛋白质的表达。结果：高糖组细胞膜和胞核部分的PKC活性较对照组明显升高(P＜0.01), MMP2,9, TIMP1,2 mRNA及蛋白质的表达较对照组明显上升(P＜0.01);而MMP-9/TIMP-1, MMP-2/TIMP-2比值较对照组明显下降(P＜0.05)。 高糖加PKC抑制剂白屈菜红碱后,MMP2,9, TIMP1 mRNA及蛋白质表达较对照组明显升高 (P＜0.01),但MMP9/TIMP1, MMP2/TIMP2比值较高糖组明显升高(分别P＜0.05,P<0.01)。PKC活性与MMP-2/TIMP-2及MMP-9/TIMP-1的蛋白质比值均呈负相关(分别r=-0.651,r=-0.702,均P＜0.05)。结论：高糖可刺激人类肾小球系膜细胞PKC活化,在DN中PKC活化与MMP2,9/TIMP1,2的表达水平有密切关系。     

胃肠安方对人胃癌生长、转移及血管生成和MMP2表达的影响

目的：观察胃肠安方对人胃癌裸鼠原位移植瘤生长和转移的作用及其对肿瘤血管生成与基质金属蛋白酶2（Matrix metalloproteinase-2,MMP2）表达的影响。方法：建立裸小鼠胃原位癌模型,随机分为胃肠安组及生理盐水对照组,观察裸小鼠胃癌种植后肿瘤生长及转移灶情况,以及肿瘤生长转移相关基因MMP2表达和肿瘤血管生成情况。结果：胃肠安方能显著抑制胃癌生长与转移。免疫组化结果表明,胃肠安组肿瘤微血管密度明显低于对照组。胃肠安方同时能抑制肿瘤组织MMP2表达。结论：胃肠安方具有抑制人胃癌裸鼠原位移植瘤生长和转移作用,其作用机制可能与抑制肿瘤新生血管生成及MMP2表达有关。     

Silica nanoparticles induced intrinsic apoptosis in neuroblastoma SH-SY5Y cells via CytC/Apaf-1 pathway

The present study was to investigate effects of Silica nanoparticles (SiNPs) on nervous system and explore potential mechanisms in human neuroblastoma cells (SH-SY5Y). Cytotoxicity was detected by cell viability and Lactate dehydrogenase (LDH) release. Flow cytometry analysis was applied to assess mitochondrial membrane potential (MMP) loss, intracellular Ca²⁺ and apoptosis. To clarify the mechanism of SiNPs-induced apoptosis, intrinsic apoptosis-related proteins were detected. Our results showed that SiNPs caused cytotoxicity, cell membrane damage and Ca²⁺ increase in a dose-dependent manner in SH-SY5Y cells. Both the mitochondrial membrane potential (MMP) loss and potential mitochondria damage resulted in Cyt C release to the cytoplasm. The elevated Cyt C and Apaf1 further triggered intrinsic apoptosis via executive molecular caspase-9 and caspase-3. The present study confirmed that SiNPs induced intrinsic apoptosis in neuroblastoma SH-SY5Y cells via CytC/Apaf-1 pathway and provided a better understanding of the potential toxicity induced by SiNPs on human neurocyte.     

Red yeast rice improves lipid pattern,high-sensitivity C-reactive protein,and vascular remodeling parameters in moderately hypercholesterolemic Italian subjects

Despite a recent health claim by the European Agency on Food Safety, the effect of high doses of dietary monacolin supplements from red yeast rice on cholesterolemia has not been tested in Italian subjects. Our aim via a crossover, double-blind, placebo-controlled randomized clinical trial was to test if a short-term treatment with 10 mg monacolins could improve lipid pattern, high-sensitivity C-reactive protein (hs-CRP), and vascular remodeling biomarkers in a small cohort of Mediterranean subjects. Thus, 25 healthy, mildly hypercholesterolemic subjects were enrolled, and after 4 weeks of a stabilization diet, subjects were randomized to the sequence placebo-washout-monacolins or monacolins-washout-placebo, with each period being 4 weeks long. At each study step, a complete lipid pattern, safety parameters, hs-CRP, and matrix metalloproteinases 2 and 9 levels were measured. When compared to the placebo group, monacolins-treated patients experienced a more favorable percent change in total cholesterol (-12.45%, 95% CI −16.19 to −8.71), low-density lipoprotein cholesterol (−21.99%, 95% CI −26.63 to −17.36), non–high-density lipoprotein cholesterol (−14.67%, 95% CI −19.22 to −10.11), matrix metalloproteinase 2 (−28.05%, 95% CI −35.18 to −20.93), matrix metalloproteinase 9 (−27.19%, 95% CI −36.21 to −18.15), and hs-CRP (−23.77%, 95% CI −30.54 to −17.01). No significant differences were observed in regards to triglycerides, high-density lipoprotein cholesterol, and safety parameters. On the basis of our data, we demonstrate that a 10-mg monacolin nutraceutical appears to safely reduce cholesterolemia, hs-CRP, and markers of vascular remodeling in Italian subjects. These results have to be confirmed in larger patient samples and longer studies.     

低强度微波对γ射线致人早幼粒白血病HL-60细胞损伤的影响

[目的]探讨低强度微波辐射对γ射线致人早幼粒白血病HL-60细胞的凋亡率、线粒体膜电位（mitochondrial membrane potential, MMP）和胞内游离Ca^2＋浓度改变的影响。[方法]将人早幼粒白血病HL-60细胞随机分为对照组、单纯微波组、单纯γ射线组和联合组（微波＋γ射线）。单纯微波组和联合组给予12gW／cm^2微波照射,每天1h,连续辐照3d;第4天给予单纯γ射线组和联合组8Gγγ射线照射。用钙离子依赖性磷脂结合蛋白-异硫氰酸荧光素／碘化丙啶（Annexin V—FITC／PI）双标记法检测细胞凋亡率,流式细胞仪检测线粒体膜电位和胞内游离Ca2＋浓度。[结果]与对照组相比,单纯γ射线组凋亡率明显增加（P〈0．05）;联合组凋亡率与单纯γ射线组相比显著降低（P〈O．05o与对照组相比,单纯γ射线组线粒体膜电位下降明显（P〈0．05）;联合组线粒体膜电位与单纯γ射线组相比显著升高（P〈0．05o与对照组相比,单纯γ射线组胞内游离Ca2＋浓度明显升高（P〈0．05）;与单纯γ射线组相比,联合组胞内游离Ca2＋浓度明显降低（P〈0．05）。[结论]本实验条件下,8Gyγ射线可以引起细胞凋亡率增加、线粒体膜电位下降和胞内游离Ca2＋浓度增高,导致细胞损伤。预先900MHz、12μW／cm^2低强度微波照射能够显著减轻γ射线引起的细胞损伤,表现为细胞凋亡率减少、线粒体膜电位升高和细胞内游离Ca^2＋浓度降低。     

MMP2及其抑制物TIMP2在宫颈癌发展中的作用

目的金属基质蛋白酶2及其组织抑制剂2(MMP2-TIMP2)在肿瘤侵袭转移过程中起决定性作用。本研究探讨MMP2及TIMP2在宫颈癌发展中的作用及意义。方法本实验采用免疫组化检测60例宫颈上皮内瘤变等CIN(Ⅱ-Ⅲ级)、79例宫颈癌组织中MMP-2与TIMP-2的基因表达情况,并与20例正常宫颈组织作对照。结果MMP2、TIMP2的阳性表达主要集中在细胞浆上,细胞膜上也有少量表达。79例宫颈癌组织中MMP2、TIMP2的阳性表达率分别为75.95%、73.42%;CIN组中MMP2、TIMP2的阳性表达率分别为83.33%、70%;正常对照组中MMP2、TIMP2的阳性表达率分别为50%、50%。MMP2的阳性表达率在CIN组与正常对照组比较,两组间差异有显著性(2=7.20,p<0.01);宫颈浸润癌与正常对照组比较,两组间差异有统计学意义(2=4.01,p<0.05);而CIN组MMP2的阳性表达率与宫颈癌组比较,则显示两组之间无差异(p>0.05)。TIMP2的阳性表达率在正常宫颈组织组与CIN组间的差异无统计学意义(p>0.05);在正常宫颈组织中,TIMP2的阳性表达率与宫颈癌组之间的差异有统计学意义(2=4.06,p<0.05);但CIN组与宫颈癌组TIMP2表达率的比较则显示无统计学差异(p>0.05)。结论MMP2、TIMP2的表达与宫颈癌的发展有关,可被视为反映宫颈癌生物侵袭性的标志物。     

盐酸利托君对先兆早产患者血清Th1/Th2细胞因子及分泌物fFN、IGFBP-1、MMP的影响

目的研究盐酸利托君对先兆早产患者血清Th1/Th2细胞因子及分泌物f FN、IGFBP-1、MMP的影响。方法选取2013年2月—2015年4月在本院进行治疗的68例先兆早产患者为研究对象,将其随机分为对照组34例和观察组34例,比较2组患者治疗前和治疗后的血清Th1/Th2细胞因子及分泌物f FN、IGFBP-1、MMP水平。结果治疗前,2组的血清Th1/Th2细胞因子及分泌物f FN、IGFBP-1、MMP水平均无显著差异(P0.05),而治疗后不同时间观察组以上指标均显著优于对照组(P0.05)。结论盐酸利托君对先兆早产患者的血清Th1/Th2细胞因子及分泌物f FN、IGFBP-1、MMP表达有积极的改善作用,因此对于此类孕妇的应用价值较高。     

Rab11a regulates MMP2 expression by activating the PI3K/AKT pathway in human hepatocellular carcinoma cells

As a member of the Rab GTPase family, Rab11a plays an important role in vesicle transport and tumor progression. However, it is not clear whether it can also be used as an oncoprotein in hepatocellular carcinoma (HCC). In this study, database and immunohistochemical analyses showed that Rab11a was highly expressed in HCC tissues, and associated with poor clinical prognosis. Rab11a overexpression promoted the proliferation, migration, invasion, and anti-apoptosis of human HCC cell lines, MHCC-97H and HCC-LM3, whereas the downregulation of Rab11a inhibited these biological tumor activities. Nude mice xenograft demonstrated that Rab11a had a positive effect on the growth of hepatocellular carcinoma cells in vivo. Further studies found that the PI3K/AKT pathway and matrix metalloproteinase 2 (MMP2) upregulation can be activated by over-expression of Rab11a. However, MMP2 upregulation induced by Rab11a can be inhibited by the PI3K/AKT pathway inhibitor, LY294002. Altogether, our study established for the first time that Rab11a can play a pro-cancer role in HCC, as a novel oncoprotein, by activating the PI3K/AKT pathway to regulate MMP2 expression.     

CXCL12/CXCR4 signaling activates Akt-1 and MMP-9 expression in prostate cancer cells: the role of bone microenvironment-associated CXCL12

BACKGROUND: Hematopoietic cells home to bone by means of chemo-attraction to marrow chemokines, and interaction of chemokines with their receptors leads to the expression/activation of adhesion molecules and proteases. Recent evidence suggests that similar mechanisms may be active in cancer metastasis. Previously, we showed that metalloproteases (MMPs), and in particular MMP-9, play a role in prostate cancer (PC) expansion in bone. METHODS: We used a variety of methods including RT-PCR, immunohistochemistry, ELISA, gelatin zymography, cellular motility and invasion, and subcellular fractionation of PC cells applied to in vivo and in vitro models. RESULTS: Here we showed that (a) CXCL12/CXCR4 axis is expressed in PC bone metastasis; (b) exogenous CXCL12 induced MMP-9 expression by PC cells; (c) bone stromal cells and bone tissue conditioned media induced the migration of PC cells in a CXCR4-dependent manner; (d) pharmacological inhibition of PI3 kinase and MAP kinase pathways abrogated CXCL12-induced MMP-9 expression and invasion of PC cells; (e) exogenous CXCL12 induced Akt1 phosphorylation is indispensable for proMMP-9 secretion, migration, and invasion of PC cells; (f) CXCR4 was localized to lipid rafts in PC cells and initiated Akt phosphorylation. CONCLUSIONS: These data suggest that chemoattractive mechanisms involve migration of cancer cells towards bone tissue, and that cell signaling induced by binding of the chemokine to its receptor leads to the activation of multiple signaling pathways and subsequent secretion of MMP-9 into the local environment. These findings provide a link between chemoattractive mechanisms, growth of tumor cells in bone, and tumor-enhanced bone matrix turnover.