Animals ; Atherosclerosis ; blood ; drug therapy ; pathology ; Dehydroepiandrosterone ; blood ; physiology ; therapeutic use ; Dehydroepiandrosterone Sulfate ; blood ; therapeutic use ; Hormone Replacement Therapy ; Humans

Animals ; Atherosclerosis ; etiology ; genetics ; pathology ; China ; Coronary Disease ; pathology ; Genes, p53 ; Humans ; Point Mutation ; Risk Factors

Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; physiology ; Neoplasm Proteins ; biosynthesis ; genetics ; physiology ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

Animals ; Cell Differentiation ; Cell Movement ; Chemokine CXCL12 ; metabolism ; Endothelial Cells ; pathology ; physiology ; Humans ; Muscle, Smooth, Vascular ; pathology ; physiology ; Receptors, CXCR4 ; metabolism ; Stem Cells ; metabolism ; pathology ; physiology ; Vascular Diseases ; metabolism ; pathology ; physiopathology

Breast Neoplasms ; classification ; genetics ; metabolism ; pathology ; Chromosome Aberrations ; Female ; Gene Deletion ; Humans ; Phyllodes Tumor ; classification ; genetics ; metabolism ; pathology ; Twist-Related Protein 1 ; metabolism ; Wnt Signaling Pathway

OBJECTIVETo identify and select smooth muscle progenitor cells from mouse bone marrow mesenchyme stem cell population and to characterize smooth muscle progenitor cells in peripheral blood.

METHODSRecombinant expression vector with the promoter of sm22alpha was constructed to have an enhancement type green fluorescent protein expression plasmid (EGFP-1). The construct was transfected into mouse bone marrow mesenchyme stem cells using Lipofectamine 2000. Morphological assessment was performed and the expressions of myocardin at protein and mRNA levels by fluorescence microscope and RT-PCR were evaluated at 3, 5, 7, and 10 d targeting CD34 positive bone mesenchyme stem cells.

RESULTSThe transfection efficiency of the positive control group was 70% +/- 1.5% (P > 0.05). Expected green fluorescent proteins expressed at 3rd day. The numbers of green fluorescent cells in experimental groups increased with the time and reached the peak at the 7th day, and declined thereafter. The shapes of the green fluorescent cells were also different from each others. The positive ratios of green fluorescent cells at different time points: 3 d: 7% +/- 0.13%, 5 d: 10% +/- 0.32%, 7 d: 20% +/- 0.26%, 10 d: 12% +/- 0.18%, P < 0.05. Myocardin mRNA expression roughly correlated with green fluorescent expressions. CD34 was expressed on the 5th day in transfected bone mesenchyme stem cells. The CD34 positive ratio was 5.2% +/- 0.21% (P > 0.05).

CONCLUSIONSThere are smooth muscle progenitor cells among mouse bone marrow mesenchyme stem cell population. Smooth muscle progenitor cells can be selected using a Psm22alpha-EGFP-1 recombinant expression approach.

Animals ; Antigens, CD34 ; immunology ; Bone Marrow Cells ; cytology ; immunology ; Cell Separation ; methods ; Cell Shape ; Green Fluorescent Proteins ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Microfilament Proteins ; genetics ; Microscopy, Fluorescence ; Muscle Proteins ; genetics ; Myocytes, Smooth Muscle ; cytology ; Nuclear Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; Recombinant Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; Transfection

Cells, Cultured ; Chemokine CCL4 ; Chemotaxis, Leukocyte ; drug effects ; Endothelial Cells ; cytology ; metabolism ; Homocysteine ; pharmacology ; Humans ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Monocytes ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

Animals ; Aorta ; pathology ; Atherosclerosis ; blood ; etiology ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Cholesterol, Dietary ; administration & dosage ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dehydroepiandrosterone ; pharmacology ; Diet, Atherogenic ; Immunohistochemistry ; Rabbits ; Random Allocation ; Triglycerides ; blood ; Vascular Cell Adhesion Molecule-1 ; metabolism

Tetramethylpyrazine (TMP), an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin II (Ang II) is highly implicated in the proliferation of vascular smooth muscle cells (VSMCs), resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang II remain to be defined. The present study was aimed to study the effect of TMP on Ang II-induced VSMC proliferation through detection of nuclear factor-kappaB (NF-kappaB) activity and bone morphogenetic protein-2 (BMP-2) expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang II group, Ang II + TMP group and TMP group. Cells in each group were harvested at different time points (15, 30 and 60 min for detection of NF-kappaB activity; 6, 12 and 24 h for measurement of BMP-2 expression). NF-kappaB activation was identified as nuclear staining by immunohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that: (1) Ang II stimulated the activation of NF-kappaB. Translocation of NF-kappaB p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min (P<0.01) and declined after 1 h. (2) TMP inhibited Ang II-induced NF-kappaB activation (P<0.01). (3) Ang II increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h (P<0.01). (4) BMP-2 expression was also kept at high level at 6 h in Ang II + TMP group but maintained at the normal level at 12 and 24 h. (5) There was no significant difference in NF-kappaB activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang II-induced VSMC proliferation through repression of NF-kappaB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-kappaB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.
Angiotensin II ; antagonists & inhibitors ; Animals ; Atherosclerosis ; drug therapy ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; antagonists & inhibitors ; Immunohistochemistry ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; analysis ; antagonists & inhibitors ; Pyrazines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; analysis ; antagonists & inhibitors

Filamin A and 14-3-3-σ are closely associated with the development of breast cancer.However,the exact relationship between them is still unknown.The present study aimed to examine the interaction of filamin A with 14-3-3-σ in the invasion and migration of breast cancer.RNA interference technology was employed to silence filamin A in MDA-MB-231 cells.Real-time PCR and Western blotting were used to detect the expression of filamin A and 14-3-3-σ at mRNA and protein levels,respectively.Double immunofluorescence was applied to show their colocalization morphologically.Wound healing assay and Trans-well assay were used to testify the migration and invasion of MDA-MB-231 cells in filamin A-silenced cells.The results showed that silencing filamin A significantly increased the mRNA and protein levels of 14-3-3σ.In addition,double immunofluorescence displayed that filamin A and 14-3-3σ were predominantly colocalized in the cytoplasm of MDA-MB-231 cells.Silencing filamin A led to the enhanced fluorescence of 14-3-3σ.Furthermore,cell functional experiments showed that silencing filamin A inhibited the migration and invasion of MDA-MB-231 cells in vitro.In conclusion,silencing filamin A may inhibit the invasion and migration of breast cancer cells by upregulating 14-3-3σ.