OBJECTIVES: To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. METHODS: Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. RESULTS: Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05). CONCLUSIONS Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.
Animals ; CCN Intercellular Signaling Proteins ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glucosamine ; pharmacology ; Intercellular Signaling Peptides and Proteins ; metabolism ; Osteoarthritis, Knee ; drug therapy ; Proto-Oncogene Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Wnt Signaling Pathway ; Wnt1 Protein ; metabolism ; beta Catenin ; metabolism

LEDGF/p75 is a newly found cell cofactor, which plays an essential role in the integration of HIV-1 cDNA into host chromosomes. LEDGF/p75 tethers HIV integrase to chromatin, protects it from degradation, and strongly influences the genome-wide pattern of HIV integration. Depleting the protein from cells or over-expressing the integrase-binding domain of LEDGF/p75 blocks viral replication. The essential role of LEDGF/p75 in HIV-1 replication makes it a new target for anti-HIV-1 drug development. This article reviews the function of LEDGF/p75, LEDGF/p75-integrase interaction and LEDGF/p75 inhibitors.
Anti-HIV Agents ; chemistry ; pharmacology ; HIV Integrase ; metabolism ; HIV-1 ; drug effects ; physiology ; Intercellular Signaling Peptides and Proteins ; metabolism ; Protein Binding

OBJECTIVETo probe matrine acting on natural killer cell (NK) activating receptor NKG2D ligands expression in CML cell line K562 and its underlying molecular mechanism.

METHODSThe expression of NKG2D ligands (major histocompatibility complex class I chain-related molecule A or B (MICA/B), UL16-binding proteins (ULBP) 1, 2, and 3 on K562 cells were analyzed before and after treated with matrine by FCM. The cytotoxic sensitivity of K562 to NK cell was detected by FCM after CFSE staining at different effect-to-target (E/T) cell ratios. The expression of signal transduction and transcriptional activator 3 (STAT3) protein as well as phosphorylated STAT3 (p-STAT3) were detected by western blot.

RESULTSAfter treatment with matrine, ULBP1 and ULBP2 expression, especially ULBP2 on K562 cells significantly increased, with mean fluorescence intensity (MFI) increasing to 615 and 1614 by 220 and 615 in the untreated cells, respectively. There was no significant change for MICA or ULBP3 expression. Matrine enhanced the susceptibility of K562 cells to NK-mediated cell lysis. At the ratio of E/T with 5:1, the proportion of the killed K562 cells increased to 32.8%, 38.1% and 40.5%, respectively (after 0.2, 0.5 and 0.8 mg/ml matrine treatment) by 29.2% in the untreated cells. The phosphorylated STAT3 protein, but not STAT3 protein, was significantly inhibited by matrine treatment in K562 cells.

CONCLUSIONMatrine induced the expression of NKG2D ligands in K562cells and enhanced the cytotoxicity of NK cells against K562, which was closely related to the inhibition of STAT3 activity in K562 cell.

Alkaloids ; pharmacology ; GPI-Linked Proteins ; immunology ; Humans ; Intercellular Signaling Peptides and Proteins ; immunology ; K562 Cells ; Quinolizines ; pharmacology ; Signal Transduction ; Up-Regulation ; drug effects

Arsenicals ; pharmacology ; DNA Methylation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Membrane Proteins ; genetics ; metabolism ; Oxides ; pharmacology

OBJECTIVETo explore the effects of berberine and insulin on adiponectin mRNA expression in 3T3-L1 adipocyte.

METHODThe 3T3-L1 adipocyte was treated with berberine and insulin for 48 hours, the level of adiponectin mRNA in 3T3-L1 adipocyte expression was determined with semiquantity RT-PCR using beta-actin as internal reference.

RESULTThe level of adiponetin mRNA in 3T3-L1 adipocyte was increased after treated with berberine only (P < 0.05), the effect of berberine was inhibited by high concentration insulin (P < 0.05).

CONCLUSIONIn vitro, berberine increases the expression of adiponectin in 3T3-L1 adipocyte, insulin inhibits the effect of berberine.

3T3-L1 Cells ; metabolism ; Adipocytes ; cytology ; metabolism ; Adiponectin ; Animals ; Berberine ; pharmacology ; Insulin ; pharmacology ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Mice ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVEThe renal disease is commonly associated with hyperlipidemia and correlates with glomerular accumulation of atherogenic lipoproteins and mesangial hypercellularity. Therefore, in this study, the authors investigated a possible growth stimulatory effect and mode of action of lipoprotein(a) [Lp(a)] in human mesangial cells HMC, and the effect of Lp(a) on adhesion and migration in human mesangial cells.

METHODSThe DNA synthesis of HMC was measured by (3)H-thymidine incorporation. The cell adhesion was detected by the expression of vinculin by means of indirect immunofluorescence. The cell migration was observed under the microscope.

RESULTSThe incubation of HMC with Lp(a) for 24 hours induced a significant dose-dependent proliferation of HMC [Lp(a): 5 microg, 10 microg, 25 microg, 50 microg/ml vs. control 0 microg/ml; (3)H-TdR incorporation (x 10(3)cpm): 1.69 +/- 0.48, 3.59 +/- 0.68, 4.14 +/- 0.78, 4.05 +/- 0.55 vs. 1.64 +/- 0.31, P < 0.01]. The vinculin staining by indirect immunofluorescence showed positive result when HMC was incubated with 10 microg/ml Lp(a) for 24 hours, while vinculin was negative when HMC was incubated with 0 microg/ml Lp(a) as the control of the study. The incubation of HMC with 10 microg/ml Lp(a) for 72 hours demonstrated significant cell migration effect compared to the control of 0 microg/ml. (16.2/LP vs. 2.4/LP, P < 0.01).

CONCLUSIONLp(a) could stimulate a proliferation, adhesion and migration effect on human mesangial cells.

Cell Adhesion ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Lipoprotein(a) ; pharmacology ; Mesangial Cells ; drug effects

OBJECTIVE: To elucidate the renoprotective effect of resveratrol (RSV) on sphingosine kinase 1 (SphK1) signaling pathway and expression of its downstream molecules including activator protein 1 (AP-1) and transformation growth factor-β1 (TGF-β1) in lipopolysaccharide (LPS)-induced glomerular mesangial cells (GMCs). METHODS: The rat GMCs line (HBZY-1) were cultured and randomly divided into 5 groups, including control, LPS (100 ng/mL), and 5, 10, 20 µmol/L RSV-treated groups. In addition, SphK1 inhibitor (SK-II) was used as positive control. GMCs were pretreated with RSV for 2 h and treated with LPS for another 24 h. GMCs proliferation was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The proteins expression of SphK1, p-c-Jun and TGF-β1 in GMCs were detected by Western blot, and DNA-binding activity of AP-1 was performed by electrophoretic mobility shift assay (EMSA). The binding activity between RSV and SphK1 protein was detected by AutoDock Vina and visualized by Discovery Studio 2016. RESULTS: LPS could obviously stimulate GMCs proliferation, elevate SphK1, p-c-Jun and TGF-β1 expression levels and increase the DNA-binding activity of AP-1 (P<0.05 or P<0.01), whereas these effects were significantly blocked by RSV pretreatment. It was also suggested that the effect of RSV was similar to SK-II (P>0.05). Moreover, RSV exhibited good binding affinity towards SphK1, with docking scores of -8.1 kcal/moL and formed hydrogen bonds with ASP-178 and LEU-268 in SphK1. CONCLUSION RSV inhibited LPS-induced GMCs proliferation and TGF-β1 expression, which may be independent of its hypoglycemic effect on preventing the development of mesangial cell fibrosis and closely related to the direct inhibition of SphK1 pathway.
Animals ; Rats ; Lipopolysaccharides/pharmacology* ; Mesangial Cells ; Resveratrol/pharmacology* ; Transcription Factor AP-1 ; Transforming Growth Factor beta1 ; Intercellular Signaling Peptides and Proteins ; Cell Proliferation ; DNA ; Cells, Cultured

OBJECTIVETo observe the effect of parathyroid hormone (PTH)(1-34) on the expression of matrix Gla protein (MGP) and Wnt/β-catenin signaling pathway and elucidate the possible molecular mechanism of PTH (1-34) in the prevention and treatment of osteoporosis.

METHODSMG63 cells treated with PTH (1-34) at 10(-9), 10(-8), and 10(-7) mol/L, alone or in combination with Wnt/β-catenin signaling pathway inhibitors DKK-1 (200 ng/ml) were examined for mRNA and protein expressions related with Wnt/β-catenin signaling with real-time PCR and Western blotting. The cell differentiation after the treatment was assessed with alkaline phosphatase (ALP) staining and cell viability assay.

RESULTSPTH (1-34) significantly increased the expression of MGP in a dose-dependent manner in MG63 cells (P<0.05 or P<0.01). PTH treatment obviously enhanced ALP activity in the cells, and this effect was suppressed by DKK-1. Combined treatment with DKK-1 partially blocked PTH-induced enhancement of ALP activity (P<0.05). PTH promoted the expression of MGP and enhanced LRP5, β-catenin, and Runx2 expressions in Wnt/β-catenin signaling pathway at both protein and mRNA levels (P<0.05 or P<0.01). DKK-1 partially blocked the effect of PTH (1-34) on Wnt/β-catenin signaling pathway (P<0.05) without affecting MGP expression.

CONCLUSIONPTH (1-34) significantly increases the expressions of MGP and proteins in the Wnt/β-catenin signaling pathway. Wnt/β-catenin signaling pathway and MGP mediate the regulation of osteogenosis by PTH.

Alkaline Phosphatase ; metabolism ; Calcium-Binding Proteins ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Cell Survival ; Extracellular Matrix Proteins ; metabolism ; Humans ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Osteogenesis ; Osteoporosis ; Parathyroid Hormone ; pharmacology ; Real-Time Polymerase Chain Reaction ; Wnt Signaling Pathway

OBJECTIVETo explore the effect of Osteoking on bone mineral density (BMD) and serum Dickkopf-1 (DKK-1) protein levels in rabbits with osteoporotic fracture (OPF).

METHODSTotally 45 female Japanese big-ear rabbits were randomly divided into the treatment group, the model group, and the blank control group (as the control group), 15 in each group. Bilateral ovaries were ectomized for 24 weeks in the treatment group and the model group. Their left radial factures were induced after confirmed osteoporosis. Rabbits in the treatment group were administered with Osteoking by gastrogavage, once per two days. Equal volume of normal saline was given to rabbits in the model group. The general BMD and serum DKK-1 protein levels were detected before ovariectomy, at week 24 and 48 after ovariectomy.

RESULTSThere was significant difference in the general BMD at week 24 after ovariectomy between the model group and the control group, and it was lower in the model group. Compared with the model group, the general BMD significantly increased and serum DKK-1 protein levels significantly decreased in the treatment group after intervention. Serum DKK-1 protein levels were significantly lower after intervention than before intervention in the treatment group.

CONCLUSIONOsteoking could improve the BMD of OPF rabbits, and reduce their serum DKK-1 protein levels as well.

Animals ; Bone Density ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Osteoporosis ; Osteoporotic Fractures ; drug therapy ; metabolism ; Ovariectomy ; Rabbits

OBJECTIVETo study the expression of Fractalkine (FKN) in the kidney tissue of rats with renal fibrosis and the effect of IL-18BP on FKN.

METHODSMale Wister rats were randomly assigned to sham-operation (n=24), unilatral ureteral obstruction (UUO, n=22), and IL-18 binding protein (IL-18BP) treatment groups (n=23). The UUO model was prepared by unilateral ureteral ligation in the later two groups. The IL-18BP treatment group received an intraperitoneal injection of IL-18BP (0.1 mg/kg) every other day after UUO inducement, for 7 times, while normal saline was administered in the other two groups. Seven or eight rats of every group were sacrificed at 3, 7 or 14 days after IL-18BP or normal saline injections. FKN levels at various times were detected by immunohistochemistry and Western blot.

RESULTSCompared with the sham-operation group, FKN levels in the kidney tissue of the untreated UUO group increased significantly at all time points (P<0.01). IL-18BP treatment decreased significantly FKN levels in the kidney tissue at all time points compared with the untreated UUO group (P<0.01).

CONCLUSIONSIL-18BP treatment may down-regulate the increased FKN levels of the rat kidney tissue caused by UUO, possibly thus delays the occurrence and development of renal fibrosis.

Animals ; Blotting, Western ; Chemokine CX3CL1 ; genetics ; Fibrosis ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Kidney ; immunology ; pathology ; Male ; Rats ; Rats, Wistar ; Ureteral Obstruction ; immunology