Applications of network pharmacology are increasingly widespread and methods abound in the field of drug development and pharmacological research. In this study, we choose rosiglitazone compound as the object to predict the targets and to discuss the mechanism based on three kinds of prediction methods of network pharmacology. Comparison of the prediction result has identified that the three kinds of prediction methods had their own characteristics: targets and pathways predicted were not in accordance with each other. However, the calcium signaling pathway could be predicted in the three kinds of methods, which associated with diabetes and cognitive impairment caused by diabetes by bioinformatics analysis. The above conclusion indicates that the calcium signaling pathway is important in signal pathway regulation of rosiglitazone compound, which provides a clue to further explain the mechanism of the compound and also provides a reference for the selection and application of methods of network pharmacology in the actual research.
Calcium Signaling ; Cognitive Dysfunction ; Computational Biology ; Diabetes Mellitus ; Humans ; Pharmacology ; methods ; Thiazolidinediones ; pharmacology

OBJECTIVETo search the effect of PPARgamma agonists for infection of RSV in vitro.

METHODSThe CPE of Hep-2 and A549 cells induced by RSV infection were observed. The effects of 15d-PGJ2 and rosiglitazone on change of CPE of A549 cells induced by RSV infection for 48 h were observed, too. MTT assay was used to detect the rate of viral suppression, and the protective effects of 15d-PGJ2 and rosiglitazone on A549 cells induced by RSV infection for 48 h.

RESULTSA549 cells interfered by 15d-PGJ2 (5 -25 micromol/L) and rosiglitazone (10-50 micromol/L) did not show obvious CPE, MTT assay also showed that the survival rate of A549 cells induced by RSV infection with PPARgamma agonists added, was significantly higher than that of RSV infection without PPARgamma agonists added, the difference was statistically significant (P < 0.01), but comparision between the two drugs showed no statistical significance. The optimal concentrations of 15d-PGJ2 and rosiglitazone were 5 micromol/L and 10 micromol/L respectively.

CONCLUSIONSPPARgamma agonist can reduce the CPE of A549 cells after RSV infection and improve the survival rate of A549 cells. PPARgamma agonist can counteract the infection of RSV in A549 cells.

Cells, Cultured ; Humans ; PPAR gamma ; agonists ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Respiratory Syncytial Viruses ; drug effects ; Thiazolidinediones ; pharmacology

Animals ; Fatty Liver ; blood ; complications ; Liver Cirrhosis ; blood ; etiology ; Male ; Mice ; Mice, Inbred C57BL ; Serum ; chemistry ; Thiazolidinediones ; pharmacology

OBJECTIVETo investigate the effect of rosiglitazone on angiotensin II (Ang II)-induced mRNA and protein expressions of toll-like receptor 4 (TLR4) and myeloperoxidase (MPO) activity in RAW264.7 cells to explore its anti- inflammatory and anti-atherosclerotic mechanisms.

METHODSMurine RAW264.7 cells were pretreated with rosiglitazone at 2.5, 5, and 10 micromol/L prior to exposure to AngII (0.1 micromol/L). TLR4 mRNA level was analyzed by RT-PCR, and TLR4 protein expression by Western blotting. MPO activity in the cell supernatant was assayed by colorimetry. In another experiment, the cells were pretreated with a neutralizing anti-TLR4 antibody (1 mg/L) for 1 h prior to rosiglitazone (10 micromol/L) treatment for 1 h, and subsequently stimulated with AngII or LPS (100 micromol/L) for 24 h to observe the change of MPO activity.

RESULTSRosiglitazone downregulated AngII-induced mRNA and protein expressions of TLR4, and inhibited MPO activity in RAW264.7 cells in a concentration-dependent manner. The TLR4 blocker partially antagonized the effect of AngII on MPO activity, and the inhibitory effect was markedly enhanced by rosiglitazone. Rosiglitazone significantly inhibited LPS (a specific TLR4 ligand)-induced MPO activity in RAW264.7 cells.

CONCLUSIONRosiglitazone downregulates Ang II-induced TLR4 expression in RAW264.7 cells and inhibits MPO secretion possibly by interfering with TLR4 to relieve the inflammatory reaction, which may be one of its anti-atherosclerotic mechanisms.

Angiotensin II ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Cell Line ; Macrophages ; cytology ; metabolism ; Mice ; Peroxidase ; metabolism ; Thiazolidinediones ; pharmacology ; Toll-Like Receptor 4 ; genetics ; metabolism

AIMTo investigate the effect of PPARgamma activators on inhibition of cardiac non myocytes (CNM) proliferation and the PPARgamma-dependent pathway possibly involved.

METHODSAngiotensin II was used to induce proliferation of primarily cultured CNM from neonatal rats, and pioglitazone or 15-deoxy-delta12,14-prostaglandin J2 (15d-PGJ2) was applied to these CNM in various dosages in vitro. MTT assay and 3H-TdR uptake were determined to estimate proliferation of CNM, and transient transfection of reporter gene containing PPRE from ACO promoter (PPRE-pGL3) with or without PPARgamma expression plasmid (PPARgamma-pSG5) to CNM was performed to determine the control of target-gene transcription.

RESULTSAngiotensin II caused a significant increase in MTT value and 3H-TdR uptake in CNM, which could be significantly reversed by pioglitazone and 15d-PGJ2 in a dose-dependent manner. Transient cotransfection of PPRE-pGL3 with PPARgamma-pSG5 to CNM resulted in significant increase in luciferase activity compared with that without PPARgamma-pSG5 cotransfection. Pioglitazone and 15d-PGJ2 induced increase in luciferase activity also in a dose-dependent manner.

CONCLUSIONPioglitazone and 15d-PGJ2, as the activators of PPARgamma, inhibit proliferation of CNM from neonatal rats, the effect may be related to the activation of PPARgamma.

Angiotensin II ; pharmacology ; Animals ; Cell Proliferation ; Cells, Cultured ; Heart ; Male ; Myocardium ; cytology ; PPAR gamma ; metabolism ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Rats ; Rats, Wistar ; Thiazolidinediones ; pharmacology

Agonists of peroxisome proliferator-activated receptor gamma (PPARgamma) are of interest as a treatment of type II diabetes, and indenone derivatives are a new class of non-TZD PPARgamma agonists. Based on existing indenone derivatives, a series of novel ones have been designed and synthesized. Meanwhile the structures have been comfirmed with 1H NMR and MS. Among them, 17b and 19 showed higher agonistic activities than rosiglitazone.
Drug Design ; Hypoglycemic Agents ; pharmacology ; Indenes ; chemical synthesis ; chemistry ; pharmacology ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; PPAR gamma ; agonists ; Structure-Activity Relationship ; Thiazolidinediones ; pharmacology

OBJECTIVESTo observe the effects of pioglitazone on morphological changes and connective tissue growth factor (CTGF) expression of the transforming growth factor beta (TGF b)-induced rat hepatic stellate cells (HSCs) in vitro, and to investigate the anti-fibrotic mechanism of pioglitazone.

METHODSCultured rat HSCs were divided into a no-treatment control group, a TGF b-treated group, and a TGFb plus different dosage pioglitazone-treated group. The morphological changes of the cultured HSCs were observed. The expression of CTGF was assessed by immunohistochemistry and flow cytometry. The level of collagen type III in the culture supernatant was measured by ELISA.

RESULTSTGFb induced morphological changes, and increased the expressions of CTGF and collagen type III of the HSCs (P less than 0.05). Pioglitazone prevented the TGFb induced morphological changes of the HSCs. The expression of CTGF and the levels of collagen type III in the pioglitazone group were lower than the TGF b-treated group (P less than 0.05). This prevention effect was dose-dependent (P less than 0.05).

CONCLUSIONPioglitazone blocks the excretion of CTGF and collagen type III of cultured HSCs, preventing the development of liver fibrosis.

Animals ; Cells, Cultured ; Collagen Type III ; secretion ; Connective Tissue Growth Factor ; metabolism ; Hepatic Stellate Cells ; drug effects ; metabolism ; Rats ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta ; pharmacology

BACKGROUNDOxidative Stress and p38 mitogen-activated protein kinase (p38MAPK) play a vital role in renal fibrosis. Pioglitazone can protect kidney but the underlying mechanisms are less clear. The purpose of this study was to investigate the effect of pioglitazone on oxidative stress and whether the severity of oxidative stress was associated with the phosphorylation level of p38MAPK.

METHODSRat mesangial cells were cultured and randomly assigned to control group, high glucose group and pioglitazone group. After 48-hour exposure, the supernatants and cells were collected. The protein levels of p22(phox), p47(phox), phosphorylated p38MAPK, total p38MAPK were measured by Western blotting. The gene expressions of p22(phox), p47(phox) were detected by RT-PCR. The levels of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The levels of superoxide dismutase (SOD) and maleic dialdehyde (MDA) in the supernatant were determined respectively.

RESULTSCompared with the control group, the expression levels of p22(phox), p47(phox), phospho-p38 and ROS significantly increased, activity of SOD decreased in high glucose group, while the level of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed p22(phox), p47(phox) expressions and oxidative stress induced by high glucose. The expressions of p22(phox), p47(phox), phospho-p38MAPK and ROS generation were markedly reduced after pioglitazone treatment (P < 0.05). The activity of SOD in the the supernatant increased (P < 0.05), while the level of MDA decreased greatly by pioglitazone (P < 0.05). The level of oxidative stress was associated with the phosphorylation level of p38MAPK (P < 0.01).

CONCLUSIONPioglitazone can inhibit oxidative stress through suppressing NADPH oxidase expression and p38MAPK phosphorylation.

Animals ; Blotting, Western ; Glucose ; pharmacology ; Mesangial Cells ; drug effects ; enzymology ; NADPH Oxidases ; genetics ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Thiazolidinediones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo investigate the effects of pioglitazone on transforming growth factor beta1 (TGFbeta1) expression in ischemia/reperfusion injury myocardium of rats.

METHODSThirty SD rats were randomly divided into five groups (n = 6): ischemia/reperfusion group, pioglitazone 5 mg/(kg x d) group, pioglitazone 10 mg/(kg x d) group, pioglitazone 20 mg/(kg x d) group and pioglitazone 20 mg/(kg x d) + peroxisome proliferator-activated receptor gamma (PPARgamma) specific antagonist GW9662 group. Left anterior descending coronary artery of rats were ligated for 30 min and reperfused for 120 min to establish the model of ischemia/reperfusion in vivo. RT-PCR was performed to detect the expression of TGFbeta1 mRNA. Western blot was performed to detect the expression of TGFbeta1 protein.

RESULTSMyocardial apoptosis was significantly suppressed by pioglitazone. Pioglitazone upregulated TGFPbeta1 expression both in mRNA and protein level. GW9662 reversed the inhibition of myocardial apoptosis and the upregulation of TGFbeta1 expression by pioglitazone.

CONCLUSIONPioglitazone can inhibit the myocardial apoptosis induced by ischemia/reperfusion. Pioglitazone may protect the myocardium from ischemia/reperfusion via upregulation of TGFbeta1. This protection may be mediated by PPARgamma.

Anilides ; pharmacology ; Animals ; Apoptosis ; Male ; Myocardial Reperfusion Injury ; metabolism ; Myocardium ; metabolism ; PPAR gamma ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effect of PPARgamma1 gene overexpression on caveolin-1 mRNA and protein expressions in a murine macrophage cell line Raw264.7.

METHODSReplication-deficient recombinant adenovirus expression vector of PPARgamma1 was constructed using the AdEasy system. Raw264.7 cells were randomly treated as follows: P group (PPARgamma1 gene overexpression), T group (Troglitazone 40 micromol/L in DMSO), PT group (PPARgamma1 gene overexpression and Troglitazone) and control group. Changes of PPARgamma1 and caveolin-1 at mRNA and protein levels were investigated.

RESULTSCaveolin-1 expression can be detected by RT-PCR in Raw264.7, by immunocytochemistry method in cell and nuclear membrane but not by immunoblotting at protein level. Caveolin-1 expression at mRNA and protein levels in Raw264.7 were significantly higher in P, T and PT groups compared to control group and the expression was also significantly higher in PT group than that in P group and T group (P < 0.05). PPARgamma expression was significantly increased in PT group and P group where remained unchanged in T group compared to control group.

CONCLUSIONPPARgamma1 overexpression can upregulate caveolin-1 expression in macrophages. Troglitazone upregulated caveolin-1 expression in the absence of increased PPARgamma1 expressions at mRNA and protein levels.

Adenoviridae ; genetics ; Animals ; Caveolin 1 ; metabolism ; Cell Line ; Chromans ; pharmacology ; Gene Expression ; Macrophages ; drug effects ; metabolism ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; metabolism ; Thiazolidinediones ; pharmacology