To explore whether PPARA is involved in the process of ferroptosis in hepatoma cells, peroxisome proliferator activated receptor (PPARA) was comprehensively analyzed in hepatocellular carcinoma (HCC) through public database and experimental data, including the expression, the functions and the potential roles of tumor progression. The research design is experimental research,data analysis based on bioinformatics and cell experiment. From January 2022 to August 2022, relevant cell experiments were conducted in the Basic Medical Laboratory of the General Hospital of the Southern Theatre of the Chinese People's Liberation Army. The expression and the correlation with clinicopathologic features of PPARA in HCC were analyzed by The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. To study the protein expression of PPARA in HCC and normal tissues through the Human Protein Atlas (HPA). The protein-protein interaction (PPI) network between PPARA and the core factor of ferroptosis was constructed based on Search Tool for the Retrival of Interacting Genes/Protein (STRING) database, then, the correlation between PPARA and the core gene Glutamate-cysteine Ligase Catalytic Subunit (GCLC) was analyzed by Gene Expression Profiling Interactive Analysis (GEPIA). Assessed the expression of PPARA in HCC cell lines SK-HEP-1, SMMC-7721, MHCC-97H, BEL-7402 and normal liver cell L02 by Western Blot (WB) and the changes of PPARA expression after 48h treatment with ferroptosis inducer Erastin were observed. Single factor analysis of variance was used to compare the expression of PPARA between groups in GEPIA database. The expression of PPARA in GSE25097 and GSE112790 data was compared by rank sum test. Survival analysis was performed using time series test method. The difference of PPARA expression between clinical and pathological features was compared using the Kruskal-Wallis test. The correlation between the expression of GCLC and PPARA was compared by the method of Spearman correlation. The expression of PPARA in cell lines was compared by paired T test. The results showed that the RNA and protein expression of PPARA in HCC was lower than that in normal tissues (P<0.05). PPARA alterations were correlated with patient clinicopathological features and prognosis (P<0.05). The PPI constructed by STRING database suggests that PPARA interact with the key factors of ferroptosis, such as NFE2 like bZIP transcription factor 2 (NFE2L2), Heme Oxygenase 1 (HMOX1), Tumor Protein P53 (TP53), GCLC, Dipeptidyl Peptidase 4 (DPP4), Citrate Synthase (CS), Arachidonate 15-Lipoxygenase (ALOX15) and Acyl-CoA Synthetase Long Chain Family Member 4 (ACSL4). Furthermore, the PPARA was significantly associated with GCLC validated via GEPIA database(R=0.6, P<0.05). The expression of PPARA increased after treatment with ferroptosis inducer Erastin for 48 h by WB. In conclusion, the expression of PPARA is lower in HCC with a poor prognosis. PPARA interacts with GCLC in regulating ferroptosis in HCC.
Humans ; Carcinoma, Hepatocellular/pathology* ; Ferroptosis ; Liver Neoplasms/pathology* ; Peroxisome Proliferator-Activated Receptors/genetics*

Animals ; Fatty Liver ; genetics ; metabolism ; Female ; Liver ; metabolism ; Liver Diseases, Alcoholic ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo explore if PPARgamma agonist rosiglitazone could enhance the anti-atherosclerotic effects of mouse peroxisome proliferator-activated receptor gamma1 (PPARgamma1) gene transfer in apolipoprotein-knock out mice.

METHODSAdult ApoE-knock out mice were fed a Western-diet for 20-weeks and then injected with PBS, Ad. PPARgamma1 (5 x 10(8)pfu) or Ad. GFP (5 x 10(8)pfu) via jugular vein. Another group of mice were intervened with rosiglitazone (dissolved in 0.5% cellulose acetate, 4 mg.kg(-1).d(-1), per gavage) 1 week before Ad. PPARgamma1 injection (n = 10, each group). Two weeks later, the lipid core and plaque composition were characterized with oil red O staining and Movat method respectively. The expression of PPARgamma, SM-actin, MOMA-2, MMP-9/TIMP-1, CD40/CD40L and TF antigens in aortic roots and plaques among four groups were compared semi-quantitatively using immunohistochemical technology.

RESULTSAll parameters were similar between AdGFP and PBS groups (P > 0.05). The area of plaque were significantly decreased and oil red O staining area significantly increased in AdPPARgamma1 [(0.86 +/- 0.12) mm(2), (150 +/- 35) x 10(3) microm(2)] and AdPPARgamma1 + RO [(0.79 +/- 0.15) mm(2), (270 +/- 49) x 10(3) microm(2)] treated mice compared with AdGFP group [(0.98 +/- 0.17) mm(2), (80 +/- 21) x 10(3) microm(2)] all P < 0.05. Elastic fiber, collagen and proteoglycan in plaques were also significantly increased in AdPPARgamma1 and AdPPARgamma1 + RO groups. Upregulation of PPARgamma, SM-actin, TIMP-1 antigen activity and downregulation of MOMA-2, MMP-9, CD40/CD40L and TF antigen activity in AdPPARgamma1 and most significantly in AdPPARgamma1 + RO group were observed (P < 0.05).

CONCLUSIONAnti-atherosclerotic effects of PPARgamma1 gene transfer in ApoE-knock out mice could be enhanced by PPARgamma agonist rosiglitazone.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; genetics ; Gene Transfer Techniques ; Male ; Mice ; Mice, Knockout ; PPAR gamma ; agonists ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Thiazolidinediones ; pharmacology ; Transfection

OBJECTIVETo investigate the association of ten SNP at peroxisome proliferator-activated receptors (PPARα, δ, γ) with hypertriglyceridemia and the gene-gene interaction.

METHODSParticipants were recruited from the Prevention of MetS and Multi-metabolic Disorders in Jiangsu province of China Study (PMMJS). A total of 820 subjects were selected from the 4083 participants who had received follow-up examination, by using simple random sampling. Participants in baseline and follow-up study surveys were both collected blood samples 11 ml in the morning after at least 8 hours of fasting. Blood samples which collected at the baseline were subjected to PPARα, PPARδ and PPARγ genotype analyses. Blood samples which collected at the follow-up were used to measure serum triglyceride levels. The logistic regression model was used to analyze the association between different SNP and hypertriglyceridemia, and the generalized multifactor dimensionality reduction (GMDR) was applied to explore the gene-gene interaction.

RESULTSThe samples included 474 in the non-hypertriglyceridemia group and 346 in the hypertriglyceridemia group. The genotype frequencies of rs1800206 in the hypertriglyceridemia group were 211 (61.0%) for LL, 132 (38.2%) for LV and 3 (0.9%) for VV, and in the non-hypertriglyceridemia group were 411 (86.7%) for LL, 59 (12.4%) for LV and 4(0.8%) for VV (χ(2) = 74.18, P < 0.01). V allele frequencies of rs1800206 in the hypertriglyceridemia group was 138(19.9%), and in the non-hypertriglyceridemia group was 67 (7.1%) (χ(2) = 60.62, P < 0.01). The genotype frequencies of rs2016520 in the hypertriglyceridemia group were 177 (51.2%) for TT, 154 (44.5%) for TC and 15 (4.3%) for CC, and in the non-hypertriglyceridemia group were 211 (44.5%) for TT, 212 (44.7%) for TC and 51 (10.8%) for CC(χ(2) = 15.93, P < 0.01). C allele frequencies of rs2016520 in the hypertriglyceridemia group was 184(26.6%), and in the non-hypertriglyceridemia group was 314 (33.1%) (χ(2) = 8.07, P < 0.01). The genotype frequencies of rs3856806 in the hypertriglyceridemia group were 149 (43.1%) for CC, 156 (45.1%) for CT and 41 (11.8%) for TT, and in the non-hypertriglyceridemia group were 269 (56.8%) for CC, 170 (35.9%) for CT and 35 (7.4%) for TT (χ(2) = 15.93, P < 0.01). T allele frequencies of rs3856806 in the hypertriglyceridemia group was 238(34.4%), and was 240 (25.3%) in the non-hypertriglyceridemia group (χ(2) = 15.96, P < 0.01). The genotype frequencies of rs1805192 in the hypertriglyceridemia group were 145 (41.9%) for PP, 158(45.7%) for PA and 43(12.4%) for AA, and in the non-hypertriglyceridemia group were 314 (66.2%) for PP, 137(28.9%) for PA and 23(4.9%) for AA (χ(2) = 50.92, P < 0.01). A allele frequencies of rs1805192 in the hypertriglyceridemia group was 244(35.2%), and was 183 (19.3%) in the non-hypertriglyceridemia group(χ(2) = 52.89, P < 0.01). After adjusting age, gender, smoking, alcohol consumption, high-fat diet, low -fiber diet and occupational physical activity factors, rs1800206, rs2016520, rs3856806 and rs1805192 were significantly associated with hypertriglyceride, while the OR (95%CI) was 3.88 (2.69 - 5.60), 0.71 (0.52 - 0.96), 1.40 (1.03 - 1.90) and 2.56 (1.88 - 3.49), respectively (P < 0.05). GMDR model analysis showed that the second-order model (rs1800206 and rs1805192) was the best model when quality traits of triglyceride was chosen as outcome (P < 0.01); while third-order model (rs1800206, rs1805192 and rs2016520) was the best model when quantitative traits of triglyceride was chosen as outcome (P < 0.01).

CONCLUSIONThe rs1800206, rs2016520, rs3856806 and rs1805192 were significantly associated with hypertriglyceridemia. There was a gene-gene interaction between multiple SNP.

Adult ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertriglyceridemia ; blood ; genetics ; Logistic Models ; Male ; Middle Aged ; Peroxisome Proliferator-Activated Receptors ; genetics ; Polymorphism, Single Nucleotide

AIMTo investigate the effect of different peroxisome proliferator-activated receptors (PPARs) activators on plasminogen activator inhibitor-1 in HepG-2 cell line and explore the effect of PPARs on PAL-1 gene expression.

METHODSStearic acid, oleic acid, linoleic acid, fenofibrate, pioglitazone were used in the treatment of HepG-2 cell culture. The level of PAI-1 and PPARs mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR) and the level of PAI-1 activity and PPARs protein was determined by colorimetric assay and western blotting respectively.

RESULTSThe mRNA and activity of PAI-1 significantly increased in the groups of oleic acid and linoleic acid compared with the control, but decreased in the group of fenofibrate. There were no significant changes in both groups of stearic acid and pioglitazone. The alterations in the level of PPARs mRNA and protein were not detected in all the treated groups compared with the control.

CONCLUSIONPeroxisome proliferator-activated receptors activators play important roles in the PAI-1 gene expression and regulation. It is likely mediated by the activation of PPARalpha, but there might be other mechanisms.

Fenofibrate ; pharmacology ; Hep G2 Cells ; Humans ; Linoleic Acid ; pharmacology ; Oleic Acid ; pharmacology ; Peroxisome Proliferator-Activated Receptors ; agonists ; metabolism ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; RNA, Messenger ; genetics

Peroxisome proliferator-activated receptors (PPARs) are nuclear transcriptional factors closely related to glucose and lipid metabolism, insulin sensitivity. Activation of PPARs targets treated type 2 diabetes, obesity, hypertension and other metabolic diseases by insulin resistance. Recently, a variety of active ingredients of traditional Chinese medicines (TCMs) have been proved to activate PPARs targets for improving insulin resistance, which has attracted widespread attention at home and abroad. In this paper, we reviewed the pathological mechanisms between insulin resistance and PPARs, and summarized the active ingredients of TCMs improved insulin resistance based on PPARs targets. This paper may provide some theoretical guidance for the development of new drugs and TCMs.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Insulin Resistance ; Metabolic Diseases ; drug therapy ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; antagonists & inhibitors ; genetics ; metabolism

OBJECTIVETo study the regulatory effect of lipi tiaozhi capsule (LTC) on the expression of peroxisome proliferator-activated receptor (PPAR) alpha and gamma mRNA in dyslipidemia rats and ApoE(-/-) mice, and to explore its mechanisms for regulating lipid metabolism. Methods 48 Wistar rats were randomly divided into the blank group, the model group, the treatment group (treated by LTC), and the control group (treated by Xuezhi-kang Capsule). After four-week modeling (except the blank group) 30 ApoE(-/-) mice were randomly divided into the blank group, the treatment group, and the control group. LTC was given by gastrogavage to rats and ApoE(-/-) mice in LTC groups while XZKC was given to XZKC groups. The medication was conducted once daily for eight weeks. The serum TC and TG contents of rats and mice were determined by enzymic method. The serum high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were detected by precipitation method. PPARalpha and gamma mRNA expressions were detected in the liver tissue of the rats and mice by fluorescent PCR.

RESULTSCompared with the model group and the blank group, the serum contents of TC, TG, and LDL-C of rats or mice in the treatment group decreased significantly (P < 0.01). The serum content of HDL-C increased significantly (P < 0.01). PPARalpha and gamma mRNA expressions of rats or mice increased significantly (P < 0.01). Compared with the control group, the serum contents of TC, TG, and LDL-C of mice and rats in the treatment group decreased (all P < 0.05), the serum content of HDL-C increased significantly (P < 0.05, P < 0.01). And PPARalpha and gamma mRNA expressions of rats or mice increased significantly (P < 0.05).

CONCLUSIONLTC could significantly increase PPARa and y mRNA expressions of experimental dyslipidemia rats and ApoE(-/-) mice, playing roles in regulating nuclear factors and further effecting lipid metabolism.

Animals ; Apolipoproteins E ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Dyslipidemias ; drug therapy ; metabolism ; Male ; Mice ; Mice, Knockout ; Peroxisome Proliferator-Activated Receptors ; genetics ; metabolism ; Phytotherapy ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVE: To investigate the effect of adipocytes in the bone marrow microenvironment of patients with multiple myeloma (MM) on the pathogenesis of MM. METHODS: Bone marrow adipocytes (BMA) in bone marrow smears of health donors (HD) and newly diagnosed MM (ND-MM) patients were evaluated with oil red O staining. The mesenchymal stem cells (MSC) from HD and ND-MM patients were isolated, and in vitro co-culture assay was used to explore the effects of MM cells on the adipogenic differentiation of MSC and the role of BMA in the survival and drug resistance of MM cells. The expression of adipogenic/osteogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4, FASN and ALP both in MSC and MSC-derived adipocytes was determined with real-time quantitative PCR. The Western blot was employed to detect the expression levels of IL-6, IL-10, SDF-1α, TNF-α and IGF-1 in the supernatant with or without PPAR-γ inhibitor. RESULTS: The results of oil red O staining of bone marrow smears showed that BMA increased significantly in patients of ND-MM compared with the normal control group, and the BMA content was related to the disease status. The content of BMA decreased in the patients with effective chemotherapy. MM cells up-regulated the expression of MSC adipogenic differentiation-related genes PPAR-γ, DLK1, DGAT1, FABP4 and FASN, but the expression of osteogenic differentiation-related gene ALP was significantly down-regulated. This means that the direct consequence of the interaction between MM cells and MSC in the bone marrow microenvironment is to promote the differentiation of MSC into adipocytes at the expense of osteoblasts, and the cytokines detected in supernatant changed. PPAR-γ inhibitor G3335 could partially reverse the release of cytokines by BMA. Those results confirmed that BMA regulated the release of cytokines via PPAR-γ signal, and PPAR-γ inhibitor G3335 could distort PPAR-γ mediated BMA maturation and cytokines release. The increased BMA and related cytokines effectively promoted the proliferation, migration and drug resistance of MM cells. CONCLUSION The BMA and its associated cytokines are the promoting factors in the survival, proliferation and migration of MM cells. BMA can protect MM cells from drug-induced apoptosis and plays an important role in MM treatment failure and disease progression.
Humans ; Osteogenesis/genetics* ; Bone Marrow/metabolism* ; Multiple Myeloma/metabolism* ; Drug Resistance, Neoplasm ; Peroxisome Proliferator-Activated Receptors/pharmacology* ; Cell Differentiation ; Adipogenesis ; Cytokines/metabolism* ; Adipocytes/metabolism* ; Bone Marrow Cells/metabolism* ; Cells, Cultured ; PPAR gamma/pharmacology* ; Tumor Microenvironment

BACKGROUNDThe chronic pathological changes in vascular walls of hypertension may exert destructive effects on multiple organ systems. Accumulating evidence indicates that inflammatory reactions are involved in the pathological changes of hypertension. Three peroxisome proliferator-activated receptors (PPARs) have been identified: PPARalpha, PPARbeta/delta, and PPARgamma, all of which have multiple biological effects, especially the inhibition of inflammation. The aim of this study was to evaluate PPAR isoforms expression profile in important organs of spontaneously hypertensive rats (SHR) and to understand the modulation of endogenous PPAR isoforms under inflammatory condition.

METHODSTissues (kidney, liver, heart, and brain) were dissected from SHR and age-matched control Wistar-Kyoto rats (WKY) to investigate the abundance of PPAR isoforms and PPAR-responsive genes (acyl-CoA oxidase and CD36). The expression of CCAAT/enhancer-binding protein delta (C/EBPdelta), which can trans-activate PPARgamma expression, was also observed. The inflammatory response was analyzed by the expression of inflammatory mediators inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, interleukin-1 beta (IL-1beta), and tumor necrosis factor alpha (TNFalpha), and formation of carbonyl and nitrated proteins.

RESULTSThe expressions of 3 PPAR isoforms and PPAR-responsive genes were markedly upregulated in SHR compared with those of WKY. Specifically, the expression of PPARalpha protein in the kidney, liver, heart and brain increased by 130.76%, 91.48%, 306.24%, and 90.70%; PPARbeta/delta upregulated by 109.34%, 161.98%, 137.04%, and 131.66%; PPARgamma increased by 393.76%, 193.17%, 559.29%, and 591.18%. In consistent with the changes in PPARgamma, the expression of C/EBPdelta was also dramatically elevated in SHR. Inflammatory mediators expressions were significantly increased in the most organs of SHR than WKY. As a consequence, increased formation of carbonyl and nitrated proteins were also observed in the most organs of SHR.

CONCLUSIONSThese findings suggest an enhanced inflammatory response in the organs of SHR, which might play a key role in pathogenesis of hypertension and secondary organ complications. Changes (increases) in PPARs expression may reflect a compensatory mechanism to the inflammatory status of hypertensive rats.

Animals ; Blood Pressure ; Blotting, Western ; E-Selectin ; genetics ; metabolism ; Gene Expression ; Hypertension ; genetics ; metabolism ; physiopathology ; Inflammation ; genetics ; metabolism ; physiopathology ; Interleukin-1beta ; genetics ; metabolism ; Male ; PPAR alpha ; genetics ; metabolism ; PPAR delta ; genetics ; metabolism ; PPAR gamma ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; genetics ; metabolism ; Plethysmography ; methods ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an important role in several pathological processes of cardiovascular diseases. In this study, the effects of XCT790, a potent and selective inverse agonist of estrogen-related receptor alpha (ERRalpha), on rat VSMCs proliferation and related signal pathways were investigated. The proliferative activity of VSMCs was determined by CCK-8 assay. The mRNA levels of ERRalpha, PGC-1alpha, OPN and MCAD were assayed by RT-PCR. The protein levels of ERRalpha, ERK2 and p-ERK1/2 were evaluated by Western blotting. ELISA was used to assess the protein expression of VEGF. The results showed that XCT790 (5-20 micromol x L(-1)) inhibited rat VSMCs proliferation, and the expression of ERRalpha and its target genes, as well as p-ERK1/2, were also inhibited. XCT790 inhibited VSMCs proliferation in a dose-dependent manner at the dose range from 5 to 20 micromol x L(-1) and in a time-dependent manner at the dose range from 10 to 20 micromol x L(-1). These findings demonstrate that XCT790 inhibits rat VSMCs proliferation by down-regulating the gene level of ERRalpha and thus inhibiting the ERK signal pathway, suggesting that ERRalpha may be a novel potential target for therapeutic approaches to inhibit VSMCs proliferation, which plays an important role in several cardiovascular diseases.
Animals ; Cadherins ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Dose-Response Relationship, Drug ; GTPase-Activating Proteins ; genetics ; metabolism ; MAP Kinase Signaling System ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Nitriles ; administration & dosage ; pharmacology ; Nuclear Proteins ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Phosphorylation ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; genetics ; metabolism ; Thiazoles ; administration & dosage ; pharmacology ; Transcription Factors ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism