Antiviral Agents/pharmacology* ; COVID-19 ; Coronavirus 3C Proteases ; Humans ; Lactams ; Leucine ; Molecular Docking Simulation ; Molecular Dynamics Simulation ; Nitriles ; Proline ; Protease Inhibitors/pharmacology* ; SARS-CoV-2

OBJECTIVETo investigate the effect of extracellular signal-regulated kinase (ERK) signaling pathway on the endothelial differentiation of periodontal ligament stem cells (PDLSC).

METHODSHuman PDLSC was cultured in the medium with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF) to induce endothelial differentiation. Endothelial inducing cells was incubated with U0126, a specific p-ERK1/2 inhibitor. PDLSC from one person were randomly divided into four groups: control group, endothelial induced group, endothelial induced+DMSO group and endothelial induced+U0126 group. The protein expression of the p-EKR1/2 was analyzed by Western blotting at 0, 1, 3, 6 and 12 hours during endonthelial induction. The mRNA expressions of CD31, VE-cadherin, and VEGF were detected by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) after a 7-day induction. The proportion of CD31(+) to VE-cadherin(+) cells was identified by flow cytometry, and the ability of capillary-like tubes formation was detected by Matrigel assay after a 14-day induction. The measurement data were statistically analyzed.

RESULTSPhosphorylated ERK1/2 protein level in PDLSC was increased to 1.24±0.12 and 1.03±0.24 at 1 h and 3 h respectively, during the endothelial induction (P<0.01). The mRNA expressions of CD31 and VEGF in induced+U0126 group were decreased to 0.09±0.18 and 0.49±0.17, which were both significantly different with those in induced group (P<0.05). The proportion of CD31(+) to VE-cadherin(+) cells of induced+U0126 group were decreased to 5.22±0.85 and 3.56±0.87, which were both significantly different with those in induced group (P<0.05). In Matrigel assay, the branching points, tube number and tube length were decreased to 7.0±2.7, 33.5±6.4, and (15 951.0±758.1) pixels, which were all significantly different with those in induced group (P<0.05).

CONCLUSIONSThe endothelial differentiation of PDLSC is positively regulated by ERK signaling pathway. Inhibition of ERK1/2 phosphorylation could suppress endothelial differentiation of PDLSC.

Antigens, CD ; genetics ; metabolism ; Butadienes ; pharmacology ; Cadherins ; genetics ; metabolism ; Cell Differentiation ; Endothelial Cells ; cytology ; physiology ; Enzyme Inhibitors ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; physiology ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; antagonists & inhibitors ; metabolism ; Nitriles ; pharmacology ; Periodontal Ligament ; cytology ; metabolism ; Phosphorylation ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Signal Transduction ; Stem Cells ; cytology ; physiology ; Time Factors ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; pharmacology

OBJECTIVE: To explore the effect of compound malt pills (CMP) on polycystic ovarian syndrome (PCOS) rat model induced by letrozole and the underlying mechanisms.
 METHODS: To establish a PCOS rat model, 48 female SD rats aged 6 weeks were randomly divided into 6 groups (n=8): A normal group, a model control group, a positive control group, a low-dose CMP group, a middle-dose CMP group, and a high-dose CMP group. Rats were treated for 21 days after the PCOS model was successfully established. Ovarian morphology changes were observed, and the expressions of ERα and ERβ was examined by immunohistochemistry, Western blot and RT-PCR, respectively.
 RESULTS: Compared with the normal group, the number of follicular cystic dilatation in the model control group was increased and the granulosa cells were decreased. After the treatment, the number of follicular cystic dilatation was reduced compared with the model control group, but the primordial follicles, corpus luteum and granulosa cells were increased. The expressions of ERα and ERβ in the model control group were significantly decreased (P<0.01), which were increased in the intervention groups (P<0.05 or P<0.01).
 CONCLUSION CMP may play a role in the treatment of PCOS by regulating the expressions of ERα and ERβ.
Animals ; Corpus Luteum ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Female ; Granulosa Cells ; drug effects ; Letrozole ; Nitriles ; adverse effects ; Ovarian Follicle ; drug effects ; Polycystic Ovary Syndrome ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triazoles ; adverse effects

OBJECTIVETo investigate the effect of Euphorbia fischeriana extract on latent HIV reactivation and the pathway involved in this process and discuss the value of Euphorbia fischeriana extract in eliminating HIV.

METHODSFresh tissues of Euphorbia fischeriana root were crushed into powder after quick freezing with liquid nitrogen and extracted with acetone followed by a three-day vacuum freeze-drying for dehydration of the extract. The extract (EFE) was separated using RP-C18 column with high-performance liquid chromatography (HPLC) and identified with mass spectrometry (MS). The activity of reactivated latent HIV was analyzed by fluorescence-activated cell sorting in a J-Lat 10.6 cell model treated with EFE (50 µg/mL) for 24 h, using TNF-α (10 ng/mL) as the positive control. The effect of a NF-κB pathway inhibitor (Bay 11-7082) on EFE activity was tested. The changes in P65 expression in the cell nuclei within 2 h and HIV protein p24 expression within 24 h were analyzed by Western blotting in cells treated with EFE.

RESULTSEFE was obtained by one-step acetone extraction, and the concentration of prostratin in the extract was around 0.53 mmol/L. About 50% of the cells showed HIV reactivation after treatment with 50 µg/mL EFE for 24 h accompanied by a significantly increased p24 expression. The activity of EFE in reactivating latent HIV was inhibited by Bay 11-7082 in a concentration-dependent manner, and p65 accumulation was detected in the cell nuclei within 2 h.

CONCLUSIONEFE we obtained contains the active compounds of prostratin and its analogues and shows a strong capacity to reactivate latent HIV through classical NF-κB pathway.

Euphorbia ; chemistry ; Flow Cytometry ; HIV ; drug effects ; HIV Infections ; Humans ; NF-kappa B ; metabolism ; Nitriles ; Phorbol Esters ; chemistry ; Plant Extracts ; pharmacology ; Signal Transduction ; Sulfones ; Tumor Necrosis Factor-alpha ; Virus Latency ; drug effects

OBJECTIVETo explore the effects of λ-cyhalothrin on hippocampus by interfering with estrogen.

METHODSThe healthy female ICR mice of postnatal 28 days were random divided into 12 groups, 4 of those were sham-operation include control, λ-cyhalothrin (LCT, 3.0 µg/g), Letrozole (Let, 1.0 µg/g), and LCT (3.0 µg/g)+Let (1.0 µg/g); and the last 8 were ovariectomized include OVX, Estradiol (E2, 10.0 µg/g), LCT, Let, E2+LCT, E2+Let, LCT+Let, E2+LCT+Let. 10 mice in every group received drugs by intraperitoneal injection for 2 days. Then half of every group initiate the ethological test (open field test and Morris water maze) 24 h later. The last half animals were sacrificed to made frozen section for immunofluorescent assay of postsynaptic density protein 95 (PSD95).

RESULTSIn ethological test, campared with Sham, OVX can lengthen incubation period in the first grid and to get on the platform (P < 0.05); campared with OVX, OVX+E2 can increase the total numbers of through grid and shorten the incubation period to get on the platform (P < 0.05); campared with OVX+E2, OVX+E2+LCT can reduce the number of grid and standing, lengthen incubation period to the platform (P < 0.05); campared with Sham, Sham+LCT can lengthen incubation period to the platform of Sham mice (P < 0.05), but campared with OVX, OVX+LCT can shoten incubation period in the first grid and to get on the platform in OVX mice (P < 0.05); campared with Sham+Let, Sham+LCT+Let can lengthen incubation period in the first grid, reduce the the number of grid and standing (P < 0.05). In the Immunohistochemical fluorescence experiment we find that, campared with Sham, OVX can reduce the expression of PSD95 in CA1,CA3 and DG (P < 0.05); however campared with OVX, E2 or LCT can both inhibit the effect of OVX (P < 0.05); campared with Sham, Sham+LCT can reduce the expression of PSD95, the same result when OVX+E2+LCT campared with OVX+E2 (P < 0.05); campared with OVX+E2+Let, OVX+E2+LCT+Let can reduce the expression of PSD95 in CA3 (P < 0.05); campared with OVX+Let, OVX+LCT+Let can increase the expression of PSD95 in DG (P < 0.05).

CONCLUSIONSWhen few estrogen exist in the body, LCT can show estrogen-like action to promote hippocampal synaptic development; but when circulating estrogen exist, LCT can inhibit synaptic development by interfering estrogen.

Animals ; Estradiol ; Estrogens ; pharmacology ; Female ; Hippocampus ; drug effects ; Humans ; Mice ; Mice, Inbred ICR ; Nitriles ; pharmacology ; Ovariectomy ; Pyrethrins ; pharmacology ; Random Allocation ; Synapses ; drug effects ; Triazoles

OBJECTIVETo investigate whether fenvalerate can induce mouse hippocampal nerve cell damage by interfering with estrogen (E2) effect.

METHODSHippocampus were dissected and cultured from Embryo 18 d ICR mice, the cells were cultured for 7 days. Fenvalerate (FEN, 0, 1, 10, 50 µg/ml), FEN (10, 50 µg/ml) and estrogen receptor antagonist ICI 182, 780 (1 µmol/L), FEN (0, 10, 50 µg/ml) and E2 (10 nmol/L) were applied to the cultured cells for 48h. Immunocytochemically stained with neurons and astrocytes to evaluate the levels respectively, and the growth of neurite. Result 1µg/ml FEN have no effect on neurons, neurites and protoplasmic astrocytes, 10 and 50 µg/ml FEN can significantly decrease the neuron viability and the length of neurite as well as increase the level of protoplasmic astrocytes (P < 0.05 vs. control group). ICI 182, 780 alone have no effect on neurons, neurites and protoplasmic astrocytes; ICI+10 µg/ml FEN significantly increase the cell viability and extend neurite length as well as decrease protoplasmic astrocytes (P < 0.05 vs. 10 µg/ml FEN alone group); ICI+50 µg/ml FEN significantly increase the cell viability and decrease protoplasmic astrocytes (P < 0.05 vs. 50 µg/ml FEN alone group). E2 alone have no effect on protoplasmic astrocytes, while can promote neuronal survival and neurite growth; E2+10 µg/ml FEN and E2+50 µg/ml FEN significantly decrease neuronal survival and neurite growth, as well as increase protoplasmic astrocytes (P < 0.05 vs. E2 alone group).

CONCLUSIONFenvalerate can induce the loss of hippocampal neurons through disrupting estrogen nuclear receptor signaling, and inhibit the length of neurite through disrupting estrogen nuclear receptor and membrane receptor signaling. The effect of estrogen disruption play an important role in developmental neurotoxicity by fenvalerate.

Animals ; Astrocytes ; drug effects ; Cells, Cultured ; Estrogens ; pharmacology ; Hippocampus ; drug effects ; pathology ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; pathology ; Nitriles ; toxicity ; Pyrethrins ; toxicity

OBJECTIVETo investigate the estrogen interference property of fenvalerate in neurodevelopmental toxicity.

METHODSThirty 4-week-old healthy female ICR mice were randomly divided into 6 groups: sham operation group, ovariectomized control group, ovariectomized with estrogen (10 µg/g) group, ovariectomized with fenvalerate (5 µg/g) group, sham operation with fenvalerate group, and ovariectomized with estrogen and fenvalerate group, with 5 mice in each group. Fenvalerate was injected intraperitoneally once a day for 7 consecutive days. Mice were sacrificed at 24 h after the last exposure to separate the hippocampus. Immunofluorescence was used to detect neuron marker (NeuN) and astrocyte marker (GFAP) in hippocampal CA1, CA3, and DG regions.

RESULTSCompared with the sham operation group (numbers of NeuN-positive cells: CA1 (54.00±1.73), CA3 (59.00 ± 1.73), DG (100.00 ± 4.58)), the sham operation with fenvalerate group (CA1 (37.67 ± 2.08), CA3 (41.33 ± 1.15), DG (80.67±0.58)) and ovariectomized control group (CA1 (44.00 ± 3.00), CA3 (51.00 ± 3.00), DG (83.00 ± 1.72)) showed significant decreases in number of neurons (NeuN-positive cells) in the hippocampus (P < 0.05). Compared with the ovariectomized control group, the ovariectomized with fenvalerate group (CA1 (47.67 ± 3.21), CA3 (49.00 ± 1.73), DG (87.33 ± 4.04)) showed no significant change in number of hippocampal NeuN-positive cells. Compared with the ovariectomized with fenvalerate group (CA1 (47.67 ± 3.21), DG (87.33 ± 4.04)), the sham operation with fenvalerate group and ovariectomized with estrogen and fenvalerate group (CA1 (40.00 ± 1.00), DG (78.67 ± 2.31)) experienced significant decreases in NeuN-positive cells (P < 0.05). Compared with the sham operation group (CA3 (11.00 ± 1.12), DG (10.67 ± 1.15)), the sham operation with fenvalerate group (CA3 (18.67 ± 2.07), DG (16.33 ± 1.53)) showed significant increase in number of astrocytes (GFAP-positive) cells (P < 0.05). Compared with the sham operation with fenvalerate group, the ovariectomized with fenvalerate group (CA3 (12.00 ± 1.00), DG (11.68 ± 1.16)) showed significant decrease in GFAP-positive cells (P < 0.05). Compared with the ovariectomized with fenvalerate group, the sham operation with fenvalerate group and ovariectomized with estrogen and fenvalerate group (CA3 (16.67 ± 2.13), DG (15.38 ± 1.42)) showed significant increases in GFAP-positive cells (P < 0.05).

CONCLUSIONThe interference with circulating estrogen is an important mechanism underlying the neurodevelopmental toxicity of fenvalerate.

Animals ; Estrogens ; pharmacology ; Female ; Hippocampus ; drug effects ; pathology ; Mice ; Mice, Inbred ICR ; Neurons ; drug effects ; pathology ; Nitriles ; toxicity ; Ovariectomy ; Pyrethrins ; toxicity

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

OBJECTIVETo evaluate the inhibitory effect of genistin combined with anastrozole on the growth and apoptosis of breast tumor tissue, and to study their anti-cancer mechanism by using the model of 7,12-dimethylbenz [alpha] anthracene (DMBA)-induced mammary tumors following ovariectomy in Sprague-Dawley (SD) rats.

METHODSThe DMBA induced postmenopausal SD rats were randomly divided into the control group, the genistein group, the anastrozole group, and the genistein combined with anastrozole group. The growth of tumors was observed in each group. The proliferation index and apoptosis index of tumor cells were determined. Moreover, estradiol (E2) and 17beta-HSD1 mRNA levels were determined by ELISA and RT-PCR respectively.

RESULTSThe tumor growth was inhibited in the genistein group and the anastrozole group. The inhibitory ratio was significantly higher in the genistein combined with anastrozole group (P < 0.05). Compared with the control group, levels of E2 and 17beta-HSD1 mRNA decreased more significantly in the genistein combined with anastrozole group (P < 0.05).

CONCLUSIONSGenistein could suppress the growth of mammary tumors in postmenopausal rats. It showed synergistic effect when combined with anastrozole, which resulted in reduced levels of E2 and 17beta-HSD1 mRNA. It had inhibitory effect on the growth of breast tumors.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; metabolism ; Female ; Genistein ; administration & dosage ; pharmacology ; Mammary Neoplasms, Experimental ; chemically induced ; pathology ; Nitriles ; administration & dosage ; pharmacology ; Ovariectomy ; Postmenopause ; Rats ; Rats, Sprague-Dawley ; Triazoles ; administration & dosage ; pharmacology

AIM: To evaluate the effect of Cocus nucifera L. flowers in reducing the major multiple symptoms of letrozole-induced polycystic ovarian disease (PCOD) in female rats. METHOD: Female, virgin Wistar rats were treated with letrozole (1 mg/kg body wt) to induce PCOD, and after 21 days of induction rats were administered orally with 100 and 200 mg·kg(-1) of Cocus nucifera flower aqueous extract, respectively. Estrus cycle and blood sugar were monitored once a week throughout the study. After scarification, various biochemical parameters, such as antioxidant status (superoxide dismutase (SOD) and glutathione reductase (GSH)) of the uterus homogenate, lipid profile (total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), and triglycerides (TG)) of the serum were determined. Weights of the uterus and ovaries were separately monitored. The characteristics of changes in the ovary were evaluated by histopathological studies. RESULTS: GC-MS analysis of the aqueous extract showed the presence of volatile and pharmacologically active phytoconstituents. C. nucifera flower extract-treated groups showed estrus cyclicity and increased uterus weight which indicates the estrogenic effect. The improved blood sugar level, ideal lipid profile, good antioxidant status, and histopathology results revealed the recovery from poly cystic ovaries. CONCLUSION The results indicate that C. nucifera flower is a potential medicine for the treatment of PCOD and this study supports the traditional uses of C. nucifera flower.
Animals ; Antioxidants ; metabolism ; Blood Glucose ; metabolism ; Cocos ; chemistry ; Estrus ; drug effects ; Female ; Flowers ; chemistry ; Gas Chromatography-Mass Spectrometry ; Hypoglycemic Agents ; pharmacology ; therapeutic use ; Letrozole ; Lipids ; blood ; Nitriles ; Oils, Volatile ; analysis ; pharmacology ; therapeutic use ; Ovary ; drug effects ; pathology ; Phytoestrogens ; pharmacology ; therapeutic use ; Phytotherapy ; Plant Extracts ; chemistry ; pharmacology ; therapeutic use ; Polycystic Ovary Syndrome ; blood ; chemically induced ; drug therapy ; pathology ; Rats, Wistar ; Triazoles ; Uterus ; drug effects