OBJECTIVE: To investigate the effect of piperlongumine(PL) on the proliferation and ferroptosis of human adriamycin-resistant chronic myeloid leukemia K562/ADR cells, and to explore its possible molecular mechanism. METHODS: CCK-8 assay was used to detect the effect of PL on the survival rate of K562/ADR cells and to screen the appropriate drug concentration. After K562/ADR cells were treated with low, medium and high concentrations of PL(2, 4, and 6 μmol/L), EdU proliferation assay and plate colony formation assay were used to detect cell proliferation and colony formation ability. CCK-8 assay was used to detect the effects of different inhibitors (Fer-1, Z-VAD, Nec-1) combined with PL on cell proliferation. The intracellular Fe²⁺, ROS, malondialdehyde(MDA) and glutathine(GSH) contents were respectively detected by iron ion colorimetry, DCFH-DA fluorescent probe, MDA and GSH kits. RT-qPCR and Western blot were respectively used to detect the expression level of GPX4 mRNA and protein in cells. Bioinformatics websites predicted miRNA that could target and regulate GPX4 . RT-qPCR was used to detect the effects of different concentrations of PL on the expression levels of the predicted miRNA. Dual luciferase gene reporter assay was used to verify the targeting relationship between miR-214-3p and GPX4 . After treating cells with PL or PL+miR-214-3p inhibitor, the Fe²⁺, ROS, MDA, GSH centents and GPX4 protein expression levels in cells were detected. RESULTS: PL inhibited K562/ADR cell proliferation in a concentration-dependent manner（r =0.979）. Compared with the blank control group, the survival rate, EdU positive cells rate in low, medium and high concentration PL groups were significantly decreased (P < 0.01). Compared with the PL group alone, the survival rate of cells in the Z-VAD+PL group was increased slightly (P < 0.05). The cell survival rate was significantly increased in medium or high concentration PL+Fer-1 group (P < 0.01). Compared with blank control group, ROS expression level in low concentration PL group was slightly increased (P < 0.05), and GSH content was slightly decreased (P < 0.05). In medium and high concentration PL groups, the contents of Fe²⁺, ROS and MDA were significantly increased (P < 0.01), while the contents of GSH, expression of GPX4 mRNA and protein were significantly decreased(P < 0.01). Bioinformatics prediction and double luciferase reporter gene experiment confirmed the targeting relationship between GPX4 and miR-214-3p. Compared with the blank control group, the expression level of miR-214-3p in cells of medium and high concentration PL groups was significantly increased (P < 0.01). Compared with PL group alone, the intracellular Fe²⁺, ROS and MDA contents in PL+miR-214-3p inhibitor group were all decreased (P < 0.01), while GSH content and GPX4 protein expression levels were significantly increased (P < 0.01). CONCLUSION Medium and high concentrations of PL can inhibit the proliferation of K562/ADR cells by inducing ferroptosis, which is related to the regulation of miR-214-3p pathway.
Humans ; Ferroptosis/drug effects* ; MicroRNAs/metabolism* ; Dioxolanes/pharmacology* ; Cell Proliferation/drug effects* ; K562 Cells ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Reactive Oxygen Species ; Doxorubicin/pharmacology* ; Signal Transduction ; Piperidones

OBJECTIVETo investigate of antiatherogenic effect and possible mechanisms of piper longuminine.

METHODThe atherosclerotic model was established by the hypercholesterol feeding rabbits. Male Mew Zealand rabbits were randomly divided into five groups: normal group, model group, the high-dose (5 mg x kg(-1) x d(-1)) and low-dose (2.5 mg x kg(-1) x d(-1) group of piperlonguminine, and simvastatin group (5 mg x kg(-1) x d(-1)). All the rabbits were fed for 60 days. Blood samples were taken from the ear edge vein of rabbits in the day before the experiment, and in the days of 20, 40 and 60 days after the experiment, respectively. All the rabbits were fasted for at least twelve hours before the blood was taken. The blood serum were analyzed for total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C). The blood serum of the 60th day were also analyzed for superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO). At last, the pathological observation of aorta and heart samples were carried out.

RESULTCompared with those in model group, the TC, TG and LDL-C levels were reduced (P < 0.05) and the HDL-C was raised in the piperlonguminine group; also, the serum SOD and NO level was raised (P < 0.05), MDA level was reduced in the piperlonguminine group (P < 0.05). Area percentage of aorta plaque was reduced (P < 0.01) in the piperlonguminine group. The aorta and heart injury was abated and coronary artery angusty extent was markedly abatement (P < 0.01). The results of observation through transmission electron microscope (TEM) indicated that the fine structure of aortal pathological degree was markedly abated.

CONCLUSIONThe piperlonguminine could inhibit the atherogenesis formation and development, which might be due to regulating the lipid metabolism and enhancing the antioxidation.

Animals ; Atherosclerosis ; blood ; chemically induced ; drug therapy ; pathology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dioxolanes ; pharmacology ; Male ; Malondialdehyde ; blood ; Nitric Oxide ; blood ; Rabbits ; Superoxide Dismutase ; blood ; Triglycerides ; blood

BACKGROUNDAstragali Radix, the root of Astragalus membranceus (Fish) Bunge Var. mongholicus (Bge), is a crude drug considered as one of the effective traditional Chinese anti-ageing material. The two isomers of 4-hydroxy-5-hydroxymethyl-[1, 3] dioxolan-2, 6'-spirane-5', 6', 7', 8'-tetrahydro-indolizine-3'-carbaldehyde (HDTIC), HDTIC-1 and HDTIC-2, were first extracted from the herb in 2002. We demonstrated previously that 0.1 micromol/L HDTIC-1 or 1.0 micromol/L HDTIC-2 strongly delay replicative senescence of human fetal lung diploid fibroblasts (2BS). In this study, we chose them to investigate their effects on the expression of senescence-associated genes to explore the mechanism of how HDTIC delays replicative senescence.

METHODSThe effects of HDTIC-1 and HDTIC-2 on the expression of p16 and p21 were observed in vitro by RT-PCR and Western blot. The anti-oxidative activities of the compounds were also observed by phenotype alteration after treatment with antioxidants.

RESULTSThere was an obvious expression of p16 in the control senescent cells. However, in the 2BS cells, after 56 population doublings (PDs) grown from PD28 in 0.1 micromol/L HDTIC-1 or 1.0 micromol/L HDTIC-2, there was a weak mRNA expression of p16 and no protein expression of p16 was observed. The expression level of p21 increased with cell ageing. Moreover, there was no difference between the expression level of p21 in the control cells and that in the same PD cells cultured with HDTIC compounds. The results also showed that 2BS cells exposed to 100 micromol/L H2O2 for 5 minutes return to their non-senescent phenotype and continue to be confluent after incubating the damaged cells with HDTIC-1 (1.0 micromol/L ) or HDTIC-2 (10 micromol/L ) for 1 hour.

CONCLUSIONSExpression of p16 by 2BS cells was strongly inhibited by HDTIC compounds, which could contribute to their delayed replicative senescence by the way of p16(INK4a)/Rb/MAPK. The anti-oxidative activities of HDTIC-1 and HDTIC-2, described in this study for the first time, might be indirectly related to their inhibition of p16 expression.

Antioxidants ; pharmacology ; Astragalus Plant ; chemistry ; Cells, Cultured ; Cellular Senescence ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; genetics ; Dioxolanes ; pharmacology ; Female ; Fibroblasts ; chemistry ; drug effects ; metabolism ; Humans ; Indolizines ; pharmacology ; Plant Roots ; chemistry ; RNA, Messenger ; analysis

OBJECTIVETo choose the most suitable concentration of 2-n-nonyl-1,3-dioxolane as a penetration enhancer in tanshinone gel preparation.

METHODIn vitro, transdermal absorption was studied using improved Frans equipment and rats skin. Tanshinone II A was tested by HPLC.

RESULTThe 4.0% concentration of 2-n-nonyl-1,3-dioxolane enhanced the transdermal absorption significantly in the preparation.

CONCLUSION2-n-nonyl-1,3-dioxolane was a new effective permeaton enhancer.

Administration, Cutaneous ; Animals ; Dioxolanes ; administration & dosage ; pharmacology ; Diterpenes, Abietane ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Female ; Gels ; In Vitro Techniques ; Male ; Mice ; Phenanthrenes ; analysis ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Skin ; drug effects ; metabolism ; Skin Absorption ; drug effects

Adenine ; analogs & derivatives ; chemistry ; pharmacology ; Animals ; Anti-HIV Agents ; chemistry ; pharmacology ; Antiviral Agents ; chemistry ; pharmacology ; Arabinofuranosyluracil ; analogs & derivatives ; chemistry ; pharmacology ; Cytosine ; analogs & derivatives ; chemistry ; pharmacology ; Dioxolanes ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Nucleosides ; chemistry ; pharmacology ; Organophosphonates ; chemistry ; pharmacology ; Purine Nucleosides ; chemistry ; pharmacology ; Tenofovir