Résumé
Objective:To investigate effect of acacetin on alveolar epithelial cell damage caused by Streptococcus pneumoniae(SP)infection by regulating sirtuin 1(Sirt1)-mediated 5'-AMP activated protein kinase(AMPK)/nuclear factor erythroid-2 related factor 2(Nrf2)signaling pathway.Methods:Alveolar epithelial cells A549 cultured in vitro were infected with SP to establish a cell damage model.After treatment with acacetin at final concentrations of 0,5,25,50,100,150,200 μmol/L,CCK-8 was performed to detect cell viability of each treatment group and optimal concentration of acacetin was screened.A549 cells cultured in vitro were ran-domly separated into five groups:control group,model group,acacetin(150 μmol/L)group,EX527(Sirt1 inhibitor,40 μmol/L)group,acacetin(150 μmol/L)+EX527(40 μmol/L)group,control group was not treated,other groups were infected with SP to establish a cell damage model,and then treated with 150 μmol/L acacetin and 40 μmol/L EX527,CCK-8 and flow cytometry were performed to measure cell viability and apoptosis rate in each group;kits were performed to measure levels of reactive oxygen species(ROS),superoxide dismutase(SOD),malondialdehyde(MDA),lactate dehydrogenase(LDH)and IL-10,IL-1β,TNF-α levels of cells in each group;Western blot was performed to measure proliferation-related proteins Ki-67,proliferating cell nuclear antigen(PCNA),apoptosis-related proteins caspase-9,Bax,Sirt1 and AMPK/Nrf2 signaling pathway proteins p-AMPK/AMPK,Nrf2 expres-sions of cells in each group.Results:Model group had decreased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and increased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels than control group(P<0.05).Compared with model group and acacetin+EX527 group,acacetin group had increased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and decreased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels(P<0.05);EX527 group had decreased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and increased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels(P<0.05).Conclusion:Acnestin can activate AMPK/Nrf2 signaling by up-regulating Sirt1 expression,thereby promoting secretion of anti-inflammatory factors,reducing production of ROS and pro-inflammatory factors,reducing inflammation and oxidative stress,and finally alleviating neuronal damage.
Résumé
Objective:To explore the proliferation inhibition and radiosensitization of Biyanqing granule on nasopharyngeal carci -noma cell line CNE-2 in vitro.Methods:CNE-2 cells were cultured in vitro.The inhibition of Biyanqing granule on the proliferation of CNE-2 cell was evaluated by MTT assay .Radiosensitization was explored by clone formation assay , and cell cycle and apopotosis were observed by flow cytometry ( FCM) .Results:Biyanqing granule could inhibit the proliferation of CNE-2 in a time-and dose-dependent manner.The IC50 in 24, 48 and 72 h was 70.79, 60.13 and 51.63 mg· ml-1(calculated according to the weight of all medicinal ma-terial), respectively.The colony formation assay showed that Biyanqing granule combined with radiation could significantly reduce the colony formation of CNE-2 cells.With the concentration increase of the main drug , the colony formation of CNE-2 cells was reduced . The number of colony formation in the negative control group , the radiation group , 10 mg· ml-1 and 20 mg· ml-1 Biyanqing combined with radiation groups (calculated according to the weight of all medicinal material ) was significant different (P<0.05).With the main drug concentration increasing , the percentage of G 2/M phase and apoptotic cells were both increased , and compared with the con-trol group, the difference was significant (P<0.05).Conclusion:Biyanqing granule can not only inhibit CNE-2 cells but also block CNE-2 cells in G2/M to improve the radiosensitization of CNE-2 cells.
Résumé
Folic acid-O-carboxymethyl chitosan ultrasmall superparamagnetic iron oxide nanoparticles (FA-OCMCS-USPIO-NPs) are a novel molecular targeting MR contrast agent. This paper reperts the pharmacokinetics and magnetic resonance response characteristics of FA-OCMCS-USPIO-NPs in normal rats and mice, and discussed its distributing regularity in animals, providing basis for tumor targeting imaging. O-phenanthroline method was used to determine iron content in rats' plasma and mice's organs following high and low doses of nanoparticles injected through tail vein, and the blood concentration-time curve was drawn, the calculated t1/2 of two groups were greater than 7 h. The results of tissue distribution showed that only a small part of nanoparticles were swallowed by the liver and spleen, while none in the heart, lung and kidney. At the same times, the phagocytosis of nanoparticles did not change with the dose. The results of MRI showed that renal excretion occurred 4 hours after injection, and signal to noise ratio (SNR) of liver and kidney returned to normal levels 24 hours after injection. There were no nanoparticles in the lungs. So a part of nanoparticles escaped from phagocytosis of liver and spleen, and it owned lower toxicity and longer half-life. indicated its use for tumor-targeting imaging. All of these indicated its use for tumor-targeting imaging.