ABSTRACT
Recombinant human interferon beta (rhIFN-β) is a glycoprotein produced by genetically engineered cells and has anti-virus, anti-tumor and immunoregulation functions. Although studies have shown that other subtypes of IFN such as IFN-γ affects cell proliferation and differentiation to some extent, the effect of rhIFN-β on chondrogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) is less known. In this study we studied the effect of rhIFN-β on the chondrogenic differentiation of hMSCs by inducing hMSCs into cartilage pellet via adding IFN-β1a into regular TGF-β3 chondrogenic differentiation medium. We collected the induced pellets and then detected GAG content, assessed pellets size, observed agreecan using alcian blue staining, and analyzed the expression of Sox and CollangenⅡusing real-time PCR and Western blotting. Addition of 100 ng/mL IFN-β1a to regular TGF-β3 chondrogenic differentiation medium could improve the concentration of GAG, increase the size of pellets, promote the formation of aggrecan and up-regulate the expression of CollangenII and Sox9. IFN-β1a combined with TGF-β3 could promote chondrogenic differentiation of hMSCs.
ABSTRACT
Objective To construct the angiotensinogen (AGT)-targeting RNA interfering plasmids, and to explore the effect of AGT gene on the differentiation of preadipocytes 3T3-L1 in the mice.Methods The small hairpin RNA (AGT-shRNA)interference plasmids,with a scrambled Non-shRNA as control,were constructed on the basis of the activity of U6 promoter-driven expression vector psiRNA-U6.1/Neo and transfected into the mouse preadipocytes 3T3-L1.Two cell lines were generated from stable transfections.RT-PCR and SDS-PAGE were performed to monitor the mRNA and protein expressions of AGT gene,respectively.The effect of adipose AGT on the differentiation of 3T3-L1 was investigated through microscopic observation and profiling of adipocyte differentiation markers. Results Both AGT-shRNA and Non-shRNA interference vectors were successfully constructed;the expression of AGT mRNA was inhibited (P < 0.05 ) and the expression of AGT protein was 43.86% of control.Both adipocyte differentiation markers,intracytoplasmic lipid levels and glycerol-3-phophate dehydrogenase (GPDH)activities were increased during the differentiation of preadipocytes 3T3-L1,but the data were lower than those in control group (P <0.05).Conclusion AGT gene silencing can significantly inhibit the differentiation of preadipocytes 3T3-L1,which demonstrates that there is important relationship between adipocyte metabolism and renin-angiotensin system (RAS)in adipose tissue.
ABSTRACT
Depression and insomnia are intimately related. Depressed patients usually manifest sleep discontinuity and early awakening, reduced or no slow wave sleep (SWS) and shortened latency of rapid eye movement (REM) sleep. These sleep abnormalities are very similar to those caused by over activated hypothalamic-pituitary-adrenal (HPA) axis with stress. Therefore, the animal models developed by post-traumatic stress disorder or chronic unpredictable mild stress could be used to evaluate drugs which have effects of both anti-depression and improvement of sleep quality, and to provide a more reliable platform for further studis on the mechanisms of depression and accompanied insomnia. This review mainly focuses on the typical features of sleep disturbance of depression, possible pathophysiological mechanisms, establishment of animal stress models and analysis of their abnormal sleep characteristics.
ABSTRACT
Histaminergic neurons solely originate from the tuberomammillary nucleus (TMN) in the posterior hypothalamus and send widespread projections to the whole brain. Experiments in rats show that histamine release in the central nervous system is positively correlated with wakefulness and the histamine released is 4 times higher during wake episodes than during sleep episodes. Endogeneous prostaglandin E2 and orexin activate histaminergic neurons in the TMN to release histamine and promote wakefulness. Conversely, prostaglandin D2 and adenosine inhibit histamine release by increasing GABA release in the TMN to induce sleep. This paper reviews the effects and mechanisms of action of the histaminergic system on sleep-wake regulation, and briefly discusses the possibility of developing novel sedative-hypnotics and wakefulness-promoting drugs related to the histaminergic system.
ABSTRACT
Dopamine(DA) modulates diverse wake-related behaviors including movement,reward, and cognition.Dopaminergic neurons are located in the substantia nigra pars compacta and ventral tegmental area.There are five distinct DA receptors(R):D_1R,D_2R(D_(2S)R and D_(2L)R), D_3R,D_4R and D_5R in the central nervous system, in which D_1R and D_2R are majorly expressed. The affinity of D_2R for endogenous DA is significantly higher than that of D_1R.Re- cently,studies by pharmacological and gene knock-out animals revealed that dopamine D_2R is essential inmaintaining wakefulness.Here,we review the progress on roles of D_2R in sleep-wake regulation.
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Aim To set up a highly effective and automatic mouse sleep-wake bioassay system,and evaluate the system through analysis of the somnogenic effects of L-stepholidine(SPD),targeting at dopamine D1/D2 receptors in mice.Methods The animals were housed in an insulated and soundproof recording chamber maintained at a constant temperature and humidity on an automatically controlled 12 h light/12 h dark cycle.The electroencephalogram and electromyogram were recorded continuously for 48 hours and analyzed by SleepSign software.Saline was administered ip to the mice at 21:00 on the first day,and SPD was given on the next day at the same hour.The vigilance state was analyzed based on the polygraphic recordings by the same software.Results The system has been demonstrated to be highly efficient and stable in recording and reliable in analyzing sleep-wake behavior in mice.With the aid of the sleep bioassay system,we found that SPD significantly increased the total time spent in sleep during dark period,and prolonged durations of non-rapid eye movement sleep episodes,with a concomitant reduction in amount and EEG power density of wakefulness.SPD rendered no effect on rapid eye movement sleep.Conclusion Through the reliable mouse sleep bioassay system,we found that SPD promotes non-rapid eye movement sleep but not rapid eye movement sleep in mice.
ABSTRACT
Pharmacokinetics of Tetramethylpyrazine Hydrochloride (TMPH) in rat was studied by using Ultraviolet Spectrophotometry. After a bolus iv injection of TMPH 30 mg/kg to rat, the pharmacokinetic characteristics are found to fit a two-compartment open model. The Pharmacokinetic parameters are: t 1/2? = 0 .1441h, t 1/2? = l .6953h, K21=2.1850h-1, K10=0.8605h-1, K12= 2 .0723h-1, AUC = 83 .3660mg?L -1h, CL = 0.3599L?kg-1h-1, Yc =0 .4182L?kg-1 , Vss =0 .7975L.kg-1