ABSTRACT
To prepare the asiaticoside nanoemulsions (ASI-NEs) and asiaticoside nanoemulsions-based gels (ASI-NBGs), compare them with the commercial cream of asiaticoside (ASI-C) in terms of transdermal characteristics, and investigate the transdermal mechanism of ASI-NEs and ASI-NBGs. Their transdermal characteristics were studied by using Franz diffusion cells. The effect of topical ASI-NEs and ASI-NBGs on ultrastructure of rabbit skin was evaluated by using HE staining method. The localization and the permeation pathway of asiaticoside were visually investigated by using laser scanning confocal microscope (CLSM). The transdermal studies in vitro showed that the cumulative amount of ASI permeated from ASI-NEs and ASI-NBGs at 12 h after application were (3 504.30±180.93), (1 187.40±128.88) μg·cm⁻² respectively, 6.57, 2.23 times of that in the control group of ASI-C; the drug deposition of ASI-NEs and ASI-NBGs in skin was (159.48±7.47), (120.53±5.71) μg·cm⁻² respectively, 5.93, 4.48 times of that of ASI-C. HE staining of the rabbit skin after application of ASI-NEs and ASI-NBGs showed that the epidermis structure was basically intact; stratum corneum was loosed and the keratin fragment was increased; at the same time, the gap of prickle cell was increased and the basal cells were arranged loosely. The study of CLSM showed that significant percutaneous enhancer effect was observed for ASI-NEs after the topical application of 6 h, as the fluorescent compound was penetrated in the dermis and diffused uniformly. The fluorescence area and the integral optical density (IOD) were 28.81, 32.51 times of that in the FITC aqueous solution group, respectively. The fluorescent preparations showed strong fluorescence in the epidermis, but weak in deeper layers; with the increase of treatment time, the fluorescence in deeper layer was increased and stronger in skin appendages. The prepared ASI-NEs and ASI-NBGs have good transdermal characteristics and the transdermal mechanism is related to breaking the ultrastructure of stratum corneum and penetrating by the path of skin adnexa.
ABSTRACT
Objective To explore whether suberoylanilide hydroxamic acid (SAHA,br.Vorinostat,a kind of histone deacetylase inhibitor) can improve the diastolic function of restrictive cardiomyopathy (RCM) mice by up-regulating the expression of wild type cardiac troponin I (WT-cTnI).Methods Sixteen male cTnI R193H mice were employed and divided into SAHA group (n=6),dimethyl sulfoxide (DMSO) group (n=5) and control groups (n=5).Mice were subcutaneously injected with 50mg/kg SAHA in SAHA group,with 1ml/kg DMSO in DMSO group and with 1ml/kg normal saline in control group,respectively.The total treatment duration lasted for 56 days.Western blotting was performed to determine the levels of acetylated histone H3 (acH3) and the expression of cTnI.The transcription levels of Tnni3 were detected with RT-qPCR.The levels of acH3 in Tnni3 promoter key area were detected with chromatin immunoprecipitation (ChIP).The data of diastolic function measurements were collected using high frequency echocardiography.Results Compared to control group,the acH3 levels in nucleus and in Tnni3 promoter key area increased significantly in SAHA group (P＜0.05).The acH3 level showed no significant difference between SAHA group and DMSO group (P＞0.0S),but the specific acH3 level in Tnni3 promoter key area increased significantly in SAHA group compared to that in DMSO group (P＜0.05).The expression of cTnI protein and Tnni3 mRNA showed no significant difference among the 3 groups.Compared to the control group,Tei index and E/A ratio increased in DMSO group.Left ventricle fractional shortening (LVFS),left ventricle ejection fraction (LVEF),stroke volume (SV),cardiac output (CO),isovolumic relaxation time (IVRT),E peak deceleration time (DT),early diastolic blood flow (E velocity) and late diastolic filling flow (A velocity) showed no significant difference between DMSO group and control group.IVRT,DT and Tei index increased,and E velocity and A velocity decreased in SAHA group compared to that in control group.Furthermore,LVFS,LVEF,SV and CO were decreased in SAHA group compared to that in control group (P＜0.05).Similarly,IVRT increased (P＜0.05) and LVFS,LVEE CO,E velocity and E/A ratio decreased in SAHA group compared to that in DMSO group.Conclusions SAHA may up-regulate the acH3 level in GATA and MEF2 binding site of Tnni3 promoter of RCM mice.However,SAHA may have no effect on the expression and transcription of Tnni3 in heart.Further studies are needed to confirm whether SAHA has any toxic effect on cardiac function.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of pseudolaric acid B (PLAB) on cell proliferation and cycle of human prostate carcinoma DU-145 cells. method: Its inhibitory effect on the cell growth was measured by MTT method. Characteristics of cell death were determined by Hoechest 33342 staining. The cell cycle was detected by flow cytometry. The expressions of cyclin B1, cyclin D1 and CDK1 were detected by Real time-PCR and Western blot, respectively.</p><p><b>RESULT</b>PLAB notably inhibited DU-145 cell growth in a dose- and time dependent manner (P < 0.05). Its IC50 values of PLAB for DU-145 cells for 24, 48 and 72 h were 4.53, 2.39 and 2.08 micromol x L(-1), respectively. Having been treated with 5 micromol x L(-1) PLAB for 24 h, the cells showed such apoptosis characteristics as nuclei chromatin condensation and apoptotic body. With the increase in PLAB concentration, the proportion of G2/M phase cells strikingly increased in a dose- and time dependent manner (P < 0.05), meanwhile cyclin B1 and CDK1 showed over-expressions (P < 0.05), and the cyclin D1 showed under-expression (P < 0.05).</p><p><b>CONCLUSION</b>PLAB can inhibit the growth of DU-145 cells and induce the cell cycle G2/M arrest, accompanied with the over-expression of cyclin B1 and CDK1, which may be related with its regulation cycle-associated protein degradation.</p>