ABSTRACT
Aim To preliminarily investigate the effect of brusatol(BRU), the monomer components fromChinese medicines on H1299 cells and its mechanisms.Methods CCK-8 and EdU staining experiment were used to detect the effect of BRU on cell prolifera-tion.Clone formation experiment was performed to measure the effect of drugs on cell clone formation.Hoechst33258 staining experiment and flow cytometry were employed to observe the cell apoptosis.Western blot assay was used to detect the protein expression levels of Bcl-xL, Bax, Bcl-2, cleaved-caspase-3, caspase-3, Gadd45α, PI3K, p-PI3K, Akt, p-Akt and NF-κB-p65.Results CCK-8 assay revealed that the inhibitory effect of H1299 cells gradually increased with the rising of BRU concentration and action time.Compared with control group, the EdU incorporation rate of the BRU treatment group decreased significantly.Treated with different concentrations of BRU for 24 h, the clone formation rate was significantly reduced in a concentration-dependent manner.Hoechst33258 experiment and flow cytometry showed that BRU could induce apoptosis in H1299 cell nucleus and increase its apoptotic rate.Western blot results revealed that BRU could significantly up-regulate the protein levels of Bax, cleaved-caspase-3, Gadd45α, and significantly down-regulate the expression of Bcl-xL, Bcl-2, caspase-3.In addition, BRU could significantly decrease the expression level of p-PI3K, p-Akt, NF-κB-p65 in cell nucleus.Conclusions BRU can inhibit the proliferation and induce apoptosis of H1299 cells in a concentration and time-dependent manner.The mechanism may be related to the inhibition of PI3K/Akt signaling pathway and the nuclear shuttle of NF-κB-p65.
ABSTRACT
Brusatol, a triterpene lactone compound mainly from Brucea javanica, sensitizes a broad spectrum of cancer cells. It is known as a specific inhibitor of nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. In this review, we provide a comprehensive overview on the antitumor effect and molecular mechanisms of brusatol in vitro and in vivo. This review also covers pharmacokinetics studies, modification of dosages forms of brusatol. Increasing evidences have validated the value of brusatol as a chemotherapeutic agent in cancers, which may contribute to drug development and clinical application.
ABSTRACT
Objective Up to now, the role of Brucea in early embryonic development of mice and its mechanism is still unclear.This paper aims to explore the role of Brusatol in mouse early embryonic development and its possible mechanism.Methods 100 kunming rats of clean grade(80 female rats and 20 male rats) were divided into 6 group: negative control group(no DMSO)、blank control group(culture in fresh CM with equal DMSO)、20nmol/L brusatol treated group、50nmol/L brusatol treated group、100nmol/L brusatol treated group、200nmol/L brusatol treated group(A solution of Brusatol was diluted in CM to concentrations of 20, 50, 100 or 200nmol/L.).Each group used an average of 20 embryos each time, repeated 4 times.Fertilized eggs after cultured 24h, 48h,72h, 96h were respectively 2-cell stage, 4-cell stage,morula and blastocyst stage..The embryo development rate was observed in the culture medium and the optimal concentration was selected, the embryos were collected to analysis the subcellular localization of the Nrf2 by immunofluorescence.The mRNA expression level of Cyclin B, CDK1 and the protein expression of Nrf2 were detected by Q-PCR and western blot respectively.Results In 4-cell stage, the embryo development rates of 20、50、100nmol/L brusatol treated groups[(75.0±2.8)%、(30.4±7.5)%、(4.2±5.9)%] significantly reduced compared with the negative control group[(93.0±2.8)%]、blank control group[(90.9±1.2)%].In morula stage, compared with blastocyst rates of negative control group、blank control group [(83.5±2.1)%、(84.2±1.2)%], 50nmol/L brusatol treated group[(19.3±13.1)%] decreased obviously [(79.00±0.06)% vs 100%, P<0.05].In the cellular immunofluorescence assay, the expression of Nrf2 protein in 50nmol/L brusatol treated group was lower than blank control group(P<0.05).We further found that 50nmol/L brusatol treated group decreased more mRNA levels of Cyclin B[(59.5±9.2)%] and CDK1[(56.0±1.4)%] than blank control group(100%) in G2/M phase(P<0.05).Conclusion In this study, Brusatol mainly affects the cell cycle transformation from G2 to M phase dependent on Cyclin B-CDK1, further inhibiting the development of the embryo through down-regulating Nrf2.
ABSTRACT
Objective To investigate the effects of brusatol on mechanical properties of the cytoskeleton as well as the invasion behavior of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). Methods Cytoskeleton staining method was used to determine the regulatory effects of brusatol on mechanical properties of the RA FLS cytoskeleton. Transwell chamber assay was used to detect the effects of brusatol on the cytoskeleton and invasion behavior of RA FLS. The effects of brusatol on the expression of matrix metalloproteinase-2/3 (MMP-2/3) of RA FLS were measured by zymography and Western blotting methods. Results Cytoskeleton staining and microscope observation showed that brusatol could significantly reduce the formation number of RA FLS pseudopodia, thus inhibited cell movement ability via regulating mechanical properties of cytoskeleton. The invasion behavior of RA FLS was inhibited by brusatol, and brusatol could down-regulate the expression of MMP-2/3. Conclusions Brusatol plays an important role in regulating mechanical properties of cytoskeleton and inhibiting the invasion behavior of RA FLS. Meanwhile, brusatol can inhibit the invasion behavior of RA FLS through down-regulating the expression of MMP-2/3. The research findings provide the corresponding experimental basis for further development of new drugs for RA treatment.