Effect Of Brucine Research Articles

Effects of brucine on mitochondrial apoptosis and expression of HSP70 in prostate cancer cells.

BackgroundMost prostate cancer patients are already in the middle/advanced stages of the disease at the time of detection. Whether brucine can regulate apoptosis in prostate cancer cells through the expression of heat shock protein 70 (HSP70) has not been reported. We preliminarily investigated the effects of brucine on the mitochondrial apoptosis and HSP70 expression of prostate cancer cells and analyzed brucine’s possible mechanisms in the hope of providing an experimental basis for its clinical application.MethodsThe effect of brucine on the activity of PC-3 cells was determined by the methodology of tetrazolium (MTT) assay method. The effect of brucine on apoptosis was measured by Hoechst 33258 staining and flow cytometry, and western blotting was used to detect the expression levels of the apoptosis-related heat shock protein 70 (HSP70), anti-apoptotic protease activating factor 1 (Apaf-1) and anti-cysteine protease-3 (caspase-3).ResultsAt 24 and 48 h, the activity of human prostate cancer PC-3 cells in the low, medium, and high dose brucine groups was significantly lower than that of the control group, with a dose-dependent difference (P<0.01). The nuclei of the control group fluoresced uniformly with intact nuclei, whereas the nuclei of the brucine group appeared crinkled, and dense granular masses of strong blue fluorescence were visible. The apoptosis rate of human prostate cancer PC-3 cells in the low, medium, and high dose brucine groups was significantly higher than that in the control group (P<0.01) and was dose-dependent. The expression levels of the HSP70 protein in the human prostate cancer PC-3 cells in the low, medium, and high dose brucine groups were significantly lower than those in the control group, while the expression levels of the Apaf-1 and caspase-3 proteins were significantly higher than those in the control group (P<0.01) and showed a dose-dependent relationship.ConclusionsBrucine downregulated HSP70 expression in human prostate cancer PC-3 cells and inhibited the mitochondrial apoptotic signaling pathway, thus acting as an anti-apoptotic agent.

Effect of Brucine on Three Chondrocyte Apoptosis Models

With the development of people’s application technology of Strychnine, Strychnine is a relatively common type of medicine in the current clinical treatment of arthritis patients. And in the actual treatment, because strychnine has an important effect on the apoptosis of three kinds of chondrocytes. Better results can often be achieved through the use of horse money, which is of great significance to patients. The analysis of the influence of strychnose on the apoptosis of the three types of chondrocytes has become the focus of research in the academic and industry circles. The purpose of this article is to analyze the impact of Strychnoma chinensis on three chondrocyte apoptosis models. In this paper, by establishing a system for culturing rat chondrocytes in vitro, using nitric oxide in vitro cultured rat chondrocytes to induce their apoptosis, and observing the effect of Strychnine, the main monomer component of Chinese medicine Strychnine, on the cellular and molecular levels. The mechanism of SD rat chondrocyte apoptosis is to explore the establishment of a system for culturing rat chondrocytes in vitro. In this paper, rat chondrocytes cultured in vitro were induced by nitric oxide to induce their apoptosis, and the mechanism of strychnine, the main monomer component of Strychnine, was observed in order to explore the mechanism of strychnine on the apoptosis of SD rat chondrocytes in vitro. The experimental results showed that strychnine induced apoptosis of chondrocytes, and the survival rate of cells during 36h and 48h after administration was 99% and 97%, respectively. Its apoptotic rate decreases with the increase of the dose and time of strychnine.

1748-P: Brucine Topical Administration Promotes Wound Healing through Downregulation of Proinflammatory Markers and Chemokine Expressions in Alloxan-Induced Diabetic Mice

Diabetes mellitus is a complex metabolic disease and delayed wound healing constitutes one of the most serious diabetes-associated complications. Diabetes can impede the wound healing process by inducing inflammation which can lead to delayed maturation of granulation tissues and reduced wound parallel tension. Brucine, one of the major constituents in Strychnos nux-vomica has antioxidant and anti-inflammatory properties. In this study, we investigated the effects of topical administration of brucine on the key mediators of wound healing, namely T cell subsets, glucocorticoid-induced tumor necrosis factor receptor expressing cells, CD4+CD25+Foxp3+ regulatory T (Treg) cells, Th17 cells, Th1 cytokines, and inflammatory mediator mRNA expression.In current study, 5 groups of mice were used (10/group): group 1, the nondiabetic normal control; group 2, wound in nondiabetic mice; group 3, wound in diabetic mice; group 4, brucine 20mg/kg treated wounds in diabetic mice. Wound size was recorded on every third day and after 14 days of treatment, the heparinized whole blood and the wound tissue of all the groups was collected and tested. Brucine administration showed a significant reduction in wound size, the levels of GITR-expressing cells, and Th1 cytokines as well as substantial down regulation of mRNA expression of the inflammatory mediators compared with the diabetic mice. Brucine also significantly up regulated the number of Tregs or also induced Th17/Treg balance and modulated various pro-inflammatory and anti-inflammatory cytokines and the mRNA expression of their mediators. Furthermore, histopathological examination showed complete re-epithelization, decreased inflammatory cells and presence of granulation tissue in the brucine treated mice. Therefore, our results suggest the beneficial effect of brucine in enhanced wound healing and rebalancing the wound environment in diabetes. Disclosure H. Madkhali: None. M. Ganaie: None. M.N. Ansari: None.

Alkaloids from nux vomica suppresses colon cancer cell growth through Wnt/β-catenin signaling pathway.

Brucine and Strychnine are alkaloids isolated from the seeds of Strychnos nux vomica L., which have long been used as a traditional medicine for the treatment of tumor. However, the effect of Brucine and Strychnine on colorectal cancer (CRC) and the underlying molecular mechanism remain unclear. In the present study, Brucine and Strychnine displayed profound inhibitory effects on the growth of human colon cancer cells. The results of flow cytometric analysis demonstrated that the two alkaloids induced cellular apoptosis. Moreover, the growth of DLD1 xenografted tumors in nude mice was significantly suppressed in the Brucine or Strychnine treated group. Mechanistically, the Wnt/β-catenin is involved in this phenomenon, which is characterized by significantly increased expression of DKK1 and APC, whereas decreased expression of β-catenin, c-Myc, and p-LRP6 in CRC cells as well as tumor tissues. Collectively, Brucine and Strychnine have targeted inhibition for colon cancer proliferation both in vitro and in vivo, and it is valuable for future exploitation and utilization as an antitumor agent of CRC.

Effects of Brucine on the OPG/RANKL/RANK Signaling Pathway in MDA-MB-231 and MC3T3-E1 Cell Coculture System.

The present study examined the effects of brucine on the OPG/RANKL/RANK signaling pathway for exploring the mechanism of brucine suppression of bone metastasis in breast cancer. MDA-MB-231 breast cancer cells and mouse osteoblast MC3T3-E1 cells were cocultured to mimic the breast cancer bone metastasis microenvironment in vitro. qRT-PCR and Western blotting were used to detect the expressions of OPG and RANKL at the mRNA and protein levels, respectively, in brucine-treated cultures and they were compared to those in untreated cultures. We aimed to understand the effect of brucine on the entire OPG/RANKL/RANK signaling pathway after analyzing these effects. Results showed that brucine treatment significantly increased both the OPG mRNA/RANKL mRNA expression ratio and the OPG protein/RANKL protein ratio in cocultures compared to those in untreated cocultures (P < 0.01). Brucine, therefore, plays a regulatory role in the OPG/RANKL/RANK signaling pathway, suggesting that it can indirectly control osteoclasts by regulating the expression and secretion of OPG and RANKL in osteoblast cells, thereby inhibiting the differentiation and bone resorption function of osteoclasts.

Binding of brucine to human serum albumin

The feature of brucine binding to human serum albumin (HSA) was investigated via fluorescence and UV/vis absorption spectroscopy. The results revealed that brucine caused the fluorescence quenching of HSA by the formation of brucine–HSA complex. The hydrophobic interaction plays a major role in stabilizing the complex; the binding site number n and apparent binding constant K A, corresponding thermodynamic parameters the free energy change (Δ G), enthalpy change (Δ H) and entropy change (Δ S) at different temperatures were calculated. The distance r between donor (HSA) and acceptor (brucine) was obtained according to fluorescence resonance energy transfer. The effect of brucine on the conformation of HSA was analyzed using synchronous fluorescence spectroscopy and UV/vis absorption spectroscopy.

Méthode nouvelle de séparation des sulfates de strychnine et brucine

The effect of brucine on the reduction of strychnine to hydrostrychnine has been studied and a new method proposed for the separation of brucine and strychnine sulphates before determination of the strychnine by reduction.