Induction of Apoptosis and Modulation of Caspase Activity on MCF-7 Human Breast Cancer Cells by Bioactive Fractionated Cocoa Leaf Extract.

BACKGROUND: The objective of this study was to investigate the potential anti-proliferative activities of a methanolic extract of cocoa leaves (CL) obtained through sequential partition and fractionation against MCF-7 breast cancer cells.  Methods: The methanolic extract of CL was partitioned in three separated solvents (hexane, dichloromethane, and methanol). Hexane partition was the most potent against MCF-7 cells growth with the lowest IC50 value. Then, it was subjected to two fractionation procedures, resulting in the identification of the CL bioactive fraction (II-F7) with potent toxicity against MCF-7 cells. RESULTS: Further investigation into CL bioactive fraction (II-F7) revealed significant dose-dependent growth inhibitory effects on MCF-7 cells, which were attributed to the induction of apoptosis, as evidenced by the presence of apoptotic bodies, fragmented DNA, and disruption of mitochondrial membrane potential. Additionally, treatment with CL bioactive fraction (II-F7) upregulated the expression of pro-apoptotic genes (DDIT3, GADD45G and HRK) and significantly increased the activities of caspase-8 and caspase-9. CONCLUSION: Overall, this study suggests that bioactive fraction (II-F7) from CL extract has significant and selective cytotoxicity against MCF-7 cells through inducing apoptosis and has potential as a therapeutic agent for breast cancer treatment.

Permixon®, hexane-extracted Serenoa repens, inhibits human prostate and bladder smooth muscle contraction and exerts growth-related functions in human prostate stromal cells.

AIMS: Recently, the European Association of Urology recommended hexane-extracted fruit of Serenoa repens (HESr) in their guidelines on management of non-neurogenic male lower urinary tracts symptoms (LUTS). Despite previously lacking recommendations, Permixon® is the most investigated HESr in clinical trials, where it proved effective for male LUTS. In contrast, underlying mechanisms were rarely addressed and are only marginally understood. We therefore investigated effects of Permixon® on human prostate and detrusor smooth muscle contraction and on growth-related functions in prostate stromal cells. MAIN METHODS: Permixon® capsules were dissolved using n-hexane. Contractions of human prostate and detrusor tissues were induced in organ bath. Proliferation (EdU assay), growth (colony formation), apoptosis and cell death (flow cytometry), viability (CCK-8) and actin organization (phalloidin staining) were studied in cultured human prostate stromal cells (WPMY-1). KEY FINDINGS: Permixon® inhibited α1-adrenergic and thromboxane-induced contractions in prostate tissues, and methacholine-and thromboxane-induced contractions in detrusor tissues. Endothelin-1-induced contractions were not inhibited. Neurogenic contractions were inhibited in both tissues in a concentration-dependent manner. In WPMY-1 cells, Permixon® caused concentration-dependent breakdown of actin polymerization, inhibited colony formation, reduced cell viability, and proliferation, without showing cytotoxic or pro-apoptotic effects. SIGNIFICANCE: Our results provide a novel basis that allows, for the first time, to fully explain the ubiquitous beneficial effects of HESr in clinical trials. HESr may inhibit at least neurogenic, α1-adrenergic and thromboxane-induced smooth muscle contraction in the prostate and detrusor, and in parallel, prostate stromal cell growth. Together, this may explain symptom improvements by Permixon® in previous clinical trials.

[Our experience of using hexane extract of Serenoa Repens fruit (Permixone) in the treatment of LUTS in clinical practice].

Benign prostatic hyperplasia (BPH) and chronic prostatitis (CP) are among the most common causes of LUTS in men. LUTS occur on average in 62.5% of males, with irritative symptoms occurring in 51%, obstructive symptoms in 26%, and postmicturic symptoms in 17% of cases. According to the literature, moderate and severe LUTS were observed in 13% of men under the age of 50 and in 28% after 70 years. It should be noted that LUTS are not BPH-specific, since may be caused by prostate inflammation. According to publications, 57% of patients examined for CP have BPH, and 39% of patients with BPH have CP. The first line of drug therapy in patients with BPH is currently considered to be alpha-blockers and inhibitors of 5-a-reductase. In addition, it is possible to use herbal preparations made from fruits, roots, seeds, pollen and plant bark. Preparations based on Serenoa repens extract are among the most studied and are widely used both in our country and worldwide for the treatment of patients with BPH and LUTS. Only lipidosterol hexane extract of Serenoa repens is recognized as a drug, the use of which has a good evidence base. The clinical examples illustrating the pharmacological properties and results of the use of the preparation of lipidosterol hexane extract Serenoa repens are presented in the article. CONCLUSION: The presented clinical cases demonstrate the efficiency of the hexane extract of Serenoa repens fruit for the treatment of LUTS associated with BPH and CP. At the same time, the drug can be effectively used both as monotherapy and in combination with alphablockers. Therefore, it is reasonable to use the hexane extract of Serenoa repens fruit in clinical practice for the treatment of LUTS associated with BPH.

[Iscussion on the use of hexane extract of Serenoa repens for combination therapy in patients with LUTS/BPH].

Based on literature data, feasibility of combined therapy with Serenoa repens extracts (including Permixon) and 1-adrenoblocators in patients with LUTS/BPH based on the pathogenesis of these drugs and clinical results was analyzed. The composition and biological activity of various SRE is influenced by the extraction method, as well as the production technology used by the manufacturer, which can lead to different results of experimental and clinical studies. The mechanism of action of SRE, including hexane extract Permixon, is multicomponent, which combines an antiandrogenic effect, influence on neurogenic regulation of urinary tract function (-adrenoreceptors, M-holinoreceptors, vanilloid receptors), anti-inflammatory and anti-edematous actions. At the same time, the effect of these preparations is not limited to the influence on the prostate gland, but can directly affect the bladder, helping to reduce functional disorders. A wider range of mechanisms of action and a possible direct effect of SRE on the bladder provides the same clinical effect as other traditionally used drugs (-blockers, 5-reductase inhibitors) with a significantly lower frequency of side effects of therapy. A number of publications demonstrate the feasibility of combined therapy with 1-adrenoblockers and SRE in patients with LUTS/BPH, especially with a high degree of urination dysfunction. Futher investigations are needed. The role of Serenoa repens extracts in the combination therapy of LUTS/BPH is still under discussion. Serenoa repens extracts monotherapy demonstrates similar effectiveness in patients with LUTS as 1-adrenoblockers and 5-reductase inhibitors. Recent studies indicate the advantages of combining Serenoa repens extracts with 1-adrenoblockers, especially in patients with moderate or severe symptoms. These advantages are associated with the multicomponent action of Serenoa repens extracts, which complements the blockade of 1-adrenergic receptors with other mechanisms of action (in particular, anti-inflammatory). The combination of drugs with different mechanisms of action, in particular, Serenoa repens extracts (Permixon) and 1-adrenoblockers has certain pathophysiologically based advantages, which allows to increase the efficiency of therapy. In addition, combination therapy is not associated with an increased rate of side effects.

Ameliorative effects of Magnolia sieboldii buds hexane extract on experimental reflux esophagitis.

Gastroesophageal reflux disease (GERD) is a disease that stomach contents continually refluxing into esophagus causes symptoms and/or complications. The study was working to find natural plant extracts with good effects and small side effects to treat reflux esophagitis (RE). The anti-inflammatory effects of hexane extract of Magnolia sieboldii (MsHE) were conducted on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The ameliorative effects of MsHE on esophageal damage in rats induced by gastric acid reflux was explored in vivo. The results showed that MsHE decreased the production of nitric oxide (NO) and expression levels of iNOS, COX-2 and TNF-α on LPS-stimulated RAW 264.7 cells and MsHE treatment ameliorated the rats' esophageal tissue damage induced by gastric acid and inhibited the increase of inflammatory mediators and pro-inflammatory cytokines by regulating NF-κB signaling pathway. In addition, MsHE protected the function of barrier of epithelial cells against inflammatory conditions through increasing the expression of tight junctions. Furthermore, liquid chromatography-mass spectrometry analysis was used for determine the active ingredients contained in MsHE. The results show that MsHE can alleviate experimental rat RE by regulating NF-κB signaling pathway. In summary, MsHE may be used as a source material of drug candidate for the treatment of RE.

Hexane fraction of Annona muricata (Sour sop) seed ameliorates testosterone-induced benign prostatic hyperplasia in rats.

BACKGROUND: Annona muricata is used in traditional African medicine to manage urinary obstruction. In this study, we hypothesized that hexane fraction of Annona muricata (HFAM) seeds will ameliorate testosterone propionate (Tp)-induced benign prostatic hyperplasia (BPh). METHODS: Castrated rats were assigned into six groups: non-castrated control, castrated control, castrated rats that received Tp (BPh group), [BPh+HFAM], [BPh+HFAM + finasteride], [BPh + finasteride]. RESULTS: The BPh rats had 3.8 and 3.9 folds increases in prostatic and organo-somatic weight, while treatment with HFAM alone and [HFAM + finasteride] decreased prostatic weight by 22% and 34%, respectively. BPh increased the activities of serum and prostatic total acid phosphatase by 95% and 121%; and activities of serum and prostatic alkaline phosphatase by 54% and 281%, respectively. Serum and prostatic lipid peroxidation were increased by 44% and 82%, respectively, in BPh rats with a concomitant decrease in prostatic superoxide dismutase by 73%. In BPh rats, serum and prostatic myeloperoxidase increased by 4.0 and 2.0 folds, while serum nitric oxide increased by 2.4 folds, respectively. Strong expression of inducible nitric oxide synthase, Bcl2, beta-catenin, androgen and estrogen receptors were observed in BPh rats. Importantly, treatment with HFAM or finasteride (or combination) attenuated prostatic weight, inflammatory and antioxidant indices in BPh rats. CONCLUSION: HFAM may serve as novel therapeutic agent against BPh.

Hematological and lipid profile evaluation of a hexane fraction of Costus afer leaves in arthritic rats.

CONTEXT: Costus afer Ker Gawl. (Costaceae) is an ethnomedical plant used as therapy against inflammatory disorders. OBJECTIVE: The objective of this study was to evaluate the hematological and lipid profile analysis of hexane fraction of C. afer leaves (CAHLF) in arthritic rats. MATERIALS AND METHODS: Male albino rats were randomly distributed into seven groups of six rats each. Rats were induced with arthritis using formaldehyde and Complete Freund's Adjuvant (CFA) for 7 and 21 d, respectively. The animals were administered orally with 50, 100, and 250 mg/kg CAHLF, 10 mg/kg diclofenac and prednisolone, 0.9% NaCl (control), and 0.9% NaCl (normal). At the end of treatment periods, blood samples were withdrawn and subjected to hematological and biochemical analysis using auto-analyzer and spectrophotometric methods. RESULTS: Hematological analysis revealed that in formaldehyde- and CFA-induced arthritic rat models, 250 mg/kg CAHLF-treated groups had significantly reduced (p < 0.05) hematocrit counts (HC) (30.98 ± 1.59% and 33.55 ± 1.10%), white blood cell counts (WBC) (5.50 ± 0.35 and 4.15 ± 0.82 × 10(9)/L), and platelet counts (PC) (401.50 ± 48.94 and 246.33 ± 5.54 × 10(9)/L) compared with control HC (46.90 ± 1.92 and 41.88 ± 2.19%), WBC (11.09 ± 0.26 and 7.37 ± 0.34 × 10(9)/L), and PC (783.67 ± 59.51 and 593.83 ± 36.3 × 10(9)/L). Furthermore, blood analysis showed that CAHLF-treated groups had reduced total cholesterol, low-density lipoprotein cholesterol, and triglycerides while they had an elevated high-density lipoprotein compared with the control group. DISCUSSION AND CONCLUSION: Findings from this study indicated that CAHLF could possess immunomodulatory and hypolipidemic properties in arthritic rats. CAHLF could be considered as a source of biopharmaceutical agents in anti-arthritis drug discovery process.

Discovery and synthesis of a new class of opioid ligand having a 3-azabicyclo[3.1.0]hexane core. An example of a 'magic methyl' giving a 35-fold improvement in binding.

In looking for a novel achiral µ opioid receptor antagonist for the treatment of pruritus, we designed and synthesised azabicyclo[3.1.0]hexane compounds as a new class of opioid ligand. During optimisation, an addition of a single methyl resulted in a 35-fold improvement in binding. An early example from the series had excellent µ opioid receptor antagonist antagonist activity and was very effective in an in vivo pruritus study.