Berberine Inhibits Human Hepatoma Cell Invasion without Cytotoxicity in Healthy Hepatocytes

Conventional chemotherapy fails to cure metastatic hepatoma mainly due to its high hepatotoxicity. Many plant-derived agents have been accepted to effectively inhibit hepatoma cell invasion. However, the investigation that whether effectual plant-derived agents against invasive hepatoma cells exert unexpected cytotoxicity in healthy hepatocytes has been ignored. This study demonstrated that berberine exhibited significant cytotoxicity in HepG2 cells mainly through upregulation of reactive oxygen species (ROS) production but was ineffective in normal Chang liver cells. Berberine exerted anti-invasive effect on HepG2 cells through suppression of matrix metalloproteinase-9 (MMP-9) expression. Moreover, berberine could significantly inhibit the activity of PI3K-AKT and ERK pathways. Combination treatment of ERK pathway inhibitor PD98059 or AKT pathway inhibitor LY294002 and berberine could result in a synergistic reduction on MMP-9 expression along with an inhibition of cell invasion. Enhancement of ROS production by berberine had no influence on its suppressive effects on the activity of PI3K-AKT and ERK pathways, as well as MMP-9 expression and HepG2 cell invasion. In conclusion, our results suggest that berberine may be a potential alternative against invasive hepatoma cells through PI3K-AKT and ERK pathways-dependent downregulation of MMP-9 expression. This study also provides a previously neglected insight into the investigation of plant-derived agents-based therapy against tumor invasion with the consideration o

Apigenin Inhibits Human SW620 Cell Growth by Targeting Polyamine Catabolism

Apigenin is a nonmutagenic flavonoid that has antitumor properties. Polyamines are ubiquitous cellular polycations, which play an important role in the proliferation and differentiation of cancer cells. Highly regulated pathways control the biosynthesis and degradation of polyamines. Ornithine decarboxylase (ODC) is the rate-limiting enzyme in the metabolism, and spermidine/spermine-N1-Acetyl transferase (SSAT) is the rate-limiting enzyme in the catabolism of polyamines. In the current study, the effect of increasing concentrations of apigenin on polyamine levels, ODC and SSAT protein expression, mRNA expression, cell proliferation and apoptosis, and the production of reactive oxygen species (ROS) was investigated in SW620 colon cancer cells. The results showed that apigenin significantly reduced cell proliferation, decreased the levels of spermidine and spermine, and increased previously downregulated putrescine contents. Apigenin also enhanced SSAT protein and mRNA levels and the production of reactive oxygen species in SW620 cells, though it had no significant effect on the levels of ODC protein or mRNA. Apigenin appears to decrease the proliferation rate of human SW620 cells by facilitating SSAT expression to induce polyamine catabolism and increasing ROS levels to induce cell apoptosis

Sinisan ameliorates colonic injury induced by water immersion restraint stress by enhancing intestinal barrier function and the gut microbiota structure

AbstractContext Sinisan (SNS) has been used to treat psychosomatic diseases of the digestive system. But little is known about how SNS affects water immersion restraint stress (WIRS).Objective To study the effects of SNS on colonic tissue injury in the WIRS model.Materials and methods Forty-eight Kunming (KM) mice were randomized into 6 groups (n = 8): The control and WIRS groups receiving deionized water; the SNS low-dose (SL, 3.12 g/kg/d), SNS middle-dose (SM, 6.24 g/kg/d), SNS high-dose (SH, 12.48 g/kg/d), and diazepam (DZ, 5 mg/kg/d) groups; each with two daily administrations for 5 consecutive days. The 5 treatment groups were subjected to WIRS for 24 h on day 6. The effects of SNS on colon tissue injury caused by WIRS were assessed by changes in colon histology, inflammatory cytokines, brain-gut peptides, and tight junction (TJ) proteins levels. 16S rRNA gene sequencing was used to detect the regulation of the gut microbiota.Results SNS pretreatment significantly reduced TNF-α (0.75- to 0.81-fold), IL-6 (0.77-fold), and IFN-γ (0.69-fold) levels; and increased TJ proteins levels, such as ZO-1 (4.06- to 5.27-fold), claudin-1 (3.33- to 5.14-fold), and occludin (6.46- to 11.82-fold). However, there was no significant difference between the levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the control and WIRS groups. SNS regulated the composition of gut microbiota in WIRS mice.Conclusion The positive effects of SNS on WIRS could provide a theoretical basis to treat stress-related gastrointestinal disorders

Neuroprotective effects of a standardized flavonoid extract from diospyros kaki leaves

Ethnopharmacological relevance: Flavonoids, extracted from the leaves of Diospyros kaki, are the main therapeutic components of NaoXingQing (NXQ), a potent and patented Chinese herbal remedy widely used in China for the treatment of apoplexy syndrome. Aim of the study: To investigate the neuroprotective effects of FLDK-P70, a standardized flavonoid extract, using in vivo rat models of both focal ischemia/reperfusion (I/R) injury induced by middle cerebral artery occlusion (MCAO) and on transient global brain ischemia induced by four-vessel occlusion (4-VO). We also aim to examine the effects of FLDK-P70 on glutamate-induced cell injury of hippocampal neurons as well as on hypoxia-induced injury of cortical neurons in primary cell culture. Materials and methods and results: Administration of FLDK-P70 for 12 days (40, 80 mg/kg body weight, p.o., 5 days before and 7 days after 4-VO) increased the survival of hippocampal CA1 pyramidal neurons after transient global brain ischemia. Similarly, administration of FLDK-P70 for 7 days (40, 80 mg/kg body weight, p.o., 3 days before and 4 days after MCAO) significantly reduced the lesion of the insulted brain hemisphere and improved the neurological behavior of rats. In primary rat hippocampal neuronal cultures, pretreatment with FLDK-P70 (5, 10 g/ml) protected neurons from glutamate-induced excitotoxic neuronal death in a dose-dependent manner. In primary rat cerebral cortical neuronal culture, pretreatment with FLDK-P70 (25, 100 g/ml) also reduced hypoxia-reoxygen induced neuronal death and apoptosis in a dose-dependent manner. Conclusions: These in vivo and in vitro results suggest that FLDK-P70 significantly protects rats fromMCAO and 4-VO ischemic injury in vivo and protects hippocampal neurons from glutamate-induced excitotoxic injury as well as cortical neurons from hypoxia-induced injury in vitro. The mechanisms of these effects may be due to the antioxidative activity of the flavonoids. These results convincingly demonstrate that FLDK-P70 may be useful for the prevention and treatment of ischemia/reperfusion injury and other related neurodegenerative diseases.No Full Tex

Radiosynthesis and Biological Evaluation of <i>N</i>-[¹⁸F]Labeled Glutamic Acid as a Tumor Metabolic Imaging Tracer

<div><p>We have previously reported that <i>N</i>-(2-[¹⁸F]fluoropropionyl)-L-methionine ([¹⁸F]FPMET) selectively accumulates in tumors. However, due to the poor pharmacokinetics of [¹⁸F]FPMET <i>in vivo</i>, the potential clinical translation of this observation is hampered. In this study, we rationally designed and synthesized [¹⁸F] or [¹¹C]labeled <i>N</i>-position L-glutamic acid analogues for tumor imaging. <i>N</i>-(2-[¹⁸F]fluoropropionyl)-L-glutamic acid ([¹⁸F]FPGLU) was synthesized with a 30±10% (n = 10, decay-corrected) overall radiochemical yield and a specific activity of 40±25 GBq/μmol (n = 10) after 130 min of radiosynthesis. <i>In vitro</i> cell experiments showed that [¹⁸F]FPGLU was primarily transported through the X_AG^– system and was not incorporated into protein. [¹⁸F]FPGLU was stable in urine, tumor tissues, and blood. We were able to use [¹⁸F]FPGLU in PET imaging and obtained high tumor to background ratios when visualizing tumors tissues in animal models.</p></div

Radio-HPLC analysis the stability of [¹⁸F]FPGLU.

<p>HPLC chromatograms of tumor tissue extract (A) and plasma (B) (S180 fibrosarcoma-bearing mice at 0.5 h after intravenous injection of [¹⁸F]FPGLU). HPLC chromatograms of urine (C) (S180 fibrosarcoma-bearing mice at 1 h after intravenous injection of [¹⁸F]FPGLU). HPLC chromatograms of [¹⁸F]FPGLU in mouse serum at 37°C for 2 h (D). (The peak at T = 5.5 min was [¹⁸F]FPGLU.).</p

Scheme of the radiosynthesis of [¹⁸F]FPGLU.

<p>Scheme of the radiosynthesis of [¹⁸F]FPGLU.</p

Small animal PET imaging and quantification.

<p>Decay-corrected whole-body PET images were acquired at different time points. (A) PET images of S180 fibrosarcoma-bearing mouse static scans at 0.5, 1, 1.5, and 2 h after the injection of [¹⁸F]FPGLU. The same S180 fibrosarcoma-bearing mouse static scan at 1 h after injection of [¹⁸F]FDG. (The red arrows indicate the tumor.) (B) PET images of SPCA-1 or LTEP-a-2 human lung adenocarcinoma-bearing nude mouse static scans at 1 h after the injection of [¹⁸F]FPGLU or [¹⁸F]FDG. (The red arrows indicate the tumor.) (C) <b>A</b> comparison of tumor uptake of [¹⁸F]FPGLU and [¹⁸F]FDG in S180 fibrosarcoma, LTEP-a-2, and SPCA-1 human lung adenocarcinoma at 1 h after injection. (n = 3 per group; bars represent means ± SD.).</p

Biodistribution of [¹⁸F]FPGLU in Normal Mice<i>^a.</i>

a<p>Means ± SD (<i>n = </i>4). Data are average % ID/g.</p