Pterostilbene suppresses gastric cancer proliferation and metastasis by inhibiting oncogenic JAK2/STAT3 signaling: In vitro and in vivo therapeutic intervention.

BACKGROUND: Gastric cancer (GC) represents a significant health burden with dire prognostic implications upon metastasis and recurrence. Pterostilbene (PTE) has been proven to have a strong ability to inhibit proliferation and metastasis in other cancers, while whether PTE exhibits anti-GC activity and its potential mechanism remain unclear. PURPOSE: To explore the efficacy and potential mechanism of PTE in treating GC. METHODS: We employed a comprehensive set of assays, including CCK-8, EdU staining, colony formation, flow cytometry, cell migration, and invasion assays, to detect the effect of PTE on the biological function of GC cells in vitro. The xenograft tumor model was established to evaluate the in vivo anti-GC activity of PTE. Network pharmacology was employed to predict PTE's potential targets and pathways within GC. Subsequently, Western blotting, immunofluorescence, and immunohistochemistry were utilized to analyze protein levels related to the cell cycle, EMT, and the JAK2/STAT3 pathway. RESULTS: Our study demonstrated strong inhibitory effects of PTE on GC cells both in vitro and in vivo. In vitro, PTE significantly induced cell cycle arrest at G0/G1 and S phases and suppressed proliferation, migration, and invasion of GC cells. In vivo, PTE led to a dose-dependent reduction in tumor volume and weight. Importantly, PTE exhibited notable safety, leaving mouse weight, liver function, and kidney function unaffected. The involvement of the JAK2/STAT3 pathway in PTE's anti-GC effect was predicted utilizing network pharmacology. PTE suppressed JAK2 kinase activity by binding to the JH1 kinase structural domain and inhibited the downstream STAT3 signaling pathway. Western blotting confirmed PTE's inhibition of the JAK2/STAT3 pathway and EMT-associated protein levels. The anti-GC effect was partially reversed upon STAT3 activation, validating the pivotal role of the JAK2/STAT3 signaling pathway in PTE's activity. CONCLUSION: Our investigation validates the potent inhibitory effects of PTE on the proliferation and metastasis of GC cells. Importantly, we present novel evidence implicating the JAK2/STAT3 pathway as the key mechanism through which PTE exerts its anti-GC activity. These findings not only establish the basis for considering PTE as a promising lead compound for GC therapeutics but also contribute significantly to our comprehension of the intricate molecular mechanisms underlying its exceptional anti-cancer properties.

Thymoquinone induces apoptosis and protective autophagy in gastric cancer cells by inhibiting the PI3K/Akt/mTOR pathway.

Gastric cancer (GC) is often diagnosed in the advanced stages with a poor prognosis. Thymoquinone (TQ) is known for its antitumor activity; however, the specific mechanism in GC remains unknown. In our study, TQ inhibited GC cell proliferation and induced apoptosis and autophagy in a concentration-dependent manner. Transmission electron microscopy showed increased autophagosome formation in GC cells treated with TQ. Meanwhile, the LC3B puncta and LC3BII protein levels were significantly increased in GC cells, while p62 expression was significantly decreased. The autophagy inhibitor, Bafilomycin A1 enhanced TQ-inhibited proliferation and TQ-induced apoptosis, suggesting that TQ-induced autophagy has a protective effect on GC cells. Furthermore, TQ decreased the phosphorylation levels of phosphatidylinositol-4,5-bisphosphate 3 kinase (PI3K), protein kinase B (Akt), and mechanistic target of rapamycin (mTOR). The PI3K agonist partially rescued TQ-induced autophagy and apoptosis. Finally, in vivo experiments showed that TQ could inhibit tumor growth and promote apoptosis and autophagy. This study provides new insights into the specific mechanism for the anti-GC effect of TQ. TQ inhibits the proliferation of GC cells and induces apoptosis and protective autophagy by inhibiting the PI3K/Akt/mTOR pathway. The results suggest that the combination of TQ and autophagy inhibitors might be a potential chemotherapeutic strategy for GC.

Adjunctive therapeutic effects of micronutrient supplementation in inflammatory bowel disease.

Growing evidence suggests that micronutrient status may have some impact on the course of inflammatory bowel disease (IBD). However, micronutrient deficiencies are easily overlooked during the treatment of IBD patients. There have been many studies on micronutrient supplementation, in which several clinical trials have been conducted targeting vitamin D and iron, but the current research is still preliminary for other vitamins and minerals. This review provides an overview of the adjunctive therapeutic effects of micronutrient supplementation in IBD, to summarize the available evidence, draw the attention of clinicians to micronutrient monitoring and supplementation in patients with IBD, and also provide some perspectives for future research directions.

Corydalis decumbens Alleviates the Migration, Phagocytosis, and Inflammatory Response of Macrophages.

Background: The role of Corydalis decumbens (CD) in macrophage activation remains unclear, particularly in the Ras homolog family member A (RhoA) signaling pathway. Therefore, the present study aimed to investigate the effect of CD on the viability, proliferation, morphological changes, migration, phagocytosis, differentiation, and release of inflammatory factors and signaling pathways in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Methods: Cell counting kit-8 and water-soluble tetrazolium salt assays were used to evaluate the viability and proliferation of RAW264.7 macrophages. A transwell assay was examined to assess cell migration. The ingestion of lumisphere assay was employed to detect the phagocytic capacity of macrophages. Phalloidin staining was performed to observe morphological changes in the macrophages. An enzyme-linked immunosorbent assay was performed to quantify inflammation-related cytokines in cell culture supernatants. Cellular immunofluorescence and western blotting were adopted to show the expression of inflammation-related factors, biomarkers of M1/M2 subset macrophages, and factors of the RhoA signaling pathway. Results: We found that CD increased the viability and proliferation of RAW264.7 macrophages. CD also impaired the migration and phagocytic capacity of macrophages, induced anti-inflammatory M2 macrophage polarization, such as M2-like morphological changes, and upregulated M2 macrophage biomarkers and anti-inflammatory factors. We also observed that CD inactivated the RhoA signaling pathway. Conclusions: CD mediates the activation of LPS-stimulated macrophages, alleviates the inflammatory responses of macrophages, and activates related signaling pathways induced by LPS.

[Effect of electroacupuncture on complement C1q and microglia phagocytosis in hippocampus of SAMP8 mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of Iba-1, complement C1q and CD68 in hippocampus of SAMP8 mice, so as to explore its mechanisms underlying improvement of Alzheimer's disease (AD). METHODS: Twenty-four male SAMP8 mice were randomly and equally divided into model and EA groups, and 12 SAMR1 mice were used as the control group. EA (2 Hz, 1.5-2.0 mA) was applied to "Baihui" (GV20), "Dazhui"(GV14) and "Shen-shu"(BL23) for 20 min once daily in the EA group, each course of treatment was 8 days, with an interval of 2 days between two courses, and the mice were treated for 3 courses. Morris water maze test was performed to assess the learning-memory ability of mice. The positive expression levels of Iba-1 and CD68 proteins in the hippocampus CA1 region were detected by immunohistochemistry. The mRNA and protein expression levels of Iba-1,C1q and CD68 in the hippocampus were detected by real-time PCR and Western blot, separately. RESULTS: Compared with the control group, the average escape latency of Morris water maze test was prolonged in the model group (P<0.01), duration of swimming in the original platform quadrant and the number of original platform crossing were significantly shorter and decreased respectively (P<0.01). Compared with the model group, the average escape latency in the EA group was shortened (P<0.05, P<0.01), the duration of swimming in the original platform quadrant and the number of original platform crossing were significantly prolonged and increased (P<0.01). The immunoactivity of Iba-1 and CD68 in hippocampal CA1 region, and mRNA and protein expression levels of hippocampal Iba-1,C1q and CD68 were significantly up-regulated in the model group in contrast to the control group (P<0.01, P<0.05), and obviously down-regulated except the mRNA expression level of hippocampal Iba-1 in the EA group relevant to the model group (P<0.01, P<0.05). CONCLUSION: EA can improve the learning and memory ability of SAMP8 mice, which may be associated with its effect in inhibiting of complement C1q-dependent microglial phagocytosis in the hippocampus.

Effects of dietary supplementation with itaconic acid on the growth performance, nutrient digestibility, slaughter variables, blood biochemical parameters, and intestinal morphology of broiler chickens.

Itaconic acid (IA) is a biologically based unsaturated dicarboxylic acid secreted by mammalian cells. While IA has potential for use in multiple applications, information regarding the influence of IA on animal production remains scarce. This study investigated the effects of dietary IA supplementation on the growth performance, nutrient digestibility, slaughter variables, blood parameters, and intestinal morphology of broiler chickens. A total of 360 one-day-old Arbor Acre broiler chicks were allotted to 6 groups, with 10 chicks per cage and 6 replicates per group in a randomized complete block design. Broiler chicks were fed a basal diet with 0 (control), 0.2, 0.4, 0.6, 0.8, or 1.0% IA. The experimental period lasted from 1 to 42 d of age. Dietary IA supplementation did not affect average daily gain (ADG) and feed/gain ratio (F/G) but quadratically increased average daily feed intake (ADFI) and linearly increased crude protein (CP) digestibility during the grower period (d 22-42). A higher breast and thigh muscle yield and a lower abdominal fat yield were observed in a linear and quadratic manner with the IA supplementation. Adding IA to the diet had significant effects on superoxide dismutase (SOD), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and catalase (CAT) levels in serum at d 21 and on total antioxidation capacity (T-AOC) at d 42. There were linear and quadratic increases in villus height and the villus height/crypt depth ratio (V/C) of the duodenum and villus height of the jejunum with the supplementation of IA. Regression analyses for ADFI, dressed yield, breast and thigh muscle yield, abdominal fat yield, serum ALT, CAT, and SOD levels, villus length of the duodenum and jejunum, and V/C of the duodenum indicated that the optimal dietary IA supplementation would be from 0.4 to 0.7%. From an economic perspective, a level of 0.4% IA in the broiler diet is recommended for improving the nutrient digestibility, slaughter performance, antioxidant ability, and intestinal morphology of broiler chickens.

Zingiberene targets the miR-16/cyclin-B1 axis to regulate the growth, migration and invasion of human liver cancer cells.

PURPOSE: Liver cancer or hepatocellular carcinoma (HCC) is considered as one of the most frequent malignancies with significantly high morbidity and mortality across the globe. MicroRNAs (miRs) are regarded as important regulators of liver cancer formation and its development. However, the full biochemical mechanism of their role is still very less understood. The main objective of the current research work was to examine the role of miR-16/cyclin-B1 axis in liver cancer regulation and how this pathway along with liver cancer migration and invasion are targeted by zingiberene molecule. METHODS: Quantitative reverse transcriptase polymerase chain reaction was used to evaluate miR-16 expression in HCC cell lines. Western blotting was performed to evaluate the expression of the miR-16 target genes. Effects on cell migration and invasion were evaluated by in vitro wound healing assay and transwell Matrigel assay, respectively. Effects of zingiberene on HCC cell viability were evaluated by MTT assay. RESULTS: Zingiberene treatment led to downregulation of miR-16 in HepG2 human hepatocellular carcinoma cells, accompanied by induction of G0/G1 cell cycle arrest targeting cyclin B1 as direct target. These effects were also accompanied by inhibition of cell migration and invasion, indicating that miR-16 can have a significant role as liver cancer suppressor after zingiberene treatment. Luciferase reporter assay confirmed that miR-16, which was one of HCC downregulated miRs, directly targeted Cyclin B1 in HCC cells. CONCLUSION: The current study indicates miR-16/cyclin B1 axis might have significant applications as a therapeutic target for patients with liver cancer.

Thymoquinone inhibits the proliferation and invasion of esophageal cancer cells by disrupting the AKT/GSK-3ß/Wnt signaling pathway via PTEN upregulation.

Although thymoquinone (TQ) has been reported to exert antitumor activity against various types of human cancers without evident toxicity, limited studies have reported the effects of TQ on esophageal cancer. Here, we showed that TQ induced cell cycle arrest in the G2/M phase and significantly inhibited cell proliferation and invasion. Further investigation of the potential mechanism revealed that TQ increased the levels of p53 and p21 but significantly reduced the expression of Cyclin B1, Cyclin A, and Cyclin E. Moreover, TQ led to a decrease in Bcl-2 and an increase in cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, and Bax, indicating that TQ induced apoptosis by activating the intrinsic mitochondrial apoptosis pathway. Western blotting showed that TQ disrupted the PI3K/AKT pathway by upregulating PTEN, thus modulating GSK-3ß activity, increasing ß-catenin degradation, and decreasing decreased MMP-2 and MMP-9 levels in Eca109 cells. However, these changes were attenuated by disrupting PTEN function (using a potent inhibitor) or downregulating PTEN expression. In addition, in vivo results showed that the efficacy of TQ as an antitumor agent in a mouse xenograft tumor model. In conclusion, TQ suppressed human esophageal cancer cells proliferation and invasion both in vitro and in vivo and could provide a novel therapeutic approach for esophageal cancer.

[Mechanisms of electroacupuncture for improving Alzheimer's disease from reducing ß amyloid protein level].

Alzheimer's disease (AD), a neurodegenerative disorder characterized by amyloid deposits and neurofibrillary degeneration, is the most common type of dementia and has no incurable therapies at the moment. Electroacupuncture (EA) therapy has been widely used in clinical treatment of AD, and has attained approving effects. This article reviews the development of researches on the mechanisms of EA underlying improving AD by diminishing ß amyloid protein （Aß） neurotoxicity, from 1) up-regulating hippocampal cellular autophagy, 2) improving cerebral energy metabolism by activating oxidation stress-related factors peroxisome proliferator-activated receptor γ coactivator 1 alpha and sirtuin 1 in the hippocampus and frontal cerebral cortex, 3) relieving inflammatory reaction by lowering expression of tumor necrosis factor-alpha and high-mobility group box 1 and increasing expression of Interleukin 10, and 4) promoting degradation of Aß1-42 by down-regulating expression of insulin degeneration enzyme, lipoprotein, transthyretin, apolipoprotein and α2 mcroglobulin. Meanwhile, a comprehensive clinical therapy of AD is proposed.

Hesperetin ameliorates DSS-induced colitis by maintaining the epithelial barrier via blocking RIPK3/MLKL necroptosis signaling.

Hesperetin, a flavonoid from citrus fruits, possess various pharmacological properties, including anti-inflammatory, anti-oxidative, anti-tumor potentials. However, the role and its mechanism in ulcerative colitis (UC) remains unclear. This study aimed to investigate the protective effects and mechanisms of hesperetin on dextran sodium sulfate (DSS) -induced colitis. Our results showed that hesperetin significantly relieved the symptoms of DSS -induced colitis and increased the expressions of zonula occludens-1 (ZO-1), occludin and mucin2 (MUC-2) as well as the decrease of tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß, IL-18, HMGB1 and IL-6. Of note, results from immunohistochemistry (IHC) and western blotting indicated that hesperetin inhibited the expressions of receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like (MLKL), the two key proteins of necroptosis pathway, and inactivated RIPK3/MLKL necroptosis signalling. Meanwhile, in the cell-coculture system between Caco-2 and RAW264.7 cells, hesperetin treatment significantly ameliorated the decrease of trans epithelial electric resistance (TEER) value while HS-173 (necroptosis inducer) could obviously influence the effect of hesperetin. In addition, hesperetin attenuated the LPS-induced increasing in 4-kDa fluorescein isothiocyanate-dextran (FD4) permeability while HS-173 could weaken the protective effect of hesperetin. Meanwhile, HS-173 reduced the changes in the expressions of phosphorylated RIPK3, phosphorylated MLKL, ZO-1, occludin and MUC-2 as well as TNF-α, IL-1ß. These findings demonstrated hesperetin ameliorated DSS-induced colitis by maintaining the epithelial barrier via blocking the intestinal epithelial necroptosis.

[Effect of early intervention of electroacupuncture on learning-memory ability and level of hippocampal phosphorylated Tau protein in SAMP8 mice].

OBJECTIVE: To explore the effect of early intervention electroacupuncture (EA) at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) on the learning-memory ability and the expression of phosphorylated Tau protein in the hippocampus of SAMP8 mice, so as to provide reference for the intervening period of EA for Alzheimer's disease (AD). METHODS: A total of 36 3-month old SAMP8 mice were randomly divided into a model group, a 3-month-old EA group and a 9-month-old EA group, 12 mice in each group. Twelve normal SAMR1 mice with the same age were taken as the control group. The mice in the 3-month-old EA group and 9-month-old EA group were treated with EA at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) separately 3 months old and 9 months old (continuous wave, 2 Hz, 1.5-2 mA), 20 min each time, once a day, 8 days as a course of treatment, with an interval of 2 days between courses, totally 3 courses of treatment were given. The mice sample in each group was collected at the age of 10 months after the learning-memory ability tested by Morris water maze. The expression of phosphorylated Tau protein in the hippocampus was detected by immunohistochemistry and Western blot, and the expression of Tau mRNA was detected by real-time PCR. RESULTS: Compared with the control group, in the model group, the escape latency was significantly increased (P<0.01), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were reduced (P<0.01), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were increased (P<0.01). Compared with the model group, in the 3-month-old EA group and 9-month-old EA group, the escape latency was significantly reduced (P<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (P<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were reduced (P<0.05). Compared with the 9-month-old EA group, in the 3-month-old EA group, the escape latency was significantly reduced (P<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (P<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA were reduced (P<0.01). CONCLUSION: The early EA intervention could more effectively improve the learning-memory ability and inhibit phosphorylation of Tau protein in the hippocampus of SAMP8 mice.

α-Hederin induces the apoptosis of gastric cancer cells accompanied by glutathione decrement and reactive oxygen species generation via activating mitochondrial dependent pathway.

α-Hederin, a monodesmosidic triterpenoid saponin, exhibited promising antitumor potential against a variety of human cancer cell lines. However, few related studies about effects of α-hederin on gastric cancer are available. Herein, our results showed that α-hederin significantly inhibited the proliferation of gastric cancer cells and arrested the cell cycle in G1 phase in vitro (p < .05). Further research of the potential mechanism reflected that α-hederin could induce intracellular glutathione decrement, adenosine triphosphate level, and mitochondrial membrane potential variation via inducing reactive oxygen species accumulation during the apoptosis of gastric cancer cells. Moreover, the detection of mitochondrial and cytosol proteins with apoptosis-inducing factor, apoptosis protease activating factor-1, and cytochrome C showed an increase in the cytosol, followed by a decrease of Bcl-2 levels and increases of caspase-3, caspase-8, caspase-9, and Bax, which revealed that α-hederin induced apoptosis via triggering activation of the mitochondrial pathway. Furthermore, the above changes were amplified when pretreated with buthionine sulfoximine, whereas attenuated in the group pretreated with NAC than α-hederin alone (p < .05). In addition, α-hederin significantly inhibited the growth of xenografted gastric tumors with favorable safety. In conclusion, α-hederin could inhibit the proliferation and induce apoptosis of gastric cancer accompanied by glutathione decrement and reactive oxygen species generation via activating mitochondrial dependent pathway.

Effect of Zinc Supplementation on Semen Quality, Sperm Antioxidant Ability, and Seminal and Blood Plasma Mineral Profiles in Cashmere Goats.

This study evaluated the effects of the different dietary zinc (Zn) levels on semen quality, on spermatozoa and seminal plasma antioxidant status, and on the seminal and blood plasma mineral status in mature male Cashmere goats during the breeding season. Twenty-eight mature male Liaoning Cashmere goats were divided into four groups based on body weight (56.2 ± 2.45 kg) and semen characteristics; these goats were fed with basal diet supplemented with 0, 20, 40, or 80 mg Zn/kg DM (zinc sulfate) for 3 months. Results showed that the Zn-supplemented diets linearly increased the semen volume (0.98, 1.04, 1.27, and 1.17 ml for the 0, 20, 40, and 80 mg Zn/kg DM supplementation, respectively) (P < 0.05) and the total sperm output (3.87, 4.52, 5.73, and 5.33 × 109/ml for the 0, 20, 40, and 80 mg Zn/kg DM supplementation, respectively) (P < 0.05); by contrast, Zn supplementation exerted no effect on sperm concentration, motility, and abnormal sperms rate. The activities of copper zinc superoxide dismutase (CuZn-SOD) (linear P < 0.05) and glutathione peroxidase (GSH-Px) (linear P < 0.05; quadratic P < 0.01) were highest in the intermediate supplementation (40 mg Zn/kg DM). Moreover, the malondialdehyde (MDA) content of spermatozoa decreased linearly (P < 0.01) with the increase in Zn supplementation. In seminal plasma, the highest GSH-Px activity was observed in 20 mg Zn/kg DM supplementation (P < 0.05). Catalase (CAT) activities both in the spermatozoa and seminal plasma showed no difference in all treatments. Seminal plasma Zn level was highest in 40 mg Zn/kg DM (linear P = 0.068), and K increased linearly (P = 0.001) with increasing Zn level. Furthermore, blood plasma Zn (linear P < 0.01; quadratic P < 0.05), Fe (linear P < 0.05; quadratic P < 0.05), and Mg (linear P < 0.05) increased with increasing Zn supplementation. These results indicated that dietary Zn supplementation in Cashmere goats during the breeding season improved the semen quality and quantity, elevated the antioxidative indices and Zn concentration, and decreased the MDA content both in spermatozoa and seminal plasma.

Electroacupuncture Improves Synaptic Function in SAMP8 Mice Probably via Inhibition of the AMPK/eEF2K/eEF2 Signaling Pathway.

Synaptic loss and dysfunction is associated with cognitive impairment in Alzheimer's disease (AD). Recent evidence indicates that the AMP-activated protein kinase (AMPK)/eukaryotic elongation factor-2 kinase (eEF2K)/eukaryotic elongation factor-2 (eEF2) pathway was implicated in synaptic plasticity in AD. Therapeutic strategies for AD treatment are currently limited. Here, we investigate the effects of electroacupuncture (EA) on synaptic function and the AMPK/eEF2K/eEF2 signaling pathway in male senescence-accelerated mouse-prone 8 (SAMP8) mice. Male 7-month-old SAMP8 and SAMR1 mice (senescence-accelerated mouse resistant 1) were randomly divided into 3 groups: SAMR1 control group (Rc), SAMP8 control group (Pc), and SAMP8 electroacupuncture group (Pe). The Pe group was treated with EA for 30 days. Transmission electron microscopy (TEM) was used to observe the structure of synapse. The protein and mRNA expression of synaptophysin (SYN) and postsynaptic density 95 (PSD95) was examined by immunohistochemistry, western blot, and real-time RT-PCR. The activity of AMPK and eEF2K was studied by western blot. Our results showed that EA ameliorated synaptic loss, increased the expression of SYN and PSD95, and inhibited AMPK activation and eEF2K activity. Collectively, these findings suggested that the mechanisms of EA improving synaptic function in AD may be associated with the inhibition of the AMPK/eEF2K/eEF2 signaling pathway.

Mitochondrial pathway mediated by reactive oxygen species involvement in α-hederin-induced apoptosis in hepatocellular carcinoma cells.

AIM: To investigate the antitumor activity of α-hederin in hepatocellular carcinoma (HCC) cells and its underlying mechanisms in vitro and in vivo. METHODS: SMMC-7721, HepG-2 and Huh-7 HCC cells were cultured in vitro and treated with α-hederin (0, 5 µmol/L, 10 µmol/L, 15 µmol/L, 20 µmol/L, 25 µmol/L, 30 µmol/L, 35 µmol/L, 40 µmol/L, 45 µmol/L, 50 µmol/L, 55 µmol/L, or 60 µmol/L) for 12 h, 24 h, or 36 h, and cell viability was then detected by the Cell Counting Kit-8. SMMC-7721 cells were treated with 0, 5 µmol/L, 10 µmol/L, or 20 µmol/L α-hederin for 24 h with or without DL-buthionine-S,R-sulfoximine (2 mmol/L) or N-acetylcysteine (5 mmol/L) pretreatment for 2 h, and additional assays were subsequently performed. Apoptosis was observed after Hoechst staining. Glutathione (GSH) and adenosine triphosphate (ATP) levels were measured using GSH and ATP Assay Kits. Intracellular reactive oxygen species (ROS) levels were determined by measuring the oxidative conversion of 2',7'-dichlorofluorescin diacetate. Disruption of the mitochondrial membrane potential was evaluated using JC-1 staining. The protein levels of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C were detected by western blotting. The antitumor efficacy of α-hederin in vivo was evaluated in a xenograft tumor model. RESULTS: The α-hederin treatment induced apoptosis of HCC cells. The apoptosis rates in the control, low-dose α-hederin (5 µmol/L), mid-dose α-hederin (10 µmol/L) and high-dose α-hederin (20 µmol/L) groups were 0.90% ± 0.26%, 12% ± 2.0%, 21% ± 2.1% and 37% ± 3.8%, respectively (P < 0.05). The α-hederin treatment reduced intracellular GSH and ATP levels, induced ROS, disrupted the mitochondrial membrane potential, increased the protein levels of Bax, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C, and decreased Bcl-2 expression. The α-hederin treatment also inhibited xenograft tumor growth in vivo. CONCLUSION: The α-hederin saponin induces apoptosis of HCC cells via the mitochondrial pathway mediated by increased intracellular ROS and may be an effective treatment for human HCC.

Electroacupuncture Upregulates SIRT1-Dependent PGC-1α Expression in SAMP8 Mice.

BACKGROUND: Abnormalities of brain energy metabolism are involved in Alzheimer disease (AD). Sirtuin 1 (SIRT1) is a class III histone deacetylase and activates peroxisome proliferator-activated receptor-γ co-activator-1α (PGC-1α), which enhances mitochondrial biogenesis and energy homeostasis. Electroacupuncture (EA) has been reported to improve brain energy metabolism in AD. However, the effect of EA on SIRT1 and PGC-1α in AD remains unclear. MATERIAL AND METHODS: ATP levels were measured using assay kits in the hippocampus and frontal cortex of senescence-accelerated mouse prone 8 (SAMP8) mice. Western blotting analysis and quantitative real-time RT-PCR were performed to measure the expression of SIRT1 and PGC-1a in the hippocampus of SAMP8 mice. PGC-1α acetylation was analyzed using immunoprecipitation. RESULTS: Compared with senescence-accelerated resistant mice 1 (SAMR1) mice, SAMP8 mice had a decline in ATP levels and the expression of SIRT1 and PGC-1α. EA treatment improved ATP levels, upregulated the expression of SIRT1 and PGC-1α, and decreased PGC-1α acetylation. CONCLUSIONS: These data suggest that EA improved brain energy metabolism, potentially associated with the upregulation of SIRT1-dependent PGC-1α expression.

Electroacupuncture Reduces Aß Production and BACE1 Expression in SAMP8 Mice.

Electroacupuncture (EA) has been reported to have beneficial effects on Alzheimer's disease (AD). BACE1 (ß-site amyloid precursor protein-cleaving enzyme 1) is involved in the abnormal production of amyloid-ß plaque (Aß), a hallmark of AD pathophysiology. Thus, the present study investigated the effects of EA on memory impairment, Aß production, and BACE1 expression in senescence-accelerated mouse prone 8 (SAMP8) mice. We found that EA improved spatial learning and memory impairment of SAMP8 mice. Furthermore, EA attenuated Aß production and repressed the expression of BACE1 in the hippocampus of SAMP8 mice. Taken together, our results suggest that EA could have a potential therapeutic application in AD and that BACE1 may be an important target of EA in the treatment of AD.

Electroacupuncture improves cognitive deficits associated with AMPK activation in SAMP8 mice.

Perturbations of brain energy metabolism are involved in Alzheimer's disease (AD). Adenosine monophosphate-activated kinase (AMPK) is a master energy sensor that monitors the levels of key energy metabolites. Electroacupuncture (EA) has demonstrated therapeutic potential for the treatment of AD. The effects of EA on cognitive functions and the changes of AMPK and its phosphorylated form (p-AMPK) expression were investigated in senescence-accelerated mouse prone 8 (SAMP8) mice. Cognitive function of SAMP8 mice was assessed using Morris water maze test after EA treatment. Then mice were sacrificed for immunohistochemistry and western blot analysis. EA stimulation significantly alleviated memory impairment of AD mice, and increased the levels of p-AMPK in the hippocampus. These results suggest that EA improved cognitive function associated with AMPK activation, AMPK may be a molecular target of EA in treating AD.

[Effects of electroacupuncture on the function of respiration chain in hippocampal mitochondria of SAMP8 mice ].

OBJECTIVE: from the perspective of mitochondria. METHODS: Tweleve SAMP8 mice aged 8 months were randomly divided into a model group (n = 6) and an electroacupuncture group (n = 6), and six SAMR1 mice aged 8 months were selected as a control group. The electroacupuncture group was treated with electroacupuncture at "Baihui" (GV 20), "Dazhui" (GV 14), "Shenshu" (BL 23) and "Taixi" (KI 3) for 20 min, once each day, 10 days for a course, and lasted for 3 courses. The other two groups were grasped and fixed in the same way at the same time without the other treatment. After the end of treatment, the learning and memory abilities of the mice were measured by Morris water maze, the activity of hippocampal mitochondrid respiratory chain enzyme complex was performed by spectrophotometry, and the levels of adenosine triphosphate (ATP) were detected by a reverse-phase high performance liquid chromatography (HPLC) method. RESULTS: Compared with the control group, the average escape latency significantly lengthened, the residence time on the plateau phase shortened, the activity of respiratory chain enzyme complexe I, II, III, and IV was decreased, and ATP concentration was decreased in the model group. Compared with the model group, the average escape latency significantly shortened, the residence time on the plateau phase lengthened, the activity of hippocampal mitochondrid respiratory chain complexes I, II, III, and IV was significantly increased, and ATP concentration was increased in the electroacupuncture group. CONCLUSION: Electroacupuncture can increase the activity of hippocampal mitochondrid respiratory chain enzyme complexe and ATP concentration and improve mitochondrial function, which may be one of underlying mechanisms of electroacupuncture in treatment of AD.

Thymoquinone prevents and ameliorates dextran sulfate sodium-induced colitis in mice.

BACKGROUND: Thymoquinone (TQ), an active ingredient of the seed oil extract of Nigella sativa Linn, has previously been shown to possess antitumor, antioxidant, and anti-inflammatory bioactivity. Whether TQ has any effect on colitis remains controversial. AIM: The aim of this study was to determine whether treatment with TQ prevents and ameliorates colonic inflammation in a mouse model of inflammatory bowel disease. METHODS: C57BL/6 murine colitis was induced by the administration of dextran sodium sulfate (DSS) (3 % W/V) in the drinking water supplied to the mice for 7 consecutive days. The mice with colitis were treated with 5, 10, or 25 mg/kg TQ orally, and changes in body weight and macroscopic and microscopic colitis scores were examined. In addition, biochemical analyses were conducted. RESULTS: The treatment of mice with TQ prevented and significantly reduced the appearance of diarrhea and body weight loss. These results were associated with amelioration of colitis-related damage, as measured by macroscopic and microscopic colitis scores. In addition, there was a significant reduction in colonic myeloperoxidase activity and malondialdehyde levels and an increase in glutathione levels. CONCLUSIONS: These results indicate that TQ administration can prevent and improve murine DSS-induced colitis. These findings suggest that TQ could serve as a potential therapeutic agent for the treatment of patients with inflammatory bowel disease.