Huangqi Fuzheng decoction exerts antitumor activity by inhibiting cell growth and inducing cell death in osteosarcoma.

Huangqi Fuzheng decoction (HFD) is a traditional Chinese medicine and has been used as adjuvant clinical therapy for breast, ovarian and thyroid cancer. However, the potential roles and molecular mechanisms of HFD in osteosarcoma remain unclear. Here, we investigated the antitumor activity of HFD in osteosarcoma and analyzed its active compounds and therapeutic targets using an integrated systems pharmacology approach. We found that HFD treatment obviously suppressed cell proliferation and induced cell death of U2OS and Saos-2 cells. In addition, HFD treatment significantly inhibited tumor growth and in combination with Cisplatin treatment without obvious serious side effects in the osteosarcoma xenograft mice. Based on systems pharmacology analysis, we identified 105 active compounds and 23 potential targets for HFD in osteosarcoma. Finally, multi-targets were validated as therapeutic targets of HFD for osteosarcoma using Western blot. Our study provides a liable strategy to explore the molecular mechanisms of traditional Chinese medicine. These findings also suggest HFD as a promising candidate medicine for osteosarcoma treatment.

Insulin degrading enzyme contributes to the pathology in a mixed model of Type 2 diabetes and Alzheimer's disease: possible mechanisms of IDE in T2D and AD.

Insulin degrading enzyme (IDE) is believed to act as a junction point of Type 2 diabetes (T2D) and Alzheimer's disease (AD); however, the underlying mechanism was not completely clear yet. Transgenic APPSwe/PS1 mice were used as the AD model and were treated with streptozocin/streptozotocin (STZ) to develop a mixed mice model presenting both AD and T2D. Morris Water Maze (MWM) and recognition task were performed to trace the cognitive function. The detection of fasting plasma glucose (FPG) and plasma insulin concentration, and oral glucose tolerance test (OGTT) were used to trace the metabolism evolution. Aß40 and Aß42 were quantified by colorimetric ELISA kits. The mRNA or protein expression levels were determined by quantitative real-time RT-PCR and Western blotting analysis respectively. T2D contributes to the AD progress by accelerating and worsening spatial learning and recognition impairments. Metabolic parameters and glucose tolerance were significantly changed in the presence of the AD and T2D. The expression levels of IDE, PPARγ, and AMPK were down-regulated in mice with AD and T2D. PPARγ activator rosiglitazone (RSZ) or AMPK activator AICAR increased the expression level of IDE and decreased Aß levels in mice with AD and T2D. RSZ or AICAR treatment also alleviated the spatial learning and recognition impairments in AD and T2D mice. Our results found that, in the mice with T2D and AD, the activators of PPARγ/AMPK signaling pathway significantly increased the expression level of IDE, and decreased the accumulation of Aß40 and Aß42, as well as alleviated the spatial learning and recognition impairments.

Requirement of AQP4 for antidepressive efficiency of fluoxetine: implication in adult hippocampal neurogenesis.

Aquaporin-4 (AQP4), a key molecule for maintaining water homeostasis in the central nervous system, is expressed in adult neural stem cells (ANSCs) as well as astrocytes. Neural stem cells give rise to new hippocampal neurons throughout adulthood, and defects in neurogenesis may predispose an individual to depression. Nevertheless, the role of AQP4 in adult hippocampal neurogenesis and chronic mild stress (CMS)-induced depression remains unknown. We herein report that AQP4 knockout disrupted 4-week fluoxetine (10 mg/kg per day i.p) treatment-induced enhancement of adult mouse hippocampal neurogenesis as well as behavioral improvement under both basal condition and CMS-evoked depressive state. Meanwhile, AQP4 knockout abolished fluoxetine-induced enhancement of hippocampal cyclic AMP-responsive element binding protein (CREB) phosphorylation. The CMS procedure inhibited hippocampal protein kinase A (PKA) activity, extracellular signal-regulated kinases (ERK1/2), and calcium/calmodulin-dependent protein kinase IV (CaMKIV) phosphorylation in AQP4(+/+) and AQP4(-/-) mice. Fluoxetine treatment could reverse CMS-induced inhibition of PKA activity and ERK1/2 phosphorylation in both genotypes. However, fluoxetine restored CMS-induced inhibition of hippocampal CaMKIV phosphorylation in AQP4(+/+) mice but failed in AQP4(-/-) mice. Notably, CMS procedure significantly increased the hippocampal AQP4 expression, which was reversed by 4-week fluoxetine treatment. Further investigation showed AQP4 knockout inhibited the proliferation of cultured ANSCs and eliminated the pro-proliferative effect of fluoxetine in vitro. Collectively, these findings suggest that AQP4 is required for the antidepressive action of fluoxetine via regulating adult hippocampal neurogenesis.

AQP4 knockout impairs proliferation, migration and neuronal differentiation of adult neural stem cells.

Aquaporin-4 (AQP4), a key molecule for maintaining water and ion homeostasis in the central nervous system, is expressed in adult neural stem cells (ANSCs) as well as astrocytes. However, little is known about the functions of AQP4 in the ANSCs in vitro. Here we show that AQP4 knockout inhibits the proliferation, survival, migration and neuronal differentiation of ANSCs derived from the subventricular zone of adult mice. Flow cytometric cell cycle analysis revealed that AQP4 knockout increased the basal apoptosis and induced a G2-M arrest in ANSCs. Using Fluo-3 Ca2+ imaging, we show that AQP4 knockout alters the spontaneous Ca2+ oscillations by frequency enhancement and amplitude suppression, and suppresses KCl-induced Ca2+ influx. AQP4 knockout downregulated the expression of connexin43 and the L-type voltage-gated Ca2+ channel CaV1.2 subtype in ANSCs. Together, these findings suggest that AQP4 plays a crucial role in regulating the proliferation, migration and differentiation of ANSCs, and this function of AQP4 is probably mediated by its action on intracellular Ca2+ dynamics.

Alterations of striatal neurotransmitter release in aquaporin-4 deficient mice: An in vivo microdialysis study.

Our previous investigation has demonstrated that the lack of aquaporin-4 (AQP4) expressions in mice is paralleled by sex- and region-specific abnormalities in neurotransmission. In the present study, we examined the effect of AQP4 deficiency on release of neurotransmitters in mouse striatum using in vivo microdialysis after high K(+) stimulus. The results showed that neurotransmitter releases under the basal and K(+)-stimulated conditions in the striatum of wildtype mice were similar to AQP4 knockout mice, except for taurine, when measured at 24h after microdialysis surgery. However, the basal extracellular levels of dopamine and its metabolites were significantly increased in knockout mice, followed by reduced or no response to depolarizing stimuli when measured at 7 d after surgery. In addition, it was found that there were higher responses of amino acids to high K(+) stimulus in knockout mice. This experiment provides the in vivo evidence that AQP4 participates in the regulation of neurotransmitter release induced by depolarizing stimuli.