ABSTRACT
The comorbidity of autism spectrum disorder and anxiety is common, but the underlying circuitry is poorly understood. Here, Tmem74-/- mice showed autism- and anxiety-like behaviors along with increased excitability of pyramidal neurons (PNs) in the prelimbic cortex (PL), which were reversed by Tmem74 re-expression and chemogenetic inhibition in PNs of the PL. To determine the underlying circuitry, we performed conditional deletion of Tmem74 in the PNs of PL of mice, and we found that alterations in the PL projections to fast-spiking interneurons (FSIs) in the dorsal striatum (dSTR) (PLPNs-dSTRFSIs) mediated the hyperexcitability of FSIs and autism-like behaviors and that alterations in the PL projections to the PNs of the basolateral amygdaloid nucleus (BLA) (PLPNs-BLAPNs) mediated the hyperexcitability of PNs and anxiety-like behaviors. However, the two populations of PNs in the PL had different spatial locations, optogenetic manipulations revealed that alterations in the activity in the PL-dSTR or PL-BLA circuits led to autism- or anxiety-like behaviors, respectively. Collectively, these findings highlight that the hyperactivity of the two populations of PNs in the PL mediates autism and anxiety comorbidity through the PL-dSTR and PL-BLA circuits, which may lead to the development of new therapeutics for the autism and anxiety comorbidity.
Subject(s)
Autism Spectrum Disorder , Autistic Disorder , Basolateral Nuclear Complex , Mice , Animals , Autistic Disorder/genetics , Autism Spectrum Disorder/genetics , Cerebral Cortex , Anxiety , Prefrontal CortexABSTRACT
BACKGROUND: Cancer related inflammation plays a fatal role in the metastatic process, which can foster tumor growth, angiogenesis and dissemination. Sparstolonin B (SsnB), derived from Chinese medicine of the tubers of Scirpus yagara, is a TLR2 and TLR4 antagonists. It has exhibited multiple activities of anti-inflammatory, anti-cancer, anti-obesity and anti-hepatitis. However, whether SsnB is involved in the regulation of inflammation-induced tumor metastasis is not well elucidated. PURPOSE: The aim of this study was to investigate the effectiveness of SsnB as a treatment of inflammation-induced tumor metastasis and identify the underlying mechanisms of its anti-tumor metastatic activity. METHOD: The anti-tumor metastatic activity in vitro was estimated by MTT, wound-healing assay, matrigel invasion analysis and extracellular matrix adhesion assay. Mice lung metastasis and hepatic metastasis experiments were performed to assess the activities in vivo. Lungs or livers were weighed and the number of metastatic nodules was determined after mice were sacrificed. The levels of pro-inflammatory cytokines in the serum, lungs and livers were detected by using enzyme-linked immunosorbent assay (ELISA). Micro-metastasis nodules in lungs or livers were analyzed by histological examination. Immunohistochemistry and western blot analysis were conducted to determine protein expression. RESULT: Herein, SsnB dose-dependently inhibited cell migration and invasion in mouse melanoma B16 cells with or without stimulation of lipopolysaccharide (LPS), Pam3csk4 or molecules from damaged tumor cells (DTC-Ms). The expression of matrix metalloproteinases (MMP)-2 was also significantly abated by SsnB in LPS-modulated B16 cells. And SsnB reduced LPS-activated B16 cells adhesion to extracellular matrix components collagen I and fibronectin in a dose-dependent manner. In vivo, SsnB obviously attenuated LPS-activated pulmonary metastasis in mice by reduction the number of metastatic nodules on the lung surfaces, lung weight and levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serums and lungs. Moreover, in experimental hepatic metastasis model mice, SsnB remarkably repressed LPS-stimulated the number of metastatic nodules along with liver weight; and SsnB significantly suppressed LPS-activated increase levels of TNF-α and IL-6 in livers. Immunohistochemistry analysis indicated that SsnB inhibited the expression of TLR4 in livers. Furthermore, SsnB remarkably blocked p38 and ERK1/2 signaling pathway in LPS-induced B16 cells. P38 and ERK1/2 signaling silencing, using BIRB0796 (small molecular inhibitor of p38 MAPK) and PD184352 (inhibitor of MEK1/2 kinases that activate ERK1/2), significantly abated LPS-induced migration and invasion of B16 cells. CONCLUSION: The present study reports a novel use of SsnB in mitigating TLRs ligands-induced melanoma metastasis by inhibition of p38 and ERK1/2 pathway.
Subject(s)
Heterocyclic Compounds, 4 or More Rings/pharmacology , Inflammation/metabolism , MAP Kinase Signaling System/drug effects , Melanoma/drug therapy , Neoplasm Metastasis/drug therapy , p38 Mitogen-Activated Protein Kinases/metabolism , Animals , Anti-Inflammatory Agents/pharmacology , Cell Line, Tumor , Cell Movement/drug effects , Female , Inflammation/drug therapy , Interleukin-6/metabolism , Lipopolysaccharides/pharmacology , Liver/drug effects , Liver/metabolism , Matrix Metalloproteinase 2/metabolism , Melanoma/metabolism , Mice , Mice, Inbred BALB C , Signal Transduction/drug effects , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
A new natural compound, dehydrophyllodulcin (1) was isolated from the tubers of Scirpus yagara, together with 11 known compounds. Among them, compounds 2, 5-8, and 10-12 were isolated from this plant for the first time. (1)H NMR, (13)C NMR, and 2D NMR data of compound 1 are first reported in this article, though it was synthesized in 1996. The structures of all compounds were determined by comprehensive analyses of their spectroscopic data and compared with literature information. Moreover, the anti-inflammatory effects of compounds 1, 3, 4, 6, and 9 against inflammatory cytokines production in Lipopolysaccharide - or Pam3csk4-stimulated macrophage RAW264.7 cells were evaluated by Enzyme-linked immunosorbent assay. And these compounds significantly inhibited the tumor necrosis factor (TNF)-α, interleukin (IL)-6 productions in RAW264.7 cells, with IC50 values less than 20 µM.
