SAG, a sonic hedgehog signaling agonist, alleviates anxiety behavior in high-fat diet-fed mice.

Anxiety is a prevalent and disabling psychiatric disorder. Mitochondrial dysfunction due to the high-fat diet (HFD) was regarded as a risk factor in the pathogenesis of anxiety. The Sonic hedgehog (SHH) pathway was known to improve mitochondrial dysfunction through antioxidant and anti-apoptotic effects on some neurological diseases. Nonetheless, its effect on anxiety has not been well studied. In this study, we aimed to explore whether SHH signaling pathway plays a protective role in anxiety by regulating mitochondrial homeostasis. SAG, a typical SHH signaling agonist, was administered intraperitoneally in HFD-fed mice. HFD-induced anxiety-like behavior in mice was confirmed using the open field and elevated plus maze tests. Immunofluorescence staining and Western blotting assays showed that the SHH signaling was downregulated in the prefrontal cortex neurons from HFD-fed mice. Electron microscopy results showed the mitochondria in the prefrontal cortex of HFD-fed mice were fragmented, which appeared small and spherical, and the area, perimeter and circularity of mitochondria were decreased. Mitofusin2 (Mfn2) and dynamin-related protein 1 (Drp1) were the key proteins involved in mitochondrial division and fusion. SAG treatment could rectify the imbalanced expression of Mfn2 and Drp1 in the prefrontal cortex of the HFD-fed mice, and alleviate the mitochondrial fragmentation. Furthermore, SAG decreased anxiety-like behavior in the HFD-fed mice. These findings suggested that SHH signal was neuroprotective in obesity and SAG relieved anxiety-like behavior through reducing mitochondrial fragmentation.

Hemolymph Node - An Immunomorphlogical Organ: Modeling the Hemolymph Node by Allografting Renal Tissue in the Rat.

There is no authoritative characterization of the attributes of the hemolymph node (HLN) since Gibbes' first description in 1884. Early reports showed that HLN are found near the kidney in human and animals with the feature of numerous erythrocytes in sinuses. Subsequent studies mainly focused on anatomy and histology, such as the source, distribution, and quantity of erythrocytes in sinuses. Recent articles mentioned that the emergence of HLN was related to immunity, but there was no strong evidence to support this hypothesis. Therefore, it is still uncertain whether the HLN is an organ of anatomy, histology, or immunology. It has been found that the development of HLN could be elicited in the parathymic area by stimuli such as Escherichia coli, allogeneic breast cancer cells, and renal tissue that were injected/transplanted into the tail of rats in our pilot studies. In this study, the model of the HLN was established by transferring allogeneic renal tissue in the rat. Intrasinusoidal erythrocytes of the node were the component for producing a red macroscopic appearance, while macrophage-erythrocyte-lymphocyte rosettes were the major immunomorphological changes, reflecting the immune activity against the invasion of the allogeneic tissue within the node. Therefore, the HLN is an immunomorphological organ.

Development and Performance Evaluation of an Intelligent Sand Control Screen Based on Polyphenylene Sulfide.

Open hole combined with a sand filter pipe is the main development mode of an unconsolidated sandstone horizontal well, which causes problems such as shaft wall collapse, sand filter pipe blockage, and even damage. At present, the polyurethane intelligent sand control screen pipe can effectively solve these problems, but its application scope is limited by the downhole temperature and material strength. Therefore, the high-strength shape memory polyphenylene sulfide (SMPPPS) was synthesized by an indoor experiment, then the shape memory performance was analyzed, and the variation law of pore size with composition and processing technology was obtained. Finally, the performance of the SMPPPS intelligent screen was evaluated. It is found that (1) with the increase of the cross-linking agent content, the melt index and melt peak temperature of SMPPPS decrease rapidly at first and then remain unchanged; the cross-linking agent content is set at 4.5 wt %. (2) The shape memory performance of SMPPPS is good, and the recovery ratio can reach 99.8% at 110 °C in 11.8 min. (3) The relationship between the pore size distribution proportion of SMPPPS and the foaming agent content, pore enhancer content, temperature, and pressure is determined through experiments; the main distribution range of pore size increases with the increase of the foaming agent content and temperature and decreases with the increase of the pore enhancer content and pressure. When the foaming agent content is 0.5 wt %, the pore enhancer concentration is 10 wt %, the molding temperature is 320 °C, the molding pressure is 10 MPa, and the main distribution range of pore size is 80-320 µm. (4) The permeability of recovery SMPPPS with a compression ratio of 300% first increases slightly and then decreases slightly with the increase of flow, with a variation range of 420.5-463 mD; the compressive strength range is 9.5-11.4 MPa; the SMPPPS has good adaptability to downhole fluid; the sand retaining accuracy is higher than that of the screen sand filter pipe, but the blockage is more serious.

Downregulation of sonic hedgehog signaling in the hippocampus leads to neuronal apoptosis in high-fat diet-fed mice.

BACKGROUND/AIMS: Obesity induces hippocampal neuronal apoptosis and leads to cognitive function deficits. Sonic hedgehog (SHH) signaling is crucial during nervous system development and is neuroprotective in many neurologic diseases. This study assessed the role of SHH signaling in the cognitive deficits in high-fat diet (HFD)-induced obese mice. METHODS: Flow cytometry assay was used to examine cell apoptosis. Tissue pathology was evaluated by Nissl staining. Immunofluorescent staining and western blotting were used to detect SHH signaling molecules and apoptosis-related proteins. The Morris water maze test was performed to evaluate mouse spatial learning and memory. RESULTS: After HFD feeding for 24 weeks, the expression of SHH signaling molecules was downregulated in the mouse hippocampus. In vitro, GANT61 inhibited SHH signaling and induced apoptosis in HT22 mouse hippocampal cells. Smoothened agonist (SAG), an agonist of SHH signaling, reduced apoptosis in GANT61-treated HT22 cells by regulating caspase-9 and caspase-3 activation. In vivo, 12-week SAG treatment also inhibited the apoptosis of hippocampal neurons in HFD-fed mice by increasing mitofusin 2 (Mfn2) and B-cell lymphoma 2 (Bcl-2) levels and decreasing dynamin-related protein 1 (Drp1) and Bcl-2 homologous antagonist/killer (Bak) levels. Behavioral testing showed that SAG administration ameliorated the cognitive impairment in HFD-fed mice. CONCLUSION: Downregulation of hippocampal SHH signaling leads to neuronal apoptosis and cognitive deficits in HFD-fed mice. These findings provide useful information for the identification of potential targets for research and therapeutic interventions for cognitive impairment in obesity.

Purple sweet potato color improves hippocampal insulin resistance via down-regulating SOCS3 and galectin-3 in high-fat diet mice.

Hippocampal insulin resistance is the key factor in cognitive deficits. The obesity induces chronic inflammation and the inflammation molecules suppressors of cytokine signaling3 (SOCS3) and galectin-3 directly impair the insulin signaling. The anti-inflammation properties of purple sweet potato color (PSPC) prompted us to investigate the effect of PSPC on cognitive impairment associated with obesity. 60 C57BL/6 mice were randomly divided into four groups: normal, high fat diets (HFD), HFD+PSPC and PSPC. The mice were fed with the HFD or normal diet for 32 weeks. The PSPC (500 mg/kg/day) was administered via oral gavage from 21 to 32 weeks. The results showed the PSPC rectified the abnormal metabolism indexes induced by HFD, including ameliorated obesity, decreased the concentration of fasting blood glucose and improved the glucose tolerance. The Morris water maze test showed the PSPC alleviated the cognitive impairment in HFD mice. The PSPC decreased the expression of Iba1, tumor necrosis factor-α, interleukin-1ß, SOCS3 and galectin-3 in hippocampus of HFD mice. The insulin signaling molecules including the p-IRS1 (Tyr608), PI3K p110α and p-AKT (Ser473) were detected and the PSPC treatment improved the insulin resistance in hippocampus of HFD mice. Furthermore, the PSPC increased Bcl-2, diminished the Bak and the cleaved-caspase3 in HFD mice hippocampus. These findings indicated that PSPC could be a potential treatment to improve the cognitive impairment associated with obesity.

GANT61 alleviates arthritic symptoms by targeting fibroblast-like synoviocytes in CIA rats.

BACKGROUND: Studies have identified that the fibroblast-like synoviocytes (FLS) exhibited tumor-like characteristics and was the key factor in the pathogenesis of Rheumatoid arthritis (RA). GANT61, an antagonist of the sonic hedgehog pathway, has been verified with inhibitory effect on many cancers. Here we investigated the effect of GANT61 on FLS and the development of collagen-induced arthritis (CIA). METHODS: 40 Sprague Dawley (SD) rats were randomly divided into four groups: normal, CIA, CIA+10 mg/kg GANT61 and CIA+20 mg/kg GANT61. CIA was induced in rat with collagen injecting. The GANT61 was administered by intraperitoneal injection every 2 days for 3 weeks. The CIA model was identified with the paw swelling, arthritis score and the pathologic changes in joint. The FLS of different group were primary cultured. The proliferative capacity of FLS was detecteded via Cell Counting Kit-8 (CCK-8) method, and the apoptosis was detecteded by flow cytometry. The Bcl-2, Bax, Caspases3 and cleaved Caspases3 in synovium and FLS were detecteded by Western Blot. RESULTS: The 20 mg/kg GANT61 treatment reduced the incidence of CIA and relieved the arthritis symptoms in CIA rats. The Bcl-2 was upregulated and the Bax was downregulated in the CIA rats synovium. The 10 mg/kg and 20 mg/kg GANT61 diminished the Bcl-2 expression, 20 mg/kg GANT61 increased the Bax and activated the Caspases3 in the CIA synovium. The proliferation of CIA-FLS was significantly higher and the apoptosis of the CIA-FLS was lower than that of the control group. The 10 mg/kg and 20 mg/kg GANT61 treatment can reduce cell proliferation and induce apoptosis by diminishing Bcl-2 and increasing the Bax in CIA-FLS. CONCLUSIONS: The GANT61 inhibit the proliferation of FLS and alleviated the arthritic symptoms in CIA rats, this implied the GANT61 may be recommended as a possible candidate for the therapy of RA.

Gabapentin regulates expression of FGF2 and FGFR1 in dorsal root ganglia via microRNA-15a in the arthritis rat model.

BACKGROUND: Arthritis is an inflammatory disease with a prevalence rate of approximately 10% in China, which commonly manifests as pain. The aim of the current study was to investigate the function of gabapentin in the dorsal root ganglion in an arthritis rat model, and assess the effect of gabapentin on the expression of fibroblast growth factor 2 (FGF2) and FGF receptor 1 (FGFR1). METHODS: A total of 30 healthy male Sprague-Dawley rats were randomly divided into the following three groups: Untreated group, control group and gabapentin group. Rats in the control and the gabapentin groups were injected with Freund's complete adjuvant to induce arthritis. A total of 7 days subsequent to model establishment, the gabapentin group was administered intraperitoneally gabapentin for 8 days. The alterations in thickness of paw pad and paw withdrawal mechanical threshold (PWMT) were detected, which indicated that the rats in the control and gabapentin groups presented with the symptoms of arthritis. RESULTS: In the control group, the PWMT value was significantly reduced (P < 0.05), whereas the PWMT value was significantly increased in the gabapentin group. Immunohistochemistry demonstrated that the expression levels of FGF2 and FGFR1 were increased in the control group compared with the untreated group, while the expression levels of FGF2 and FGFR1 were reduced in the gabapentin group. Moreover, the FGF2 antagonist PD173074 partially improved the plantar thickness and PWMT of the arthritic rats. Bioinformatics analysis predicted microRNA-15a binding sites in the 3'untranslated regions (UTR) of FGF2 and FGFR1. Furthermore, the expression of microRNA-15a was reduced in the control group compared with untreated rats, whereas microRNA-15a in the gabapentin group was upregulated compared with the control. Additionally, the luciferase reporter assay confirmed that microRNA-15a could inhibit the protein expression through pairing with the 3'UTR of FGF2 and FGFR1 mRNAs. CONCLUSION: Gabapentin may relieve arthritis pain and reduce the expression of FGF2 and FGFR1 in dorsal root ganglia. Furthermore, microRNA-15a may be involved in the regulatory process.

[Sonic hedgehog (SHH) promotes the proliferation of synovial fibroblasts of rats with collagen-induced arthritis].

OBJECTIVE: To investigate the effect of sonic hedgehog (SHH) on the proliferation of synovial fibroblasts (SFs). METHODS: The serum samples were collected from 30 rheumatoid arthritis (RA) patients, 30 systemic lupus erythematosus (SLE) patients, 30 ankylosing spondylitis (AS) patients and 30 healthy subjects. The concentrations of serum SHH were detected by ELISA. Collagen induced arthritis (CIA) were developed by type 2 collagen in Sprague-Dawley rats. The SFs were isolated from knee synovial tissues of CIA rats, and then identified by the detection of vimentin by immunofluorescence technique. Before and 72 hours after blocking SHH-glioma-associated oncogene 1 (Gli-1) signaling pathway with GANT61, the expression level of SHH in SFs was detected by Western blotting, and the proliferation of SFs was examined with CCK-8 assay. RESULTS: The level of serum SHH in the RA patients was remarkably higher than that in the SLE, AS patients and the healthy controls. In the CIA rats, the expression of SHH in SFs in vitro was higher than that in the healthy control rats. After 72-hour treatment of GANT61 to block SHH-Gli-1 signaling pathway, the expression level of SHH protein in SFs from CIA rats was reduced, and meanwhile the proliferation of the SFs was inhibited. CONCLUSION: SHH plays an important role in the proliferation of SFs and could be used as a potential therapeutic target for RA.

The Effect of SHH-Gli Signaling Pathway on the Synovial Fibroblast Proliferation in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by chronic synovitis. This study aims to investigate the role of sonic hedgehog (SHH)-Gli signaling pathway in synovial fibroblast proliferation in rheumatoid arthritis. The expression of serum SHH in RA patients group was significantly increased compared with the systemic lupus erythematosus (SLE), ankylosing spondylitis (AS), and healthy subject (healthy control, HC) groups, respectively; serum SHH expression of RA patients was positively correlated with rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibodies (anti-CCP Ab), while there was no significant correlation between SHH expression and erythrocyte sedimentation rate (ESR). SHH, Ptch, Smo, and Gli molecules were highly expressed in rat RA-synovial fibroblast (RA-SF); after blocking the SHH-Gli signaling pathway with a Gli specific inhibitor, Gli-antagonist 61 (GANT61), RA-SF proliferation was inhibited in a dose-dependent manner and the apoptosis rate of RA-SF was increased as well; the expression levels of fibroblast growth factor receptor 1 (FGFR1) and FGFR3 declined in SF cells after GANT61 treatment. Our results suggest that SHH-Gli pathway is involved in the pathogenesis of RA, and blocking SHH-Gli pathway inhibits RA-SF cell proliferation and increases cell apoptosis, which may shed light on developing new ideas for RA treatment.