Multiple Intraperitoneal Injections Research Articles

Sestrin2 remedies neuroinflammatory response by inhibiting A1 astrocyte conversion via autophagy.

Most central nervous diseases are accompanied by astrocyte activation. Autophagy, an important pathway for cells to protect themselves and maintain homeostasis, is widely involved in regulation of astrocyte activation. Reactive astrocytes may play a protective or harmful role in different diseases due to different phenotypes of astrocytes. It is an urgent task to clarify the formation mechanisms of inflammatory astrocyte phenotype, A1 astrocytes. Sestrin2 is a highly conserved protein that can be induced under a variety of stress conditions as a potential protective role in oxidative damage process. However, whether Sestrin2 can affect autophagy and involve in A1 astrocyte conversion is still uncovered. In this study, we reported that Sestrin2 and autophagy were significantly induced in mouse hippocampus after multiple intraperitoneal injections of lipopolysaccharide, with the elevation of A1 astrocyte conversion and inflammatory mediators. Knockdown Sestrin2 in C8-D1A astrocytes promoted the levels of A1 astrocyte marker C3 mRNA and inflammatory factors, which was rescued by autophagy inducer rapamycin. Overexpression of Sestrin2 in C8-D1A astrocytes attenuated A1 astrocyte conversion and reduced inflammatory factor levels via abundant autophagy. Moreover, Sestrin2 overexpression improved mitochondrial structure and morphology. These results suggest that Sestrin2 can suppress neuroinflammation by inhibiting A1 astrocyte conversion via autophagy, which is a potential drug target for treating neuroinflammation.

Гистологические и иммуногистохимические изменения префронтальной коры головного мозга мышей под действием дексаметазона

Introduction. The effects of the synthetic glucocorticoid dexamethasone on target organs are mediated by the interaction with the glucocorticoid receptor, but the effects of the drug on the intact brain have been poorly studied. We aimed to look at a one-time and repeated exposure to a two-dose dexamethasone on the expression of the glucocorticoid receptor and cellular changes in the prefrontal cortex over time. Materials and methods. Male C57Bl/6 mice aged 7–8 weeks received a one-time or multiple intraperitoneal injections of dexamethasone in doses of 1.0 or 2.5 mg/kg. Structural changes in the prefrontal cortex were analyzed on paraffin sections stained by hematoxylin and eosin 90 days after the injection. The expression of the glucocorticoid receptor was studied with immunohistochemistry using monoclonal antibodies and quantitative analysis. Results. A histological study of the prefrontal cortex exposed to a two-dose dexamethasone revealed the absence of an obvious dose dependence and the reversibility of dystrophic and hydropic changes. The most pronounced changes were revealed with a one-time administration of the drug (on day 7) and repeated administration (on day 15) while normal tissue architecture returned to that of the control groups. The expression level of the glucocorticoid receptor after a one-time dexamethasone injection at a dose of 2.5 mg/kg showed a significant growth (р<0.05) on days 3 and 10 compared to that of the control group, while after multiple dexamethasone injections, the GR expression decreased on day 15, but by day 30 it returned to its initial level. Conclusion. Stereotypical dystrophic and hydropic changes in the prefrontal cortex during exposure to two different regimens and dexamethasone doses developed on days 7 and 15. We revealed an opposite effect of 2.5 mg/kg dexamethasone administration regimens on the expression of the glucocorticoid receptor in the prefrontal cortex of mice relative to the control group: at a one-time administration the expression increased by 8.1% on day 10 and at multiple administration the expression grew by 9.9% on day 15. Keywords: dexamethasone, glucocorticoids, brain, prefrontal cortex, glucocorticoid receptor, immunohistochemistry

Antinociceptive Effect of Dendrobii caulis in Paclitaxel-Induced Neuropathic Pain in Mice.

Paclitaxel-induced neuropathic pain (PINP) is a serious adverse effect of chemotherapy. Dendrobii caulis (D. caulis) is a new food source used as herbal medicine in east Asia. We examined the antinociceptive effects of D. caulis extract on PINP and clarified the mechanism of action of transient receptor potential vanilloid 1 receptor (TRPV1) in the spinal cord. PINP was induced in male mice using multiple intraperitoneal injections of paclitaxel (total dose, 8 mg/kg). PINP was maintained from D10 to D21 when assessed for cold and mechanical allodynia. Oral administration of 300 and 500 mg/kg D. caulis relieved cold and mechanical allodynia. In addition, TRPV1 in the paclitaxel group showed increased gene and protein expression, whereas the D. caulis 300 and 500 mg/kg groups showed a significant decrease. Among various substances in D. caulis, vicenin-2 was quantified by high-performance liquid chromatography, and its administration (10 mg/kg, i.p.) showed antinociceptive effects similar to those of D. caulis 500 mg/kg. Administration of the TRPV1 antagonist capsazepine also showed antinociceptive effects similar to those of D. caulis, and D. caulis is thought to exhibit antinociceptive effects on PINP by modulating the spinal TRPV1.

BrdU does not induce hepatocellular damage in experimental Wistar rats

Bromodeoxyuridine (BrdU) is used in studies related to cell proliferation and neurogenesis. The multiple intraperitoneal injections of this molecule could favor liver function profile changes. In this study, we evaluate the systemic and hepatocellular impact of BrdU in male adult Wistar rats in 30 %-partial hepatectomy (PHx) model. The rats received BrdU 50 mg/Kg by intraperitoneal injection at 0.5, 1, 2, 3, 6, 9 and 16 days after 30 %-PH. The rats were distributed into four groups as follows, control, sham, PHx/BrdU(-) and PHx/BrdU(+). On day 16, we evaluated hepatocellular nuclei and analyzed histopathological features by haematoxylin-eosin stain and apoptotic profile was qualified by caspase-3 presence. The systemic effect was evaluated by liver markers such as alanine transferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (AP), bilirubin, total proteins and serum albumin content. The statistical analysis consisted of a student t-test and one-way ANOVA. BrdU did not induce apoptosis or hepatocellular damage in male rats. Multiple administrations of BrdU in male rats did not induce significant decrease body weight, but increased serum ALT and LDH levels were found. Our results show that the BrdU does not produce hepatocellular damage.

The role of the SGK3/TOPK signaling pathway in the transition from acute kidney injury to chronic kidney disease.

Introduction: Profibrotic phenotype of renal tubular epithelial cells (TECs) featured with epithelial to mesenchymal transition (EMT) and profibrotic factors secretion, and aberrant accumulation of CD206+ M2 macrophages are the key points in the transition from acute kidney injury (AKI) to chronic kidney disease (CKD). Nevertheless, the underlying mechanisms involved remain incompletely understood. Serum and glucocorticoid-inducible kinase (SGK) is a serine/threonine protein kinase, required for intestinal nutrient transport and ion channels modulation. T-LAK-cell-originated protein kinase (TOPK) is a member of the mitogen activated protein kinase family, linked to cell cycle regulation. However, little is known about their roles in AKI-CKD transition. Methods: In this study, three models were constructed in C57BL/6 mice: low dose and multiple intraperitoneal injection of cisplatin, 5/6 nephrectomy and unilateral ureteral obstruction model. Rat renal tubular epithelial cells (NRK-52E) were dealt with cisplatin to induce profibrotic phenotype, while a mouse monocytic cell line (RAW264.7) were cultured with cisplatin or TGF-β1 to induce M1 or M2 macrophage polarization respectively. And co-cultured NRK-52E and RAW264.7 through transwell plate to explore the interaction between them. The expression of SGK3 and TOPK phosphorylation were detected by immunohistochemistry, immunofluorescence and western blot analysis. Results: In vivo, the expression of SGK3 and p-TOPK were gradually inhibited in TECs, but enhanced in CD206+ M2 macrophages. In vitro, SGK3 inhibition aggravated epithelial to mesenchymal transition through reducing the phosphorylation state of TOPK, and controlling TGF-β1 synthesis and secretion in TECs. However, SGK3/TOPK axis activation promoted CD206+ M2 macrophage polarization, which caused kidney fibrosis by mediating macrophage to myofibroblast transition (MMT). When co-cultured, the TGF-β1 from profibrotic TECs evoked CD206+ M2 macrophage polarization and MMT, which could be attenuated by SGK3/TOPK axis inhibition in macrophages. Conversely, SGK3/TOPK signaling pathway activation in TECs could reverse CD206+ M2 macrophages aggravated EMT. Discussion: We revealed for the first time that SGK3 regulated TOPK phosphorylation to mediate TECs profibrotic phenotype, macrophage plasticity and the crosstalk between TECs and macrophages during AKI-CKD transition. Our results demonstrated the inverse effect of SGK3/TOPK signaling pathway in profibrotic TECs and CD206+ M2 macrophages polarization during the AKI-CKD transition.

Autophagy-mediated circHIPK2 promotes lipopolysaccharide-induced astrocytic inflammation via SIGMAR1.

Circular RNAs (circRNAs) are a subclass of noncoding RNAs and widely involve in the occurrence of multiple human diseases. It is an urgent task to clarify circRNA upstream regulation mechanism and seek their biofunction. Our previous study has confirmed that circular RNA HIPK2 (circHIPK2) promotes astrocyte activation via SIGMAR1, sigma non-opioid intracellular receptor 1, in a mouse model of single high-dose lipopolysaccharide (LPS) injection. However, what mechanism circHIPK2 is regulated by and whether it is involved in the inflammatory response of astrocytes remain unclear. In this study, we reported that circHIPK2 and SIGMAR1 were significantly increased in mouse prefrontal cortex after multiple intraperitoneal injection of LPS, with the elevation of inflammatory mediators. Knockdown circHIPK2 in primary astrocytes suppressed the SIGMAR1 expression and inflammation. Pretreatment of autophagy inducer rapamycin on astrocytes suppressed the circHIPK2 expression and inactivated inflammatory response. These results implied that autophagy inducer rapamycin could suppress astrocytic inflammation by inactivating circHIPK2-SIGMAR1 axis. Autophagy may be a promising upstream administrator of circHIPK2 and therapeutic target for central nervous system inflammation.

Red Ginger Extract Prevents the Development of Oxaliplatin-Induced Neuropathic Pain by Inhibiting the Spinal Noradrenergic System in Mice.

Oxaliplatin is a well-known chemotherapeutic drug that is widely used to treat colorectal cancer. However, it can induce acute side effects in up to 90% of patients. Serotonin and norepinephrine reuptake inhibitors (SNRIs) are used as first-choice drugs; however, even SNRIs are known to be effective only in treatment and not for prevention. Therefore, finding a drug that can prevent the development of cold and mechanical forms of allodynia induced by oxaliplatin is needed. This study demonstrated that multiple oral administrations of 100 mg/kg and 300 mg/kg of red ginger extract could significantly prevent pain development in mice. The role of the noradrenergic system was investigated as an underlying mechanism of action. Both the spinal α1- and α2-adrenergic receptors were significantly downregulated after treatment. Furthermore, the noradrenaline levels in the serum and spinal cord were upregulated and downregulated after treatment with paclitaxel and red ginger, respectively. As the active sub-component of red ginger, ginsenoside Rg3 (Rg3) was identified and quantified using HPLC. Moreover, multiple intraperitoneal injections of Rg3 prevented the development of pain in paclitaxel-treated mice, suggesting that RG3 may induce the effect of red ginger extract.

Assessment of mouse VEGF neutralization by ranibizumab and aflibercept.

To assess the interaction between ranibizumab, aflibercept, and mouse vascular endothelial growth factor (VEGF), both in vivo and in vitro. In vivo, the effect of intravitreal injection of ranibizumab and aflibercept on oxygen induced retinopathy (OIR) and the effect of multiple intraperitoneal injections of ranibizumab and aflibercept on neonatal mice were assessed. In vitro, the interaction of mouse VEGF-A with aflibercept or ranibizumab as the primary antibody was analyzed by Western blot. In both experiments using intravitreal injections in OIR mice and multiple intraperitoneal injections in neonatal mice, anti-VEGF effects were observed with aflibercept, but not with ranibizumab. Western blot analysis showed immunoreactive bands for mouse VEGF-A in the aflibercept-probed blot, but not in the ranibizumab-probed blot. Aflibercept but not ranibizumab interacts with mouse VEGF, both in vivo and in vitro. When conducting experiments using anti-VEGF drugs in mice, aflibercept is suitable, but ranibizumab is not.

Secondary iron overload induces chronic pancreatitis and ferroptosis of acinar cells in mice.

Disruption of iron homeostasis is associated with multiple diseases. It has been found that patients with genetic iron overload develop massive iron deposition in the pancreas. However, few studies have focused on the effect of secondary iron overload on the pancreas. The objective of the present study was to investigate the pathogenic consequences of secondary iron overload in mice. An iron overload mouse model was constructed by intraperitoneal injection of 120mg/kg body weight of iron dextran every other week for 12weeks. Iron deposition, immunocyte infiltration, fibrosis, oxidative stress and ferroptosis were assessed using Prussian blue staining, immunohistochemical analysis, Masson staining, Sirius red staining, RT‑qPCR analysis and western blot analysis. It was found that iron‑overloaded mice showed pancreatic iron overload, together with elevated gene expression of the iron storage factor ferritin H, and decreased expression of the iron transportation mediator divalent metal transporter 1, ferroportin 1 and transferrin receptor. Iron‑overloaded mice developed mild pancreatitis with increased serum amylase and lipase activities, as well as elevated gene expression levels of pro‑inflammatory cytokines, including interleukin (IL)‑1β, IL‑6 and inducible nitric oxide synthase. Acinar atrophy, massive immunocyte infiltration and pancreatic fibrosis were noted in the iron‑overloaded mice. As an underlying mechanism, iron‑overloaded mice showed increased pancreatic oxidative stress, with an elevated malondialdehyde level, and decreased SOD and glutathione peroxidase activity. Furthermore, iron overload led to ferroptosis with promoted expression of cytochrome c oxidase subunit II, and decreased transcripts of glutathione peroxidase 4 and solute carrier family 7 member 11. These results provided evidence that multiple intraperitoneal injections of iron dextran in mice lead to iron overload‑induced chronic pancreatitis, which suggested that secondary iron overload is a risk factor for pancreatitis and highlights the importance of iron in maintaining the normal functions of the pancreas.

Positive regulation of endothelial Tom70 by metformin as a new mechanism against cardiac microvascular injury in diabetes

Microvascular protection is the main mechanism of metformin against diabetic complications. Cardiac microvascular endothelial cells (CMECs) are the basic component of cardiac microvessels, and they suffer from oxidative stress and mitochondrial dysfunction under type 2 diabetes mellitus (T2DM). Translocase of the outer mitochondrial membrane 70 (Tom70) improves mitochondrial dysfunction, but its role in the hearts of T2DM patients remains unclear. The purpose of this study was to demonstrate the protective effect of metformin on diabetic cardiac microvascular injury and to identify the role of Tom70 in this effect. T2DM mice were established by multiple intraperitoneal injections of low-dose streptozotocin and 12-week high-fat feeding. CMECs were isolated and cultured with normal glucose (NG), high glucose (HG), and HG plus high fat (HG-HF) media. The results indicated that long-term metformin treatment partly reversed cardiovascular complication and mitigated cardiac microvascular injury in T2DM. In addition, exposure to HG-HF led to CMEC damage, aggravated oxidative stress, aggravated mitochondrial dysfunction, and reduced mitochondrial Tom70 expression, whereas upregulation of Tom70 significantly ameliorated these injuries. Furthermore, metformin treatment promoted Tom70 expression and effectively reversed CMEC injury induced by HG-HF. However, all of these effects were interrupted after Tom70 was knocked down. In conclusion, T2DM damages microvascular integrity by activating a cycle of decreased Tom70 expression, mitochondrial dysfunction, and reactive oxygen species (ROS) overload in CMECs. However, metformin suppresses oxidative stress, relieves mitochondrial dysfunction, and promotes the expression of Tom70, ultimately ameliorating diabetic microvascular injury and heart complications.

Knockdown of IFNAR2 reduces the inflammatory response in mouse model of type 1 diabetes

Backgroundand Purpose: To investigate the biological role of interferon α/β receptor 2 (IFNAR2) in type 1 diabetes (T1D). MethodsFirst, IFNAR2 mRNA and protein expression levels in serum of T1D patients and healthy controls were detected by RT-qPCR and Western blot. For experimental studies, 80 male C57BL/6 mice were randomly divided into 4 groups with 20 mice in each group: the control group, the T1D group, the T1D + ad-con group and the T1D + ad-si-IFNAR2 group. The T1D mouse model was generated by multiple intraperitoneal injections of small doses of streptozotocin (STZ). Body weight and blood glucose levels were measured weekly until 6 weeks. After 6 weeks, all mice were sacrificed and the levels of insulin (Ins), tumor necrosis factor α (TNF-α), interleukin 4 (IL-4), IL-6, and type I interferon γ (IFN-γ), IFNAR2 protein expression, the number of dendritic cells (DCs), and changes in islet β cells were assessed. ResultsIFNAR2 mRNA and protein expression levels in serum of T1D patients were significantly higher than those in healthy controls (P < 0.05). Furthermore, IFNAR2 protein expression, number of DCs, and IFNAR2 mRNA, blood glucose, TNF-α, and IFN-γ levels were significantly upregulated in T1D mice compared with the control group (P < 0.05), while weight, and Ins, IL-6, and IL-4 levels were decreased (P < 0.05). However, knockdown of IFNAR2 reversed these trends. There was no significant difference in markers between the T1D + ad-con group and the T1D group (P > 0.05). ConclusionsKnockdown of IFNAR2 reduced the inflammatory response and improved islet function of T1D mice.

In Vitro and In Vivo Analysis of Extracellular Vesicle-Mediated Metastasis Using a Bright, Red-Shifted Bioluminescent Reporter Protein.

Cancer cells produce heterogeneous extracellular vesicles (EVs) as mediators of intercellular communication. This study focuses on a novel method to image EV subtypes and their biodistributionin vivo. A red-shifted bioluminescence resonance energy transfer (BRET) EV reporter is developed, called PalmReNL, which allows for highly sensitive EV tracking in vitroandin vivo.PalmReNL enables the authors to study the common surface molecules across EV subtypes that determine EV organotropism and their functional differences in cancer progression. Regardless of injection routes, whether retro-orbital or intraperitoneal, PalmReNL positive EVs, isolated from murine mammary carcinoma cells, localized to the lungs. The early appearance of metastatic foci in the lungs of mammary tumor-bearing mice following multiple intraperitoneal injections of the medium and large EV (m/lEV)-enriched fraction derived from mammary carcinoma cells is demonstrated. In addition, the results presented here show that tumor cell-derived m/lEVs act on distant tissues through upregulating LC3 expression within the lung.

The Effect of Coffee Arabica Gayo Leaf Extract (Coffea arabica L.) in Increasing Phosphoinositide 3-kinase and Glucose Transporter-4 Expression in the Skeletal Muscle

BACKGROUND: Diabetes mellitus (DM) is a chronic and progressive metabolic disease characterized by hyperglycemia due to impaired insulin secretion associated with a lack of phosphoinositide 3-kinase (PI3K) and glucose transporter-4 (GLUT-4) expression in the skeletal muscle membrane. AIM: The aim of the study is to understand the effect of coffee Arabica Gayo leaf extract (Coffea arabica L.) in increasing PI3K expression and GLUT-4 expression in the skeletal muscle membrane. METHODS: Thirty-five male Wistar rats with Type 2 DM (T2DM) induced using a combination of a high-fat diet for 5 weeks followed by multiple intraperitoneal injections of low-dose streptozotocin (30 mg/kg). Divided into seven groups as such two groups that did not receive treatment and five groups that received treatment. The dosage administered was 150, 200, and 250 mg/kg/day through the nasogastric tube for 30 days. PI3K and GLUT-4 expression in the skeletal muscle membrane was evaluated by Immunohistochemistry in their gastrocnemius muscles. RESULTS: The study showed an increased expression of PI3K and GLUT-4 expression. There was a significant difference between coffee Arabica Gayo leaf extract and Metformin in increasing GLUT-4 expression (p = 0.036) and PI3K expression between coffee Arabica Gayo dose 250 mg/kg/day and group without treatment (p = 0.008). CONCLUSION: Coffee Arabica Gayo leaf extract (C. arabica L) at a dose of 250 mg/kg/day can increase PI3K expression in skeletal muscle and a dose of 200 mg/kg/day and 250 mg/kg/day can increase the expression of GLUT-4 in the skeletal muscle membrane greater than metformin.

The Decreasing of Homeostatic Model Assessment – Insulin Resistance Levels after Given Coffee Arabica Gayo Leaf Extract (Coffea arabica L.) to Type 2 Diabetes Mellitus Rats

BACKGROUND: Diabetes mellitus (DM) continues to increase with currently more than 463 million people in the world live with DM. One of the causes of Type 2 DM (T2DM) is insulin resistance. Determining laboratory-based tests for early identification of T2DM is important. One of the tests developed was the detection of homeostatic model assessment – insulin resistance (HOMA-IR) levels. AIM: Using chlorogenic acid found in coffee leaves as antidiabetic agent for alternative treatments in DM, this research is conducted to understand the effect of Coffee Arabica Gayo Leaf Extract (Coffea arabica L.) in decreasing HOMA-IR levels in T2DM rats. MATERIALS AND METHODS: Thirty-five male Wistar rats (Rattus norvegicus) had T2DM induced using a combination of feeding a high-fat diet for 5 weeks followed by multiple intraperitoneal injections of low-dose streptozotocin (30 mg/kgBW), divided into 7 groups (2 groups that did not receive treatment [K1 and K3] and 5 groups that received treatment [K2, K4, K5, K6, K7]). The extract was administered with dosage 150, 200, and 250 mg/kgBW/day through a nasogastric tube for 30 days. Then, the HOMA-IR value will be obtained by multiplying fasting plasma insulin by fasting plasma glucose, then divide by a constant of 22.5. RESULTS: The study showed a significant difference before and after treatment, p value was &lt; 0.05, which a decrease HOMA-IR levels (p = 0.002) after given Coffee Arabica Gayo Leaf Extract at 200 and 250 mg/kgBW/day to Wistar rats for 30 days. The decrease HOMA-IR levels are greater than The T2DM group that received metformin and group without treatment. CONCLUSION: Coffee Arabica Gayo Leaf Extract can significantly decrease HOMA-IR levels at a dose of 200 and 250 mg/kgBW. The decrease of HOMA-IR levels is greater than The T2DM group that received metformin and group without treatment.

Pronociceptive Roles of Schwann Cell-Derived Galectin-3 in Taxane-Induced Peripheral Neuropathy.

Chemotherapy-induced peripheral neuropathy (CIPN) is a severe dose-limiting side effect of taxanes such as paclitaxel and docetaxel. Despite the high medical needs, insufficient understanding of the complex mechanism underlying CIPN pathogenesis precludes any endorsed causal therapy to prevent or relieve CIPN. In this study, we report that elevation of plasma galectin-3 level is a pathologic change common to both patients with taxane-treated breast cancer with CIPN and a mouse model of taxane-related CIPN. Following multiple intraperitoneal injections of paclitaxel in mice, galectin-3 levels were elevated in Schwann cells within the sciatic nerve but not in other peripheral organs or cells expressing galectin-3. Consistent with this, paclitaxel treatment of primary cultures of rat Schwann cells induced upregulation and secretion of galectin-3. In vitro migration assays revealed that recombinant galectin-3 induced a chemotactic response of the murine macrophage cell line RAW 264.7. In addition, perineural administration of galectin-3 to the sciatic nerve of naive mice mimicked paclitaxel-induced macrophage infiltration and mechanical hypersensitivity. By contrast, chemical depletion of macrophages by clodronate liposomes suppressed paclitaxel-induced mechanical hypersensitivity despite the higher level of plasma galectin-3. Deficiency (Galectin-3 -/- mice) or pharmacologic inhibition of galectin-3 inhibited paclitaxel-induced macrophage infiltration and mechanical hypersensitivity. In conclusion, we propose that Schwann cell-derived galectin-3 plays a pronociceptive role via macrophage infiltration in the pathogenesis of taxane-induced peripheral neuropathy. Therapies targeting this phenomenon, which is common to patients with CIPN and mouse models, represent a novel approach to suppress taxane-related CIPN. SIGNIFICANCE: These findings demonstrate that the elevation of plasma galectin-3 is a CIPN-related pathologic change common to humans and mice, and that targeting galectin-3 is a therapeutic option to delay CIPN progression.

Systemic Injection of Thalidomide Prevent and Attenuate Neuropathic Pain and Alleviate Neuroinflammatory Response in the Spinal Dorsal Horn.

Background and objectiveThalidomide (Tha) has been shown to exert immunomodulatory and anti-inflammatory properties. Whether Tha can alleviate spinal nerve ligation (SNL)-induced neuropathic pain (NP) is still unclear. This study aimed to investigate the therapeutic effect of Tha on the SNL-induced NP and further explore the potential analgesic mechanisms of Tha.MethodsThe effects of Tha on SNL-induced mechanical allodynia were assessed by pain behavioral testing. The expressions of the astrocyte marker glial fibrillary acidic protein (GFAP) and the microglia marker Iba1 in the spinal dorsal horn were evaluated by immunofluorescence histochemistry. Protein expressions of the tumor necrosis factor alpha (TNF-α) in the spinal dorsal horn were tested by Western blot assay. Data were analyzed using one-way ANOVA or two-way ANOVA.ResultsBy the pretreatment with a single intraperitoneal injection, the PWMT in SNL+Tha group was significantly increased from day 1 to day 2 after SNL (P < 0.05 compared with SNL+Veh group). By the posttreatment with a single intraperitoneal injection, the PWMT in SNL+Tha group was also significantly increased from day 3 to day 4 after SNL (P < 0.05 compared with SNL+Veh group). By the posttreatment with multiple intraperitoneal injection, both the PWMT and the PWTL in SNL+Tha group were similarly significantly increased from day 3 to day 14 after SNL (P < 0.05 compared with SNL+Veh group). Furthermore, the GFAP and Iba1 expressions and TNF-α levels of the ipsilateral spinal dorsal horn in SNL+Tha group were significantly weaker from day 3 to day 14 after SNL than those in SNL+Veh group (P < 0.05).ConclusionTha can significantly alleviate NP induced by SNL. The analgesic mechanism may be related to inhibition of astrocyte and microglia activation as well as down-regulation of TNF-α levels in the spinal dorsal horn.

Clinicopathological studies on the use of laser-activated adipose-derived stromal vascular fraction in treatment of streptozotocin-induced diabetes in rats

Diabetes mellitus (DM) is associated with severe progressive degenerative complications in many organs especially diabetic nephropathy (DN). Adipose-derived stromal vascular fraction (SVF) is easily obtained and has abundant viable stem cell number obviating extensive expansion in culture; thus, SVF utilization provides promising anticipation. Low-intensity laser irradiation (LILI) was found to strengthen the therapeutic effect of non-expanded SVF through enhancing viability, protein expression, and migration of stem cells. The current study aimed to demonstrate the effect of transplanted laser-activated SVF in streptozotocin (STZ)-induced diabetic rats. Forty-five male Sprague Dawley rats were used in this experiment and divided randomly into four groups: (I) control group, (II) diabetic untreated group, (III) and (IV) diabetic-treated groups in which laser-activated SVF transplantation performed as a single and multiple IP injections, respectively, (V) cell tracking group. DM was induced by a single intraperitoneal (IP) injection of STZ at a dose of 55 mg/kg. Rats received single and multiple IP SVF injections, at a dose of 1.5 × 106 nucleated cells/rat on the 7th day of DM induction and a second dose injected in group IV on the 21th day of DM induction (2-week interval). Insulin gene expression quantification; glycemic profile evaluation (blood glucose, C-peptide, and glycosylated hemoglobin); biochemical, histopathological, and immunohistochemical parameters; and microalbuminuria were assessed in all experimental groups. Both SVF-treated groups exhibited improvement in glycemic profile which was confirmed by the significant increase of insulin gene expression and C-peptide levels. Liver transaminases, lipid profile, and microalbuminuria were normalized while serum creatinine and urea concentrations were ameliorated in treated groups compared to diabetic untreated rats. In conclusion, laser-activated SVF transplantation in diabetic rats triggered improvement of the diabetic state and ameliorated some of its complications with regard to the early interference. The second SVF treatment showed more improvement regarding the blood glucose, C-peptide, histopathology, and immunohistochemical results of the pancreas in diabetic states.

Losartan, an Angiotensin II Type 1 Receptor Antagonist, Alleviates Mechanical Hyperalgesia in a Rat Model of Chemotherapy-Induced Neuropathic Pain by Inhibiting Inflammatory Cytokines in the Dorsal Root Ganglia.

Chemotherapy-induced peripheral neuropathy (CIPN) adversely impacts quality of life and a challenge to treat with existing drugs used for neuropathic pain. Losartan, an angiotensin II type 1 receptor (AT1R) antagonist widely used to treat hypertension, has been reported to have analgesic effects in several pain models. In this study, we assessed losartan's analgesic effect on paclitaxel-induced neuropathic pain (PINP) in rats and its mechanism of action in dorsal root ganglion (DRG). Rats received intraperitoneal injections of 2mg/kg paclitaxel on days 0, 2, 4, and 6 and received single or multiple intraperitoneal injections of losartan potassium dissolved in phosphate-buffered saline at various times. The mechanical thresholds, protein levels of inflammatory cytokines, and cellular location of AT1R and interleukin 1β (IL-1β) in the DRG were assessed with behavioral testing, Western blotting, and immunohistochemistry, respectively. Data were analyzed by two-way repeated-measures analysis of variance for the behavioral test or the Mann-Whitney U test for the Western blot analysis and immunohistochemistry. Single and multiple injections of losartan ameliorated PINP, and losartan delayed the development of PINP. Paclitaxel significantly increased, and losartan subsequently decreased, the expression levels of inflammatory cytokines, including IL-1β and tumor necrosis factor α (TNF-α), in the lumbar DRG. AT1R and IL-1β were expressed in both neurons and satellite cells and losartan decreased the intensity of IL-1β in the DRG. Losartan ameliorates PINP by decreasing inflammatory cytokines including IL-1β and TNF-α in the DRG. Our findings provide a new or add-on therapy for CIPN patients.

The Effect of Puguntano Leaf Extract (Curanga Fel - Terrae Merr.) On P38 Mapk Levels and Glut-4 Expression in Type 2 Diabetic Rat Muscle.

BACKGROUND:Puguntano (Curanga feel-terrae Merr.) contains flavonoids, saponins, tannins, and steroids/ terpenoids which improved post-receptor insulin signalling in rats model of type 2 diabetes mellitus (T2DM).AIM:This study aimed to determine the effect of puguntano leaf extract on p38 mitogen-activated protein kinase (MAPK) levels and glucose transporter-4 (GLUT-4) expression in diabetic rats muscle.METHODS:Forty-eight male Wistar rats had T2DM induced using a combination of feeding a high-fat diet for 5 weeks and multiple intraperitoneal injections of low-dose streptozotocin (30 mg/kg). The diabetic rats were randomly divided into control and treatment groups, and 200 mg/kg/day puguntano extract was administered orally for 10 days to treatment group. Subsequently, p38 MAPK levels were measured by Sandwich Elisa and plasma membrane GLUT-4 expression was evaluated by Immunohistochemistry in their gastrocnemius muscles.RESULTS:There were significantly higher p38 MAPK levels and GLUT-4 expression in the treatment group than in the control group.CONCLUSION:These data suggest that a puguntano leaf extract can improve post-receptor insulin signalling by enhancing p38 MAPK levels and GLUT-4 expression in a rat model of T2DM.

Breast Tumor Microcalcification Induced by Bone Morphogenetic Protein-2: A New Murine Model for Human Breast Tumor Diagnosis.

Widespread use of screening mammography has recently increased the detection of breast microcalcifications. These nonpalpable microcalcifications with specific features in breast tissues are clinically considered an early indicator of breast carcinoma. Our goal in this study was to develop a murine breast microcalcification model for optimizing in vivo imaging. Recombinant human BMP-2 was expressed in E. coli, and the purified bioactive protein was used as inducing factor for the production of breast microcalcifications in a murine animal model. Syngeneic breast tumors were obtained by injection of MDA-MB-231 human breast cancer cells with Matrigel into the mammary fat pad of female nude mice. Different doses of bioactive rhBMP-2 were administered either as single or multiple intraperitoneal injections or directly into tumor on a weekly basis. Three weeks after the first injection of rhBMP-2, the microcalcification of breast tumor was detected by microcomputed tomography followed by intravenous injection of radiotracer [18F] Sodium fluoride for positron emission tomography imaging. Our findings indicate that rhBMP-2 induced microcalcifications of breast tumor by both systemic and direct injection of rhBMP-2 into tumors in a dose-dependent manner. Although little is known about the molecular mechanism of microcalcification, here we report a new murine model of human breast tumor induced microcalcification by rhBMP-2 to optimize in vivo imaging methods and to study the role of BMP-2 as a mediator of pathological mineralization and bone-like microcalcification formation in breast tumor. This BMP-2-induced microcalcification model may allow us to discriminate the type of microcalcification in tumors and to perform quantitative analysis on the calcification as a new detection strategy for early identification of pathological mineralization of breast tissues in women.