IL-6 In Rats Research Articles

Tetramethylpyrazine reduces the consequences of nitric oxide inhibition in pregnant rats.

Pre-eclampsia (PE) is closely associated with perinatal morbidity and mortality and we want to investigate tetramethylpyrazine (TMP)'s effects on PE. Pregnant Sprague-Dawley rats were randomly divided into five groups: normal pregnant (PC), PE, PE+TMP 20 mg/kg, PE+TMP 40 mg/kg, and PE+TMP 60 mg/kg group. The PE rat model was established via L-NAME treatment. Systolic blood pressures (SBP) and urinary protein concentration were detected via the tail-cuff method and CBB kit, respectively. mRNA levels of key genes were analyzed via quantitative PCR and protein levels of key genes were measured by ELISA or western blot. TMP decreased SBP and urinary protein concentration of PE rats. TMP inhibited L-NAME-induced decrease in pups alive ratio, pups weight, and the ratio of pups/placenta weight and reversed L-NAME induced changes in placental histology, whereas it had little effect on placental weight. Urinary nephrin and podocin expressions were enhanced and serum placental growth factor level was decreased in PE rats, whereas TMP inhibited the above phenomena. TMP suppressed L-NAME-induced sFlt-1 upregulation in serums and kidneys of PE rats, whereas it downregulated IL-6 and MCP-1 expression in PE rats' serums, placentas and kidneys. TMP also suppressed the increase in placental sFlt-1 and vascular endothelial growth factor level caused by L-NAME. In addition, TMP inhibited CHOP and GRP78 expressions and decreased the ratio of p-elF2α/elF2α in PE rats. TMP attenuated the consequences of NO inhibition in pregnant rats.

N-acetylcysteine and alpha-lipoic acid improve antioxidant defenses and decrease oxidative stress, inflammation and serum lipid levels in ovariectomized rats via estrogen-independent mechanisms

Sexual hormone deficiency has been associated with metabolic changes, oxidative stress and subclinical inflammation in postmenopausal women. Hormone replacement therapies are effective in many instances, even though some patients either do not respond or are not eligible. The aim of this study was to evaluate the impact of short- (15 days) versus long-term (60 days) sexual hormone depletion and whether antioxidant supplementation with N-acetylcysteine (NAC) and alpha-lipoic acid (LA) improves oxidative stress, metabolic, and inflammatory parameters in ovariectomized (OVX) rats. Short-term OVX rapidly depleted circulating estrogen, causing uterine atrophy and body weight gain without affecting oxidative damage, inflammatory and lipid metabolism markers. In contrast, long-term OVX augmented oxidative damage in serum and peripheral tissues as well as increased serum total cholesterol, TNF-α and IL6 levels. Triglycerides, glucose and HDL cholesterol were not altered. Long-term OVX-induced oxidative stress was associated with depletion of GSH and total non-enzymatic antioxidants as well as decreased activity of Glutathione Peroxidase (GPx) and Glutathione Reductase (GR), but not Superoxide Dismutase (SOD) and Catalase (CAT). NAC and LA supplementation prevented GSH and total non-enzymatic antioxidants depletion as well as restored GPx and GR activities, TNF-α, IL6 and cholesterol in OVX rats. NAC and LA effects appear to be independent on NRF2 activation and estrogen-like activity, since NAC/LA did not promote NRF2 activation and were not able to emulate estrogen effects in OVX rats and estrogen-receptor-positive cells. The herein presented data suggest that NAC and LA may improve some deleterious effects of sexual hormone depletion via estrogen-independent mechanisms.

MicroRNA-340-5p relieved chronic constriction injury-induced neuropathic pain by targeting Rap1A in rat model.

Neuropathic pain (NP) is one of the main challenges towards NP syndrome treatment. miR-340-5p exhibit different expression levels in NP models. Its effects on NP remained unclear. The objective of this study was to explore the potential regulation mechanisms of miR-340-5p in NP. Rat model of chronic constriction injury (CCI) was established to induce NP in vivo. NP levels were assessed using mechanical withdrawal threshold (MWT). The inflammation response in CCI rats were determined by HE staining and ELISA assay. The target genes of miR-340-5p were verified by luciferase report assays. In CCI rats, level of miR-340-5p was down-regulated both in spinal cord tissues and isolated microglial cells. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were decreased in CCI rats, which were restored upon miR-340-5p overexpression. miR-340-5p overexpression also decreased inflammation as well as expression levels of COX-2, IL-1β, TNF-α and IL-6 in CCI rats. Luciferase report assays revealed Rap1A was a target gene of miR-340-5p in the experimental model. Elevated miR-340-5p decreased Rap1A expression level in vitro and in vivo. Overexpression of Rap1A protein restored expression levels of COX-2, IL-1β, TNF-α and IL-6, reduced the PWT and PWL and increased inflammation response in CCI rats. miR-340-5p alleviated CCI-induced NP by targeting Rap1A. miR-340-5p and Rap1A may be the potential treatment targets for NP therapeutics.

Protective effects of luteolin on injury induced inflammation through reduction of tissue uric acid and pro-inflammatory cytokines in rats

Background and aimLuteolin belongs to flavone group of flavonoids, present in many plants with potent antioxidant, anti-inflammatory and anti-proliferative effects. The objective of present study was to investigate protective effect of luteolin on injury induced inflammation via Monosodium urate (MSU) crystals induced and Acetaminophen (AMP) induced liver injury in rats. Experimental procedureProtective effect of luteolin was observed by measurement of rat paw edema, lysosomal enzymes, antioxidants status and cytokine level. Measurement of uric acid level and neutrophil infiltration were done in AMP induced liver injury in rats. Luteolin was tested at 30 and 50 mg/kg doses and compare with colchicine. Results and conclusionLuteolin significantly decreases paw edema in dose dependent manner compare to control group in MSU crystal-induced rats. Luteolin (50 mg/kg) was showed significant decrease in serum level of oxidative and lysosomal enzymes, proinflammatory cytokines i.e. tumor necrosis factor (TNF)-α (39.28 ± 3.17), interleukin (IL)-1β (12.07 ± 1.24), and IL-6 (24.72 ± 2.52) in MSU crystal-induced rats. In AMP induced liver injury, tissue uric acid level and myeloperoxidase were decreased significantly after treatment with luteolin as well as N-acetylcysteine. Serum level of liver enzymes was significantly reduced after treatment with luteolin. Histological observation of ankle joints and liver was support to protective effect of luteolin at both doses. In conclusion, luteolin showed anti-inflammatory effect through restoration of cytokine level, lysosomal enzymes level and antioxidants status. The reduction of liver tissue uric acid content may be one of the mechanisms for protective effect of luteolin. It can contribute to reduce injury induced inflammation.

Does aerobic exercise associated with tryptophan supplementation attenuates hyperalgesia and inflammation in female rats with experimental fibromyalgia?

The objective of this study was to verify the effects of aerobic exercise associated with tryptophan (TRP) supplementation on hyperalgesia, as well as on cortisol, IL-6 and TNF concentrations in female rats with experimental fibromyalgia (FM). Female Wistar rats (initial body weight: ~ 350 g; age: 12 months) were randomly divided into 5 groups: CON (Control); F (Fibromyalgia induced); FE (Fibromyalgia induced plus exercise); FES (Fibromyalgia induced plus exercise and TRP supplementation) and FS (Fibromyalgia induced plus TRP supplementation). Fibromyalgia was induced with two injections (20 μL) of acidic saline (pH 4.0) into the right gastrocnemius muscle with a 3-day interval. Control animals received the same doses of neutral saline (pH 7.4). The exercised animals underwent progressive low-intensity aerobic exercise (LIAE) on a treadmill (10–12 m/min, 30–45 min/day, 5 days/week) for three weeks. During this period, the supplemented animals received a TRP supplemented diet (210 g/week), while the others received a control diet. Mechanical hyperalgesia was evaluated weekly and serum cortisol and muscle IL-6 and TNF concentrations were assessed after three weeks of interventions. Experimental FM caused bilateral hind paw hyperalgesia and augmented serum cortisol and muscle IL-6 concentrations. After 3 weeks of interventions, LIAE alone reduced hyperalgesia (151%) and reduced serum cortisol concentrations (72%). Tryptophan supplementation itself diminished hyperalgesia (57%) and reduced serum cortisol concentrations (67%). Adding TRP supplementation to LIAE did not further reduce hyperalgesia significantly (11%), which was followed by an important decrease in muscle IL-6 concentrations (68%), though reduction in serum cortisol pulled back to 45%. Muscle TNF concentrations were not affected. In conclusion, the association of TRP supplementation to LIAE does not potentiate significantly the reduction of bilateral mechanical hyperalgesia promoted by LIAE in female rats with experimental FM, however an important decrease in IL-6 is evident.

Anti-inflammatory Effect of Juniperus Procera Extract in Rats Exposed to Streptozotocin Toxicity.

Background: Chronic inflammation is a critical health issue and implicated in several chronic health problems such as tumors, auto-immune disorder, hypertension or diabetes. However, Juniperus procera is one of the famous ancient plants that has been traditionally used to treat several diseases such as hyperglycemia, hepatitis, jaundice, bronchitis, and pneumonia.Objective: Current study is an attempt to investigate the anti-inflammatory effect of Juniperus procera extract on rats exposed to cytotoxicity caused experimentally by streptozotocin injections.Methods: Five groups of adult Wistar rats (10 rats each) were examined as (Normal control, Normal rats treated with Juniperus procera extract, rats administrated with streptozotocin, rats administrated with streptozotocin and treated with insulin and, rats administrated with streptozotocin and Juniperus procera extract). At the end of the experiment, blood was collected from experimented rats. Animals then were killed and small parts of both pancreas and liver were collected for gene expression and histopathological examination.Results: Serum analysis showed a significant increase in glucose, IL-6, IL-2 and TNF-α levels in rats exposed to streptozotocin. That change was reduced in rats cotreated with insulin or Juniperus procera extract. Moreover, streptozotocin showed a significant upregulation of IL-6, TNF-α and A2M genes, while, either insulin or Juniperus procera treatment was restored to normal status. Streptozotocin induced inflammation within hepatic tissues which clearly reduced in hepatic tissues of both insulin and junipers cotreated groups.Conclusion: Streptozotocin toxicity induces acute inflammation and increases serum glucose, IL-6, IL-2 and TNF-α levels. However, Juniperus procera extract was found to significantly prevent that reaction within four weeks experimented frame time.

In vivo, Extract from Withania somnifera Root Ameliorates Arthritis via Regulation of Key Immune Mediators of Inflammation in Experimental Model of Arthritis.

BackgroundRheumatoid Arthritis (RA) is a devastating disease characterized by continual addition of leukocytes and T cells within the articular cavity causing inflammation and cartilage destruction. Withania somnifera is one of the most precious medicinal herbs, reported to have antioxidant, anti‐inflammatory, and immunomodulatory properties.ObjectiveThe purpose of this study was to evaluate anti-inflammatory activity of aqueous extract of Withania somnifera roots (WSAq) in Collagen Induced Arthritic (CIA) rats.MethodsTo achieve this, we assessed the level of inflammatory cytokines such as Tumor Necrosis Factor (TNF)-α, IL-1β, IL-6 and IL-10 in CIA rats. Further, transcription factor, oxidative stress parameters and CD+8 expressions were also analyzed in CIA rats.ResultsArthritic rats showed a greater increase in the levels of pro inflammatory cytokines such as TNF-α, IL-1β, IL-6, transcription factor NF-κB and a decrease in IL-10 concentration than controls rats. Oral administration of WSAq at a dose of 300mg/kg.wt. (WSAq300) appreciably attenuated the production of these pro inflammatory cytokines. This anti-inflammatory activity of WSAq300 might be partly mediated through an increase in the secretion of IL-10 and inhibition of NF-κB activity. Further, arthritic rats also show increased oxidative stress as compared to control rats. This increased oxidative stress in the arthritic rats appears to be the outcome of both an activated pro-oxidant and a poor antioxidant defense system. Treatment with WSAq300 strongly ameliorates all these ROS parameters significantly to near normal. Additional, metalloproteinase MMP-8 levels were also measured and found to be increased in CIA rats, which after treatment with WSAq300 came down to near normal.ConclusionFrom the above results, it can be concluded that the use of WSAq300 may be a valuable supplement which can improve human arthritis.

Curculigoside exerts significant anti‑arthritic effects invivo and invitro via regulation of the JAK/STAT/NF‑κB signaling pathway.

The present study aimed to investigate the anti-arthritic effects of curculigoside isolated from the rhizome of Curculigo orchioides Gaertn in vivo and in vitro, as well as to determine the potential underlying mechanisms. A rat model of arthritis was induced with type II collagen. Arthritic rats were treated with curculigoside (50 mg/kg) and blood samples were collected to determine serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-10, IL-12 and IL-17A. Furthermore, indices of the thymus and spleen were determined. The anti-proliferative effects of curculigoside were detected with Cell Counting kit-8 assays in rheumatoid arthritis-derived fibroblast-like synoviocyte MH7A cells. In addition, expression levels of Janus kinase (JAK)1, JAK3, signal transducer and activator of transcription (STAT)3, nuclear factor (NF)-κB p65 and its inhibitor (IκB) were determined by western blotting. The results revealed that curculigoside inhibited paw swelling and arthritis scores in type II collagen-induced arthritic (CIA) rats. Additionally, curculigoside decreased serum levels of TNF-α, IL-1β, IL-6, IL-10, IL-12 and IL-17A in CIA rats. Curculigoside also significantly inhibited MH7A cell proliferation in a time and concentration-dependent manner. Furthermore, treatment downregulated the expression of JAK1, JAK3 and STAT3, and upregulated cytosolic nuclear factor (NF)-κB p65 and IκB. In conclusion, the results of the present study indicated that curculigoside exhibited significant anti-arthritic effects in vivo and in vitro, and the molecular mechanism may be associated with the JAK/STAT/NF-κB signaling pathway.

Dexmedetomidine alleviates lung ischemia-reperfusion injury in rats by activating PI3K/Akt pathway.

This research aims to investigate the role and mechanism of PI3K/Akt pathway in the pathological process of lung ischemia-reperfusion injury in dexmedetomidine-treated rats. Forty-five healthy male Sprague-Dawley rats were divided into three groups: sham operation group, lung ischemia-reperfusion group (IR group) and dexmedetomidine pretreatment group (Dex group). Rats in the sham operation group did not receive other procedures except for opening left chest. The left lung hilar of rats in the IR group was clamped with non-invasive vascular clamp after anesthesia to establish an ischemic model. After 1 hour, the vascular clamp was released and the rats were reperfused for 2 hours. As for rats in the Dex group, 3 μg/kg of dexmedetomidine (pumping time of 10 min) was pumped through the tail vein before releasing the left hilar clamp. After the experiment, blood samples and lung tissues were collected. Serum levels of interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), IL-10, and IL-1 in rats were examined. Activities of malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD) and catalase (CAT) in rat lung tissues were also detected. Besides, the expressions of hypoxia-inducible factor-la (HIF-la), p-Akt, Caspase-3, and Caspase-9 in lung tissues were detected by Western blot. The mRNA expression levels of HIF-1a, p-Akt, Caspase-3, and Caspase-9 in lung tissues were evaluated by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Lung ischemia-reperfusion markedly increased the levels of IL-6, TNF-α, IL-10, and IL-1 in the IR group. In contrast, dexmedetomidine pretreatment decreased the expression levels of IL-6, TNF-α, IL-10, and IL-1in the Dex group. Also, the activities of MDA and MPO in lung tissues of rats in the IR group significantly increased after lung ischemia-reperfusion injury, whereas dexmedetomidine pretreatment reversed the elevated activities of MDA and MPO in the Dex group. Furthermore, dexmedetomidine pretreatment also improved the activities of SOD and CAT in rat lung tissues compared with rats with lung ischemia-reperfusion injury. In addition, dexmedetomidine pretreatment increased the expression levels of HIF-1α, p-Akt and HIF- in the Dex group when compared to those in the IR group. The mRNA expressions of HIF-1a, p-Akt, Caspase-3, and Caspase-9 in lung tissue of rats was significantly reduced after dexmedetomidine pretreatment. Rat lung ischemia-reperfusion can induce severe lung injury. Dexmedetomidine treatment can attenuate lung ischemia-reperfusion injury by activating the PI3K/Akt signaling pathway at the transcriptional level.

Beneficial effects of andrographolide in a rat model of autoimmune myocarditis and its effects on PI3K/Akt pathway.

The study is to investigate effects of andrographolide on experimental autoimmune myocarditis (EAM). Lewis rats were immunized on day 0 with porcine cardiac myosin to establish EAM. The EAM rats were treated with either andrographolide (25, 50, 100 mg/kg/day) or vehicle for 21 days. An antigen-specific splenocytes proliferation assay was performed by using the cells from control rats immunized with cardiac myosin. Survival rates, myocardial pathology and myocardial functional parameters (left ventricle end-diastolic pressure, ± dP/dt and left ventricular internal dimension) of EAM rats received andrographolide were significantly improved. Andrographolide treatment caused an decrease in the infiltration of CD3+ and CD14+ positive cells in myocardial tissue. Moreover, andrographolide treatment caused a reduction in the plasma levels of tumor necrosis factor-alpha, interleukin-17 (IL-17) and myosin-antibody, and an increase in the level of IL-10 in EAM rats. Oral administration of andrographolide resulted in the decreased expression of p-PI3K, p-Akt without any change of PI3K and Akt. Further results indicate andrographolide significantly inhibited myosin-induced proliferation in splenocytes, and this effect was inhibited by co-treatment of SC79 (Akt activator). Our data indicate andrographolide inhibits development of EAM, and this beneficial effect may be due to powerful anti-inflammatory activity and inhibitory effect on PI3K/Akt pathway.

Effects of Ginsenoside Rg1 on JAK2/STAT3 Signaling Pathway in Rats with Hepatic Fibrosis Induced by Concanavalin a

Background: Ginsenoside Rg1, the active ingredient of natural plants, has been proved to have anti-liver fibrosis activity. However, the specific mechanism behind these effects has not been determined. This study aims to inverstigate the specific mechanism of the antihepatic fibrosis of ginsenoside Rg1. Methods: The rat model of hepatic fibrosis induced by Concanavalin A (Con A) were established and hepatic stellate cells were isolated. Then hepatic fibrosis and cell damage were evaluated and inverstigated the mechanism. Findings: We found that the anti-hepatic fibrosis effect of ginsenoside Rg1 was related to JAK2/STAT3 signaling pathway and IL-1β, IL-6, IL-10. In addition, the expression of IL-1β, IL-6, IL-10, α-SMA, TGF-β, CollaⅠ and CollaⅡ mRNA in rats treated with different doses of ginsenoside Rg1 for 8 weeks were decreased compared with model group, and mRNA expression of IL-10 was increased, which was associated with drug dosage. Forthermore, the expression of p-JAK2, p-STAT3, Caspase3, α-SMA, TGF-β, CollaⅠ and CollaⅡ protein decreased, and the degree of decrease was correlated with the dosage of administration. Similar results were detected by Immunohistochemistry. The expression of JAK2 and STAT3 protein were not changed significantly. Besides, flow cytometry revealed that ginsenoside Rg1 could promote hepatic stellate cell (HSC) apoptosis. Interpretation: TTaken together, our results indicate that the anti-fibrotic effect of ginsenoside Rg1 is related to JAK2/STAT3 signaling pathway and IL-1β, IL-6, IL-10, and ginsenoside Rg1 is a potential drug against liver fibrosis. Funding Statement: This study was supported by the National Science Foundation of China (grant 81270498) and the National Science Foundation of China (grant 81500473). Declaration of Interests: The authors declare no conflict of interest Ethics Approval Statement: During these studies, all animal procedures were conducted in accordance with the Anhui Medical University Policy on the Care, Welfare and Treatment of Laboratory Animals and were reviewed the Institutional Animal Care and Use Committee either at Anhui Medical University or by the ethical review process at the institution where the work was performed. The experimental protocol for this study was approved by the Anhui Medical University of Animal welfare, Ethics and Strategy and Anhui Province Key Laboratory of Major Autoimmune Diseases, Hefei, China.

Resveratrol exhibits an effect on attenuating retina inflammatory condition and damage of diabetic retinopathy via PON1

Diabetic retinopathy (DR), an obstacle of the visual microvascular system, is a serious complication of diabetic patients. Paraoxonase 1 (PON1) has been extensively evaluated as a genetic candidate for diabetic microvascular complications, and PON1 is associated with DR. In this study, the biological functions of PON1 and its related proteins were determined via gene ontology (GO) enrichment analysis; we demonstrated that treatment with resveratrol alleviated retinal inflammatory activities to evaluate its protective effects on streptozotocin (STZ)-induced diabetic rats and high-glucose (HG) stimulated rat retinal endothelial cells (RRECs). The GO enrichment analysis suggested that PON1 may regulate inflammatory responses and microvascular complications in DR. In an in vivo study, resveratrol significantly recovered the insulin level and PON1 expression and activity, as well as clearly reduced the retinal vascular permeability, retinal AGEs, LDL, Ox-LDL, caspase3 activity, retinal damage, IL-1β, IL-6, TNFα, VEGF, IFNγ and MCP-1 in STZ-diabetic rats. Moreover, resveratrol reduced the caspase3 activity and Ox-LDL expression in HG stimulated RRECs. However, its protective effect was a deficiency in PON1-silenced RRECs. PON1 is a pivotal modulator in the role of resveratrol in reversing the RREC damage induced by HG. Furthermore, we found that resveratrol exhibits an effect on attenuating the retinal inflammatory condition and damage of DR via PON1. Our study suggests that resveratrol-induced PON1 in the retina may be a promising therapeutic strategy to prevent diabetes-related retinopathy.

Changes in TNF-α, IL-6, IL-10 and VEGF in rats with ARDS and the effects of dexamethasone.

Changes in tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and vascular endothelial growth factor (VEGF) in serum and bronchoalveolar lavage fluid (BALF) in rats with acute respiratory distress syndrome (ARDS) and the intervention effect of dexamethasone were observed to explore the theoretical basis of dexamethasone in the treatment of ARDS. Seventy-two rats were randomly divided into normal control group (group N, n=24), ARDS model group (group L, n=24) and dexamethasone group (group D, n=24). The ARDS rat model was established by jointly injecting oleic acid and lipopolysaccharide via the caudal vein, while rats in group D received intervention with dexamethasone. The wet/dry weight ratios of lung tissues were measured, and the levels of TNF-α, IL-6, IL-10 and VEGF in serum and BALF were measured via enzyme-linked immunosorbent assay. The wet/dry weight ratio of lung tissues of rats in group D was significantly decreased compared with that in group L (P<0.05 or P<0.01). The levels of TNF-α, IL-6 and VEGF in serum and BALF of rats in group L and D were obviously increased compared with those in group N at each time point (P<0.01). The levels of TNF-α, IL-6 and VEGF in serum and BALF of rats in group D were significantly decreased compared with those in group L (P<0.01). In conclusion, there is a serious imbalance between anti-inflammatory response and inflammatory response in rats with ARDS induced by oleic acid combined with lipopolysaccharide of Escherichia coli, whereas dexamethasone can alleviate lung injury through inhibiting expression levels of inflammatory factors and promoting expression levels of anti-inflammatory factors.

Interleukin-6 Serum Levels Are Elevated in Individuals with Degenerative Cervical Myelopathy and Are Correlated with Symptom Severity.

BackgroundFew studies have investigated systemic inflammation levels in degenerative cervical myelopathy (DCM). This study evaluated the concentration of inflammatory cytokines in DCM patients and assessed whether they can predict symptom severity.Material/MethodsA total of 40 consecutive DCM patients and 10 healthy volunteers were included in this study. Concentrations of interleukin (IL)-1β, IL-6, interferon-γ, and tumor necrosis factor-α were compared between DCM patients and normal controls. Spearman’s correlation coefficient was used to examine relationships of cytokines with age, body mass index (BMI), symptom duration, and symptom severity. A DCM compression rat model was established to examine the levels of inflammatory cytokines in serum and cerebrospinal fluid (CSF).ResultsSerum level of IL-6 is significantly higher in DCM patients compared with normal people (0.8±0.5 pg/ml vs. 0.5±0.3 pg/ml, P=0.036). Positive correlations were found between IL-6 levels with BMI (ρ=0.519; P=0.001) and symptom severity (ρ=−0.556, P<0.001). In DCM compression model rats, IL-6 was elevated in CSF (40.5±3.3 vs. 13.2±2.4 pg/ml, P<0.001) and serum (7.1±1.7 vs. 2.9±1.6 pg/ml, P<0.001) samples from rats in the compression operation group compared with the sham operation group. Infusion of IL-6 in rats receiving the sham operation also led to motor function damage and mechanical allodynia threshold decline.ConclusionsSerum IL-6 level was elevated in DCM patients and its concentration can predict symptom severity. Local infusion of IL-6 led to myelopathy symptoms in model rats, which suggests that anti-inflammation can effectively treat DCM.

Effects of short- and long-term regular exercise on reproductive tissue in streptozotocin-induced diabetic male Wistar rats.

Reproductive disorders are one of the complications of diabetes mellitus. Since conflicting results have been obtained from different studies, which examined serum levels of cytokines in patients with diabetes, and considering the fact that the origin of cytokines cannot be accurately determined from their serum changes, attempts were made in the present study to study histological changes and testicular tissue levels of TNF-α and IL-1 in rats treated with exercise. Considering the effects of exercise in reducing blood sugar level and its complications, two types of short-term and long-term regular exercises were also considered to evaluate their effects on male reproductive tissues. In this study, 60 male rats with the weight range of 250±50 g were used and were randomly divided into six groups (10 rats each). Healthy groups included sedentary control group, and groups treated with two and eight weeks of exercise. Rats with type 1 diabetes (induced by streptozotocin) included sedentary control group, groups treated with two and eight weeks of exercise (six groups). All groups were evaluated in terms of testicular tissue levels of TNF-α and IL-1 using ELISA and the histometry of spermatogonia, primary spermatocytes, Sertoli cells, epithelial thickness, diameter of veins, and thickness of the seminiferous tubule. Histological changes resulting from diabetes, particularly in the diameter of testicular veins and a number of cells, including Sertoli, highlights the important fact that tissue perfusion in patients with diabetes is especially crucial, in a way that exercise proved useful for tissue structures by offsetting this complication. Measurement of the cytokines IL-1 and TNF-α in the current study showed that perfusion problems are more important in diabetic complications than inflammatory factors. The main result of this research is recommendation of investigating the tissue of interest for diagnosis of diabetes complications, measuring inflammatory mediators of tissue rather than evaluating their serum concentrations, and focusing on vascular complications as a major complication of diabetes. Furthermore, regular exercise could help improve the function of reproductive organs in healthy groups and prevent diabetes infertility complications to an acceptable degree in diabetic groups.

Influences of hydrogen-rich saline on acute kidney injury in severely burned rats and mechanism

Objective: To explore the influences of hydrogen-rich saline on acute kidney injury in severely burned rats and to analyze the related mechanism. Methods: Fifty-six Sprague Dawley rats were divided into sham injury group (n=8), burn group (n=24), and hydrogen-rich saline group (n=24) according to the random number table. Rats in sham injury group were treated by 20 ℃ water bath on the back for 15 s to simulate injury, and rats in burn group and hydrogen-rich saline group were inflicted with 30% total body surface area (TBSA) full-thickness scald (hereinafter referred to as burns) by 100 ℃ water bath on the back for 15 s. Immediately after injury, hydrogen-rich saline at the dose of 10 mL/kg were intraperitoneally injected to the rats in hydrogen-rich saline group at one time, while normal saline with the same dose were intraperitoneally injected to the rats in sham injury group and burn group. At post injury hour (PIH) 6, rats in the 3 groups were intraperitoneally injected with 4 mL·kg(-1)·%TBSA(-1) lactated Ringer's solution for resuscitation. Eight rats from sham injury group at PIH 72 and eight rats from burn group and hydrogen-rich saline group at PIH 6, 24, and 72 were sacrificed respectively after their blood samples from abdominal aorta were collected. Then their kidney tissue was harvested for histopathological observation and renal tubular injury scoring by hematoxylin and eosin staining, serum creatinine and blood urea nitrogen were detected by the clinical blood biochemical analyzer, expression distribution and mRNA expressions of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 in renal tissue were evaluated by immunohistochemical staining and real time fluorescent quantitive reverse transcription polymerase chain reaction respectively, and protein expression of high mobility group protein 1 (HMGB1) was detected by Western blotting. Data were processed with Kruskal-Wallis H test, Dunn test, one-way analysis of variance, Bonferroni test. Results: (1) The renal tubular structure of rats in sham injury group at PIH 72 was complete with no inflammatory cell infiltration and no cellular degeneration or necrosis. Since PIH 6, the changes such as vacuolation and shape change of cells and aggregation of broken protein in renal tubules were observed in rats of burn group, and all these changes deteriorated with time. The renal injury of rats in hydrogen-rich saline group at different post injury time points were relieved compared with those of rats in burn group at the corresponding time points. The renal tubular injury scores of rats in burn group and hydrogen-rich saline group at PIH 6, 24, and 72 were significantly higher than the score in sham injury group at PIH 72 (P 0.05), the levels of serum creatinine of rats in burn group at all the time points and hydrogen-rich saline group at the other time points were significantly higher than the level of serum creatinine of rats in sham injury group at PIH 72 (P 0.05), and the mRNA expression levels of TNF-α, IL-1β, and IL-6 at the other time points in hydrogen-rich saline group were significantly decreased (P 0.05). Compared with those in burn group, the protein expressions of HMGB1 of rats in hydrogen-rich saline group at PIH 6, 24, and 72 were significantly decreased (P<0.05). Conclusions: Hydrogen-rich saline can alleviate the acute kidney injury in severely burned rats through regulating the release of inflammatory cytokines in renal tissue.

Effects of Shugan-Jianpi Recipe on the Expression of the p38 MAPK/NF-κB Signaling Pathway in the Hepatocytes of NAFLD Rats.

Background: In traditional Chinese medicine, the Shugan-Jianpi recipe is often used in the treatment of nonalcoholic fatty liver disease (NAFLD). This study aimed to explore the mechanism of the Shugan-Jianpi recipe in relation to rats with NAFLD induced by a high-fat diet. Methods: Rats were randomly divided into eight groups: normal group (NG), model group (MG), low-dose Chaihu–Shugan–San group (L-CG), high-dose Chaihu–Shugan–San group (H-CG), low-dose Shenling–Baizhu–San group (L-SG), high-dose Shenling–Baizhu–San group (H-SG), low dose of integrated-recipes group (L-IG), and high dose of integrated-recipes group (H-IG). After 26 weeks, a lipid profile, aspartate, and alanine aminotransferases in serum were detected. The serum levels of inflammatory factors including interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) were analyzed using the enzyme linked immunosorbent assay (ELISA) method. Hepatic pathological changes were observed with hematoxylin-eosin (HE) and oil red O staining. The expression of the p38 mitogen-activated protein kinases (MAPK)/nuclear factor-κB (NF-κB) pathway was detected by quantitative real-time PCR and Western blotting. Results: A pathological section revealed that NAFLD rats have been successfully reproduced. Compared with the model group, each treatment group had different degrees of improvement. The Shugan-Jianpi recipe can inhibit the serum levels of IL-1β, IL-6, and TNF-α in NAFLD rats. The expression of mRNA and a protein related to the p38 MAPK/NF-κB signaling pathway were markedly decreased as a result of the Shugan-Jianpi recipe. Conclusions: The Shugan-Jianpi recipe could attenuate NAFLD progression, and its mechanism may be related to the suppression of the p38 MAPK/NF-κB signaling pathway in hepatocytes.

MiR-134-5p attenuates neuropathic pain progression through targeting Twist1.

Neuropathic pain is a kind of chronic pain because of dysfunctions of somatosensory nerve system. Recently, many studies have demonstrated that microRNAs (miRs) play crucial roles in neuropathic pain development. This study was designed to investigate the effects of miR-134-5p on the process of neuropathic pain progression in a rat model established by chronic sciatic nerve injury (CCI). First, we observed that miR-134-5p was significantly decreased in CCI rat models. Overexpression of miR-134-5p strongly alleviated neuropathic pain behaviors including mechanical and thermal hyperalgesia. Meanwhile, inflammatory cytokine expression, such as IL-6, IL-1β and TNF-α in CCI rats were greatly repressed by upregulation of miR-134-5p. Twist1 has been widely regarded as a poor prognosis biomarker in diverse diseases. Here, by using bioinformatic analysis, 3'-untranslated region (UTR) of Twist1 was predicted to be a downstream target of miR-134-5p in our study. Here, we found that overexpression of miR-134-5p was able to suppress Twist1 dramatically. Furthermore, it was exhibited that Twist1 was increased in CCI rats time-dependently and Twist1 was inhibited in vivo. Subsequently, downregulation of Twist1 in CCI rats could depress neuropathic pain progression via inhibiting neuroinflammation. In conclusion, our current study indicated that miR-134-5p may inhibit neuropathic pain development through targeting Twist1. Our findings suggested that miR-134-5p might provide a novel therapeutic target for neuropathic pain.

Detection of IL-1β, IL-6 and TNF-α in Sprague-Dawely rats' atrophic thymus induced by lipopolysaccharide.

This study aimed to investigate developmental changes of the thymus and intra- thymic IL-1β, IL-6 and TNF-α expression in weaned Sprague-Dawley rats induced by lipopolysac- charide. Forty healthy weaned rats aged 26 days and weighing 83±4 g were randomly and equally divided into two groups. The lipopolysaccharide group was treated daily with a single injection of lipopolysaccharide for 10 consecutive days, and the saline group was treated with an equal volume of sterilized saline. On the 1st, 4th, 7th and 10th day, histological changes and distribu- tion of IL-1β-, IL-6- and TNF-α-positive cells were detected in the thymus by hematoxylin-eosin and immunohistochemistry staining, respectively. Subsequently, the expression levels of IL-1β, IL-6 and TNF-α were evaluated in the thymus by the ELISA method. Thymus weight and index were significantly smaller in lipopolysaccharide-treated rats than in saline-treated rats (p⟨0.05), but no substantial changes were found in the thymus microstructure after lipopolysaccharide induction. Moreover, a large number of IL-1β-, IL-6- and TNF-α-positive cells were observed with brownish-yellow color and mainly distributed in the thy- mus parenchyma, both integrated optical density and average optical density increased signifi- cantly in lipopolysaccharide-treated rats than those in saline-treated rats. Compared with the saline group, most of the thymic homogenates had higher levels of IL-1β, IL-6 and TNF-α in the lipopolysaccharide group on different days. These findings indicate that the thymus atrophied after lipopolysaccharide induction in weaned Sprague-Dawley rats, and excessive production of intrathymic IL-1β, IL-6 and TNF-α was probably involved in the atrophic process.

Osthole alleviated diabetic neuropathic pain mediated by the P2X4 receptor in dorsal root ganglia

Diabetes mellitus (DM) is considered the primary cause of neuropathic pain. Osthole (7-methoxy-8[3-methylpent 2-enyl]coumarin) is a component extracted from Cnidium monnieri (L.) cusson plant seeds and has anti-inflammatory and anti-oxidative properties. The aim of the present study was to investigate the effects of osthole on diabetic neuropathic pain (DNP) involving the P2X4 receptor on satellite glial cells (SGCs) in the dorsal root ganglia (DRG) of type 2 diabetic rats. These data showed that the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in DM rats were lower than those in control rats. MWT and TWL in DM rats treated with osthole were higher compared with those in untreated DM rats. The expression levels of P2X4 mRNA and protein in the DRG of DM rats were higher compared with those in the control rats, while those in DM rats treated with osthole were significantly lower compared with those in the untreated DM rats. Osthole treatment decreased the co-expression levels of P2X4 and glial fibrillary acidic protein (GFAP) and reduced the up-regulated expression of interleukin-1 beta (IL-1β), tumour necrosis factor-α (TNF-α), brain-derived neurotrophic factor (BDNF) and phosphorylated-p38MAPK and enhanced the down-regulation of IL-10 in DM rats. Thus, osthole treatment may act on the P2X4 receptor to alleviate the mechanical and thermal hyperalgesia in DM rats.