TNF-α Immunostaining Research Articles

Repurposing simvastatin for treatment of Klebsiella pneumoniae infections: in vitro and in vivo study

Simvastatin had minimum inhibitory concentrations of 32 to 128 µg/mL against Klebsiella pneumoniae isolates and hindered the biofilm-formation ability of 58.54% of the isolates. It considerably diminished the bacterial cell counts in the biofilms as revealed by scanning electron microscope. Also, qRT-PCR revealed a downregulation of the biofilm genes (bcsA, wza, and luxS) by simvastatin in 48.78% of the isolates. Moreover, simvastatin has significantly improved the survival of mice and decreased the burden of bacteria in the infected lungs. Also, the histological architecture was substantially improved in the simvastatin-treated group, as the alveolar sacs and bronchioles appeared normal with minimal collagen fiber deposition. The immunohistochemical studies exposed that the TNF-α, NF-kβ, and COX-2 immunostaining considerably declined in the simvastatin-treated group. Furthermore, ELISA exposed that both IL-1β and IL-6 were considerably diminished in the lungs of the simvastatin-treated group.

Influence of melatonin supplementation on tissue response of endodontic sealers in Wistar rats.

This study evaluated the influence of melatonin supplementation on tissue's response of endodontic sealers in Wistar rats. Forty-eight rats received subcutaneous implants of four polyethylene tubes: one empty (control) and three filled with endodontic sealers (AH Plus, Endofill and Sealapex). Half of the animals were supplemented with melatonin (ME) and the remaining treated with water (WA) for 15days before the implantation until euthanasia, forming the groups: control-WA, AH Plus-WA, Endofill-WA, Sealapex-WA, Control-ME, AH Plus-ME, Endofill-ME and Sealapex-ME. After 5, 15 and 30days, (n = 8) tubes were removed and evaluated in H&E., immunohistochemistry, PSR, VK and POL. The results were statistically analyzed (p < 0.05). In animals treated with water, Endofill-WA evoked more intense inflammatory infiltrate compared to AH Plus-WA and Control-WA in a 30-day period (p < 0.05). In animals supplemented with melatonin, there was any difference among endodontic sealers' response in any period of analysis (p > 0.05). Comparing the individual response of each sealer, over a 30-day period, Endofill-ME and Sealapex-ME showed less inflammatory infiltrate compared to Endofill-WA and Sealapex-WA, respectively (p < 0.05). Immunostaining for IL-6 and TNF-α was less intense for all groups in animals supplemented with melatonin, in most periods, except for the Endofill sealer (p < 0.05). Furthermore, Endofill-ME at 5days and AH-Plus-ME at 30days showed a higher percentage of mature collagen fibers compared to the Endofill-WA and AH Plus-WA, respectively (p < 0.05). Positive structures for von Kossa staining and birefringent to polarized light were observed only for Sealapex-WA and Sealapex-ME in all periods. It can be concluded that melatonin influences the tissue response to endodontic sealers, modulating the inflammatory and reparative process.

Evaluation of the Therapeutic Potential of Amantadine in a Vincristine-Induced Peripheral Neuropathy Model in Rats.

This study aimed to evaluate the therapeutic potential of amantadine in a vincristine-induced peripheral neuropathy model in rats. Forty-eight male Wistar rats were used. The treated groups received oral amantadine at doses of 2, 5, 12, 25 and 50 mg/kg, with daily applications for 14 days. The mechanical paw withdrawal threshold was measured using a digital analgesimeter. Immunohistochemical analysis of IL-6, TNFα, MIP1α, IL-10, CX3CR1, CXCR4, SOD, CAT and GPx, and enzymatic activity analysis of CAT, SOD and GPx were performed, in addition to quantitative PCR of Grp78, Chop, Ho1, Perk, Bax, Bcl-xL, Casp 3, Casp 9, IL-6, IL-10, IL-18 and IL-1β. The results showed an increase in nociceptive thresholds in animals that received 25 mg/kg and 50 mg/kg amantadine. Immunohistochemistry showed a decrease in the immunostaining of IL-6, TNFα, MIP1α and CX3CR1, and an increase in IL-10. CAT and SOD showed an increase in both immunochemistry and enzymatic analysis. qPCR revealed a reduced expression of genes related to endoplasmic reticulum stress and regulation in the expression of immunological and apoptotic markers. Amantadine demonstrated antinociceptive, anti-inflammatory and antioxidant effects in the vincristine-induced peripheral neuropathy model in rats, suggesting that amantadine may be considered an alternative approach for the treatment of vincristine-induced peripheral neuropathic pain.

Extract of Actaea racemosa Protects Mice Ovarian Follicles Against Doxorubicin-induced Toxicity

Background: Given the cytotoxicity of chemotherapy drugs used in cancer treatment, there is a need to develop alternative agents to protect female fertility. This study investigated the effect of Actaea racemosa (A. racemosa) extract on mice ovarian cells and the damage caused by doxorubicin (DOX) to the mice ovaries. Methods: We evaluated the effects of A. racemosa extract on mice ovaries (n=42) after DOX treatment. The mice were pre-treated with saline solution (controls) or with 0.5 or 5 mg/kg A. Racemosa extract. Afterward, during a period of 10 days, they were treated daily with one of the six protocols: (i) saline solution (control), (ii) 10 mg/kg DOX, (iii) 0.5 mg/kg A. racemosa extract, (iv) both DOX and 5 mg/kg A. racemosa extract, (v) A. racemosa extract (5 mg/kg), and (vi) both DOX and 0.5 mg/kg A. racemosa extract. At the end of these treatments, the ovaries were fixed for histopathological examinations. Ovarian follicular morphology, stromal cell density, collagen fibers, and TNF-α expression were evaluated. Some ovaries were fixed for transmission electron microscopy or stored at -80oC to study the mRNA expression for Caspase-3 and TNF-α. Results: The Mice treated with A. racemosa extract had reduced follicular degeneration and cell death after exposure to DOX. Ovaries of mice treated with 0.5 mg/kg A. racemosa extract had granulosa cells and oocytes with preserved ultrastructure, decreased immunostaining for TNF-α, and reduced Caspase-3 mRNA. Conclusion: The A. racemosa extract supported follicular survival and protected the ovarian follicles and stromal cells against DOX-induced cytotoxicity.

Differences in Stromal Immunostaining of Tumour Necrosis Factor-alpha and its Receptors in Neoplastic and Non Neoplastic Ovarian Lesions: A Cross-sectional Study

Introduction: Ovarian cancer has an unknown pathogenesis, and cytokines may play an important role in the aetiology and prognosis. Tumour Necrosis Factor-alpha (TNF-α) and its receptors (TNFR1 and TNFR2) are involved in the biology of ovarian cancer, tumour pathogenesis, and their relationship with prognostic factors. They are involved in biological processes such as immunoregulation, growth modulation, and cell differentiation. Aim: To evaluate stromal immunostaining of TNF-α and its receptors (TNFR1 and TNFR2) in malignant ovarian neoplasms, comparing it with benign ovarian neoplasms and non neoplastic ovarian lesions. Materials and Methods: A cross-sectional study was carried out at the Department of Gynaecology and Obstetrics, Federal University of Triângulo Mineiro, Uberaba, Minas Gerais, Brazil, from January 1997 to December 2020. Patients with ovarian lesions who underwent surgical treatment according to preestablished criteria (n=95) were included in the study. Patients with benign (n=37) and malignant (n=43) ovarian epithelial neoplasms, and non neoplastic ovarian lesions (n=15) were included. Data evaluated included age, parity, hormonal status (menarche or menopause), histological grade, and staging. Immunohistochemical study was performed to evaluate stromal TNF-α, TNFR1, and TNFR2. Data were analysed by GraphPad Prism 6 and IBM Statistical Package for the Social Sciences (SPSS) Statistics 20.0 software. The comparison between non neoplastic tumours, benign and malignant neoplasms was performed by the Fisher’s exact test with a significance level below 0.05. Results: Considering staining intensity 0 and 1 as “weak immunostaining” and 2 and 3 as “strong immunostaining,” TNF-α stromal immunostaining was stronger (2/3) in benign ovarian neoplasms compared to non neoplastic tumours (p-value=0.0016) and in malignant neoplasms compared to non neoplastic tumours (p-value&lt;0.0001). TNFR1 immunostaining was stronger (2/3) in the stroma of malignant neoplasms compared to benign neoplasms (p-value&lt;0.0001) and stronger (2/3) when comparing benign neoplasms with non neoplastic ovarian lesions (p-value=0.0002). For TNFR2, stromal immunostaining was stronger (2/3) in malignant neoplasms compared to benign neoplasms (p-value =0.0091) and stronger in malignant neoplasms compared to non neoplastic lesions (p-value=0.0004). Conclusion: A stronger immunostaining for TNF-α and its receptors was found in ovarian cancer, suggesting that they may be targets for further studies to verify their role in carcinogenesis and the progression of ovarian neoplasms. A better understanding of the role of TNF-α and its receptors in the tumour stroma of ovarian tumours may lead to future studies that may clarify the mechanisms of carcinogenesis and tumour progression.

Rosavin improves insulin resistance and alleviates hepatic and kidney damage via modulating the cGAS-STING pathway and autophagy signaling in HFD/STZ-induced T2DM animals.

Background: Inflammation-mediated insulin resistance in type 2 diabetes mellitus (T2DM) increases complications, necessitating investigation of its mechanism to find new safe therapies. This study investigated the effect of rosavin on the autophagy and the cGAS-STING pathway-related signatures (ZBP1, STING1, DDX58, LC3B, TNF-α) and on their epigenetic modifiers (miR-1976 and lncRNA AC074117.2) that were identified from in silico analysis in T2DM animals. Methods: A T2DM rat model was established by combining a high-fat diet (HFD) and streptozotocin (STZ). After four weeks from T2DM induction, HFD/STZ-induced T2DM rats were subdivided into an untreated group (T2DM group) and three treated groups which received 10, 20, or 30 mg per kg of R. rosea daily for 4 weeks. Results: The study found that rosavin can affect the cGAS-STING pathway-related RNA signatures by decreasing the expressions of ZBP1, STING1, DDX58, and miR-1976 while increasing the lncRNA AC074117.2 level in the liver, kidney, and adipose tissues. Rosavin prevented further weight loss, reduced serum insulin and glucose, improved insulin resistance and the lipid panel, and mitigated liver and kidney damage compared to the untreated T2DM group. The treatment also resulted in reduced inflammation levels and improved autophagy manifested by decreased immunostaining of TNF-α and increased immunostaining of LC3B in the liver and kidneys of the treated T2DM rats. Conclusion: Rosavin has shown potential in attenuating T2DM, inhibiting inflammation in the liver and kidneys, and improving metabolic disturbances in a T2DM animal model. The observed effect was linked to the activation of autophagy and suppression of the cGAS-STING pathway.

CD20 + cells blockage by rituximab delays wound healing in oral traumatic ulcers in rats

ObjectiveWound healing of oral traumatic ulcers (OTU) is strongly associated with cytokines and inflammatory cells, and the reduction of anti-inflammatory cells, such as lymphocyte B, may interfere with OTU repair. We aimed to evaluate the role of CD20 + cells in the healing process of OTU in rats. DesignWistar male rats were divided into four groups: a control group (treated with 0.1 mL/kg of saline) and three groups treated with anti-CD20 rituximab (RTX) at 2.5, 10, or 40 mg/kg 24 h before OTU production. The animals were weighed (day 0) and euthanized on days 3, 7, 14, and 21 after ulceration. With Blood cells (hematological analysis) and the traumatically induced ulcers were clinically measured. The mucosal samples were histologically (scores 0–4), histochemically (collagen assay (picrosirius)), histomorphometrically (cell counting), and immunohistochemically (CD20+, Tumor Necrosis Factor alpha(TNF-α), Interleukin(IL)− 1β, IL-6 and α-smooth-muscle-actin (α-SMA)) analyzed. ANOVA-1–2-way/Bonferroni, Kruskal-Wallis/Dunn, and correlation analyses were performed (GraphPad Prism 5.0, p < 0.05). ResultsRTX leads to leukopenia, lymphocytopenia, and neutropenia (p < 0.001), and high doses reduced the OTU area (p = 0.001), impaired histologic scores (p < 0.05), and delayed polymorphonuclear (p < 0.001) and mononuclear (p < 0.001) cells, and total (p = 0.011), type-I (p = 0.008), and type-III (p = 0.021) collagen. ConclusionRTX treatment reduced CD20+ cells in OTU (p = 0.001), TNF-α (p = 0.006), and α-SMA (p = 0.022) immunostaining and delayed IL-6 reduction (p = 0.006), with no influence in IL-1β immunostaining. CD20 + cell blockage by RTX reduced cell migration, acute inflammation, and wound healing in OTU.

Thymoquinone attenuates doxorubicin-induced lung damage via heat shock proteins, inflammation and endoplasmic reticulum stress

The aim of this study is to determine possible therapeutic effects of THQ on doxorubicin induced lung damage in rats. Rats were divided into five groups (n = 8): control, THQ, olive oil, DOX (a single dose of 15 mg/kg intraperitoneally (i.p.) on seventh day of the experiment), and DOX+THQ (10 mg/kg THQ per day and 15 mg/kg DOX i.p. on seventh day). Animals were euthanized, and lung tissues were evaluated histopathologically. Caspase3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα immunostaining were performed to determine the expression levels of these proteins among groups. TUNEL method was used for evaluation of apoptotic index. Moreover, total antioxidant status (TAS), total oxidant status (TOS) and IL6 in lung tissue were measured by ELISA assay. The DOX group had histopathological deterioration compared to the control group. There was an increase in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα expressions in the DOX group. While TAS level of the DOX group decreased, IL6 and TOS level increased when compared with the other groups. However, there was improvement in lung tissue in DOX + THQ group compared to the DOX group. There was a decrease in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2, TNFα expressions and TOS, IL6 in DOX+THQ group compared to the DOX group. We suggest that THQ can be used as a protective agent to reduce the toxic effects of DOX.

Anti-TPO-mediated specific features of the placenta immunohistochemical profile and possible mechanisms for fetal loss.

Passive transfer of antithyroid antibodies in mice leads to reproductive disorders. The purpose was to assess the placental tissue of experimental animals under the influence of the circulating thyroperoxidase antibodies. We performed an immunohistochemical examination of murine placentae after a passive transfer of thyroperoxidase antibodies. Placentae of mice that passively transferred IgG from healthy donors were used as control samples. For histological examination, 30 placental samples were selected from mice from the anti-TPO group and 40 placental samples were taken from mice from the IgG group. Immunostaining for VEGFR1, THBS 1, Laminin, CD31, CD34, FGF-β, CD56, CD14, TNF-α, kisspeptin, MCL 1, and Annexin V was performed. There is a significant decrease in the relative area of the expression of VEGFR1 (23.42 ± 0.85 vs. 33.44 ± 0.35, P < 0.01), thrombospondin 1 (31.29 ± 0.83 vs. 34.51 ± 0.75, P < 0.01), CD14 (25.80 ± 0.57 vs. 32.07 ± 0.36, P < .01), CD56 (30.08 ± 0.90 vs. 34.92 ± 0.15, P < 0.01), kisspeptin (25.94 ± 0.47 vs. 31.27 ± 0.57, P < 0.01), MCL 1 (29.24 ± 1.06 vs. 38.57 ± 0.79, P < 0.01) in the labyrinth zone of the placentae of mice from the anti-TPO group compared with control group. A significant increase in the relative expression of laminin and FGF-β was noted in the group of mice to which antibodies to thyroperoxidase were transferred, compared with the control group (36.73 ± 1.38 vs. 29.83 ± 0.94, P < 0.01 and 23.26 ± 0.61 vs. 16.38 ± 1.01, P < 0.01respectively). Our study exposed an imbalance of pro- and anti-angiogenic factors, decreased representation of placental macrophages and NK cells, abnormal trophoblast invasion processes, and insufficient expression of antiapoptotic factors in the placentae of mice in which anti-TPO antibodies were passively transferred.

The Effect of Alpha-Lipoic Acid against Methotrexate on Testicular Damage in Rats

Aim: The toxic effects of methotrexate, a chemotherapeutic, on the testicles is an important side effect. Methotrexate impairs spermatogenesis and fertility and causes oligospermia. In this study, we aimed to minimize the testicular toxicity, those being the side effects of methotrexate, by using the probable protective effects of α-lipoic acid, a potent antioxidant.&#x0D; Materials and Methods: Twenty-eight male Sprague Dawley rats that we employed in this research were separated into three groups as control (0.09% PS) (n=8), methotrexate (20 mg/kg) (n= 10), and methotrexate (20 mg/kg) + α-lipoic acid (100 mg/kg) (n= 10). We performed a histochemical analysis on the testicular tissue of rats using hematoxylin-eosin and Masson’s trichrome. We performed an immunohistochemical analysis using inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-α) primer ab.&#x0D; Results: The histochemical evaluation revealed a significant decrease in the methotrexate-induced testicular toxicity in the α-lipoic acid-treated groups. On the other hand, TNF-α and iNOS immunostaining results were also observed to support these results.&#x0D; Conclusion: The treatment use of α-lipoic acid succeeded in protecting against methotrexate-induced testicular damage through an α-lipoic acid-mediated antioxidant and anti-inflammatory mechanisms. α-lipoic acid can be used in combination with methotrexate as a protector against side effects during anticancer therapy. In the present study, it was shown that α-lipoic acid can be used in combination with methotrexate as a protector against side effects during anticancer treatment.

Obesity and gestational diabetes independently and collectively induce specific effects on placental structure, inflammation and endocrine function in a cohort of South African women.

Maternal obesity and gestational diabetes mellitus (GDM) are associated with insulin resistance and health risks for mother and offspring. Obesity is also characterized by low-grade inflammation, which in turn, impacts insulin sensitivity. The placenta secretes inflammatory cytokines and hormones that influence maternal glucose and insulin handling. However, little is known about the effect of maternal obesity, GDM and their interaction, on placental morphology, hormones and inflammatory cytokines. In a South African cohort of non-obese and obese pregnant women with and without GDM, this study examined placental morphology using stereology, placental hormone and cytokine expression using real-time PCR, western blotting and immunohistochemistry, and circulating TNFα and IL-6 concentrations using ELISA. Placental expression of endocrine and growth factor genes was not altered by obesity or GDM. However, LEPTIN gene expression was diminished, syncytiotrophoblast TNFα immunostaining elevated and stromal and fetal vessel IL-6 staining reduced in the placenta of obese women in a manner that was partly influenced by GDM status. Placental TNFα protein abundance and maternal circulating TNFα concentrations were reduced in GDM. Both maternal obesity and, to a lesser extent, GDM were accompanied by specific changes in placental morphometry. Maternal blood pressure and weight gain and infant ponderal index were also modified by obesity and/or GDM. Thus, obesity and GDM have specific impacts on placental morphology and endocrine and inflammatory states that may relate to pregnancy outcomes. These findings may contribute to developing placenta-targeted treatments that improve mother and offspring outcomes, which is particularly relevant given increasing rates of obesity and GDM worldwide. KEY POINTS: Rates of maternal obesity and gestational diabetes (GDM) are increasing worldwide, including in low-middle income countries (LMIC). Despite this, much of the work in the field is conducted in higher-income countries. In a well-characterised cohort of South African women, this study shows that obesity and GDM have specific impacts on placental structure, hormone production and inflammatory profile. Moreover, such placental changes were associated with pregnancy and neonatal outcomes in women who were obese and/or with GDM. The identification of specific changes in the placenta may help in the design of diagnostic and therapeutic approaches to improve pregnancy and neonatal outcomes with particular significant benefit in LMICs.

Anti-inflammatory and anti-apoptotic potential of beta-glucan on chemotherapy-induced nephrotoxicity in rats

Background/Aim: Cyclophosphamide (CP) is an anti-cancer agent that mediates nephrotoxicity. Beta (β)-glucan has restorative effects on kidney toxicities through its antioxidant potential; however, the effects of β-glucan on CP-induced renal injury remain unknown. In an experimental nephrotoxicity model using rats, we sought to examine the potential protective action of β-glucan on kidney histomorphology, apoptosis, and TNF-α expression. Methods: Male albino Wistar rats were divided equally into four groups: control, CP, β-glucan, and CP+β-glucan. The kidney tissues of the rats were examined for TNF-α and caspase-3 immunostaining to evaluate inflammation and apoptosis, respectively. Hematoxylin and eosin (H&amp;E) and periodic acid–Schiff (PAS) staining were used for histopathological analyses. Results: The CP group showed severe histopathological damage in the renal tissues of rats. In the renal tissue of the CP group, immunoreactivities for TNF-α (1.25 [0.079] and caspase-3 (1.506 [0.143] were also higher than the control group (0.117 [0.006] and 0.116 [0.002], respectively; P&lt;0.001). In the CP+β-glucan group, the histopathological changes significantly improved. Conclusion: Beta-glucan has therapeutic potential against CP-induced nephrotoxicity in rat kidney.

Interleukin-17 plays a role in dental pulp inflammation mediated by zoledronic acid: a mechanism unrelated to the Th17 immune response?

To evaluate the influence of RORγT inhibition by digoxin on inflammatory changes related to interleukin-17 (IL-17) in the pulp of rats treated with zoledronate (ZOL). Forty male Wistar rats were divided into a negative control group (NCG) treated with saline solution, a positive control group (PCG) treated with ZOL (0.20 mg/kg), and three groups treated with ZOL and co-treated with digoxin 1, 2, or 4 mg/kg (DG1, 2, and 4). After four intravenous administrations of ZOL or saline solution in a 70-day protocol, the right molars were evaluated by histomorphometry (number of blood vessels, blood vessels/µm2, cells/µm2, total blood vessel area, and average blood vessel area) and immunohistochemistry (IL-17, TNF-α, IL-6, and TGF-β). The Kruskal-Wallis/Dunn test was used for statistical analysis. PCG showed an increase in total blood vessel area (p=0.008) and average blood vessel area (p=0.014), and digoxin treatment reversed these changes. DG4 showed a reduction in blood vessels/µm2 (p<0.001). In PCG odontoblasts, there was an increase in IL-17 (p=0.002) and TNF-α (p=0.002) immunostaining, and in DG4, these changes were reversed. Odontoblasts in the digoxin-treated groups also showed an increase in IL-6 immunostaining (p<0.001) and a reduction in TGF-β immunostaining (p=0.002), and all ZOL-treated groups showed an increase in IL-17 (p=0.011) and TNF-α (p=0.017) in non-odontoblasts cells. ZOL induces TNF-α- and IL-17-dependent vasodilation and ectasia, and the classical Th17 response activation pathway does not seem to participate in this process.

Emamectin Benzoate-Induced Hepatotoxicity in Rats with Special Reference to Protective Potential of Nigella sativa Oil

This study was designed to explore the hepatotoxicity of emamectin in male rats and the possible effect of Nigella sativa oil (NSO) in ameliorating this. Twenty-eight male rats were used in this study. They were divided into four groups, Control group: rats orally administered distilled water; NSO group: rats administered NSO orally; EMB group: rats administered emamectin benzoate orally; and EMB+NSO group: rats orally co-administered NSO with EMB, with the administrations being performed every other day for 6 weeks. Body weight was measured, liver alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) activities were determined, and total protein and albumin levels were recorded. Histopathological examination of the liver was also performed, along with caspase-3 and TNF-α immunostaining of liver tissue. EMB treatment resulted in decreased body weight, while the co-administration of NSO modulated the EMB-induced alterations in body weight. There were also increases in the activities of serum ALT, AST, and ALP and decreases in total protein and albumin levels in the EMB group. Co-treatment with NSO significantly reduced serum ALT, AST, and ALP and improved total protein and albumin levels. Histopathological examination of the liver in the EMB group revealed the presence of different histopathological alterations that were improved by the co-administration of NSO. Immunostaining of caspase-3 and TNF-α in the liver revealed strong expression in the EMB-treated group. Meanwhile, the EMB+NSO group showed weak positivity for immunoreactive cells.

Thymoquinone alleviates doxorubicin induced acute kidney injury by decreasing endoplasmic reticulum stress, inflammation and apoptosis

ABSTRACT Doxorubicin (DOX) is used as an anticancer drug despite its many side effects. Thymoquinone (THQ) is a plant-derived substance that exhibits antioxidant and anti-inflammatory properties. We investigated the protective effects of THQ on DOX induced nephrotoxicity in rats. Rats were divided into five groups of eight: group 1, untreated control; group 2, olive oil group given olive oil intraperitoneally (i.p.) for 14 days; group 3, THQ group given 10 mg/kg THQ i.p. for 14 days; group 4, DOX group given a single dose of 15 mg/kg DOX i.p. on day 7 of experiment; group 5, DOX + THQ given 10 mg/kg THQ i.p. for 14 days and 15 mg/kg DOX i.p. on day 7. Kidney tissues were evaluated for histopathology. Caspase-3, IL-17, GRP78 and TNF-α immunostaining was used to determine the expression levels of these proteins among the groups. The TUNEL method was used to determine the apoptotic index. Total antioxidant status (TAS), total oxidant status (TOS), and TNF-α and TGF-β1 levels in kidney tissue were measured using ELISA assay. Histopathologic damage, caspase-3, IL-17, GRP78 and TNF-α immunoreactivity, TUNEL positive cells, TOS, TNF-α and TGF-β1 levels were increased in group 4 compared to group 1. The TAS of group 4 decreased compared to group 1. We found decreased caspase-3, IL-17, GRP78 and TNF-α expressions and TUNEL positive cells in group 5 compared to group 4. In rats given DOX, THQ reduced kidney damage by suppressing endoplasmic reticulum stress, inflammation and apoptosis pathways.

TUMOR NECROSIS FACTOR (TNF)-α PLAYS A ROLE IN ORAL ULCERATION IN RATS

<h3>Objectives</h3> Evaluate the role of tumor necrosis factor (TNF)-α in the healing process of oral traumatic ulcers (OTUs) in rats. <h3>Study Design</h3> The animals were randomly divided into 6 experimental groups: 1 treated intravascularly with saline solution and 5 treated with infliximab at Infliximab (INF)-1, INF-INF-3, INF-5, INF-7, or INF-10 mg/kg. The animals were euthanized on days 1, 3, 7, 14, and 21 after ulceration. The ulcers were clinically measured (wound area) and the mucosa samples were histologically (scores 0-4), histomorphometrically (cell counting polymorphonuclear cells), and immunohistochemically (scores 0-3 for TNF-α and alfa-smooth muscle actin (α-SMA)) analyzed. Evans blue assay was performed to measure vascular permeability. One- and two-way analyses of variance and Kruskal-Wallis tests were performed (GraphPad Prism 5.0, <i>P</i> < .05). <h3>Results</h3> There was a reduction in the OTU area of the infliximab-treated groups (INF-5, INF-7, and INF-10) on day 1 (<i>P</i> = .043) and Evan blue extravasation (<i>P</i> = .032), polymorphonuclear cell counts (<i>P</i> = .001), and TNF-α immunostaining (<i>P</i> = .025) were significantly reduced in the dose INF-5 group compared to the saline group on the same day. In day 3, all INF-treated groups showed lower scores than the saline group (<i>P</i> = .023), but α-SMA immunostaining was reduced in day 7 in the INF-treated groups (<i>P</i> = .006). <h3>Conclusions</h3> TNF-α blockade reduced acute inflammatory process and delay wound healing in OTU.

Tumor necrosis factor alpha (TNF-α) blockage reduces acute inflammation and delayed wound healing in oral ulcer of rats.

Oral traumatic ulcers (OTU) are common in dental routine, and the control of proinflammatory cytokines, such as the tumor necrosis factor-alpha (TNF-α), may interfere with OTU repair. Our aim was to evaluate the role of TNF-α in the healing process of OTU in rats. Wistar male rats were divided into six groups: a control-group (treated with 0.1mL/kg of saline) and five groups treated with anti-TNF-α infliximab (INF) at 1, 3, 5, 7, and 10mg/kg immediately before OTU production. The animals were weighed (day 0) and euthanized on days 1, 3, 7, 14 and 21 after ulceration. The ulcers were clinically measured, and the mucosa samples were histologically (scores 0-4), histochemically (collagen assay (pircrosirius)), histomorphometrically (cell counting), and immunohistochemically (TNF-α, α-smooth-muscle-actin (α-SMA), monocyte-chemoattractive-protein-1 (MCP-1), interleukin-8 (IL-8), and fibroblast-growth-factor (FGF)) analyzed. The Evans blue assay was used to measure the vascular permeability. ANOVA-1-2-way/Bonferroni, Kruskal-Wallis/Dunn, and correlation analyses were performed (GraphPad Prism 5.0, p < 0.05). High doses of INF reduced the OTU area (p = 0.043), body mass loss (p = 0.023), vascular permeability (p < 0.001), and reduced delayed histologic scores (p < 0.05), polymorphonuclear (p < 0.001) and mononuclear (p < 0.001) cells, blood vessel counting (p = 0.006), and total (p < 0.001), type-I (p = 0.018), and type-III (p < 0.001) collagen. INF treatment reduced TNF-α immunostaining and delayed MPC-1, FGF, and α-SMA expression, with little/none influence in IL-8 immunostaining. TNF-α blockage by INF reduced acute inflammation in OTU but delayed cell migration and wound healing.

Bariatric Surgery-induced High-density Lipoprotein Functionality Enhancement Is Associated With Reduced Inflammation.

Emerging evidence suggests an association between impaired high-density lipoprotein (HDL) functionality and cardiovascular disease (CVD). HDL is essential for reverse cholesterol transport (RCT) and reduces inflammation and oxidative stress principally via paraoxonase-1 (PON1). RCT depends on HDL's capacity to accept cholesterol (cholesterol efflux capacity [CEC]) and active transport through ATP-binding cassette (ABC) A1, G1, and scavenger receptor-B1 (SR-B1). We have studied the impact of Roux-en-Y gastric bypass (RYGB) in morbidly obese subjects on RCT and HDL functionality. Biomarkers associated with increased CVD risk including tumour necrosis factor-α (TNF-α), high-sensitivity C-reactive protein (hsCRP), myeloperoxidase mass (MPO), PON1 activity, and CEC in vitro were measured in 44 patients before and 6 and 12 months after RYGB. Overweight but otherwise healthy (mean body mass index [BMI] 28 kg/m2) subjects acted as controls. Twelve participants also underwent gluteal subcutaneous adipose tissue biopsies before and 6 months after RYGB for targeted gene expression (ABCA1, ABCG1, SR-B1, TNF-α) and histological analysis (adipocyte size, macrophage density, TNF-α immunostaining). Significant (P < 0.05) improvements in BMI, HDL-cholesterol, hsCRP, TNF-α, MPO mass, PON1 activity, and CEC in vitro were observed after RYGB. ABCG1 (fold-change, 2.24; P = 0.005) and ABCA1 gene expression increased significantly (fold-change, 1.34; P = 0.05). Gluteal fat adipocyte size (P < 0.0001), macrophage density (P = 0.0067), and TNF-α immunostaining (P = 0.0425) were reduced after RYBG and ABCG1 expression correlated inversely with TNF-α immunostaining (r = -0.71; P = 0.03). RYGB enhances HDL functionality in association with a reduction in adipose tissue and systemic inflammation.

A Novel Murine Model of a High Dose Brachytherapy-Induced Actinic Proctitis.

BackgroundRadiation proctitis affects 1-20% of cancer patients undergoing radiation exposure due to pelvic malignancies, including prostate, gynecological and rectum cancers. The patients manifest rectal discomfort, pain, discharge, and bleeding. Notably, the efficacy of prophylactic measures remains controversial due to the lack of adequate animal models that mimic this condition.ObjectiveThe present study then aimed to develop a murine model of high-dose-rate (HDR) brachytherapy-induced proctitis.Material/MethodsC57BL/6 male mice were subjected to HDR (radiation source: iridium-192 [Ir-192]) through a cylindrical propylene tube inserted 2 cm far from the anal verge into the rectum. The animals received radiation doses once a day for three consecutive days (fractions of 9.5 Grays [Gy]), 3.0 mm far from the applicator surface. The sham group received only the applicator with no radiation source. The survival rate was recorded, and a colonoscopy was performed to confirm the tissue lesion development. Following euthanasia, samples of the rectum were collected for histopathology, cytokines dosage (IL-6 and KC), and immunohistochemical analysis (TNF-α and COX-2).ResultsHDR significantly reduced animals’ survival ten days post first radiation exposure (14% survival vs. 100% in the non-irradiated group). Day seven was then used for further investigation. Mice exposed to radiation presented with rectum injury confirmed by colonoscopy and histopathology (P < 0.05 vs. the control group). The tissue damage was accompanied by an inflammatory response, marked by increased KC and IL-6 tissue levels, and immunostaining for TNF-α and COX-2 (P < 0.05 vs. control group).ConclusionsWe established a novel animal model of actinic proctitis induced by HDR brachytherapy, marked by inflammatory damage and low animal mortality.

Elucidation of the Metabolite Profile of Yucca gigantea and Assessment of Its Cytotoxic, Antimicrobial, and Anti-Inflammatory Activities.

The acute inflammation process is explained by numerous hypotheses, including oxidative stress, enzyme stimulation, and the generation of pro-inflammatory cytokines. The anti-inflammatory activity of Yucca gigantea methanol extract (YGME) against carrageenan-induced acute inflammation and possible underlying mechanisms was investigated. The phytochemical profile, cytotoxic, and antimicrobial activities were also explored. LC-MS/MS was utilized to investigate the chemical composition of YGME, and 29 compounds were tentatively identified. In addition, the isolation of luteolin-7-O-β-d-glucoside, apigenin-7-O-β-d-glucoside, and kaempferol-3-O-α-l-rhamnoside was performed for the first time from the studied plant. Inflammation was induced by subcutaneous injection of 100 μL of 1% carrageenan sodium. Rats were treated orally with YGME 100, 200 mg/kg, celecoxib (50 mg/kg), and saline, respectively, one hour before carrageenan injection. The average volume of paws edema and weight were measured at several time intervals. Levels of NO, GSH, TNF-α, PGE-2, serum IL-1β, IL-6 were measured. In additionally, COX-2 immunostaining and histopathological examination of paw tissue were performed. YGME displayed a potent anti-inflammatory influence by reducing paws edema, PGE-2, TNF-α, NO production, serum IL-6, IL-1β, and COX-2 immunostaining. Furthermore, it replenished the diminished paw GSH contents and improved the histopathological findings. The best cytotoxic effect of YGME was against human melanoma cell line (A365) and osteosarcoma cell line (MG-63). Moreover, the antimicrobial potential of the extract was evaluated against bacterial and fungal isolates. It showed potent activity against Gram-negative, Gram-positive, and fungal Candida albicans isolates. The promoting multiple effects of YGME could be beneficial in the treatment of different ailments based on its anti-inflammatory, antimicrobial, and cytotoxic effects.