Subplantar Injection Research Articles

Extrato de tarântula cubensis (Theranekron®) inibe a inflamação no edema induzido por algas vermelhas na pata de ratos

ABSTRACT The aim of this study was to investigate the anti-inflammatory effect of alcoholic extract of Tarantula cubensis alcoholic extract (TCAE) in experimentally induced inflammation in rats. Fifty-four adult Sprague-Dawley male rats were randomly divided into nine groups. Paw edema was induced by 0.2mL subplantar (s.p.) injection of 1% carrageenan (CAR) into the right hind paw. Rats were treated with the nonsteroidal anti-inflammatory drug (NSAID) indomethacin (INDO) (10mg/kg, p.o.) or TCAE at different doses (1, 10 or 100µg/kg) injected s.c. for systemic or s.p. for local anti-inflammatory effect. Saline was used as control. Changes in paw thickness, volume, and weight were calculated as percentages. Formalin-fixed paws were used for histopathological examination. We detected that TCAE applied s.c. at 10µg/kg and 100µg/kg doses resulted in thinner paw thickness, lower paw volume, and lower paw weights four hours after the induction of inflammation when compared with the INDO group (p<0.05). The paw edema inhibitory effect of TCAE applied at a dose of 10µg/kg, s.c. was 68% when compared with the INDO which had an inhibitory effect of 56%. These results were verified with similar histopathological findings. The anti-inflammatory feature of 10µg/kg of TCAE given systematically was similar to the effects of INDO. Our results suggest that TCAE has anti-inflammatory effects by reducing edema and decreasing inflammatory reaction. These results may be attributed to the inhibition of the production of proinflammatory mediators. Thus, TCAE may be considered as a potential anti-inflammatory agent for treating acute inflammatory conditions.

Study of novel 1,4-dihydropyridine derivatives as prospective anti-inflammatory remedies: a randomised controlled trial

Background. Over the past decades, scientific community is motivated on finding new anti-inflammatory agents with a safe and high-effective profile to manage pathology.Objectives. A study of the anti-inflammatory action of novel compounds, 1,4-dihydropyridine derivatives, in a classical formalin-induced paw oedema test in white rats.Methods. Originally synthesised 1,4-dihydropyridine derivatives were preliminarily subjected to virtual screening using the SwissTargetPrediction toolkit. White laboratory rats (130 animals) were divided into a control (formalin oedema) and intact group, 4 comparison (meloxicam, sodium metamizole, sodium diclofenac and indomethacin) and 7 experiment groups by the number of 1,4-dihydropyridine derivatives studied. The samples anti-inflammatory efficacy was evaluated in acute formalin-induced paw oedema model simulated by right hind limb subplantar injection of 0.1 mL 2% formalin. The studied substances were administered intragastrically at 5 mg/kg 1.5 h prior to oedema induction. Oncometry was assessed quantitatively by limb circumference. Animals were managed in compliance with GOST 33044–2014 “Principles of Good Laboratory Practice” at all experiment steps. Experimental data were analysed statistically to describe quantitative variability with variance σ2, mean limb girth a and standard deviation σ. Data homogeneity and reliability were estimated by variation coefficient V and the Wilcoxon T(W) criterion.Results. As the most anti-inflammatory effective, partially hydrogenated mar-040 pyridines (ethyl 4-({[5-cyano-6-{[2-(diphenylamino)-2-oxoethyl]thio}-4-(2-furyl)-2-methyl-1,4-dihydropyridin-3-yl]carbonyl}amino) benzoate) were shown 33-fold superior to indomethacin, 26-fold — to sodium diclofenac, 25-fold — to meloxicam and 30-fold — to sodium metamizole; mar-037 pyridines (ethyl 4-[({[3-cyano-5-({[4-(ethoxycarbonyl)phenyl]amino}carbonyl)-4-(2-furyl)-6-methyl-1,4-dihydropyridin-2-yl]thio}acetyl)amino] benzoate) — 17–23-fold superior vs. reference drugs. We also show that mаr-014 (ethyl 4-({[5-cyano-6-({2-[(3,5-dichlorophenyl) amino]-2-oxoethyl}thio)-4-(2-furyl)-2-methyl-1,4-dihydropyridin-3-yl]carbonyl}amino)benzoate) and mar033 (ethyl 2-[({[3-cyano-5-({[4-(ethoxycarbonyl)phenyl]amino}carbonyl)-4-(2-furyl)-6-methyl-1,4-dihydropyridin-2-yl]thio}acetyl)amino]-4,5,6,7-tetrahydro-1-benzothiophene-3-carboxylate) compounds are 2.7-fold more effective vs. reference drugs.Conclusion. The synthesised 1,4-dihydropyridine compounds reveal high efficacy in experimental assays. Selected novel 1,4-dihydropyridine derivatives exhibit a marked anti-inflammatory activity and offer value in future preclinical trials.

Analgesic properties of the dry combined extract BNO 1016 in leukotriene depended hyperalgesia.

Combined dry extract BNO 1016 is a herbal drug commonly used for rhinosinusitis therapy. It is known that a significant role in rhinosinusitis pathogenesis is played by an inflammatory reaction in maxillary sinuses of the nasal cavity. At the initial stage of any inflammation the largest impact is caused by leukotrienes. Major chronic rhinosinusitis (CRS) according to the guideline EPOS-2020 include facial pain as clinical diagnostic criteria for rhinosinusitis. Therefore, it is reasonable to study the analgesic effects of the combined dry extract BNO 1016 on the model of zymozan- induced hyperalgesia. The tested drug is the combined dry extract BNO 1016 ("Bionorica SE", Germany) at different dose levels. The reference drug is ibuprofen. Leukotriene hyperalgesia was induced by subplantar injection of zymozan into the paw of Wistar rats. Pain threshold was evaluated with Ugo-Basil analgesimeter and analgesic activity was calculated together with mean effective dose (ED50) with help of the Probit Analysis. The highest analgesic activity was observed at a dose level of 500 mg/kg of BNO 1016 during all time-points of the experiment and reached 169.5% 3h after hyperalgesia onset. The analgesic activity of BNO 1016 at doses of 50 mg/kg in 1h, 2h, 3h after induction hyperalgesia, 150 mg/kg and 500 mg/ kg was credibly higher than that of Ibuprofen at all time-points of the experiment. Based on the experimental data, the index of ED50 for analgesic activity of BNO 1016 was calculated with Probit Analysis. The combined dry extract BNO 1016 shows a high level of analgesic activity in the leukotriene-depended hyperalgesia model, which is very promising for the improvement of treatment of patients with CRS, but additional preclinical and clinical researches are reasonable.

CHARACTERISTICS OF THE MECHANISMS OF CYTOPROTECTIVE ACTIVITY OF CRYOPRESERVED PLACENTA EXTRACT ACCORDING TO MORPHOLOGICAL AND BIOCHEMICAL STUDIES OF THE GASTRIC MUCOSA IN THE EXPERIMENT

Relevance. Non-steroidal anti-inflammatory drugs (NSAIDs) are used as antipyretics and analgesics. In this case, NSAIDs have a side effect - ulcerogenic. Therefore, it is important to find means to correct the ulcerogenic action of NSAIDs. Potential antiulcerogenic action may be in cryopreserved human placenta extract "Cryocell-placental cryoextract".
 Objective: to investigate the effect of cryopreserved placenta (CEP) extract on the morphological state of the stomach and the processes of protein synthesis in the gastric mucosa with the introduction of NSAIDs diclofenac sodium in rats with experimental rheumatoid arthritis.
 Materials and methods. The study was performed on 28 nonlinear male rats weighing 200-220 g. The animals were divided into 4 groups: I (n = 7) – intact rats; II (n = 7) – rats with adjuvant arthritis (AA), this is a treatment without treatment; III (n = 7) – rats with AA, treated with diclofenac sodium (DN), 8.0 mg / kg, intragastrically (i/g); IV (n = 7) – rats with AA, treated with DN, 8.0 mg / kg, i/g and CEP, 0.16 ml / kg body weight, intramuscularly (i/m).
 AA was simulated by subplantar injection of complete Freund's adjuvant into the hind right limb at a rate of 0.1 ml per rat. The day of adjuvant administration was considered as the "0" day of the experiment. AA treatment was performed from 14 to 28 days. DN was applied daily, and CEP – with an interval of 3 days: for 14, 17, 20, 23 and 26 days. On day 28, the animals were removed from the experiment by cervical dislocation under inhalation anesthesia.
 To assess morphological changes, gastric fragments were fixed in 10.0% neutral formalin solution. Histological sections 5–7 μm thick were stained with hematoxylin and eosin, as well as Schiff's reagent (fuchsin sulfuric acid) with iodic acid. The concentration of total protein (TP) was determined spectrophotometrically by light absorption at a wavelength of 546 nm. The content of RNA and DNA in the homogenates of the gastric mucosa was determined spectrophotometrically by reaction with perchloric acid.
 Results. In rats with AA, the mucous membrane (MM) had a preserved structure in all animals. There were no signs of hyper- or hyposecretion of mucins when stained with a chic reaction with alcyan blue. There were foci of desquamation of the superficial epithelium and single lymphocytes. The introduction of DN in rats with AA led to signs of erosive changes in MM. Deep erosions and single ulcerative defects were found in 28.6% of rats, reaching the depth of the lesion to the muscle plate. There is a small amount of mucopolysaccharides with a predominance of acidic glycoproteins. Reduced content of neutral glycoproteins. Single desquamated epitheliocytes were detected. Focal lympho-plasmacytic infiltration with exudate with eosinophilic leukocytes was noted in the submucosal layer of MM. Against the background of the combined use of DN and CEP in rats with AA in any animal, no significant violations of the histoarchitectonics of the MM, except for isolated foci of desquamation of the superficial epithelium.
 In gastric MM homogenates in rats with AA there was a decrease (p <0.01) in the level of TP by 14.1% and a decrease (p <0.05) in the content of RNA and DNA by 27.5% and 14.3%, respectively, relative to indicators of intact animals.
 Against the background of DN monotherapy, there was an even greater decrease (p <0.001) in the level of TP: by 26.2% (40.3 ± 1.43 μg / mg) relative to intact animals (54.6 ± 1.39 μg / mg). And the content of RNA and DNA was 0.59 ± 0.57 μg / ml and 2.21 ± 0.27 μg / ml, respectively.
 Against the background of the combined use of DN and CEP, comparable to the level of intact animals indicators of TP and DNA were noted. Thus, the level of TP was 53.0 ± 2.09 μg / mg, which was only 2.9% lower than intact animals (54.6 ± 1.39 μg / mg), and the DNA content by 5.1% (p = 0.3) was lower than intact animals (3.1 ± 0.12 μg / ml) and was respectively 2.94 ± 0.08 RNA level (1.72 ± 0.12 μg / ml) on 17.6% exceeded (p = 0.1) intact animals (1.46 ± 0.08 μg / ml).
 Conclusions. The combined use of placental cryoextract and diclofenac sodium eliminates the negative effect of the latter on protein metabolism in the gastric mucosa of rats with adjuvant arthritis. Thus, the level of total protein was 53.0 ± 2.09 μg / mg, which is only 2.9% lower than intact animals. Restoration of diclofenac sodium-induced mucin secretion by the integumentary epithelium of the gastric mucosa is one of the mechanisms of cytoprotective activity of cryopreserved placenta extract in experimental ulcerogenesis.

Hydrogen Sulfide Inhibits Inflammatory Pain and Enhances the Analgesic Properties of Delta Opioid Receptors.

Chronic inflammatory pain is present in many pathologies and diminishes the patient’s quality of life. Moreover, most current treatments have a low efficacy and significant side effects. Recent studies demonstrate the analgesic properties of slow-releasing hydrogen sulfide (H2S) donors in animals with osteoarthritis or neuropathic pain, but their effects in inflammatory pain and related pathways are not completely understood. Several treatments potentiate the analgesic actions of δ-opioid receptor (DOR) agonists, but the role of H2S in modulating their effects and expression during inflammatory pain remains untested. In C57BL/6J male mice with inflammatory pain provoked by subplantar injection of complete Freund’s adjuvant, we evaluated: (1) the antiallodynic and antihyperalgesic effects of different doses of two slow-releasing H2S donors, i.e., diallyl disulfide (DADS) and phenyl isothiocyanate (P-ITC) and their mechanism of action; (2) the pain-relieving effects of DOR agonists co-administered with H2S donors; (3) the effects of DADS and P-ITC on the oxidative stress and molecular changes caused by peripheral inflammation. Results demonstrate that both H2S donors inhibited allodynia and hyperalgesia in a dose-dependent manner, potentiated the analgesic effects and expression of DOR, activated the antioxidant system, and reduced the nociceptive and apoptotic pathways. The data further demonstrate the possible participation of potassium channels and the Nrf2 transcription factor signaling pathway in the pain-relieving activities of DADS and P-ITC. This study suggests that the systemic administration of DADS and P-ITC and local application of DOR agonists in combination with slow-releasing H2S donors are two new strategies for the treatment of inflammatory pain.

An Experimental Study of the Cryopreserved Placenta Extract Effect On the Sodium Diclofenac Anti-Inflammatory Activity

Background. As a means of correcting the ulcerogenic effect of nonsteroidal anti-inflammatory drugs, our attention was attracted by a cryopreserved extract of the human placenta, which has a multivector spectrum of biological activity. To date, there is no information about its effect on the specific activity of this class of medicines (anti-inflammatory, analgesic, etc.) in published sources. Objective. We are aimed to characterize the effect of cryopreserved placenta extract on the anti-inflammatory activity of diclofenac sodium when administered separately in a model of acute exudative inflammation. Methods. Experimental studies in vivo were conducted on 28 nonlinear male laboratory rats. The model of acute exudative inflammation was reproduced by subplantar injection of 0.1 ml of 1.0% aqueous solution of λ-karagenin into the right hind limb of rats. Cryopreserved placenta extract was administered intramuscularly at a dose of 0.16 ml/kg 60 minutes before diclofenac sodium (8 mg/kg). Results. Preventive administration of diclofenac sodium caused an antiexudative effect as early as 30 minutes after administration of λ-karagenin – its anti-inflammatory activity was 11.0%, which is 4.6 times higher than similar indicators at the same time in rats injected with placental cryoextract. At 60 minutes of observation, diclofenac sodium was comparable in anti-inflammatory activity with cryopreserved placenta extract: 28.6% and 22.2%, respectively, but at 120 and 180 minutes, diclofenac sodium exceeded the studied cryoextract in antiphlogistic effect by 1.6 times in both periods of observation. The anti-inflammatory effect of the combined separate administration of placenta cryoextract and diclofenac sodium before λ-karagenin for 30 and 60 minutes was 12.7% and 32.3%, respectively, which is comparable with analogous indicators against the background of diclofenac sodium monotherapy. However, at 120 minutes of observation, the group of combined use of placenta cryoextract and diclofenac sodium showed the greatest anti-inflammatory effect among rats of all the studied groups – 52.6%, which was 2.2 times higher than the indicators of the placenta cryoextract monotherapy group and 1.4 times lower than the indicators of the rats of the diclofenac sodium monotherapy group. Conclusions. 4 hours after administration, placental cryoextract had a suppressive effect on kinins like diclofenac sodium, and in the prostaglandin period of caragenin-induced inflammation against the background of combined use of the studied cryoextract and diclofenac sodium, the anti-inflammatory activity was 46.4 %. This suggests a suppressive effect on the production of prostaglandins as a possible mechanism of anti-exudative action of cryopreserved placenta extract.

The effect of complex herbal extract and methotrexate on suppressing adjuvant arthritis in rats

The present study evaluated the therapeutic benefits of complex herbal preparation named CBMDS, consisting of turmeric (Curcuma longa), Boswellia (Boswellia serrata), Methylsulphonylmethane, Devil’s Claw (Harpagophytum procumbens) and Silymarin, using it in combination with methotrexate, in order to suppress adjuvant arthritis in rats, and to attenuate methotrexate-induced liver damage. Adjuvant arthritis was induced in 28 rats by a single subplantar injection of complete Freund’s adjuvant (0.1 mL) into the left hind paw. The animals were divided into four groups (with seven animals in each). Group I received CBMDS, Group II - CBMDS in combination with methotrexate, and Group III just methotrexate. The treatment lasted from day 0 to day 17 (CBMDS was given daily except weekends in a dose of 160 mg/kg, methotrexate - 2 mg/kg once a week). Group IV was the control group. Clinical (body weight, hind paw volume, erythrocyte sedimentation rate, leukocyte count), biochemical (serum pro-/antioxidant activity markers), immunological (serum interleukin levels) and histological changes in joint and liver tissues were evaluated. CBMDS significantly alleviated arthritis and reduced hepatic damage, which was more evident in the methotrexate group. The combined treatment also markedly reduced arthritic symptoms and levels of malondialdehyde. Antioxidant activity was significantly higher in treated Groups I and II. CBMDS and its combination with methotrexate promoted anti-arthritic action, reduced histological changes in the joint tissues, and minimized methotrexate-induced liver toxicity.

Effect of a Selection of Skin Penetration Enhancers on Topical Anti-Inflammatory Effect of Boswellic Acids in Carrageenan-Induced Paw Edema in Rats.

Background:Boswellia species have been used for treatment of chronic inflammatory disease. Several studies have documented the anti-inflammatory effect of Boswellic acids (BAs) after systemic administration. This study was aimed to evaluate the effect of some skin penetration enhancers on topical anti-inflammatory effect of BAs in rats.Materials and Methods:Male Wistar rats weighting 180–220 were used. Anti-inflammatory activity was assessed using carrageenan test. BAs dissolved in ethanol, propylene glycol 2%, 5%, olive oil and applied topically. Menthol, D-limonene, or eucalyptus oil 0.5%, 1% were also tested as other skin penetration enhancers and applied topically 30 min prior to subplantar injection of carrageenan into the right hind paw of rats. The volume of the paw was measured at 0 and 4 h after carrageenan with a digital plethysmometer and the difference was used as an index of inflammation. Piroxicam gel was used as a standard drug.Results:A 4% ethanolic solution of BAs showed significant anti-inflammatory effect. Propylene glycol (2% and 5%) in alcohol did not change the effect. Olive oil also enhanced penetration of BAs. Menthol 0.5%, 1% and D-limonene 0.5%, 1% did not show any significant change compared to olive oil alone. In the present study, eucalyptus oil 1% in olive oil was known as the best carrier for transdermal delivery of BAs.Conclusion:BAs have considerable topical anti-inflammatory effects and olive oil alone or especially in combination with eucalyptus oil can be promising vehicles for skin penetration of topical BAs.

ОЦЕНКА АНАЛЬГЕТИЧЕСКОЙ И ПРОТИВОВОСПАЛИТЕЛЬНОЙ АКТИВНОСТИ АДДУКТА 1-ГИДРОКСИ-1,1-ЭТИЛИДЕНДИФОСФОНОВОЙ КИСЛОТЫ И БИС(2-ПИРИДИЛ-1,2,4-ТРИАЗОЛИЛ-3)ПРОПАНА ПРИ КУРСОВОМ ВВЕДЕНИИ У САМЦОВ КРЫС

The article presents the results of evaluation of analgesic and anti-inflammatory activity of the adduct of 1-hydroxy-1,1-ethylidendiphosphonic acid and bis(2-pyridyl-1,2,4-triazolyl-3)propane (HEA+BPP) in male rats with a 21-day course administration. The aim of the study was to evaluate analgesic and anti-inflammatory activity in male rats with 21-day course administration of HEA+BPP course administration with 21-day course administration. The animals were divided into 7 groups of 10 individuals. Analgesic activity of HEA+BPP was tested on 40 males, and anti – inflammatory activity was tested on 30 others. In studies of analgesic activity of HEA+BPP, males of one group were control and received intraperitoneal injections of 0.2 ml of saline solution every day at 10.00 for 21 days, the others groups received 0.2 ml of intraperitoneal injections of HEA+BPP at doses of 5, 50 and 100 mg/kg at the same time for 21 days. In analgesic studies, groups of animals were similarly formed, when testing the anti-inflammatory effect of HEA+BPP, only without injecting a dose of 100 mg/kg. HEA+BPP synthesized at the Department of General and inorganic chemistry of the faculty of biology and chemistry of the Tauride Academy (structural division) of the V. I. Vernadsky Crimean Federal University. The chemical purity of HEA+BPP was at least 98 %. The analgesic activity of HEA+BPP was evaluated by the parameters of pain sensitivity of animals in the background and 1 hour after intraperitoneal injections on the 1st, 6th, 10th, 14th and 21st days of the experiment in the models of acute pain stress «tail-flick», Randall-Sellitto and «hot plate». Before the tail-flick and Randall-Sellitto tests were performed, the animals were placed in special rat retainers after injection. The anti-inflammatory activity of HEA+BPP was evaluated in the formalin paw edema test in rats. Acute inflammatory response was reproduced by subplantar injection of 0.1 ml of 2 % formalin solution in the form of an aqueous solution into the left hind leg of a rat, and then the value of limb edema was measured using the method of water plethysmometry on the 10th and 21st days of course administration of HEA+BPP in doses of 5 and 50 mg/kg. The data are presented as mean and standard error mean, statistical analysis performed using the software packages StatSoft/STATISTICA 8 and Graph Pad Prism 7.0. The significance of differences between groups was determined using one-way analysis of variance (ANOVA) with post hoc test Tukey and non-parametric criterion of multiple comparisons of Dunn, according to official methodological recommendations on statistical treatment of the results of preclinical studies of medicines, given in the literature. During course administration to male rats of HEA+BPP for 21 days in the dose range from 5 to 100 mg / kg, it was found that this compound mainly demonstrates effectiveness as an analgesic with a single injection, only for mechanical pain, this substance is effective with a course administration at a dose of 5 mg/kg: The «tail-flick» test showed, that using of HEA+BPP as an analgesic involved in the spinal mechanism of pain sensitivity regulation is advisable in doses of 5, 50 and 100 mg/kg only with a single injection: latent period of the reaction discharge of the tail increased relative to control at a dose of 5 mg/kg on the 1st day of administration of HEA+BPP by 32.7 % (p≤0.05), at a dose of 50 mg/kg on the 1st and 21st days of administration – by 40 % (p≤0.01) and 67.1 % (p≤0.001) and at a dose of 100 mg/kg on the 1st day of administration – by 80.0 % (p≤0.01), respectively. At a dose of 5 mg/kg on the 10th day of administration, on the contrary, was discovered algogenic effect of HEA+BPP (latent period of the reaction discharge of the tail was significantly decreased relative to control by 24.1 % (p≤0.01). In the test of Randall-Selitto the efficiency of single use HEA+BPP as an analgesic in mechanical pain at doses of 50 and 100 mg/kg (pain threshold was significantly increased relative to control at 25.2 % (p≤0.01) and 80 % (p≤0.01), and in course administration only at a dose of 5 mg/kg (pain threshold was significantly increased relative to control at 55.3 % (p≤0.001), 72.5 % (p≤0.001) and 49.6 % (p≤0.01) on the 6th, 14th and 21st day of the course introduction). The «hot plate» test shows the feasibility of only a single use of HEA+BPP as an analgesic in doses of 5 and 50 mg/kg, since these effects were unstable and unstable when administered on a course basis. The «formalin test» revealed the inflammatory and exudative effect of HEA+BPP at a dose of 5 mg/kg, in which HEA+BPP on the 21st day of administration increased the formalin edema of the rat paw by 70.9 % (p≤0.01).

Active Cousinia thomsonii Extracts Modulate Expression of Crucial Proinflammatory Mediators/Cytokines and NFκB Cascade in Lipopolysaccharide-Induced Albino Wistar Rat Model.

IntroductionChronic inflammation is implicated in a multitude of diseases, including arthritis, neurodegeneration, autoimmune myositis, type 2 diabetes, rheumatic disorders, spondylitis, and cancer. Therefore, strategies to explore potent anti-inflammatory regimens are pivotal from a human-health perspective. Medicinal plants represent a vast unexplored treasure trove of therapeutically active constituents with diverse pharmacological activities, including anti-inflammatory properties. Herein, we evaluated Cousinia thomsonii, an edible medicinal herb, for its anti-inflammatory/immunomodulatory properties.MethodsSoxhlet extraction was used to obtain different solvent extracts (hexane, ethyl acetate, ethanol, methanol, and aqueous extract) in increasing order of polarity. In vitro anti-inflammatory assays were performed to investigate the effects of extracts on protein denaturation, proteinase activity, nitric oxide surge, and erythrocyte-membrane stabilization. The most effective extracts, ie, ethyl acetate (CTEA) and ethanol (CTE) extracts (150–200 g) were selected for further in vivo analysis using albino Wistar rats. Wistar rats received varying concentrations of CTEA and CTE (25, 50, and 100 mg/kg) for 3 weeks, followed by a single subplantar injection of lipopolysaccharide. Dexamethasone served as positive control. Blood was obtained from the retro-orbital plexus and serum separated for estimation of proinflammatory cytokines (IL6, IL1β, IFNγ and TNFα). Western blotting was performed to study expression patterns of crucial proteins implicated in the NFκB pathway, ie, NFκB p65, NFκB1 p50, and NFκB2 p52. Histopathological examination was done and gas chromatography–mass spectrometry (GC-MS) carried out to reveal the identity of compounds responsible for ameliorating effects of C. thomsonii.ResultsAmong five tested extracts, CTEA and CTE showed marked inhibition of protein denaturation, proteinase activity, nitric oxide surge and erythrocyte-membrane hemolysis at 600 μg/mL (P<0.001). Both these extracts showed no toxic effects up to a dose of 2,500 mg/kg. Extracts exhibited concentration-dependent reductions in expression of IL6, IL1β, IFNγ, TNFα, NFκB-p65, NFκB1, and NFκB2 (P<0.05). Healing effects of extracts were evident from histopathological investigation. GC-MS analysis revealed the presence of important anti-inflammatory compounds, notably stigmast-5-en-3-ol, oleate, dotriacontane, ascorbic acid, n-hexadecanoic acid, and α-tocopherol, in C. thomsonii.ConclusionC. thomsonii possesses significant anti-inflammatory/immunomodulatory potential by virtue of modifying levels of proinflammatory cytokines/markers and NFκB proteins.

Pain influences food preference and food-related memory by activating the basolateral amygdala in rats.

The amygdala has been demonstrated to contribute to pain-related behavior and food preference. Here, the effect of pain on food preference and food-matched visual-cue memory, in the presence or absence of a basolateral amygdala (BLA) lesion, has been evaluated using a novel innovative apparatus and protocol. Forty adult male Wistar rats wererandomly divided intofive groups (n = 8) as follows: control, pain, ibuprofen + pain, BLA lesion, BLA lesion + pain groups. Bilateral lesions of the BLA were produced by passing a current of 1.5mA for 7s. Pain was induced ontheright hind pawof theratsby sub-plantar injection of 50μl of 2.5% formalin. The animals were encountered with four different meals including wholemeal, wholemeal + sugar, white flour, and biscuit. Each test sessionconsisted of six trials with inter-trial intervals of 15min. The number of visits to each meal zone and port, the amount oftime spentin each food zone and port, traveled distance in each food zone, food consumption per each visit and the total food consumption were recorded. The control group showed a high biscuit preference and low white flour preference. Rats suffering BLA lesion and rats in the BLA lesion + pain group exhibited a shifted preference curve. They had a bias toward eating wholemeal + sugar rather than white flour and biscuit. This group also showed an impaired spatial memory. In conclusion, our findings suggest that the BLA may be involved in pain-induced food preference and food-matched visual-cue memory.

Retraction Note: Release-Active Dilutions of Diclofenac Enhance Anti-inflammatory Effect of Diclofenac in Carrageenan-Induced Rat Paw Edema Model.

This article has been retracted. Please see the Retraction Notice for more detail: https://doi.org/10.1007/s10753-020-01359-x.

Anti-Inflammatory Activity of Rhaphidophora pinnata (L.F) Schott Leaf Extract

BACKGROUND: Cancer growth is influenced by many factors and in general it is an interaction between gene factors and environmental factors, especially the microenvironment that exists around cancer. The inflammatory response plays a decisive role in various stages of cancer growth.&#x0D; AIM: The aim of this study was to determine the anti-inflammatory activity of ethanol extract of Rhaphidophora pinnata leaves.&#x0D; METHODS: R. pinnata leaf extract was obtained by percolation method using 96% ethanol as the solvent at room temperature. Anti-inflammatory activity was determined based on the paw edema method. Thirty male albino mice were treated orally with sodium carboxyl cellulose suspension (as negative control group), R. pinnata leaf extract (35, 70, 140, and 280 mg/kgBW), and diclofenac (as positive control group), 60 min before 0.2 mL 1% carrageenan injection. The paw thickness was measured using plethysmometer before injecting the carrageenan and after 1, 2, 3, 4, 5, and 6 h.&#x0D; RESULTS: The subplantar injection of carrageenan caused a time-dependent paw edema in the mice. Oral administration of R. pinnata leaf extract inhibited paw swelling at 1, 2, 3 4, 5, and 6 h after carrageenan injection. R. pinnata leaf extracts doses of 35, 70, 140, and 280 mg/kgBW gave a percentage inhibition of 56.56%, 56.18%, 62.77%, and 49.30%, respectively. The effective dose of R. pinnata leaf extract as an anti-inflammatory was 140 mg/kgBW.&#x0D; CONCLUSION: Ethanol extract of R. pinnata leaf has anti-inflammatory activity in male albino mice.

Astragalin attenuates oxidative stress and acute inflammatory responses in carrageenan-induced paw edema in mice.

Astragalin is a flavonoid existed in several edible and medicinal plants and was recorded to have multiple biological and pharmacological significances. This work aimed to assess the possible protective effect of astragalin administration against oxidative tension, acute inflammation and histopathological deformations in a mouse paw edema model induced following intra sub-plantar injection of carrageenan. Thirty-six male Swiss mice were divided into four groups: control, carrageenan, astragalin (75mg/kg) + carrageenan, and indomethacin (10mg/kg) + carrageenan. Astragalin administration for five consecutive days to carrageenan injected mice showed a significant reduction in the development of paw in a time dependent effect, inhibited lipoperoxidation by-product, malondialdehyde and increased superoxide dismutase and catalase activities. Astragalin was found also to suppress the inflammatory signaling in the inflamed tissue as exhibited by the decreased myeloperoxidase activity along with the decreased protein and transcriptional level of pro-inflammatory cytokines including tumor necrosis factor-alpha, interleukin-1 beta and interleukin-6. Moreover, inducible nitric oxide synthase and cyclooxygenase-2 expressions and their products (nitric oxide and prostaglandin E2) were downregulated. Additionally, astragalin decreased monocyte chemoattractant protein-1 and nuclear factor kappa B expression in the inflamed paw tissue. The recorded findings provide evidences for the potential application of astragalin as a plant-derived remedy for the treatment of acute inflammation due to its promising antioxidant and anti-inflammatory activities along with its ameliorative impact against the histopathological changes in the paw tissue.

Cashew (Anacardium occidentale L.) Nuts Counteract Oxidative Stress and Inflammation in an Acute Experimental Model of Carrageenan-Induced Paw Edema.

Background: Anacardium occidentale L. is a medicinal plant with powerful anti-oxidative and anti-inflammatory properties. Acute inflammatory events cause tissue alterations, decrease of anti-oxidative endogenous enzymes such as superoxide dismutase, catalase and glutathione, neutrophils infiltration, increase in the activities of myeloperoxidase, malondialdehyde, and pro-inflammatory release. Methods: Paw edema was induced by subplantar injection of carrageenan into the right hind paw in rats, but 30 min before a group of animals were orally treated with 100 mg/kg of cashew nuts to evaluate the anti-inflammatory and anti-oxidative response. Results: In the present work, we found that (1) cashew nuts reduced the development of carrageenan-induced paw edema limiting the formation of edema and pain; (2) cashew nuts ameliorated the diminutions of the anti-oxidative enzymes caused by carrageenan injection; (3) cashew nuts decreased myeloperoxidase malondialdehyde activity induced by carrageenan; and (4) cashew nuts acted by blocking pro-inflammatory cytokines response and nitrate/nitrite formation stimulated by carrageenan injection. Conclusions: The mechanisms of anti-inflammatory and analgesic effects exerted by cashew nuts were relevant to oxygen free radical scavenging, anti-lipid peroxidation, and inhibition of the formation of inflammatory cytokines.

Mechanisms Underlying the Anti-Inflammatory Activity of Bergamot Essential Oil and Its Antinociceptive Effects.

Renewed interest in natural products as potential source of drugs led us to investigate on both the anti-inflammatory and anti-nociceptive activity of Citrus bergamia Risso et Poiteau (bergamot) essential oil (BEO). Carrageenan-induced paw edema in rats was used as an experimental model of inflammation. Because of the toxicity of furocoumarins, we performed our study by using the BEO fraction deprived of these compounds (BEO-FF). Treatment with BEO-FF led to a significant inhibition of paw edema induced by a sub-plantar injection of carrageenan. Moreover, histological examination of BEO-FF-treated rat paw biopsies showed a reduction of pathological changes typical of edema. Pre-treatment with BEO-FF significantly reduced interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α levels in the paw homogenates, as well as nitrite/nitrate and prostaglandin E2 (PGE2) content in exudates. In addition, BEO-FF possesses antioxidant properties, as determined by cell-free assays. Furthermore, results of the writhing test showed that BEO-FF elicited a pronounced analgesic response, as demonstrated by a significant inhibition of constrictions in mice receiving acetic acid, with respect to control animals, whereas the results of the hot plate test suggested that the supra-spinal analgesia participates in the anti-nociceptive effect of BEO-FF. Our study indicates that BEO-FF exerts anti-inflammatory and anti-nociceptive effects, and suggests its potential role as an anti-edemigen and analgesic drug.

Analgesic, anti-inflammatory and hypouricemic effects of GT1 film-coated tablets on experimental animals

Aim: To evaluate pain relief, anti-inflammatory and hypouricemic effects of GT1 tablets on experimental animals.&#x0D; Method: GT1 at the doses of 22.32 g/kg/day and 66.96 g/kg/day were evaluated for its analgesic effect in three models (hot plate, pain threshold, and acetic acid-induced writhing), its chronic anti-inflammatory effect in the granulomatous reaction model, and its hypouricemic effect in potassium oxonate-induced hyperuricemic mice. Acute anti-inflammatory effects of GT1 at the doses of 11.16 g/kg/day and 33.48 g/kg/day were evaluated in rats with two models: carrageenin-induced paw edema and peritonitis.&#x0D; Results: GT1 prolonged the temperature reaction time on the hot plate (22.73 s and 20.37 s at both doses of 22.32 g/kg and 66.96 g/kg, respectively, compared to 16.96 s in control group), reduced the number of acid acetic-induced writhing effects, decreased the weight of granulomas, and decreased the level of acid uric in blood and urine (p &lt; 0.05). GT1 caused a significant reduction in paw edema after subplantar injection of carrageenan in rats (p &lt; 0.05). Moreover, there was a substantial decline of GT1 at the dose of 11.16 g/kg/day in terms of the volume and the quantity of protein in the inflammation fluid of the peritonitis model (p &lt; 0.05).&#x0D; Conclusion: GT1 at both doses of 11.16 g/kg/day and 33.48 g/kg/day posed acute anti-inflammatory effects on rats. GT1 at both doses of 22.32 g/kg/day and 66.96 g/kg/day exerted analgesic, chronic anti-inflammatory and hypouricemic effects on mice.

Amelioration of adjuvant-induced arthritis by exposure to low dose gamma radiation and resveratrol administration in rats

Purpose: Low dose radiation has been reported as an effective treatment for rheumatoid arthritis via multiple dose exposures. The present study was designed to increase the therapeutic efficacy of low dose radiation with the minimum exposure level in arthritic rats by concurrent administration of resveratrol (RSV) as an adjunctive therapy with anti-inflammatory properties.Materials and methods: Rats were rendered arthritic by sub-plantar injection of Freund’s complete adjuvant (FCA) and exposed to low dose radiation at a total exposure level of 0.5 Gy (2 × 0.25). During the exposure course, RSV (50 mg/kg) was orally administered once daily for two weeks. Diclofenac (3 mg/kg) was administered as a standard anti-inflammatory drug. Paw volume was measured every 4 days. After 28 days of induction, rats were sacrificed and serum was collected for estimation of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), thiobarbituric acid reactive substances (TBARS), and total nitrate/nitrite (NOx). Furthermore, paws were dissected for histopathological examinations and immuno-histochemical estimation of nuclear factor-kappa B p65 (NF-κB p65) expression.Results: Administration of RSV during the low dose radiation exposure course produced a significant decrease in the paw swelling and a potentiated inhibition in the serum levels of TNF-α, IL-1β, TBARs, and NOx. The dual treatment strategy alleviated the histopathological damage to a greater extent than that produced by each treatment. Moreover, a pronounced suppression of NF-κB p65 expression in the synovial tissue was observed in the combination group. The combination treatment showed a nearly similar potency to that observed in the diclofenac treated group.Conclusion: Administration of RSV augmented the modulatory activity of low dose radiation with minimum exposure level on the disease progression.

Изучение противовоспалительной активности экстракта змееголовника молдавского

Змееголовник молдавский (Dracocephalum moldavica L.) в народной медицине используется в качестве противовоспалительного, ранозаживляющего, отхаркивающего и седативного средства. В ФГБНУ ВИЛАР разработан змееголовника молдавского травы экстракт сухой под условным названием «Люкатил» (сумма фенольных соединений 64,12% в пересчёте на цинарозид). Цель работы - изучение острой токсичности и противовоспалительной активности экстракта змееголовника для разработки на его основе лекарственного препарата. Методика. Проведено определение параметров острой токсичности и противовоспалительной активности экстракта. При изучении острой токсичности экстракта по методу Кербера использованы белые нелинейные мыши-самцы в количестве 30 особей. «Люкатил» вводили животным внутрижелудочно в дозах 500, 1000, 1500 и 2000 мг/кг. Для выявления противовоспалительной активности экстракта змееголовника молдавского использована in vitro ферментная биотест-система на основе индуцибельной NO-синтазы. Для выявления противовоспалительной активности экстракта in vivo использованы нелинейные мыши-самцы. Оценку влияния экстракта в дозе 200 мг/кг на экссудативную стадию воспаления проводили на модели 1% формалинового отёка. В качества препарата сравнения использовали индометацин (5 мг/кг). Формалиновый отёк вызывали однократным субплантарным введением под апоневроз задней правой лапки мыши 0,05 мл 1% формалина в качестве флогогенного агента. Величину отёка определяли по разнице в массе лапок контрольных и опытных животных и рассчитывали процент снижения степени отёка. Результаты. При однократном введении экстракт «Люкатил» не приводил к гибели животных, изменения внешнего вида и поведенческих реакций мышей не наблюдалось. В соответствии с классификацией токсичности химических веществ по ГОСТ 12.1.007-76 «Люкатил» является малотоксичным веществом. In vitro установлена высокая противовоспалительная активность экстракта, при этом остаточная активность iNOS снижалась до 25%. Экстракт в дозе 200 мг/кг in vivo обладал статистически значимым противовоспалительным эффектом. Он подавлял развитие экссудативной фазы воспаления на 33,7%, по сравнению с контрольной группой животных, уступая противовоспалительному эффекту индометацина. Заключение. Змееголовника молдавского травы экстракт сухой под условным названием «Люкатил» является малотоксичным веществом, обладает выраженным противовоспалительным эффектом в опытах in vitro, in vivo и является перспективным объектом для дальнейшего фармакологического изучения в качестве противовоспалительного лекарственного средства.Moldavian dragonhead (Dracocephalum moldavica L.) is used in traditional medicine as an anti-inflammatory, wound-healing, expectorant, and sedative means. In our Institute, a Moldavian dragonhead herb dry extract (total phenolic content, 64.12% in cynaroside equivalent) was developed and conventionally named Lyukatil. Objective. To study acute toxicity and anti-inflammatory activity of the dragonhead extract for developing a drug based on this extract. Method. Parameters of acute toxicity and anti-inflammatory activity of the extract were assessed. The study of acute toxicity of the extract was performed using the Kerber method on male white mongrel mice (n=30). Lyukatil was administered to the animals intragastrically at doses of 500 mg/kg, 1000 mg/kg, 1500 mg/kg, and 2000 mg/kg. Anti-inflammatory activity of the Moldavian dragonhead extract was determined in vitro using an enzyme Biotest system based on inducible NO synthase. Mongrel male mice were used to study the anti-inflammatory activity of the extract in vivo. The effect of the extract at a dose of 200 mg/kg on the exudative phase of inflammation was evaluated on a model of 1% formalin-induced edema. Indomethacin 5 mg/kg was used as a reference drug. Formalin edema was induced by a single subplantar injection of 0.05 ml of 1% formalin as a phlogogenic agent under the aponeurosis of the right hind leg. The degree of edema was determined by the difference in leg weights in control and experimental animals; then the decrease in edema was calculated in per cent. Results. A single administration of the extract Lyukatil did not cause death of animals, changes in the appearance or in behavioral responses, shortness of breath, or drowsiness. In accordance with the toxicity classification for chemical substances as per GOST Standard 12.1.007-76, Lyukatil is a low-toxic substance. The extract at a dose of 200 mg/kg exerted a significant anti-inflammatory effect as shown by suppression of the exudative phase of formalin-induced inflammation by 33.7% compared to the control group. However, this effect was inferior to the anti-inflammatory effect of indomethacin. Conclusions. The Moldavian dragonhead herb dry extract under the conventional name of Lyukatil is a low-toxic substance that has a significant anti-inflammatory effect both in vitro and in vivo and is a promising target for further pharmacological studies as an anti-inflammatory drug.

Anti-Inflammatory Potential Effect of Flavonoid Rich Ethyl Acetate Fraction of Methanolic Extracts of Stereospermum Suaveolens DC (Bignoniaceae) Leaves in Experimental Animals

Objective: To assess anti-inflammatory potential effect of flavonoid rich ethyl acetate fraction of methanolic extract of Stereospermum suaveolens DC (Bignoniaceae) leaves. Method: The phytochemical investigation and TLC analysis was studied as per standard phytochemical method. The inflammation was induced in wistar rats by sub-plantar injection of 0.1 ml of 1 % solution of carrageenan. The flavonoid rich ethyl acetate fraction of methanolic extract of Stereospermum suaveolens DC (Bignoniaceae) leaves was treated at different doses of 125mg/kg, 250mg/kg and 500mg/kg (p.o.). The rat paw volume was measured at 1h, 2h, 3h, 4h, 5h using Digital plethysmometer (VJ instruments - VJDP-01). The percentage inhibition of paw edema was calculated. Result: The phytochemical investigation revealed presence of flavonoid, saponins, alkaloids, carbohydrates and phenolic components.The TLC study shown confirmation of presence of flavonoid. The significant paw edema inhibition and percentage of inhibition obtained in a dose of 250mg/kg is high as compared to 125mg/kg and 500mg/kg dose fraction in experimental model of inflammation. Conclusion: From the present obtained study it was concluded that flavonoid rich ethyl acetate fraction of methanolic extract of Sterospermum suaveolens DC leaves has potential anti-inflammatory activity.