miR-155调控前列腺癌细胞NF-κB通路机制研究

目的探讨miR-155调控前列腺癌细胞核因子转录κB(NF-κB)通路的分子机制。方法前列腺癌DU145和PC-3细胞株经Anti-miRTM miR-155抑制剂(anti-miR-155)转染后(实验组),采用蛋白印迹法(Western blot)和逆转录聚合酶链反应(RT-PCR)分别检测转染后细胞中RelA和RelB蛋白及mRNA表达水平,同时检测NF-κB活性及细胞因子水平。以转染AntimiR~(TM)阴性对照的DU145和PC-3细胞株为对照组。结果与对照组相比,实验组细胞RelA、白细胞介素(IL-6)、白细胞介素(IL-21)表达水平,以及NF-κB活性显著降低(P0.05),RelB表达水平比较差异无统计学意义(P0.05)。结论降低人前列腺癌细胞NF-κB活性和RelA表达水平,可能是miR-155参与调控前列腺癌生物学行为的分子机制之一。     

The correlation between NF-κB inhibition and disease activity by coadministration of silibinin and ursodeoxycholic acid in experimental colitis

NF‐κB is one of the most important nuclear factors responsible for overexpression of proinflammatory cytokines. This is demonstrated by increased NF‐κB activity and other dependent immune factors in inflammatory bowel disease (IBD). Anti‐inflammatory effects of silibinin and ursodeoxycholic acid (UDCA) along with their NF‐κB inhibitory property are thought to be beneficial in colitis. Trinitrobenzene sulfonic acid was used to induce colitis rat models. After instillation, 48 rats were treated with oral silibinin, UDCA alone or a combination of both. Intraperitoneal dexamethasone was used in the control group. After 12 days of treatment, colonic samples were tested for the severity of mucosal damage macroscopically and microscopically. The levels of activated NF‐κB, IL‐1β, TNF‐α, myeloperoxidase, thiobarbituric acid reactive substances (TBARS), protein carbonyl, and the antioxidant power of the bowel homogenates were determined. The results indicated a significant reduction in NF‐κB activity as well as the levels of IL‐1β, TNF‐α, TBARS, protein carbonyl, myeloperoxidase activity, and an improvement in antioxidant power of colitis in treated rats. Combination therapy resulted in a more prominent improvement in bowel antioxidant power and myeloperoxidase activity. In conclusion, combination of silibinin and UDCA by inhibition of NF‐κB and other relevant inflammatory factors of colitis is a good candidate for management of Crohn’s disease.     

Gastroprotective effect of esculin on ethanol‐induced gastric lesion in mice

The gastroprotective effect of esculin was investigated in a mouse model of ethanol‐induced gastric lesion. Administration of esculin at doses of 5, 10, and 20 mg/kg body weight prior to ethanol ingestion led to significant gastroprotection compared with untreated mice. Gastric mucosal lesions were evaluated by macroscopic and histopathological alterations, lesion index, and myeloperoxidase (MPO) activity. Pretreatment with esculin significantly reduced macroscopic and histopathological damage, gastric lesion index, and MPO activity in a dose‐dependent manner. Moreover, esculin significantly reduced nitric oxide (NO) production, inducible NO synthase (iNOS) levels, and nuclear factor‐kappa B (NF‐κB) p65 protein expression in gastric tissues after ethanol challenge. Analysis of inflammatory cytokines indicated that esculin pretreatment markedly suppressed the increased expression of tumor necrosis factor‐alpha (TNF‐α) and interleukin‐6 (IL‐6) in ethanol‐treated mice. The results demonstrate a protective effect of esculin against gastric injury and suggest that the underlying mechanism might be associated with inhibition of NF‐κB activation, which subsequently reduces expression of iNOS, TNF‐α, and IL‐6.     

Anti‐inflammatory effect of 4‐methylcyclopentadecanone in rats submitted to ischemic stroke

This study aimed to investigate the anti‐inflammatory effect of 4‐methylcyclopentadecanone (4‐MCPC) in rats suffering from a cerebral ischemia/reperfusion (I/R) injury. In this study, the focal cerebral ischemia in rats was induced by middle cerebral artery occlusion (MCAO) for 2 h, and the rats were treated with 4‐MCPC (8 mg/kg) just 0.5 h before reperfusion. The ischemic infarct volume was recorded 24 h after the MCAO. In addition, myeloperoxidase (MPO) activity and TNF‐α and IL‐1β levels in the ischemic cerebral cortex were determined by ELISA, while nuclear translocation of NF‐κB p65 subunit and expression of p‐IκBα were investigated by Western blotting. Our results showed that 4‐MCPC treatment decreased infarct volume significantly, compared with I/R group (16.8%±7.5% vs. 39.7%±10.9%); it reduced MPO activity (0.43 ± 0.10 vs. 1.00 ± 0.51 U/g) and expression levels of TNF‐α (18.90 ± 3.65 vs. 35.87 ± 4.87 ng/g) and IL‐1β (1.68 ± 0.23 vs. 2.67 ± 0.38 ng/g) in ischemic brain tissues of rats. Further study revealed that 4‐MCPC treatment markedly reduced nuclear translocation of NF‐κB p65 subunit and expression of p‐IκBα in ischemic cerebral cortex. Taken together, our results suggest that 4‐MCPC protects against cerebral I/R injury and displays anti‐inflammatory actions through inhibition of the NF‐κB signal pathway.     

右美托咪定对骨科下肢手术患者止血带诱发炎症的影响及机制研究

目的探讨右美托咪定对骨科下肢手术患者止血带导致炎症过程的影响。方法将2013年6月到2015年3月该院骨科病房收治的48例需行下肢手术的患者按照随机数字表法分为右美托咪定组及对照组,每组24例。所有患者均采用蛛网膜下腔阻滞联合硬膜外麻醉,右美托咪定组于穿刺后给予负荷量1μg/kg,维持量0.5μg/(kg·h)。对照组给予同等容量生理盐水。于使用止血带前(T1),松开止血带时(T2),松开止血带后30min(T3)时,采用酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),细胞间黏附分子-1(ICAM-1)、核转录因子(NF-κB)的变化情况。结果右美托咪定组患者心动过缓发生率显著高于对照组,差异具有统计学意义(P0.05);两组患者血清各炎症因子水平在T0时差异无统计学意义(P0.05);与T0相比,两组患者血清各炎症因子水平在T1和T2时均显著升高,且差异具有统计学意义(P0.05);右美托咪定组患者血清TNF-α、IL-6、ICAM-1、NF-κB水平在T1和T2时均显著低于对照组,差异有统计学意义(P0.05)。结论右美托咪定可能通过降低血清NF-κB水平来减轻骨科下肢手术患者使用止血带诱发的炎性反应过程。     

女性海洛因依赖者子宫、卵巢变化的B超检查及探讨

阿片类药物可以使人体许多系统受累,海洛因对内分泌器的影响已经有多方面的报道。在戒毒工作中,女性吸毒者月经紊乱、闭经、流产很常见,除内分沁的改变外,子宫、卵巢有否变化引起我们的注意。为此,我们从３１８例女性吸毒者中,选择海洛因依赖者１０５例进行Ｂ超观察...     

Geniposide reduces development of streptozotocin‐induced diabetic nephropathy via regulating nuclear factor‐kappa B signaling pathways

Renal pathology was a commonly seen complication in patients with diabetes. Geniposide (GPO) was previously demonstrated to modulate glucose metabolism in diabetes. This study was to investigate effects of GPO in streptozotocin‐induced diabetic rats and its underlying mechanism. Renal function in diabetic rats was evaluated by levels of serum creatinine (Scr), blood urea nitrogen (BUN), and urinary albumin. Renal inflammation was appraised by inflammatory cells infiltration and pro‐inflammatory cytokines production. Renal monocytes, T lymphocytes infiltration, and intercellular adhesion molecule‐1 (ICAM‐1) expression were quantitated by immunohistochemistry. Moreover, renal nuclear factor‐kappa B (NF‐κB) was assayed by Western blotting. Diabetic rats showed renal dysfunction as evidenced by increased levels of Scr, BUN, urinary albumin, and elevator renal index. Histological examination revealed significant glomerular basement membrane (GBM) thickening. However, GPO notably improved renal function and diabetes‐induced GBM changes. Additionally, diabetic rats showed noteworthy renal inflammation,as reflected by enhancement of monocytes and T lymphocytes infiltration, increased expression of ICAM‐1, tumor necrosis factor‐α, interleukin‐1 (IL‐1), and IL‐6. Interestingly, the level of monocytes infiltration positively correlated with the severity of GBM. Further study indicated diabetic rats displayed increased activation of NF‐κB, indicated by increased expression of NF‐κB p65, IKKα, and p‐IκBα in renal tissue. However, all the changes in renal inflammation and NF‐κB pathway were obviously reversed in GPO‐treated diabetic rats. Our works indicate GPO ameliorates structural and functional abnormalities of kidney in diabetic rats, which is associated with its suppression of NF‐κB‐mediated inflammatory response.     

Anticonvulsive and neuroprotective effects of synergetic combination of phenytoin and gastrodin on the convulsion induced by penicillin in mice

Phenytoin (PHT) is a commonly prescribed first‐line antiepileptic drug. However, long‐term administration of PHT can cause memory loss and balance disturbance. Gastrodin (GD) is the major bioactive component in Tianma and has sedative, anticonvulsive, memory strengthening, and neuroprotective effects. To combine the two drugs seems attractive; however, little was known about the efficacy of combination therapy. In this study, convulsive attack was successfully induced by penicillin. Isobolographic analysis, memory and balance behavior test, histopathological examination, and Western blot analysis were used to investigate whether the combination therapy of GD and PHT can enhance anticonvulsive effect and reduce the side effects associated with PHT. The GD alone (950.60 mg/kg) and the PHT alone (45.50 mg/kg) could produce an anticonvulsive effect, while comparable effect could be produced by PHT : GD = 1 : 50 (8.59 : 429.27 mg/kg), which reduce the dose of PHT by 81% and GD by 55%. After the chronic anticonvulsive experiments of 16 days, the balance disturbance and short‐/long‐term memory loss were observed in the PHT group, while the PHT + GD therapy can protect the normal balance and memory function. The neuron morphology of hippocampus was preserved, and the number of surviving neurons after combination therapy was more than the model group. The amount of NF‐κB (p65) expression was increased in combination group. All above suggested the potential of the combination of PHT and GD enhances the anticonvulsive effect and the neuroprotective effect and reduces the PHT‐associated memory and balance disturbance. The PHT + GD strategy would provide new possibilities as a novel promising methodology to treat epileptic patients.     

Protective effect of metformin on a rat model of lipopolysaccharide‐induced preeclampsia

Recent in vitro and clinical studies have found that metformin (MET) may play a preventive or therapeutic role in preeclampsia (PE) and may be a candidate drug for the prevention and/or treatment of PE. In this study, we used lipopolysaccharide (LPS) to induce a PE‐like rat model and investigated the intervention effect of MET from the perspectives of clinical manifestations, placental morphology, serum marker for placental injury, systemic inflammatory response and oxidative/nitrative stress, and placental nuclear factor‐κB (NF‐κB) signaling. The results showed that MET improved LPS‐induced hypertension, proteinuria, fetal growth restriction (FGR) and stillbirth, alleviated placental injury and decreased maternal serum marker alpha‐fetoprotein (MS‐AFP) level; MET suppressed LPS‐induced TNF‐α and IL‐6 productions, reduced oxidative/nitrative stress as evidenced by increased superoxide dismutase (SOD) activity, decreased inducible nitric oxide synthase (iNOS) activity, and decreased levels of malondialdehyde (MDA) and nitric oxide (NO); MET inhibited LPS‐induced NF‐κB activation in placentas. Based on these findings, it can be concluded that MET is beneficial to the PE‐like rat model by protecting placentas from injury, suppressing systemic inflammatory response and oxidative/nitrative stress, and inhibiting placental NF‐κB signaling pathway. MET is a promising drug for prevention and/or treatment of PE.     

Adjuvant effect of caffeine on acetylsalicylic acid anti‐nociception: Prostaglandin E2 synthesis determination in carrageenan‐induced peripheral inflammation in rat

In the present study, we report a synergistic interaction between acetylsalicylic acid (ASA) and caffeine (CAF) on the inhibition of nociception in a model of peripheral inflammation in rat; on the contrary no interaction have been found on anti‐inflammatory effects and peripheral prostaglandin E2 (PGE‐2) synthesis inhibition. Acute inflammation was induced by the subplantar injection of carrageenan into the right hind paw, and the effects of the drugs were evaluated from 0 to 5h. Nociception was assessed using the Randall & Selitto test, and the inflammatory response by plethismometry. Oral administration of ASA (10–400mg/kg) induced dose‐related anti‐nociceptive and anti‐inflammatory effects. On the other hand, oral CAF administration (5–50mg/kg) did not show a dose‐related inhibitory effect, neither on the inhibition of nociception nor on the inflammatory response. To analyze a possible interaction between both drugs a dose—response curve to ASA plus a fixed dose of CAF (5mg/kg) was obtained 3h after the injection of carrageenan, when the inflammatory pain peaked. A fixed dose of CAF was able to improve the anti‐nociceptive, but not the anti‐inflammatory, effects of ASA. We also assessed, by enzyme immunoassay, the effects of the combination on peripheral PGE‐2 levels: CAF did not alter the inhibitory effect of ASA on PGE‐2 synthesis. Our results corroborate the well‐known clinical effects of combining ASA and CAF; on the other hand, we rule out that prostaglandin synthesis inhibition at peripheral sites would be the mechanism responsible of the adjuvant anti‐nociceptive effect of CAF.     

Levobupivacaine attenuates lipopolysaccharide‐induced acute lung injury

Levobupivacaine (LB), a kind of local anesthetic, possesses anti‐inflammatory properties. High‐mobility group box 1 (HMGB1), a nuclear DNA‐binding protein, plays a key role in the development of acute lung injury (ALI). The aim of this study was to investigate whether LB attenuates ALI by the inhibition of HMGB1 expression and to investigate the molecular mechanisms. ALI in male rats was induced by an intratracheal instillation of LPS (5 mg/kg), and male rats received mini‐osmotic pumps containing LB 30 min after LPS exposure. A549 alveolar epithelial cells were incubated with LPS in the presence or absence of LB. An enzyme‐linked immunosorbent assay was used to detect the levels of inflammatory cytokines. Western blotting was used to detect the changes in the expression of toll‐like receptor 2/4 (TLR2/4) and the activation of NF‐κB. The results showed that LB significantly protected animals from LPS‐induced ALI as evidenced by a decrease in the ratio of lung wet to dry weight, total cells, neutrophils, macrophages, and myeloperoxidase activity, associated with a reduced lung histological damage. We also found that LB post‐treatment markedly inhibited the release of HMGB1 and other pro‐inflammatory cytokines. Furthermore, LB significantly inhibited LPS‐induced TLR2/4 protein overexpression and NF‐κB activation in the lung tissues and in LPS‐stimulated A549 alveolar epithelial cells in vitro. These data indicate that LB attenuated LPS‐induced ALI by the inhibition of HMGB1 expression in rats. These benefits were associated with the inhibition of TLR2/4‐NF‐κB pathway by LB.     

Pam 3CysS K4对糖尿病小鼠肾功能及肾小管上皮细胞 TLR2表达的影响

【目的】探讨 Toll 样受体（Toll‐like receptors ,TLR）的特异激动剂 Pam3CysSK4对糖尿病小鼠肾功能及肾小管上皮细胞 Toll 样受体2（TLR2）表达的影响。【方法】选取 C57小鼠24只,随机分为正常组、糖尿病模型组（模型组）和干预组,每组8只,模型组采用链脲佐菌素（STZ）诱导建立小鼠糖尿病模型,干预组建立糖尿病小鼠模型后并给予股静脉注射 Pam3CysSK4,每周给予100μg ,正常组小鼠给予与其他两组等量枸橼酸缓冲液注射。三组喂养8周后,收集24 h 尿液,检测24 h 尿蛋白;称小鼠体重、肾重,计算肾重／体重;检测血清肌酐（Cr）,血尿素氮（BUN）、血糖及白细胞介素‐1（IL‐1）评估肾小球硬化指数,检测肾小管上皮细胞 TLR2、核转录因子‐κB （NF‐κB）及髓样分化因子88（MyD88）、单核细胞趋化蛋白‐1（MCP‐1）表达水平。【结果】干预组和模型组血糖均高于正常组;干预组小鼠体重和24 h 尿量低于模型组,而模型组低于正常组;干预组小鼠肾重和肾重／体重高于模型组,而模型组高于正常组,其差异均有统计学意义（ P ＜0．05）。干预组和模型组 Cr 、BUN 、IL‐1 和24尿蛋白含量、肾小球硬化指数及 TLR2、NF‐κB 、MyD88和 MCP‐1均高于正常组,而干预组高于模型组,其差异均有统计学意义（ P ＜0．05）。【结论】高糖能够上调小鼠肾小管上皮细胞 TLR2水平,激活 TLR 信号转导通路,促进炎症因子的释放,增加对肾脏的损伤作用。     

A novel mechanism of cytokine release in phagocytes induced by aggretin,a snake venom C‐type lectin protein,through CLEC‐2 ligation

Summary Background: Macrophages are major immune cells and play an important role in modulating homeostasis and the immune defense mechanism. In inflammatory responses to the infection of pathogens, macrophages are activated, producing various inflammatory mediators. Snake venom C‐type lectin proteins (snaclecs) have diverse targets, including platelet GPVI, GPIb, integrin α2β1 or CLEC‐2 expressed in platelets, endothelial cells or myeloid cells. Methods: In this study, murine macrophages (RAW 264.7 cells) and human monocytes (THP‐1) were treated with different snaclecs, including aggretin, gramicetin, trowaglerix and convulxin, in the absence or presence of LPS for 24 h. Results: The production of cytokines, such as tumor necrosis factor‐α (TNF‐α) and interleukin‐6 (IL‐6), in supernatants was measured by ELISA. Aggretin increased the production of TNF‐α and IL‐6 in both RAW264.7 and THP‐1 cells; however, the other snaclecs did not. Aggretin induced extracellular signal‐regulated kinase 1/2 (ERK1/2) and c‐Jun N‐terminal kinase (JNK) tyrosine phosphorylation of RAW264.7 cells. Pretreatments with inhibitor of ERK, JNK, p38 or NF‐κB abolished cytokine release caused by aggretin. Aggretin bound to THP‐1 cells in a concentration‐dependent manner and it displaced the CLEC‐2 mAb binding to THP‐1 cells and the immobilized aggretin selectively bound to CLEC‐2 of both platelets and THP‐1 cell lysates. Furthermore, aggretin elevated the plasma level of IL‐6 in ICR mice as it was administered intramuscularly. Conclusion: These results indicate that aggretin may induce cytokine TNF‐α/IL‐6 release via interacting with CLEC‐2 receptor and the subsequent MAPK and NF‐κB activation in monocytes/macrophages.     

Curcumin and its analogues: Potential anticancer agents

This review chronicles the exploration of the curcumin in terms of development of analogues for the anticancer activity over the last century. Curcumin is a natural phytochemical obtained from dried root and rhizome of Turmeric (Curcuma Longa). It has been shown to interfere with multiple cell signaling pathways, including apoptosis (activation of caspases and downregulation of antiapoptotic gene products), proliferation (HER‐2, EGFR, and AP‐1), angiogenesis (VEGF), and inflammation (NF‐κB, TNF, IL‐6, IL‐1, COX‐2, and 5‐LOX). In the last decade it has been much explored and various synthetic analogues have been prepared and evaluated for various pharmacological activities. Most of the analogues have shown very good anticancer activity in various models and various cell lines. However, some analogues have also shown antioxidant, anti‐HIV, antimutagenic, antiangiogenic, antimalarial, antitubercular, antiandrogenic, COX inhibitory activities. Few analogues have shown very potent results and may be considered as clinical candidates for the development of future anticancer agent. This review contains 728 curcumin analogues and covers the literature from 1815 to mid 2009 and 93 references are cited. © 2009 Wiley Periodicals, Inc. Med Res Rev, 30, No. 5, 818–860, 2010     

Serotonin depletion leads to cortical hyperexcitability and trigeminal nociceptive facilitation via the nitric oxide pathway

(Headache 2011;51:1152‐1160) Objective.— To investigate the role of nitric oxide (NO) in the development of cortical hyperexcitability and trigeminal nociceptive facilitation induced by serotonin (5‐HT) depletion. Background.— Nitric oxide and 5‐HT are important in the pathogenesis of primary headaches. An increase in cortical excitability and trigeminal nociception has been demonstrated in animals with low 5‐HT levels. Although the mechanism underlying this increase is unclear, an alteration of the NO system is one possible explanation. Methods.— Male Wistar rats were divided into control and 5‐HT‐depleted groups. 5‐HT was depleted by i.p. injection of parachlorophenylalanine (100 mg/kg). Three days after injection, a microelectrode was inserted into the cerebral cortex for electrocorticograph recording and waves of cortical spreading depression (CSD) were triggered with KCl application. N‐nitro‐L‐arginine methyl ester (L‐NAME; 10 mg/kg by i.v. injection) or saline was given after the second CSD wave. Following the experiment, the cerebral cortex and brain stem were removed for anti‐neuronal nitric oxide synthase (nNOS) and anti‐Fos immunohistochemistry. Results.— Relative to the control group, the 5‐HT‐depleted group exhibited a higher frequency of CSD waves, more nNOS‐immunoreactive cells in both the cerebral cortex and brainstem and more Fos‐immunoreactive cells in the trigeminal nucleus caudalis (TNC). In the control group, L‐NAME application led to fewer nNOS‐immunoreactive cells in the cerebral cortex and TNC, and fewer Fos‐immunoreactive cells in the TNC; however, L‐NAME was without effect on the CSD pattern. By contrast, in addition to decreased nNOS and Fos expression, L‐NAME significantly reduced the frequency of CSD events in the 5‐HT‐depleted group. Conclusions.— Inhibition of NO production can counter both the cortical hyperexcitability and facilitation of trigeminal nociception that develop in the depleted 5‐HT state. Therefore, NO is likely involved in the increase in both CSD events and CSD‐evoked trigeminal nociception under decreased 5‐HT conditions.     

Targeting Oxidative Injury and Cytokines' Activity in the Treatment with Anti‐Tumor Necrosis Factor‐α Antibody for Complex Regional Pain Syndrome 1

Cytokines and oxygen free radicals have been implicated in the potential pathogenic development of complex regional pain syndrome (CRPS). We aimed to analyze the relationship between clinical status, circulating levels of cytokines, and markers of oxidative damage during the treatment with anti‐TNFα antibodies. The patient chosen for treatment had not had improvement through a number of conventional therapies and fulfilled the current diagnostic criteria for CRPS‐1. We investigated the clinical variables before and after systemic administration of 1.4 mg/kg anti‐TNFα antibody (infliximab), repeated after 1 month in a dose of 3 mg/kg. Blood samples were collected before and after anti‐TNFα antibodies administration, and plasma was analyzed for 8‐isoprostane‐prostaglandin F2α (8‐iso‐PGF2α, a marker of oxidative injury) and cytokines (TNF‐α, IL‐4, IL‐6, IL‐7, IL‐8, IL‐10, IL‐17A). Plasma concentrations of 8‐iso‐PGF2α were measured with radioimmunoassay (RIA), and the kinetics of cytokines were detected in plasma by antibody‐based proximity ligation (PLA). Pathologically high levels of 8‐iso‐PGF2α were found in the patient. Immediately after each administration of infliximab, the levels of 8‐iso‐PGF2α decreased. Although the patient showed an improvement of the cutaneous dystrophic symptoms and diminished pain associated with these lesions, the levels of circulating TNFα increased after the administration of anti‐TNFα antibodies. In a patient with CRPS‐1 treated with anti‐TNFα antibodies, we report increased levels of circulating TNFα and a temporary mitigation of oxidative stress as measured by plasma F₂‐isoprostane. This case report provides evidence 2 supporting the indication of monitoring the oxidative stress biomarkers during treatment with anti‐TNFα antibodies in CRPS 1.     

Therapeutic effect of long‐interval repeated intravenous administration of human umbilical cord blood‐derived mesenchymal stem cells in DBA/1 mice with collagen‐induced arthritis

Rheumatoid arthritis (RA) is a common inflammatory chronic disease. It has been reported that mesenchymal stem cells (MSCs) have the effect of immune suppression in collagen‐induced arthritis (CIA) mice model. However, the in vivo therapeutic effect from the long‐interval repeated intravenous administration of human umbilical cord blood‐derived (hUCB)‐MSCs had not been investigated in CIA mice model. This study was undertaken to investigate the effects of long‐interval repeated intravenous administration of hUCB‐MSCs at different doses in CIA mice model. Mice were intravenously injected with three different doses of hUCB‐MSCs once every 2 weeks for three times. RA severity was assessed by clinical joint score and histologic analysis including hematoxylin and eosin staining, safranin‐O staining, and toluidine blue staining. We used real‐time polymerase chain reaction and flow cytometry to quantify differences in inflammatory cytokines and Tregs. Mice treated with hUCB‐MSCs showed significant improvement in clinical joint score. Histologic analysis revealed that hUCB‐MSCs definitely reduced joint inflammation, cartilage damage, and formation of pannus in multimedium and multihigh groups. These hUCB‐MSCs also significantly decreased IL‐1 beta protein levels in multimedium and multihigh groups and IL‐6 protein levels in all hUCB‐MSCs‐treated groups. Furthermore, mRNA levels of IL‐1 beta and IL‐6 were decreased significantly in all hUCB‐MSCs‐treated groups, whereas the expression of anti‐inflammatory cytokine IL‐10 was increased in the multihigh group. Tregs known as suppressor T cells were also significantly increased in the multihigh group. Our findings suggest that long‐interval repeated intravenous administration of hUCB‐MSCs has therapeutic effects by improving symptoms of RA in CIA mice model in a dose‐dependent manner.     

5‐HT3 receptors antagonists reduce serotonin‐induced scratching in mice

Serotonin (5‐hydroxytryptamine, 5‐HT) acts as a pruritogen in humans and animals, but the mechanisms of action through that serotonin induces itch response have not been extensively discovered. In our study, we attempted to investigate the role of 5‐HT3 receptors in scratching behavior due to intradermal serotonin injection. Intradermal injection of serotonin (14.1–235 nmol/site) into the nape of the neck of mice was performed to elicit itch. Scratching behavior was evaluated by measuring the number of bouts during 60 min after injection. We evaluated the effect of intraperitoneal pretreatment with ondansetron and tropisetron (0.1, 0.3, and 1 mg/kg) on itch induced by serotonin. Also, intradermal ondansetron and tropisetron at doses 50, 100, and 200 nmol/site were concurrently administrated with serotonin. Serotonin produced a significant enhancement in scratching at dose 141 nmol/site. Concurrent administration of ondansetron (50, 100, and 200 nmol/site) and tropisetron (100 and 200 nmol/site) with serotonin reduced scratching activity compared to the animals that only received serotonin. Also, pretreatment with intraperitoneal ondansetron and tropisetron (0.3 and 1 mg/kg) 30 min before serotonin attenuated the itch response. We showed that the scratching induced by intradermal serotonin is mediated by 5‐HT3 receptors subtype. It can be concluded that 5‐HT3 may play a role in mediating serotonin‐associated itch responses, and we introduce 5‐HT3 receptors as possible targets for antipruritic agents.