Carbenoxolone enhances inhibitory effect of RSL3 against cisplatin-resistant testicular cancer cells by promoting ferroptosis / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of RSL3 on the proliferation, invasion and migration of cisplatinresistant testicular cancer cells (I-10/DDP) and the effect of carbenoxolone on the activity of RSL3 against testicular cancer.@*METHODS@#MTT assay was used to evaluate the survival rate of I-10/DDP cells following treatment with RSL3 (1, 2, 4, 8, 16 or 32 μmol/L) alone or in combination with carbenoxolone (100 μmol/L) or after treatment with Fer-1 (2 μmol/L), RSL3 (4 μmol/L), RSL3+Fer-1, RSL3+carbenoxolone (100 μmol/L), or RSL3+Fer-1+carbenoxolone. Colony formation assay was used to assess the proliferation ability of the treated cells; wounding-healing assay and Transwell assay were used to assess the invasion and migration ability of the cells. The expression of GPX4 was detected using Western blotting, the levels of lipid ROS were detected using C11 BODIPY 581/591 fluorescent probe, and the levels of Fe2+ were determined with FerroOrange fluorescent probe.@*RESULTS@#RSL3 dose-dependently decreased the survival rate of I-10/DDP cells, and the combined treatment with 2, 4, or 8 μmol/L RSL3 with carbenoxolone, as compared with RSL3 treatment alone, resulted in significant reduction of the cell survival rate. The combination with carbenoxolone significantly enhanced the inhibitory effect of RSL3 on colony formation, wound healing rate (P=0.005), invasion and migration of the cells (P < 0.001). Fer-1 obviously attenuated the inhibitory effects of RSL3 alone and its combination with carbenoxolone on I-10/DDP cells (P < 0.01). RSL3 treatment significantly decreased GPX4 expression (P=0.001) and increased lipid ROS level (P=0.001) and Fe2+ level in the cells, and these effects were further enhanced by the combined treatment with carbenoxolone (P < 0.01).@*CONCLUSION@#Carbenoxolone enhances the inhibitory effect of RSL3 on the proliferation, invasion and migration of cisplatin-resistant testicular cancer cells by promoting RSL3-induced ferroptosis.

Sex-Dependent Glial Signaling in Pathological Pain: Distinct Roles of Spinal Microglia and Astrocytes / 神经科学通报·英文版

Increasing evidence suggests that spinal microglia regulate pathological pain in males. In this study, we investigated the effects of several microglial and astroglial modulators on inflammatory and neuropathic pain following intrathecal injection in male and female mice. These modulators were the microglial inhibitors minocycline and ZVEID (a caspase-6 inhibitor) and the astroglial inhibitors L-α-aminoadipate (L-AA, an astroglial toxin) and carbenoxolone (a connexin 43 inhibitor), as well as U0126 (an ERK kinase inhibitor) and D-JNKI-1 (a c-Jun N-terminal kinase inhibitor). We found that spinal administration of minocycline or ZVEID, or Caspase6 deletion, reduced formalin-induced inflammatory and nerve injury-induced neuropathic pain primarily in male mice. In contrast, intrathecal L-AA reduced neuropathic pain but not inflammatory pain in both sexes. Intrathecal U0126 and D-JNKI-1 reduced neuropathic pain in both sexes. Nerve injury caused spinal upregulation of the astroglial markers GFAP and Connexin 43 in both sexes. Collectively, our data confirmed male-dominant microglial signaling but also revealed sex-independent astroglial signaling in the spinal cord in inflammatory and neuropathic pain.

Regulation of Tumor Necrosis Factor-alpha-induced Airway Mucin Production and Gene Expression by Carbenoxolone, Prunetin, and Silibinin / 결핵및호흡기질환

BACKGROUND: In this study, we tried to investigate whether carbenoxolone, prunetin, and silibinin affect tumor necrosis factor (TNF)-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells. METHODS: Confluent NCI-H292 cells were pretreated with each agent (carbenoxolone, prunetin, and silibinin) for 30 min and then stimulated with TNF-alpha for 24 hours. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. RESULTS: Carbenoxolone, prunetin and silibinin inhibited the production of MUC5AC mucin protein induced by TNF-alpha; the 3 compounds also inhibited the expression of MUC5AC mucin gene induced by TNF-alpha. CONCLUSION: This result suggests that carbenoxolone, prunetin and silibinin can inhibit mucin gene expression and production of mucin protein induced by TNF-alpha, by directly acting on airway epithelial cells.

Effect of central microinjection of carbenoxolone in an experimental model of focal cerebral ischemia

Previous experimental studies have shown the protective effects of CBX on brain ischemic injures in global and in vitro models of ischemia. However, effects of CBX in temporary model of focal cerebral ischemia are not clear. Hence, the aim of this study was to investigate the effects of central micro injection of CBX on post-ischemic reperfusion injuries in a temporary model of focal cerebral ischemia. Transient focal cerebral ischemia was induced in rats by 60 min middle cerebral artery occlusion [MCAO], followed by 23 h reperfusion. CBX was administered into the right ventricle at doses of 1, 12, 25, 50 and/or 100 micro g/kg at the beginning of MCAO. Cortical and striatal infarct volumes and motor dysfunctions were assessed 24 h after MCAO. Administration of CBX at doses of 1, 12, 25 and/or 50 micro g/kg significantly reduced cortical infarct volumes by 35%, 49%, 41% and 43%, respectively [P<0.001]. In addition, CBX only at dose of 25 micro g/kg significantly reduced striatal infract volume and improved neurological dysfunctions [P<0.01]. Our findings indicated that central microinjection of CBX has protective effect on against ischemic reperfusion injuries in a transient model of focal cerebral ischemia

Hypertonic stimulation induces synthesis and release of glutamate in cultured rat hypothalamic astrocytes and C6 cells / 神经科学通报·英文版

<p><b>OBJECTIVE</b>To investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line.</p><p><b>METHODS</b>Astrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 min; carbenoxolone (CBX)+IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca(2+)+HS group, astrocytes were pre-incubated with Ca Ca(2+) (1,000 micromol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM).</p><p><b>RESULTS</b>(1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 min (P < 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P< 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca(2+)+HS group were significant lower than that in the HS group (P < 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells.</p><p><b>CONCLUSION</b>HS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.</p>

Studies on analgesic effects and sites of oxymatrine-carbenoxolone sodium complex / 中国中药杂志

<p><b>OBJECTIVE</b>To study the analgesic effects and sites of oxymatrine-carbenoxolone sodium complex (OCSC).</p><p><b>METHOD</b>Adopting formalin test, warm water tail-flick test and intracerebroventricularly (icv) injection to observe the analgesic effects of OCSC in mice.</p><p><b>RESULT</b>Intraperitoneally injecting (ip) OCSC (75, 150 mg x kg(-1)) remarkedly inhibited the pain of mice in the formalin test and prolonged latent phases of tail-shrinking of mice, icy OCSC (1.875, 3.75, 7.5 mg x kg(-1)) significantly prolonged latent phases of tail-shrinking of mice, it had dose-dependent effect with concentration.</p><p><b>CONCLUSION</b>The result indicated that OCSC has obvious analgesic effects and its mechanism may be involved in central nervous system (CNS).</p>

Effect of carbenoxolone on expression of Fos, NMDAR2 and GFAP in the hippocampus of pentylenetetrazo-kindled epileptic rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Gap junctions, the clusters of intercellular channels, play an important role in synchronizing electrical activity. This study investigated the effect of gap junction blocker carbenoxolone (CBX) on epileptic activity in pentylenetetrazo (PTZ)-kindled rats.</p><p><b>METHODS</b>Thirty adult male SD rats were randomly divided into three groups: control, PTZ-kindled and CBX-treated groups (n=10 each). The rats from the PTZ-kindled and the CBX-treated groups were intraperitoneally injected with PTZ (35 mg/kg x d) to induce epilepsy. After epilepsy kindling, they were intraperitoneally injected for 3 days with CBX (10 mg/kg) (CBX-treated group) or with normal saline (PTZ-kindled group). The control group received intraperitoneal injections of normal saline. Anti-GFAP, anti-Fos, and anti-NMDARZ immunohistochemical ABC methods were used to detect the expression of GFAP-Li, Fos-Li and NMDAR2-Li in the hippocampus respectively.</p><p><b>RESULTS</b>Spontaneous seizures occurred in PTZ-kindled epileptic rats. CBX administration reduced spontaneous seizures. The NMDAR2-Li and Fos-Li neurons as well as GFAP-Li astrocytes in hippocampi increased in PTZ-kindled epileptic rats compared with controls. The numbers of Fos-Li (93.75 +/-7.94 vs 165.25 +/-15.87, P < 0.05) and NMDAR2-Li neurons (61.47 +/-3.62 vs 148.72 +/-14.53, P < 0.01) in the CBX-treated group were significantly less than in the PTZ-kindled group. There were no significant differences in the GFAP-Li expression between the CBX-treated and the PTZ-kindled groups.</p><p><b>CONCLUSIONS</b>CBX may inhibit spontaneous seizures and decrease the numbers of Fos-Li and NMDARZ-Li neurons, thus providing anti-epileptic effects.</p>

Influence of Sex on Gap Junctional Distribution and Vascular Dilatation in the Rat Mesenteric Arteries / 대한마취과학회지

BACKGROUND: Besides prostacyclin and nitric oxide, the endothelium-derived hyperpolarizing factor (EDHF), which is another distinct endothelium-dependent vasodilator, is involved in relaxing the vascular smooth muscle cells. The myoendothelial gap junction (MEGJ) and female sex hormone play important roles in the EDHF-mediated responses. Therefore, this study was designed to determine the influence of gender on the gap junctional distribution and endothelium-dependent vasodilation in the rat mesenteric arteries. METHODS: Male and female Sprague-Dawley rat were euthanized and the tertiary branch of the mesenteric artery was harvested. Immunohistochemistry and confocal microscopic examination of the arterial wall were performed after treating them with specific antibodies to delineate the distribution of connexin 43, a gap junctional protein. Segments of the mesenteric artery, 5 mm in length, were connected to two tungsten wires under isometric tension. The arterial segments were suspended in a modified Krebs solution (37 degrees C) aerated with 95% O2 and 5% CO2 in a vertical water-jacketed temperature-controlled tissue bath. The standard dose-response curve for acetylcholine (10(-9)-10(-5) M) was drawn in the presence of the NO synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME; 10(-4) M) plus indomethacin (10(-5) M) and/or gap junctional inhibitor, carbenoxolone (10(-4) M). RESULTS: In the female rat mesenteric artery, the gap junctional plagues were more prevalent particularly along the endothelial layer. The inhibition of the relaxation response to acetylcholine was depressed in the presence of L-NAME plus indomethacin and augmented in the presence of carbenoxolone when compared with the male rat mesenteric arteries (P < 0.05). CONCLUSIONS: Gender differences in the rat mensenteric arteries have an effect on the expression of connexin 43 and the release of EDHF through MEGJ may play a key role in controlling the female arterial tone.

Renal humoral modulation of skeletal muscle tone in mice: implications for 'the pulmonary-renal cascade'.

The pulmonary-renal cascade may regulate the respiration and skeletal muscle contractility. To evaluate this working hypothetical model, we conducted experiments to ascertain the skeletal muscle tone of the Swiss mice (20-35 g). The animals were evaluated for their skeletal muscle tone via several techniques i.e. inclined plane test, grip strength test and swim test. Groups of mice (n=6) were pre-treated with mefenamic acid (60 mg/kg, i.p), carbenoxolone (100 mg/kg i.p) or vehicle only 15 minutes before the treatment with heparin (500 U/kg, i.v), urokinase (5500 U/kg, i.v) and erythropoietin (150 U/kg, i.v). Heparin potentiated the loss of skeletal muscle tone induced by mefenamic acid and carbenoxolone while urokinase & erythropoietin significantly enhanced the skeletal muscle tone as evaluated by all or one of the tests. Other groups of mice (n=6) were pretreated with mefenamic acid (1 mg i.c.v), carbenoxolone (160 microg i.c.v) or minoxidil (30 microg i.c.v) and the effects of heparin & urokinase and erythropoietin on skeletal muscle tone were evaluated. To study the effects of heparin and urokinase on nerve regeneration, two groups of mice underwent a sham and sciatic nerve crush procedure. The mice treated with urokinase recovered much faster as compared to those treated with heparin or saline. These experimental results suggest that gap junction blockers and potassium channel openers interact with heparin, urokinase and erythropoietin to control the skeletal muscle tone.

Influence of Myoendothelial Gap Junction on the Regulation of Vasomotor Tone / 대한마취과학회지

BACKGROUND: In addition to nitric oxide (NO) and prostacyclin (PGI2), there is another endothelium-derived mechanism of smooth muscle relaxation, which is associated with an endothelium-derived hyperpolarizing factor (EDHF). To assess the role of gap junctions in endothelium dependent hyperpolarization, we investigated the relationship between distribution of myoendothelial gap junction (MEGJ) and relative importance of the EDHF pathway in the regulation of vascular tone. METHODS: Immunohistochemistry and confocal microscopic examination of the mesenteric arterial wall of male Sprague-Dawley rat following treatment with specific antibodies were performed to delineate the distribution of connexin 43, a gap junctional protein. The standard dose-response curve for acetylcholine (10-9-10-5 M) of the mesenteric artery was regarded as the release of EDHF in the presence of the NO synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME, 10-4 M) and indomethacine (10-5 M). The effects of the gap junction inhibitors such as 18alpha-glycyrrhetinic acid (18alpha-GA; (2 x 10-4 M)) and carbenoxolone (3 x 10-4 M) was assessed regarding relaxtion to acetylcholine, contraction to phenylephrine (5 x 10-6 M) in the proximal and distal mesenteric arteries. RESULTS: In the distal artery, gap junctional plaques were more prevalent, and the relaxation response to acetylcholine was augmented and the contraction response to phenylephrine was depressed compared with the proximal artery. In both the proximal and distal mesenteric arteries, acetylcholine-induced relaxations attributable to EDHF were near completely blocked by 18alpha-GA and carbenoxolone to the same degree. Regardless of the presence of L-NAME plus indomethacin, 18alpha-GA significantly augmented the contraction response to phenylephrine. CONCLUSIONS: The vasomotor regulatory response by EDHF in the rat mesenteric arteries may be explained by extensive heterocellular coupling through MEGJs. Moreover, the release of EDHF through MEGJ may have a essential role in the regulation of resistor arterial tone.

Conduta na doença ulcerosa / Peptic ulcer managment

Relata sobre as causas e formas de tratamentos da úlcera gastroduodenal. Aconselha avaliaçäo criteriosa nos casos cronicos visando uma opçäo entre o tratamento continuo de manutençäo ou a opçäo cirurgica

Farmacoterapia de la úlcera péptica / Pharmacotherapy in peptic ulcer

Siendo la úlcera péptica una enfermedad de alta prevalencia, la industria farmacéutica ha introducido en los últimos años un gran número de fármacos potencialmente útiles en su tratamiento. En este artículo se revisan los fármacos actualmente disponibles y otros en fase de investigación en las variedades de antisecretores o protectores de mucosa. Se discute las ventajas, desventajas, dosis propuestas y eficacia de cada uno de ellos. Finalmente se plantea algunas interrogantes de problemas aún no resueltos en la fármacoterapia de esta enfermedad