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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.
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Humains , Mâle , Carbénoxolone/pharmacologie , Lignée cellulaire tumorale , Prolifération cellulaire , Cisplatine/pharmacologie , Ferroptose , Colorants fluorescents/pharmacologie , Lipides , Tumeurs embryonnaires et germinales , Espèces réactives de l'oxygène , Tumeurs du testiculeRésumé
Objective To observe the regulation of Panx1 on ATP/IP3 signaling pathway and its mechanism during cisplatin-induced apoptosis of lung adenocarcinoma cells. Methods Human lung adenocarcinoma cell line A549 was used as the research object and carbenoxolone (CBX) was used as a drug interference tool. A549 cells were divided into normal control group, the CBX group, the cisplatin (DDP) group and the CBX+DDP group. MTT assay and Annexin V/PI assay were used to detect the survival and apoptosis rates of A549 cells. The relative concentrations of extracellular adenosine triphosphate (ATP) and intracellular inositol triphosphate (IP3) were measured by Chemiluminescence and ELISA. Results Compared with DDP group, the cell survival rate of CBX+DDP group increased, while the early and late apoptotic rates and the release concentration of extracellular ATP and intracellular IP3 decreased (all P < 0.01). Conclusion Panx1 can increase the sensitivity of lung adenocarcinoma cells to cisplatin by regulating the ATP/IP3 signaling pathway.
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Aim Toinvestigatetheantagonisticeffect of intrathecal injection of carbenoxolone (CBX ) on neuropathic pain and its underlying mechanism.Meth-ods SixtymaleSprague-Dawleyratswererandomly divided into five groups (n =12 ):group I received sham surgery then treated with saline;group Ⅱ re-ceived SNT then treated with saline;groupⅢreceived SNT then treated with 0. 05 μg CBX;group Ⅳ re-ceived SNT then treated with 0. 5 μg CBX;group Ⅴreceived SNT then treated with 5 μg CBX.Treatment was undertaken with 10 μl volume as a single intrathe-cal injection on postoperative day 10.Mechanical with-drawl thresholds were measured 1 d before operation, 1,3,5,7 and 10 d after surgery,1 h before intrathe-cal administration,and 1 ,2,4,6 h after intrathecal administration.Lumbar spinal cord was obtained 2 h after intrathecal administration to determine the expres-sions of GFAP by immunohistology and TNF-α,IL-1βby ELISA in bilateral spinal dorsal horns.Results Comparedwiththeshamgroup,thebilateralMWTin group Ⅱ ~Ⅴ was significantly decreased.Compared with the MWT 1 h before intrathecal administration on day 10,the values at 1 ,2,4,6 h after administration of group Ⅱ and Ⅲ had no marked difference.The ip-silateral MWT in groupⅣhad no significant difference at 1,2,4 h after administration,the contralateral MWT was significantly increased,whereas GFAP and TNF-α,IL-1βwas significantly decreased in the spinal cord .In group Ⅴthe bilateral MWT was significantly improved at 1 ,2,4 h after administration,whereas GFAP and TNF-α,IL-1βwere significantly decreased inthespinalcord.Conclusions IntrathecalCBXcan inhibit the development of bilateral MWT.The analge-sic effect of CBX is implemented partly via suppressing the actation of GFAP and the realease of TNF-α,IL-1βin the spinal doral horn.
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Objective To observe the expression of pannexin1(PX1) in the dorsal horn of spinal cord in model ratwith neu-ropathipain afteselective ligation of sciatinerve branche.Method50 male SD ratwere randomly divided into 3 group,inclu-ding the control group(Wgroup ,n= 10) ,sham operation group(sham group ,n= 10) and sciatinerve branch selective injury group(SNI group ,n=30) .30 ratwere killed on postoperative 3 ,5 ,7 ,14 d and the lumbasegmenof the spinal cord wataken fodetecting the expression of PX1 by using Western blo.Othe20 ratwere killed on 7 d afteSNI and the expression of glial fibril-lary acidiprotein(GFAP) in the spinal cord wadetected with immunohistology .Among them ,10 ratin the SNI group were trea-ted with intrathecal intubation before operation and administrated with saline 20 μL ocarbenoxolone(CBX) 20 μL by intrathecal injection on postoperative 7 d fodetermining the expression of GFAP by the immunohistology .ResultThe expression of PX1 in the SNI group waincreased and enhanced with time ,which wasignificantly highethan thain the Wgroup and the sham group (P<0 .05);the GFAP expression on 7 d in the SNI group waobviously increased compared with the Wgroup and the sham group(P<0 .05);afteintrathecal injection of CBX ,the expression of GFAP wasignificantly decreased compared with thain the normal saline group(P<0 .05) .No statistically significandifferencein the expression of PX1 and GFAP were found in the Wgroup and the sham group .Conclusion PX1 may be involved in the activation of astrocyte,prompting thaPX1 playan importanrole in the neuropathipain caused by the peripheral nervel injury .
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Background: Gap junctions (GJs) are clusters of channels that connect the interiors of adjoining neurons and mediate electrical/electrotonic coupling by transfer of ions and small molecules. Electrotonic coupling between principal neurons via GJs is increasingly recognized as one of the mechanisms in the pathogenesis of the abnormal neuronal synchrony that occurs during seizures. Carbenoxolone the succinyl ester of glycyrrhetinic acid obtained from liquorice has been shown to have the property of blocking gap junctional intercellular communication. The objectives were to study if carbenoxolone has in-vivo anticonvulsive activity in pentylenetetrazole (PTZ) and maximal electroshock (MES) seizure models and to probe the functional role of GJs in seizures. Methods: Carbenoxolone was tested for anticonvulsive effect in albino rats subjected to seizures by the PTZ and MES at three doses 100 m/kg, 200 m/kg, 300 m/kg. In the PTZ model parameters observed were seizure protection, seizure latency and seizure duration. In the MES model parameters observed were seizure protection and seizure duration. Results: The results showed that the carbenoxolone has anticonvulsant activity in both PTZ and MES induced seizures with better protection in the PTZ induced seizures. In the PTZ model carbenoxolone produced a statistically significant increase in seizure latency, decrease in seizure duration and seizure protection. In the MES model carbenoxolone produced a statistically significant decrease in seizure duration. Conclusions: Carbenoxolone has in-vivo anticonvulsive effect and could be useful in both petitmal (absence) seizures and grand mal (generalized tonic-clonic epilepsy) seizures. The protective effect of carbenoxolone could be due to blockade of GJ channels that mediate electro tonic coupling and thereby prevent the neural synchronization that is characteristic of seizures. The study also supports the view that GJs have a functional role in the electrophysiology of seizures and GJ blockers have potential as a new class of antiepileptic drugs.
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Objective To investigate the efficacy of connexin43 (Cx43) in trigeminocervical complex on allodynia in rat models of migraine.Methods SD rats were randomly divided into 9 groups (n=6):blank group,sham-operated group,saline group,improved inflammatory soup for 3 d group,improved inflammatory soup for 7 d group,carbenoxolone (CBX) prevention group,CBX prevention control group,CBX treatment group and CBX treatment control group.Improved inflammatory soup was used to stimulate the dual matter adjacent to the superior sagittal sinus of rats repeatedly to induce migraine models in the later 7 groups.The pain threshold of periorbital skin was measured by Von-Frey hairs.On the 11th postoperative day,the spinal cord was removed for detection of Cx43 expression in the trigeminocervical complex by immunofluorescence.Results Von-Frey hairs study showed that:the pain threshold of improved inflammatory soup groups had a daily drop; rats in the improved inflammatory soup for 3 d group suffered allodynia,while those in the CBX prevention group did not; the pain threshold in the CBX treatment group ascended after using CBX.Immunofluorescence of nucleus and Cx43 showed that:the mean optical density (OD) of improved inflammatory soup for 3 d group and for 7 d group significantly incresed (P<0.05); nucleus and Cx43 abundantly expressed in lamina Ⅰ and Ⅱ of dorsal horn in the improved inflammatory soup for 3 d group and lamina Ⅰ-Ⅳ of dorsal horn in improved inflammatory soup for 7 d group; significant difference was noted on the mean OD values between of CBX prevention/treatment groups and their control groups (P<0.05).Conclusion Repeatedly infusing improved inflammatory soup to dual matter can effectively induce allodynia in rat models of migraine and Cx43 abundantly expresses in trigeminocervical complex after being given the soup; allodynia in migraine model of rats can be prevented and relieved by intraperitoneal administrating of CBX,which suggests that Cx43 may play an important role in allodynia of migraine.
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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.
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Humains , Carbénoxolone , Test ELISA , Cellules épithéliales , Expression des gènes , Isoflavones , Mucine-5AC , Mucines , Nécrose , Silymarine , Facteur de nécrose tumorale alphaRésumé
0.05). Conclusions The gap junction formed by CX43 should be an important role in switching and developing the epilepsy. The synthetic polypeptide corresponding to amino acid sequence of CX43 would be a kind of new type antiepileptic drug, which might have less side effects and more potent pertinency.
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Objective To observe the behavior changes and connexin 32(CX32),connexin 43(CX43)expressions in hippocampus and the effect of carbenoxolone on their expression in epileptic immature rats induced by lithium-pilocarpine.Methods Seventy-two SD immature rats of 21 d were randomly divided into control group(n=24),lithium-pilocarpine kindled group(n=24)and carbenoxolone treated group(n=24),each group by 24 h,3 d,7 d and 30 d were subdivided into 4 groups(n=6).Immuno-histochemisty was used to observe the expressions of CX32 and CX43 in hippocampus areas of immature rats,and to observe their behavior changes.Results The scores of the severe elileptiform seizures(Racine Ⅳ/Ⅴlevel)in lithium-pilocarpine group were significantly higher than those in carbenoxolone treated group;The latency in carbenoxolone treated group was prolonged significantly(P
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Objective To study the role of gap junctions in epileptiform activity. Methods The epileptiform activity was induced by zero-Mg 2+ medium in cultured hippocampal neurons of newborn rats. Immunocytochemistry and real time RT-PCR were introduced to evaluate the expression of gap junction Cx32 and Cx43. Results The level of Cx32 mRNA increased quickly one hour after the neurons were treated with zero-Mg 2+ medium and was raised by 10 times 5 hours later, while Cx32 protein began to develop at the 2nd hour (21.80?1.74) and was raised by 5 times at the 8th hour (47.30?5.75). The expression of Cx43 mRNA went up obviously 5 hours later, and Cx43 protein developed visibly 8 hours later. Carbenoxolone depressed the expressions of Cx32 and Cx43. Conclusions The expression of Cx32 and Cx43 increases dramatically after epileptic discharges and carbenoxolone inhibits both the discharges and the expression of gap junctions, which indicates that gap junction could contribute to epileptogenesis.
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Objective Complex of oxymatrine(OMT)-carbenoxolone sodium(CS)was prepared,and the acute toxicity of the OMT-CS complex and their protective effects on injured liver induced by carbon tetrachloride(CCl_4)were also studied.Methods The OMT-CS complex was formed after preparing OMT and CS solutions individually then mixing them in ratio,the constant was detemined by UV-vis,the complex was characterized by powder X-ray diffraction,IR,and NMR.The sequence method was employed to test LD_50 of the complex by iv injection.The aspartate transaminase(AST)and alanine aminotransferase(ALT)in serum were measured in acute hepatic injuried models by CCl_4 proportionally im injected the complex in mice.Results The ratio of OMT and CS in the complex appeared to be 1∶1.The results of powder X-ray diffraction measurement indicated that a novel crystalline structure of complex was formed.The acute toxicities of LD_50 of the complexes in 1∶1,and 2∶1 were lower than that of both OMT and CS only.Compared with the model group by CCl_4,AST and ALT levels in serum were lower than that in the complex at 1∶1 and 2∶1 ratios.Conclusion The toxicity of the complex in different proportions on acute hepatic injuried mice by CCl_4 decreases obviously,compared with their precursor drugs.