Exploratory Development Of Changing On Gap Junctional Intercellular Communication In Neurosurgery

Part OneEffects of hypothermia on gap junctional intercellularcommunication of gliacyteObjective: To observe effects of hypothermia on gap junctional intercellularcommunication (GJIC) in rat's glial cells.Methods and materials: After glial cells(GCs) had grown to confluence,they. were divided into five groups and were exposed to different conditions. GCs ingroup one were incubated in nomoxia and normothermic culture condition for 24hours. GCs in group two were exposed to nomoxia and hypothermia for 24 hours.GCs in group three were incubated in mild hypoxia and normothermic culturecondition tbr 24 hours. GCs in group four were exposed to mild hypoxia andhypothermia for 24 hours. GCs in group five were exposed to nomoxia for 22 hoursand then were incubated in TPA. for 2 hours. The effects of hypothermia on GJIC inglial cells were determined by scrape—loading and dye transfer(SLDT) technique ofusing a laser scanning confocal microscope (LSCM). PCR technique and immuocytochemistry were employed to detect the expression of Cx43 in the culturedcells.esults: In group one, the fluorescence distance of GCs was 160.5±8.6μm, Ingroup two, the fluorescence distance of GCs was 126.24±10.4μm, In group three,the fluorescence distance of GCs was 154.1±6.9μm, In group four, the fluorescencedistance of GCs was 122.8±6.0μm. There was significantly difference in fluorescencedistance between group one and group two (p＜0.01), between group three and groupfour(p＜0.01). There was no difference in fluorescence distance between group one andgroup three (p＞0.05), between group two and group four(p＞0.05).GCs of hypothermia group for 24 hours: Red fluorescence intensity of rhodaminemarking Cx43 was 81.7±2.6, an average correspondence area of fluorescent light spotwas 67.82±14.16; GCs of normothermic group for 24 hours: Red fluorescenceintensity of rhodamine marking Cx43 was 85.04±7.9, an average correspondence areaof fluorescent light spot was 234.85±90.99. There was no difference in redfluorescence intensity of rhodamine between normothermic group and hypothermiagroup (p＞0.05), but an average correspondence area of fluorescent light spot innormnothermic group was larger than fluorescent light spot in hypothermia group, therewas significantly difference in both (p＜0.01).Real-time PCR result: The expression of Cx43 mRNA in normothermic groupwas higher than that in hypothermia group (p＜0.05). there was significantly differencein both (p＜0.01).Conclusions: Under the condition of this experiment, we found GCs'GJIC,whether in nomoxia or in mild hypoxia, was much weaker than that amongnormothermic cells after applying hypothermia to these cultured cells. Hypothermiacould inhibit the GJIC founction. This suggests that hypothermia may have certaineffect on CNS's recovery fiom trauma via depressing GJIC founction. Part TwoEffects of cerebroprotein hydrolysate injection on gap junctionalintercellular communication of rat's glial cellsObjective: To observe effects of cerebroprotein hydrolysate injection(CHI) ongap junctional intercellular communication (GJIC) in rat's glial cells.Methods and materials: After glial cells(GCs) had grown to confluence,they.were divided into three groups:CHI group, nomal control group and TPA group.The effects of CHI on GJIC in glial cells were determined by scrape—loading and dyetransfer(SLDT) technique of using a laser scanning confocal microscope(LSCM).RT-PCR technique was employed to detect the expression of Cx43 mRNA inthe cultured cells, glial cells(GCs) were cultured in dishes until nearly confluent. Thenthey were also divided into the following:④Nomoxia control group⑤NomoxiaDrug group⑥Mild hypoxia control group⑦Mild hypoxia drug group⑧TPA groupto detect GJIC.Results: In CHI group, the fluorescence distance of GCs was 189.8±6.6μm, Innomal control group, the fluorescence distance of GCs was 154.7±6.0μm, There wassignificantly difference in fluorescence distance between CHI group and nomalcontrol group (p＜0.05), The expression of Cx43 mRNA in CHI group was higher thanthat in normal control group (p＜0.05). The distances of groups were④157.5±5.9μm,⑤198.2±8.8μm,⑥153.8±7.6μm,⑦195.9±12.9μm. There wassignificantly difference in fluorescence distance between Nomoxia drug and control,between Mild hypoxia drug group and Mild hypoxia control group (p＜0.01). Therewas no difference in fluorescence distance compared with nomoxia cotrol and hypoxiacontrol group.Conclusions: Under the condition of this experiment, we found GCs' GJIC andthe expression of Cx43 mRNA, whether in nomoxia or in mild hypoxia, werenotably enhanced after applying CHI to these cultured cells. This suggests that CHI may have certain effect on CNS via enhancing GJIC founction.Part ThreeInhibitive effect of hypothermia on gap junctional intercellularcommunication in rabbit's vascular smooth muscle cellsPurpose: To observe effects of sludged blood compound (SBC) on gap junctionalintercellular communication (GJIC) in rabbit's vascular smooth muscle cells(VSMs)and illuminate inhibitive effects of hypothermia on this GJIC.Methods and materials: After VSMs had grown to confluence, they weredivided into three groups and were exposed to different conditions. VSMs in groupone were incubated in SBC and normothermic culture condition for 36 hours. VSMsin group two were exposed to normothermic culture condition for 36 hours. VSMs ingroup three were incubated in SBC and hypothermia culture condition for 36 hours.The effects of SBC or hypothermia on GJIC in VSMs were determined byscrape—loading and dye transfer(SLDT) technique of using a fluorescent microscope.Results: In group one, the fluorescent diffusion length of VSMs was longer thangroup two,but, in group three, the fluorescent diffusion length of VSMs was shorterthan group one. There was significantly difference in fluorescent diffusion lengthbetween group one and group two (P＜0.01), between group three and groupone(P＜0.01 ).Conclusions: Under the condition of this experiment, we found VSMs' GJIC wasnotably enhanced after applying SBC into these cultured cells. Hypothermia couldinhibit this GJIC founction. Maybe such effects would be one of mechanism of actionabout hypothermia could relieve CVS after SAH.