本文選題：微波 + γ射線　； 參考：《蘇州大學(xué)》2015年碩士論文

【摘要】：目的:以原代小鼠骨髓基質(zhì)細(xì)胞為研究對象,建立低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)細(xì)胞模型,分析低劑量微波輻射與ROS產(chǎn)生及NF-κB活化的相關(guān)性,探討低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)的可能機(jī)理。方法:1.低劑量微波輻射誘導(dǎo)小鼠骨髓基質(zhì)細(xì)胞適應(yīng)性反應(yīng)模型的建立制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為6組:對照組、微波照射組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、假照射組(細(xì)胞置于微波輻照裝置中,不給予微波照射)、γ射線照射組(1.5 Gy 60Co-γ射線照射,劑量率0.376 Gy/min)、微波復(fù)合組(900MHz,120μW/cm2微波照射,4h/d,連續(xù)5d,照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線照射)、假照射復(fù)合組(假照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線照射)。γ射線照射結(jié)束后,立即收集細(xì)胞,用堿性單細(xì)胞凝膠電泳實(shí)驗(yàn)和γ-H2AX焦點(diǎn)形成試驗(yàn)檢測細(xì)胞DNA單、雙鏈損傷。2.ROS在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用(1)低劑量微波輻射誘導(dǎo)小鼠骨髓基質(zhì)細(xì)胞產(chǎn)生ROS:制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為6組:對照組、褪黑素組(500n M褪黑素處理細(xì)胞)、微波照射組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、假照射組(細(xì)胞置于微波輻照裝置中,不給予微波照射)、褪黑素+微波組(500n M褪黑素處理細(xì)胞,4小時(shí)后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、H2O2組【培養(yǎng)液中1:1000(V/V)濃度加入H2O2,處理細(xì)胞20~30分鐘】,處理結(jié)束后立即收集細(xì)胞,用試劑盒測定細(xì)胞內(nèi)ROS水平。(2)ROS在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用:制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為6組:對照組、微波照射組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、褪黑素+微波組(500n M褪黑素處理細(xì)胞,4小時(shí)后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、γ射線照射組(1.5 Gy 60Co-γ射線照射,劑量率0.376Gy/min)、微波復(fù)合組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d,照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線)、褪黑素+微波+γ射線組(500n M褪黑素處理細(xì)胞,4小時(shí)后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d,微波照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線)。照射結(jié)束后,立即進(jìn)行堿性單細(xì)胞凝膠電泳實(shí)驗(yàn)和γ-H2AX焦點(diǎn)形成試驗(yàn),檢測細(xì)胞DNA單、雙鏈損傷。3.NF-κB活化在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用(1)低劑量微波輻射誘導(dǎo)NF-κB活化:制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為3組:對照組、微波照射組(900MHz,120μW/cm2的微波輻照,4h/d,連續(xù)5d)、假照射組(細(xì)胞置于微波輻照裝置中,不給予微波照射)。于微波照射結(jié)束后0、0.5、1、2、4、6小時(shí)收集細(xì)胞,提取細(xì)胞胞漿和胞核蛋白,用Wsetern Blot檢測NF-κB表達(dá)水平。(2)NF-κB活化在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用:制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為6組:對照組、微波照射組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、BAY 11-7082+微波組(10μM BAY 11-7082處理30分鐘,然后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、γ射線照射組(1.5 Gy 60Co-γ射線照射,劑量率0.376 Gy/min)、微波復(fù)合組(900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d,照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線)、BAY 11-7082+微波+γ射線組(10μM BAY11-7082處理細(xì)胞,30分鐘后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d,微波照射結(jié)束4小時(shí)后給予1.5 Gy 60Co-γ射線照射)。照射結(jié)束后立即進(jìn)行堿性單細(xì)胞凝膠電泳實(shí)驗(yàn)和γ-H2AX焦點(diǎn)形成試驗(yàn)檢測細(xì)胞DNA單、雙鏈斷裂。4.ROS在低劑量微波輻射誘導(dǎo)的NF-κB活化中的介導(dǎo)作用:制備小鼠骨髓基質(zhì)細(xì)胞,隨機(jī)分為3組:對照組、褪黑素+微波組(500n M褪黑素處理細(xì)胞,4小時(shí)后給予900MHz,120μW/cm2的微波照射,4h/d,連續(xù)5d)、假照射組(細(xì)胞置于微波照射裝置中,不給予微波照射)。微波照射結(jié)束后0、0.5、1、2、4、6小時(shí)收集細(xì)胞,提取細(xì)胞胞漿和胞核蛋白,Wsetern Blot檢測NF-κB表達(dá)水平。結(jié)果:1.低劑量微波輻射誘導(dǎo)小鼠骨髓基質(zhì)細(xì)胞適應(yīng)性反應(yīng)模型的建立(1)與對照組相比,假照射組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均無顯著變化(P0.05);與單純?chǔ)蒙渚�照射組相比,假照射與γ射線復(fù)合組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均無顯著變化(P0.05),說明微波假照射對DNA損傷無明顯影響。(2)與對照組相比,微波照射組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均無明顯變化(P0.05),γ射線照射組細(xì)胞尾長、尾矩明顯增大(P0.0001),γ-H2AX焦點(diǎn)形成數(shù)量明顯增加(P0.0001),說明微波照射對DNA損傷無明顯影響,而γ射線暴露可導(dǎo)致明顯的DNA單鏈和雙鏈損傷。(3)與單純?chǔ)蒙渚�照射組相比,微波復(fù)合組細(xì)胞彗星尾長、尾距明顯減小(P0.0001),γ-H2AX焦點(diǎn)形成數(shù)量明顯降低(P0.0001),說明低劑量微波輻射能夠誘導(dǎo)適應(yīng)性反應(yīng),拮抗γ射線暴露導(dǎo)致的DNA單鏈和雙鏈損傷。2.ROS在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用(1)與對照組相比,褪黑素組和假照射組細(xì)胞內(nèi)ROS水平均無顯著變化(P0.05),說明褪黑素和微波假照射對細(xì)胞內(nèi)ROS水平無明顯影響。(2)與對照組相比,微波照射組和H2O2組細(xì)胞內(nèi)ROS水平均顯著升高(P0.01),說明微波照射和H2O2處理可誘導(dǎo)細(xì)胞內(nèi)ROS水平顯著升高。(3)與微波組相比,褪黑素+微波組細(xì)胞內(nèi)ROS水平顯著降低(P0.05),說明褪黑素預(yù)處理能明顯清除細(xì)胞內(nèi)微波誘導(dǎo)產(chǎn)生的ROS。(4)與對照組相比,微波照射組、褪黑素+微波組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均無顯著變化(P0.05),γ射線照射組細(xì)胞尾長、尾矩明顯增大(P0.01),γ-H2AX焦點(diǎn)形成數(shù)量明顯增多(P0.0001),說明微波照射和褪黑素對DNA損傷無明顯影響,而γ射線暴露可導(dǎo)致明顯的DNA單鏈和雙鏈損傷。(5)與單純?chǔ)蒙渚�照射組相比,微波與γ射線復(fù)合照射組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均有顯著變化(P0.0001),說明低劑量微波輻射能夠誘導(dǎo)適應(yīng)性反應(yīng),拮抗γ射線暴露導(dǎo)致的DNA單鏈和雙鏈損傷。(6)與微波復(fù)合照射組相比,褪黑素+微波+γ射線組細(xì)胞彗星尾長、尾距明顯增大(P0.01),γ-H2AX焦點(diǎn)形成數(shù)量明顯增多(P0.0001),說明褪黑素預(yù)處理后再給予微波+γ射線照射可抑制微波誘導(dǎo)適應(yīng)性反應(yīng)產(chǎn)生。3.NF-κB活化在低劑量微波輻射誘導(dǎo)適應(yīng)性反應(yīng)中的作用(1)與對照組相比,假照射組細(xì)胞胞漿和胞核中NF-κB水平均無顯著變化(P0.05),說明微波假照射對NF-κB活化無顯著影響。(2)與對照組相比,微波照射組胞核內(nèi)NF-κB隨照射后時(shí)間增加而升高(P0.01),胞漿內(nèi)NF-κB表達(dá)水平隨時(shí)間增加而降低,說明微波誘導(dǎo)NF-κB的活化,活化后的NF-κB由細(xì)胞漿移位進(jìn)入細(xì)胞核。(3)與微波組相比,BAY 11-7082+微波組胞漿和胞核內(nèi)NF-κB水平均明顯降低(P0.01),說明NF-κB抑制劑BAY 11-7082預(yù)處理可抑制微波誘導(dǎo)NF-κB活化。(4)與對照組相比,微波照射組、BAY 11-7082+微波組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均無顯著差異(P0.05),γ射線照射組細(xì)胞尾長、尾矩明顯增大(P0.0001),γ-H2AX焦點(diǎn)形成數(shù)量明顯增多(P0.0001),說明微波照射和NF-κB抑制劑BAY 11-7082對DNA損傷無明顯影響,而γ射線暴露可導(dǎo)致明顯的DNA單鏈和雙鏈損傷。(5)與單純?chǔ)蒙渚�照射組相比,微波復(fù)合照射組細(xì)胞彗星尾長、尾距及γ-H2AX焦點(diǎn)形成數(shù)量均有顯著差異(P0.0001),說明低劑量微波輻射能夠誘導(dǎo)適應(yīng)性反應(yīng),拮抗γ射線暴露導(dǎo)致的DNA單鏈和雙鏈損傷。(6)BAY 11-7082+微波+γ射線組與微波復(fù)合照射組相比,細(xì)胞彗星尾長、尾距明顯增大(P0.01),γ-H2AX焦點(diǎn)形成數(shù)量明顯增多(P0.0001),說明NF-κB抑制劑預(yù)處理后再給予微波+γ射線照射可抑制微波誘導(dǎo)適應(yīng)性反應(yīng)產(chǎn)生。4.ROS在低劑量微波輻射誘導(dǎo)的NF-κB活化中的介導(dǎo)作用(1)與對照組相比,假照射組細(xì)胞胞漿和胞核中NF-κB水平均無顯著變化(P0.05),說明微波假照射對NF-κB活化無明顯影響。(2)與對照組相比,褪黑素+微波組胞核內(nèi)NF-κB隨照射后時(shí)間增加并無顯著變化(P0.05),說明褪黑素預(yù)處理后再給予微波照射可通過抑制微波誘導(dǎo)ROS產(chǎn)生,繼而抑制NF-κB活化。結(jié)論:1.預(yù)先給予900MHz,120μW/cm2低劑量微波輻射能夠誘導(dǎo)小鼠骨髓基質(zhì)細(xì)胞產(chǎn)生適應(yīng)性反應(yīng),減輕γ射線致小鼠骨髓基質(zhì)細(xì)胞DNA單鏈和雙鏈損傷。2.900MHz,120μW/cm2微波輻射能夠誘導(dǎo)細(xì)胞產(chǎn)生ROS,ROS誘導(dǎo)細(xì)胞內(nèi)NF-κB活化,進(jìn)而誘導(dǎo)適應(yīng)性反應(yīng)。
[Abstract]:Objective : To study the correlation between low dose microwave radiation and ROS production and NF - 魏B activation in mouse bone marrow stromal cells . The cells were collected immediately after 緯 - ray irradiation , the cells were collected immediately after irradiation with alkaline single - cell gel electrophoresis and the 緯 - H2AX focus formation assay . ROS were induced in mouse bone marrow stromal cells by low - dose microwave radiation . ( 2 ) The effect of ROS in the adaptive response induced by low - dose microwave radiation : The mouse bone marrow stromal cells were randomly divided into 6 groups : control group , microwave irradiation group ( 900 MHz , 120 渭W / cm2 microwave irradiation , 4h / d , continuous 5d ) , melatonin + microwave group ( 500 n M melatonin treatment cells , dose rate of 0.376Gy / min ) , microwave irradiation group ( 1.5 Gy 60Gy / min ) , microwave irradiation group ( 900MHz , 120渭W / cm2 microwave irradiation , 4h / d , continuous 5d , microwave irradiation at 120 渭W / cm2 , 4h / d , continuous 5d , microwave irradiation at 900 MHz , 120渭W / cm2 , 4h / d , continuous 5d , microwave irradiation at 900 MHz , 120 渭W / cm2 for 4 hours , irradiation at 4 h / d , 5 days for 5 days , microwave irradiation for 4 hours , 1.5 Gy 60 Co - 緯 rays ) . Activation of NF - 魏B in low - dose microwave radiation induced by low - dose microwave radiation ( 1 ) low - dose microwave radiation induced NF - 魏B activation : the preparation of mouse bone marrow stromal cells was randomly divided into three groups : control group , microwave irradiation group ( 900 MHz , 120 渭W / cm2 microwave irradiation , 4h / d , continuous 5d ) , sham irradiation group ( cells were placed in microwave irradiation device , microwave irradiation was not given ) . Cells were collected at 0 , 0.5 , 1 , 2 , 4 and 6 hours after microwave irradiation , and the expression level of NF - 魏B was detected by Western Blot . The effects of ROS on NF - 魏B activation induced by low - dose microwave radiation were studied . The effects of ROS on NF - 魏B activation induced by low - dose microwave radiation were as follows : control group , melatonin + microwave group ( 500n M melatonin treatment cells , 900 MHz , 120 渭W / cm2 microwave irradiation , 4 h / d , 5 days ) , sham irradiation group ( cells were placed in microwave irradiation apparatus , no microwave irradiation ) . Results : Compared with the control group , the amount of ROS in the comet tail length , tail distance and 緯 - H2AX were significantly increased compared with the control group ( P0.05 ) . ( 4 ) Compared with the control group , there was no significant change in the number of focal forms of the comet tail , tail distance and 緯 - H2AX in the microwave irradiation group ( P0.01 ) . ( 6 ) The effect of microwave irradiation on NF - 魏B activation was significantly increased compared with the control group ( P0.05 ) .

【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R594.8

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