中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2008年
4期
301-303
,共3页
洪涛%段剑%汪阳%冯九庚%江志群
洪濤%段劍%汪暘%馮九庚%江誌群
홍도%단검%왕양%풍구경%강지군
外膜成纤维细胞%脑血管痉挛%共培养%氧合血红蛋白
外膜成纖維細胞%腦血管痙攣%共培養%氧閤血紅蛋白
외막성섬유세포%뇌혈관경련%공배양%양합혈홍단백
Adventitial fibroblasts%Cerebral vasospasm%Co-culture%Oxyhemoglobin
目的 在体外模拟血管壁外层结构,建立外膜成纤维细胞(AF)和平滑肌细胞(SMC)共培养,研究血管AF在蛛网膜下腔出血后脑血管痉挛中的作用.方法 (1)以多聚碳酸酯膜(PET膜)作为载体,将AF和SMC进行共培养.(2)模拟体内出血环境,在PET膜一侧接种AF细胞并加入含10-6 mol/L OxyHb的培养液,分别共培养24h、48h、72h,应用电镜观察SMC的长度.(3)提取各组AF缝隙连接蛋白Cx43进行RT-PCR半定量分析.结果 (1)扫描电镜测量发现,加入OxyHb处理的AF能使PET膜对侧未经OxyHb处理的SMC产生收缩(P＜0.001),且收缩程度与OxyHb处理AF细胞时间成正比.(2)AF缝隙连接蛋白Cx43半定量RT-PCR分析显示:24h、48h、72h组中Cx43mRNA平均相对含量较正常组上调(P＜0.01).结论 血管壁外层的AF受到OxyHb作用后可以引起未直接接触OxyHb的SMC发生持续收缩且缝隙连接蛋白Cx43mRNA表达上调,提示AF在蛛网膜下腔出血引起的脑血管痉挛中可能发挥重要作用.
目的 在體外模擬血管壁外層結構,建立外膜成纖維細胞(AF)和平滑肌細胞(SMC)共培養,研究血管AF在蛛網膜下腔齣血後腦血管痙攣中的作用.方法 (1)以多聚碳痠酯膜(PET膜)作為載體,將AF和SMC進行共培養.(2)模擬體內齣血環境,在PET膜一側接種AF細胞併加入含10-6 mol/L OxyHb的培養液,分彆共培養24h、48h、72h,應用電鏡觀察SMC的長度.(3)提取各組AF縫隙連接蛋白Cx43進行RT-PCR半定量分析.結果 (1)掃描電鏡測量髮現,加入OxyHb處理的AF能使PET膜對側未經OxyHb處理的SMC產生收縮(P＜0.001),且收縮程度與OxyHb處理AF細胞時間成正比.(2)AF縫隙連接蛋白Cx43半定量RT-PCR分析顯示:24h、48h、72h組中Cx43mRNA平均相對含量較正常組上調(P＜0.01).結論 血管壁外層的AF受到OxyHb作用後可以引起未直接接觸OxyHb的SMC髮生持續收縮且縫隙連接蛋白Cx43mRNA錶達上調,提示AF在蛛網膜下腔齣血引起的腦血管痙攣中可能髮揮重要作用.
목적 재체외모의혈관벽외층결구,건립외막성섬유세포(AF)화평활기세포(SMC)공배양,연구혈관AF재주망막하강출혈후뇌혈관경련중적작용.방법 (1)이다취탄산지막(PET막)작위재체,장AF화SMC진행공배양.(2)모의체내출혈배경,재PET막일측접충AF세포병가입함10-6 mol/L OxyHb적배양액,분별공배양24h、48h、72h,응용전경관찰SMC적장도.(3)제취각조AF봉극련접단백Cx43진행RT-PCR반정량분석.결과 (1)소묘전경측량발현,가입OxyHb처리적AF능사PET막대측미경OxyHb처리적SMC산생수축(P＜0.001),차수축정도여OxyHb처리AF세포시간성정비.(2)AF봉극련접단백Cx43반정량RT-PCR분석현시:24h、48h、72h조중Cx43mRNA평균상대함량교정상조상조(P＜0.01).결론 혈관벽외층적AF수도OxyHb작용후가이인기미직접접촉OxyHb적SMC발생지속수축차봉극련접단백Cx43mRNA표체상조,제시AF재주망막하강출혈인기적뇌혈관경련중가능발휘중요작용.
Objective To study the pathological role of the adventitial fibroblasts in the experimenlal cerebral vasospasm through the co-culture of adventitial fibroblasts and smooth muscle cells in vitro,imitating the outer layer of vascular wall.Methods (1)Smooth muscle cells(SMC)and adventitial fibroblasts(AF)were grown each on opposite sides of polyethylene terephthalate(PET).(2)The AF were incubated in the medium containing 10-6 mol/L oxyhemoglobin.When the two types of the cells were cocuhured for 24h,48h and 72h.the length changes of the smooth muscle cells were examined with electron microscopy.(3)The total RNA of the adventitial fibroblasts in the co-culture system was extracted.Semiquantitative RT-PCR was performed to measure the relative contain of mRNA for connexins in those cells.Results (1)The AF incubated with oxyhemoglobin induced the contraction of the cocuhured smooth muscle cells,which incubated without oxyhemoglobin(P＜0.001).(2)The results of semiquantitative RT-PCR showed that the average relative contain of mRNA for Cx43 at 24h,48h and 72h groups was distinctly higher than that in the normal group(P＜0.01).Conclusion The AF incubated with oxyhemoglobin can produce the contraction of the cocuhured SMC,even the SMC is not derectly stimulated by oxyhemoglobin.The expression of mRNA for connexin43 in adventitial fibroblasts was upregulated in the cell model of cerebral vasospasm.This phenomenon suggests that the AF play an important role in cerebral vasospasm,the mechanism of the smooth muscle cells contraction.
