【摘要】 目的探讨脑出血患者血肿周围组织细胞因子表达和细胞凋亡的关系。方法对30例脑出血患者采取非功能区漏斗式入颅,从入颅路径过程中必须切除的脑组织里取靠近血肿旁约1cm脑组织作为实验组,按发病到手术的时间将实验组分为6h以内组6例、6～12h组7例、12～24h组5例、24～72h组6例、72h以上组6例;取7例远离血肿的脑组织作为对照组(6h以内6例,6～12h1例)。应用HE染色、免疫组化染色及逆转录-聚合酶链反应(RT-PCR)观察组织病理,肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)等细胞因子和Tunel、促凋亡相关基因(Bax)、抑凋亡相关基因(Bcl-x)的表达情况。结果HE染色光镜观察显示:对照组、6h以内组血肿周围组织基本正常,6～12h组损伤较轻,12～24h组损伤较重,24～48h组损伤严重,以后逐渐好转。免疫组化显示:TNF-α蛋白和mRNA及IL-1β和IL-6mRNA表达于12～72h达高峰(P<0.01),以后逐渐降低。Tunel、Bax蛋白和mRNA表达在12～72h达峰值(P<0.05,P<0.01)。Bcl-x蛋白及mRNA表达在12～72h有升高趋势,但无统计学差异。相关分析显示:TNF-α蛋白和mRNA、IL-1β及IL-6mRNA的表达与Tunel、Bax蛋白和mRNA的表达呈显著正相关(P<0.01),与Bcl-x蛋白和mRNA表达无相关性(P>0.05)。结论脑出血后血肿周围组织炎性细胞因子的表达与细胞凋亡密切相关,并与组织病理损伤程度相一致。更多还原

【Abstract】 Objective To study the relationship between the expressions of cytokines and apoptosis-related genes in the perihematoma tissues of intracerebral hemorrhage(ICH)patients.Methods Using cone-shaped cut into the non-functional area in the brain of 30 ICH patients during operation,a few brain tissues about 1 cm away from the hematoma were obtained from those that must be cut on the way into the brain.The 30 subjects were categorized into 5 groups based on the time from the onset of illness to operation:6 in < 6 h group,7 in 6~12 h group,5 in 12~24 h group,6 in 24~72 h group,and 6 in > 72 h group.The brain tissues of the control group were obtained on the way into the cranium,but far from the hematoma,in the 7 patients chosen from the first 2 groups(6 in < 6 h group,1 in 6~12 h group).The HE staining,the immunohistochemical analysis and reverse transcription-polymerase chain reaction(RT-PCR)were performed on each tissue to detect the pathological changes,the expressions of cytokines [tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)],the Tunel-positive cells,the apoptosis-related gene expressions(Bax,Bcl-x).Results The tissues near to hematoma were nearly normal in control group and < 6 h group,and damaged slightly in 6~12 h group,relatively severely in 12~24 h group and the most severely in 24~48 h group.However,the other groups got better later and lookedsimilar to the control group on the 8th day.TNF-α,IL-1β and IL-6 mRNA levels,as well as the protein expressions increased gradually in ICH patients,to the maximum between 12~72 h(P<0.01)and then decreased gradually.The Tunel-positive cells and the expression of Bax mRNA and protein reached the peak level between 12~72 h(P<0.05,P<0.01).The expression of Bcl-x protein and mRNA tended to increase between 12~72 h but there was no significant difference between them.The correlation analysis showed that the expressions of TNF-α protein and mRNA,IL-1β and IL-6 mRNA were strongly positively related to those of Tunel,Bax protein and mRNA(P<0.01),but not related to Bcl-x protein and mRNA expression(P>0.05).Conclusion The expressions of cytokines are closely related to the apoptosis in the tissues surrounding the hematoma in ICH patients and consistent with the degree of histopathological damage.更多还原