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目的探讨脑出血患者血肿周围组织炎性反应与细胞因子表达的相关性。方法将30例脑出血患者作为出血组,于血肿旁约1 cm处取少许脑组织。按发病到手术时间,将出血组分为<6 h组(6例),6~12 h组(7例),12~24 h组(5例),24~72 h组(6例),>72 h组(6例)。从前两组中选7例患者,在手术入颅路径上远离血肿处取少许脑组织为对照组。应用HE染色、免疫组化及逆转录-聚合酶链反应(RT-PCR)等技术观察炎性细胞浸润、胶质细胞增生、肿瘤坏死因子(TNF)-α的蛋白及mRNA、白细胞介素(IL)-1β及IL-6 mRNA的表达情况。结果光镜观察显示:对照组、<6 h组血肿周围组织基本正常,6~12 h组损伤较轻,12~24 h组损伤较重,24~48 h组损伤严重,以后逐渐好转,8 d时与对照组相似。免疫组化显示:中性粒细胞、淋巴细胞、巨噬细胞浸润从6~12 h逐渐明显,12~72 h达高峰(P<0.05),胶质细胞增生从12~24 h开始增加,24 h以后明显增加(P<0.01),TNF-α蛋白表达从6~12 h开始增加,12~24 h达高峰(P<0.05),以后逐渐减少;RT-PCR显示:TNF-αmRNA与其蛋白表达基本一致,IL-1β和IL-6 mRNA的表达于12~72 h达高峰(P<0.01),以后有下降趋势。相关分析显示:中性粒细胞、淋巴细胞、巨噬细胞的浸润和小胶质细胞的增生与TNF-α蛋白和mRNA、IL-1β、IL-6的mRNA表达呈显著正相关(P<0.05),星形胶质细胞的增生与TNF-α蛋白和mRNA表达无相关性,与IL-1β、IL-6 mRNA表达呈显著正相关(P<0.05)。结论脑出血后血肿周围组织炎性反应与细胞因子的表达有密切关系。
Objective To investigate the correlation between the inflammatory reaction and the expression of cytokines in the perihematoma of patients with intracerebral hemorrhage. Methods Thirty patients with cerebral hemorrhage were selected as bleeding group, and a little brain tissue was taken at about 1 cm beside the hematoma. According to the time from onset to operation, the bleeding was divided into six groups (6 cases), 6-12 hours group (7 cases), 12-24 hours group (5 cases), 24-72 hours group (6 cases) > 72 h group (6 cases). In the previous two groups, 7 patients were selected, and a little brain tissue was taken away from the hematoma as the control group in the route of operation into the skull. HE staining, immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) were used to observe the inflammatory cell infiltration, glial cell proliferation, tumor necrosis factor (TNF) -α protein and mRNA, interleukin IL) -1β and IL-6 mRNA expression. Results The light microscope showed that in the control group, the tissue around the hematoma was normal in <6 h group, the lesion was lighter in 6 ~ 12 h group, the heavier in 12 ~ 24 h group and the worse in 24 ~ 48 h group, Similar to the control group. Immunohistochemistry showed that the infiltration of neutrophils, lymphocytes and macrophages gradually increased from 6 to 12 hours and reached the peak at 12 to 72 hours (P <0.05), while glial cell proliferation increased from 12 to 24 hours (P <0.01). The expression of TNF-αprotein increased from 6 to 12 h and peaked at 12 to 24 h (P <0.05), and then decreased gradually. RT-PCR showed that the expression of TNF-αmRNA and its protein The expression of IL-1β and IL-6 mRNA reached the peak at 12 to 72 hours (P <0.01), and then decreased. Correlation analysis showed that the infiltration of neutrophils, lymphocytes and macrophages and the proliferation of microglial cells were positively correlated with the mRNA expressions of TNF-α, IL-1β and IL-6 (P <0.05 ). There was no correlation between the proliferation of astrocytes and the expression of TNF-α protein and mRNA, but positively correlated with the expression of IL-1β and IL-6 mRNA (P <0.05). Conclusion The inflammatory reaction in the perihematomal tissue after intracerebral hemorrhage is closely related to the expression of cytokines.