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目的研究脑出血患者血肿周围组织补体激活与炎性反应的关系。方法对30例脑出血患者(实验组)在手术时于血肿旁约1cm处取少许脑组织,按发病到手术的时间将实验组分为<6h组(6例),6~12h组(7例),12~24h组(5例),24~72h组(6例),>72h组(6例)。从前2组中选7例患者(对照组)在手术入颅路径上远离血肿处取少许脑组织。应用免疫组化染色,逆转录-聚合酶链反应(RT-PCR)分别观察补体C3和补体抑制剂(Clusterin)的表达与炎性细胞的浸润、胶质细胞增生、炎性细胞因子的表达情况。结果与对照组相比,脑出血患者C3表达高峰在12~72h(P<0.05~0.01),Clusterin表达于12~72h有增高趋势,但无统计学意义;炎性细胞浸润从6~12h逐渐明显,12~72h达高峰(P<0.05~0.01);小胶质细胞、星形胶质细胞于24~72h后增生明显(均P<0.01);肿瘤坏死因子-α(TNF-α)表达于12~72h达高峰(P<0.05~0.01),后逐渐降低。RT-PCR显示:TNF-αmRNA的表达与蛋白表达基本一致;白细胞介素(IL)-1β(IL-1β)和IL-6mRNA表达于12~72h达峰值(均P<0.01)。相关因素分析显示:C3表达与炎性细胞浸润和胶质细胞增生及与细胞因子表达呈显著正相关(r=0.42~0.80;P<0.05~0.01),而Clusterin与上述表达无相关性。结论脑出血患者血肿周围组织补体激活与炎性反应有密切关系。
Objective To study the relationship between complement activation and inflammatory reaction in the perihematoma of patients with intracerebral hemorrhage. Methods Thirty patients with intracerebral hemorrhage (experimental group) underwent surgery at about 1 cm beside the hematoma. The experimental groups were divided into 6 hours (6 cases), 6 to 12 hours (7 cases) ), 12 ~ 24h group (5 cases), 24 ~ 72 hours group (6 cases) and> 72 hours group (6 cases). Seven patients from the previous two groups (control group) received a small amount of brain tissue at a distance from the hematoma on the approach to the skull. The expressions of complement C3 and clusterin, inflammatory cell infiltration, glial cell proliferation and the expression of inflammatory cytokines were observed by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) . Results Compared with the control group, the peak of C3 expression in ICH patients was between 12 and 72 hours (P <0.05 and 0.01). Clusterin expression increased from 12 to 72 hours, but there was no statistical significance. Inflammatory cell infiltration gradually decreased from 6 to 12 hours (P <0.05 ~ 0.01). The microglia and astrocyte proliferated significantly after 24 ~ 72h (all P <0.01). The expression of tumor necrosis factor-α (TNF-α) Reached the peak at 12 ~ 72h (P <0.05 ~ 0.01), then decreased gradually. RT-PCR showed that the expression of TNF-α mRNA was consistent with the protein expression; the expression of IL-1β and IL-6 mRNA reached the peak at 12-72h (all P <0.01). Correlation analysis showed that there was a significant positive correlation between C3 expression and inflammatory cell infiltration, glial cell proliferation and cytokine expression (r = 0.42-0.80; P <0.05-0.01), while Clusterin had no correlation with the above expression. Conclusion There is a close relationship between complement activation and inflammatory response in the perihematoma of patients with intracerebral hemorrhage.