Zhang Pan, Guo Cong-fang, Luo Ning, Wang Bing, Xu Xin-Nü
Tianjin First Central Hospital, Tianjin 300192, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2009 Dec;21(12):726-8.
To observe the change in number of spleen T lymphocytes and B lymphocytes and apoptosis of splenocytes in rats with sepsis, and explore the mechanism of immune imbalance in sepsis.
Eighty healthy male Wistar rats were randomly divided into two groups: sham operation (n=30) and sepsis model (n=50). Sepsis model was reproduced by cecal ligation and puncture (CLP). The rats were sacrificed and their spleens were obtained at 6, 12, 24, 48, 96 hours after the model was reproduced. The pathological changes in spleen were observed by optical microscopy. Immunohistochemistry method was used to examine the positive expression of CD4(+)/CD8(+)T lymphocyte and B lymphocyte, Bax and Bcl-2. Apoptotic cells of spleen were detected by means of terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL).
By optical microscopy, white pulps were atrophied and splenic nodules were destroyed after CLP. At 6, 12, 24, 48, 96 hours after CLP, the number of CD(+)T lymphocyte, B lymphocyte and Bcl-2 positive expressed cells in sepsis model group were obviously reduced as compared with sham operation group (all P<0.01). CD8(+) positive lymphocytes were not significant changed (all P>0.05). Apoptotic splenocytes and Bax positive cells were obviously increased compared with those of sham operation group (all P<0.01). Correlation analysis showed that Bcl-2 expression and cell apoptosis was negatively correlated (r=-0.659, P<0.01), but Bax positive cells and apoptosis showed positive correlation (r=0.522, P<0.01).
Disturbance of immunological function exists in the rats with sepsis at 24 hours after CLP. The number of CD4(+) T lymphocyte, B lymphocyte was obviously reduced, apoptotic splenocytes were obviously increased. Bax and Bcl-2 play an important role in apoptosis of splenocytes in sepsis.
观察脓毒症大鼠脾脏T淋巴细胞和B淋巴细胞数量变化及脾细胞凋亡情况,探讨脓毒症免疫失衡机制。
80只健康雄性Wistar大鼠随机分为两组:假手术组(n = 30)和脓毒症模型组(n = 50)。采用盲肠结扎穿孔术(CLP)制备脓毒症模型。造模后6、12、24、48、96小时处死大鼠,取脾脏。光镜观察脾脏病理变化。采用免疫组织化学方法检测CD4(+)/CD8(+)T淋巴细胞、B淋巴细胞、Bax和Bcl-2的阳性表达。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测脾脏凋亡细胞。
光镜下,CLP术后白髓萎缩,脾小结破坏。CLP术后6、12、24、48、96小时,脓毒症模型组CD(+)T淋巴细胞、B淋巴细胞及Bcl-2阳性表达细胞数量均明显低于假手术组(均P < 0.01)。CD8(+)阳性淋巴细胞差异无统计学意义(均P > 0.05)。与假手术组相比,脓毒症模型组脾细胞凋亡及Bax阳性细胞明显增多(均P < 0.01)。相关性分析显示,Bcl-2表达与细胞凋亡呈负相关(r = -0.659,P < 0.01),而Bax阳性细胞与凋亡呈正相关(r = 0.522,P < 0.01)。
CLP术后24小时脓毒症大鼠存在免疫功能紊乱。CD4(+)T淋巴细胞、B淋巴细胞数量明显减少,脾细胞凋亡明显增加。Bax和Bcl-2在脓毒症脾细胞凋亡中起重要作用。