电针足三里穴对严重腹腔感染大鼠胸腺细胞凋亡的保护作用

Mechanism ud protection of electro-acupuncture at Zusanli points(足三星穴) on the apoptosis of thymocytes in rats with severe abdominal infection

目的探讨电针足三里穴对腹腔感染大鼠胸腺细胞凋亡的影响及机制。方法将40只SD大鼠随机均分成正常对照组、模型组、非经非穴组、足三里组。采用盲肠结扎穿孔(CLP)致腹腔感染动物模型,用流式细胞仪以膜联蛋白-碘化丙啶(Annexin V-PI)双染色法及光镜、透射电镜下观察CLP致腹腔感染后36 h胸腺细胞的凋亡情况,并观察血浆皮质酮浓度和胸腺细胞中Bcl-2蛋白含量的变化。结果CLP致腹腔感染可导致胸腺细胞凋亡增多,模型组[(44.7±3.3)%]、非经非穴组[(42.7±3.0)%]、足三里组[(32.6±3.3)%]胸腺细胞凋亡率显著高于正常对照组[(21.2±2.3)%,P均<0.05];而足三里组胸腺细胞凋亡率显著低于模型组和非经非穴组(P均<0.05);电镜和光镜下观察胸腺细胞凋亡呈典型的形态学特征。模型组[(353.1±75.8)μg/L]、非经非穴组[(370.7±81.0)μg/L]、足三里组[(348.6±77.9)μg/L]血浆皮质酮浓度显著高于正常对照组[(161.2±46.5)μg/L,P均<0.05];足三里组与模型组和非经非穴组比较差异无统计学意义(P均>0.05)。模型组(71.2±5.6)、非经非穴组(73.5±5.9)、足三里组(82.4±6.8)胸腺细胞中Bcl-2蛋白含量显著低于正常对照组(95.3±6.3.P均<0.05),而足三里组胸腺细胞中Bcl-2蛋白含量相对值显著高于模型组和非经非穴组(P均<0.05)。结论电针足三里穴可以减轻CLP致腹腔感染大鼠胸腺细胞凋亡,其机制可能与提高胸腺细胞中Bcl-2蛋白含量有关。

Objective To study the influence of electro-acupuncture （EA） at Zusanli points （足三里穴） on the apoptosis of thymocytes in rats with abdominal infection and its mechanism. Methods ： A total of 40 Sprague-Dawley （SD） rats were randomly divided into four groups, including normal control group, model group, non-acupoint group and Zusanli group. The abdominal infection model of rat was made by cecal ligation and puncture （CLP）. After abdominal cavity infection for 36 hours, the apoptosis of thymoeytes was observed under electron microscope and light microscope, and the apoptosis ratio of thymocytes was determined by Annexin V-PI method with flow cytometry technique. The content of Bcl-2 protein of thymocytes and concentration of corticosterone in plasma were determined. Results Abdominal infection resulted from CLP could significantly increase the apoptosis of thymocytes and lead to the typical histopathological changes of apoptosis of thymocytes under electron microscope and light microscope. Apoptosis ratios of thymocytes in model group [（44.7±3.3）%], non-acupoint group [（42.7±3.0）%] and Zusanli group [（32.6±3.3）%] were significantly higher than the ratio in the control group [（21.2±2.3）%, all P〈0. 053. And apoptosis ratio of thymocytes in Zusanli group was significantly lower than the ratios in model group and non-acupoint group （both P〈0.05）. Abdominal infection resulted from CLP could increase the plasma concentration of corticosterone. The plasma concentrations of corticosterone in model group [（353.1 ± 75.8）μg/L], non-acupoint group [（370.7±81.0） μg/L] and Zusanli group [（348.6±77.9） μg/L] were significantly higher than the concentration in control group [（161.2±46.5） μg/L, all P〈0. 053. But there was no significant difference among model group, non-acupoint group and Zusanli group （all P〉0.05）. Abdominal infection resulted from CLP also could reduce the content of Bcl-2 protein of thymocytes. The content of Bcl-2 protein of thymocytes in model group （71.2±5.6）, non-acupoint group （73.5±5.9） and Zusanli group （82.4±6.8） were significantly lower than normal control group （95.3±6.3, all P〈0. 05）. And the content of Bcl-2 protein of thymocytes in Zusanli group was significantly higher than those in model and non-acupoint groups （both P〈0. 05）. Conclusion EA at Zusanli points can significantly reduce the apoptosis of thymocytes in rats with abdominal infection resulted from CLP and the mechanism may be closely associated with the increasing content of Bcl-2 protein of thymocytes.