南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1789-1795.doi: 10.12122/j.issn.1673-4254.2023.10.18

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低强度脉冲超声抑制脓毒症大鼠脾脏淋巴细胞的凋亡

唐文韬，邓 娟，贺思程，李君粉，周艺情，王 嫣   

  1. 重庆医科大学生物医学工程学院、超声医学工程国家重点实验室，重庆 400016；重庆市生物医学工程学重点实验室，重庆 400016
  • 出版日期:2023-10-20 发布日期:2023-11-02

Inhibitory effect of low-intensity pulsed ultrasound on apoptosis of splenic lymphocytes in septic rats

TANG Wentao, DENG Juan, HE Sicheng, LI Junfen, ZHOU Yiqing, WANG Yan   

  1. State Key Laboratory of Ultrasound in Medicine and Engineering, College of Biomedical Engineering, Chongqing Medical University, Chongqing 400016, China; Chongqing Key Laboratory of Biomedical Engineering, Chongqing 400016, China
  • Online:2023-10-20 Published:2023-11-02

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摘要： 目的 探讨低强度脉冲超声（LIPUS）对脓毒症SD大鼠脾脏淋巴细胞凋亡的抑制效应及可能机制。方法 通过盲肠结扎穿刺术建立脓毒症SD大鼠模型，将78只雌性SD大鼠随机分为超声组（LIPUS，n=26）、盲肠结扎穿刺组（CLP，n=26）和假手术组（Sham，n=26）。超声参数如下：声强200 mW/cm2，辐照时间20 min，频率0.37 MHz。观察各组大鼠术后72 h内的存活情况；苏木精-伊红染色法观察脾脏淋巴细胞的数量变化；脱氧核糖核苷酸末端转移酶介导的缺口末端标记法和流式细胞术检测脾脏淋巴细胞凋亡情况；免疫组化、蛋白质印迹法和实时荧光定量PCR检测线粒体凋亡通路B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）和半胱氨酸蛋白酶3（Caspase 3）的蛋白及mRNA的表达水平。结果 Sham组72 h内全部存活，LIPUS组的存活率显著高于CLP组（P<0.05）；与CLP组相比，LIPUS组脾脏淋巴细胞凋亡率显著降低（P<0.05），Bcl-2的蛋白及mRNA表达水平显著升高（P<0.05），Bax及Caspase 3的蛋白及mRNA表达水平显著降低（P<0.05）。 结论 LIPUS可能通过调节线粒体通路上的关键分子（Bcl-2、Bax及Caspase 3）抑制脓毒症SD大鼠脾脏淋巴细胞凋亡，提高其72 h内存活率、延长存活时间。

关键词: 低强度脉冲超声；脓毒症；凋亡；线粒体通路

Abstract: Objective To investigate the inhibitory effect of low- intensity pulsed ultrasound (LIPUS) on apoptosis of splenic lymphocytes in rats with sepsis and explore its possible mechanism. Methods Seventy-eight female SD rats were randomly divided into LIPUS group, cecal ligation and puncture (CLP) group and sham-operated group (Sham) (n=26), and in the former two groups, rat model of sepsis were established by CLP. Immediately after the operation, the rats in LIPUS group received pulsed ultrasound therapy with an ultrasound intensity of 200 mW/cm2, irradiation time of 20 min, and frequency of 0.37 MHz. The survival of the rats in each group was observed within 72 h after CLP. The changes in splenic lymphocyte counts were observed using HE staining, and apoptosis of the splenic lymphocytes was detected using TUNEL assay and flow cytometry. The expression levels of Bcl-2, Bcl2-associated X protein (Bax) and caspase-3 were detected by immunohistochemistry, Western blotting and RT-qPCR. Results All the rats in the sham-operated group survived for over 72 h. The survival rates of the rats was significantly higher in LIPUS group than in CLP group (P<0.05). Compared with those in CLP group, the apoptosis rate of the splenic lymphocytes in LIPUS group was significantly decreased (P<0.05), the protein and mRNA expression levels of Bcl-2 were increased (P<0.05), and the protein and mRNA expression levels of Bax and caspase-3 were decreased (P<0.05). Conclusions LIPUS inhibits apoptosis of splenic lymphocytes in septic SD rats possibly by regulating the key molecules in the mitochondrial pathway, thereby improving the survival rate and prolonging the survival time of the rats.

Key words: low-intensity pulsed ultrasound; sepsis; apoptosis; mitochondrial pathway