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[低强度脉冲超声对脓毒症大鼠脾脏淋巴细胞凋亡的抑制作用]

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

作者信息

Tang W, Deng J, He S, Li J, Zhou Y, Wang Y

机构信息

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.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2023 Oct 20;43(10):1789-1795. doi: 10.12122/j.issn.1673-4254.2023.10.18.

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) (=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/cm, 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 (<0.05). Compared with those in CLP group, the apoptosis rate of the splenic lymphocytes in LIPUS group was significantly decreased (<0.05), the protein and mRNA expression levels of Bcl-2 were increased (<0.05), and the protein and mRNA expression levels of Bax and caspase-3 were decreased (<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.

摘要

目的

探讨低强度脉冲超声(LIPUS)对脓毒症大鼠脾脏淋巴细胞凋亡的抑制作用,并探讨其可能机制。

方法

78只雌性SD大鼠随机分为LIPUS组、盲肠结扎穿孔(CLP)组和假手术组(Sham)(每组 = 26只),前两组采用CLP法建立脓毒症大鼠模型。术后立即对LIPUS组大鼠进行脉冲超声治疗,超声强度为200 mW/cm²,照射时间为20 min,频率为0.37 MHz。观察CLP术后72 h内各组大鼠的存活情况。采用HE染色观察脾脏淋巴细胞计数变化,采用TUNEL法和流式细胞术检测脾脏淋巴细胞凋亡情况。采用免疫组织化学、蛋白质印迹法和RT-qPCR检测Bcl-2、Bcl-2相关X蛋白(Bax)和半胱天冬酶-3(caspase-3)的表达水平。

结果

假手术组大鼠全部存活超过72 h。LIPUS组大鼠存活率显著高于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蛋白和mRNA表达水平升高(P<0.05),Bax和caspase-3蛋白及mRNA表达水平降低(P<0.05)。

结论

LIPUS可能通过调节线粒体途径中的关键分子抑制脓毒症SD大鼠脾脏淋巴细胞凋亡,从而提高大鼠存活率,延长其生存时间。

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The immunology of sepsis.脓毒症的免疫学。
Immunity. 2021 Nov 9;54(11):2450-2464. doi: 10.1016/j.immuni.2021.10.012.

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