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| 清胰汤靶向Bcl-2/Bax/Caspase-3通路介导线粒体凋亡减轻重症急性胰腺炎相关心肌损伤的实验研究 |
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李霄1, 李彩霞2, 崔立华2, 乔媛媛1, 李静1, 潘建红1
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1.天津市中西医结合医院(天津市南开医院)超声科, 天津 300100;2.天津市中西医结合医院(天津市南开医院), 天津市急腹症器官损伤与中西医修复重点实验室, 天津 300100
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| 摘要: |
| [目的] 探究细胞凋亡在重症急性胰腺炎（SAP）心肌损伤中的作用及清胰汤干预SAP心肌损伤大鼠的作用机制。[方法] 将24只雄性SD大鼠随机分为4组，分别为假手术组、模型组与清胰汤低、高剂量组（QYD-L、QYDH），每组6只。留取相应组织、血液标本进行相应检测。采用HE染色法评估胰腺与心肌组织的病理学改变；通过生化试剂盒测定血清中淀粉酶（AMY）与脂肪酶（LIP）的活性。使用全自动生化分析仪对血清中乳酸脱氢酶（LDH）、肌酸激酶（CK）及其同工酶（CK-MB）的水平进行检测。采用ELISA法定量分析血清中炎症因子包括白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）及白细胞介素-1β（IL-1β）的浓度；TUNEL检测心脏细胞凋亡情况；透射电镜观察心肌组织线粒体超微结构；通过RT-PCR技术分析心肌组织中B淋巴细胞瘤-2（Bcl-2）与Bcl-2相关X蛋白（Bax）的mRNA转录水平；利用Western blot法测定Bax、Bcl-2及剪切型半胱氨酸-天冬氨酸蛋白酶-3（Cleaved Caspase-3）在大鼠心肌组织中的表达情况。[结果] 相较于假手术组，模型组大鼠的胰腺与心肌组织出现显著病理损伤，同时血清中AMY、LIP以及心肌损伤标志物CK-MB、CK和LDH和炎性因子IL-6、TNF-α、IL-1β的含量均明显上升（P<0.05）；模型组大鼠表现出严重的线粒体病理损伤与心肌细胞凋亡，而分子水平上，促凋亡因子Bax的mRNA及蛋白表达均上调，同时其关键执行蛋白Cleaved Caspase-3的表达也显著增加；与之相反，抗凋亡蛋白Bcl-2的转录与表达水平则显著下调，上述差异均具有统计学意义（P<0.05）。与模型组比较，清胰汤干预组（QYD-L、QYD-H）大鼠心肌组织损伤程度得到改善，血清AMY、LIP活性显著降低，CK-MB、CK、LDH水平降低，炎性因子IL-6、TNF-α、IL-1β水平显著降低；线粒体病理损伤得到明显改善，细胞凋亡水平受到显著抑制。在分子机制上，促凋亡因子Bax的mRNA及蛋白表达均下降，其下游的Cleaved Caspase-3蛋白表达也随之减少；抗凋亡因子Bcl-2的基因转录与蛋白表达水平则显著回升（P<0.05）。[结论] 清胰汤能够改善SAP诱导的心肌损伤，其机制可能与调控Bcl-2/Bax/Caspase-3信号通路，抑制细胞凋亡有关。 |
| 关键词: SAP心肌损伤 清胰汤 细胞凋亡 Caspase-3 Bcl-2/Bax |
| DOI：10.11656/j.issn.1672-1519.2026.09.12 |
| 分类号:R285.5 |
| 基金项目:天津市卫生健康委员会天津市中医药重点领域科研项目（2024018）。 |
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| Experimental study on Qingyi Decoction targeting the Bcl-2/Bax/Caspase-3 pathway to mediate mitochondrial apoptosis and alleviate severe acute pancreatitis-associated myocardial injury |
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LI Xiao1, LI Caixia2, CUI Lihua2, QIAO Yuanyuan1, LI Jing1, PAN Jianhong1
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1.Department of Ultrasound, Tianjin Hospital of ITCWM Nankai Hospital(Tianjin Nankai Hospital), Tianjin 300100, China;2.Tianjin Hospital of ITCWM Nankai Hospital(Tianjin Nankai Hospital), Tianjin Key Laboratory of Acute Abdomen Disease Associated Organ Injury and ITCWM Repair, Tianjin 300100, China
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| Abstract: |
| [Objective] This study aims to examine the role of apoptosis in myocardial injury caused by severe acute pancreatitis（SAP） and to elucidate the mechanism by which Qingyi Decoction（QYD）intervenes in SAP-induced myocardial injury in rats. [Methods] Twenty-four male SD rats were randomly assigned to four groups：a sham-operated group，a model group，a low-dose QYD group（QYD- L），and a high-dose QYD group（QYD-H），with six rats in each group. Tissue and blood samples were collected for analysis. Pathological alterations in pancreatic and myocardial tissues were identified through HE staining. Biochemical measurements were conducted to assess serum amylase and lipase activities. The activities of Lactate Dehydrogenase（LDH），Creatine Kinase（CK），and Creatine Kinase-MB（CK-MB）were measured utilizing an automatic biochemical analyzer. Serum concentrations of the inflammatory cytokines IL-6，TNF-α，and IL-1β were quantified using enzyme-linked immunosorbent assay（ELISA）. The assessment of cardiac cell apoptosis was conducted utilizing the TUNEL assay. The ultrastructure of myocardial mitochondria was examined using transmission electron microscopy. RT-PCR was utilized to detect the mRNA expression levels of B-cell lymphoma-2（Bcl-2）and Bcl-2-associated X （Bax）protein. [Results] In comparison to the sham-operated group，the model group demonstrated significant damage to pancreatic and myocardial tissues，elevated serum levels of amylase，lipase，and myocardial injury markers（CK-MB，CK，LDH，P<0.05）， increased levels of inflammatory cytokines（IL-6，TNF-α，IL-1β，P<0.05），pronounced mitochondrial pathological damage，heightened myocardial apoptosis，upregulated mRNA and protein expression of Bax，and significantly increased protein expression of cleaved Caspase-3. The expression levels of Bcl-2 mRNA and protein were significantly decreased（P<0.05）. In comparison to the model group， the QYD intervention groups exhibited enhanced myocardial tissue integrity，a significant reduction in serum amylase and lipase activities，decreased levels of CK-MB，CK，and LDH，as well as markedly lower concentrations of inflammatory cytokines（IL-6，TNF-α， IL-1β）. Mitochondrial pathological damage was mitigated，myocardial apoptosis was markedly reduced，and there was a downregulation in the mRNA and protein expression of Bax，alongside a significant decrease in the protein expression of cleaved Caspase-3. Simultaneously，there was a significant increase in the mRNA and protein expression of Bcl-2（P<0.05）. [Conclusion] QYD can improve myocardial injury induced by SAP，and its mechanism may be related to regulating Bcl-2/Bax/Caspase-3 signaling pathway and inhibiting apoptosis. |
| Key words: SAP-induced myocardial injury Qingyi Decoction apoptosis Caspase-3 Bcl-2/Bax |
