Department of Emergency, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China; Department of Intensive Care Unit, JingJiang Chinese Medicine Hospital, Jingjiang, 214500, China.
Department of Intensive Care Unit, Jingjiang People's Hospital, Jingjiang, 214500, China.
J Ethnopharmacol. 2020 Oct 28;261:113068. doi: 10.1016/j.jep.2020.113068. Epub 2020 Jun 24.
Shenfu injection (SFI) is a well-known Chinese herbal medicine widely used in the treatment of septic shock in China.
The aims of this study are to investigate the protective effects of SFI on sepsis-induced myocardial injury in mice and to identify the underlying mechanism of action.
Seventy-two male C57/B6J mice (5-6 weeks old) were randomly divided into five groups: control (NC), sham sepsis (sham), sepsis (Lipopolysaccharide- LPS), sepsis treated with a low dose SFI, and sepsis treated with a high dose SFI. Sepsis was induced in mice by intraperitoneal injection of LPS. Myocardial tissue samples were collected from different groups at 6 h, 12 h, and 24 h post-LPS injection. Myocardial injury was examined using hematoxylin-eosin (H&E) and TUNEL staining. Western-blot analysis was performed to determine the protein expression of B-cell lymphoma 2 (Bcl-2), BH3 interacting-domain death agonist (Bid), truncated-Bid (t-Bid) and caspase-9 in all the groups. Moreover, the structural changes in the mitochondria of cardiomyocytes were also observed by transmission electron microscopy.
H&E staining revealed structural damage, local necrosis, interstitial edema, inflammatory cell infiltration and vacuolar changes in the myocardial tissue in the sepsis (LPS) group; almost intact myocardial tissue was observed in the high dose SFI group with improvements in interstitial edema and inflammatory cell infiltration. We observed that LPS-induced cardiomyocyte apoptosis was significantly improved with high dose SFI as compared with sepsis (LPS) group (P ˂ 0.05). LPS was found to decrease the protein expression of Bcl-2 and increase the level of Bid, t-Bid and caspase-9. Treatment with SFI significantly increased the Bcl-2 protein expression (P ˂ 0.05) and decreased the protein expression of Bid, t-Bid and caspase-9 as compared with LPS group (P ˂ 0.05). Markedly swollen myocardial mitochondria with partial vacuolation were observed in LPS treated mice while SFI treatment was found to significantly improve the LPS-induced morphological damage of the mitochondria.
In conclusion, we demonstrate that SFI protects against sepsis-induced myocardial injury in mice through the suppression of myocardial apoptosis. It upregulates the protein expression of Bcl-2 and downregulates the protein expression of Bid, t-Bid and caspase-9, and alleviates sepsis-induced mitochondrial damage.
参附注射液（SFI）是一种在中国广泛用于治疗脓毒性休克的著名中药。
本研究旨在探讨 SFI 对脓毒症诱导的小鼠心肌损伤的保护作用，并确定其作用机制。
72 只雄性 C57/B6J 小鼠（5-6 周龄）随机分为五组：对照组（NC）、假脓毒症（假）、脓毒症（脂多糖- LPS）、低剂量 SFI 治疗的脓毒症和高剂量 SFI 治疗的脓毒症。通过腹腔注射 LPS 诱导小鼠脓毒症。在 LPS 注射后 6 h、12 h 和 24 h 从不同组收集心肌组织样本。使用苏木精-伊红（H&E）和 TUNEL 染色检查心肌损伤。通过 Western blot 分析确定所有组中 B 细胞淋巴瘤 2（Bcl-2）、BH3 相互作用域死亡激动剂（Bid）、截断 Bid（t-Bid）和半胱天冬酶-9 的蛋白表达。此外，还通过透射电子显微镜观察心肌细胞中线粒体的结构变化。
H&E 染色显示，在脓毒症（LPS）组的心肌组织中观察到结构损伤、局部坏死、间质水肿、炎症细胞浸润和空泡变化；高剂量 SFI 组几乎完整的心肌组织，间质水肿和炎症细胞浸润得到改善。我们观察到与脓毒症（LPS）组相比，高剂量 SFI 显著改善 LPS 诱导的心肌细胞凋亡（P ˂ 0.05）。LPS 降低 Bcl-2 蛋白表达，增加 Bid、t-Bid 和半胱天冬酶-9 水平。与 LPS 组相比，SFI 治疗显著增加 Bcl-2 蛋白表达（P ˂ 0.05），并降低 Bid、t-Bid 和半胱天冬酶-9 蛋白表达（P ˂ 0.05）。在 LPS 处理的小鼠中观察到明显肿胀的心肌线粒体伴部分空泡化，而 SFI 治疗可显著改善 LPS 诱导的线粒体形态损伤。
总之，我们证明 SFI 通过抑制心肌细胞凋亡来保护小鼠免受脓毒症引起的心肌损伤。它上调 Bcl-2 蛋白表达，下调 Bid、t-Bid 和半胱天冬酶-9 蛋白表达，并减轻脓毒症引起的线粒体损伤。
