Department of Neurology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou 310000, Zhejiang, China.
College of Pharmacy, Hangzhou Medical College, Hangzhou 310000, Zhejiang, China.
J Chem Neuroanat. 2023 Nov;133:102338. doi: 10.1016/j.jchemneu.2023.102338. Epub 2023 Sep 12.
Mesenchymal stem cells (MSCs) and Salvianolic acid B (SAB) are known to exert potent anti-inflammatory and anti-oxidative properties. But the effect of SAB and MSCs combination treatment on the cerebral ischemia/reperfusion injury (CI/RI) is not clear.
After the CI/RI animal model established, rats were administered with MSCs and SAB individually or combination treatment. To evaluate the therapeutic potential, behavioral tests, TTC staining, Hematoxylin-eosin (HE) staining, and immunofluorescence assays were performed to evaluate the neuroprotection and endogenous neurogenesis. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and enzyme linked immunosorbent assay (ELISA) were performed to evaluate the anti-apoptosis and anti-inflammatory effect. Meanwhile, the expression of the TLR4/NF-ĸB/MYD88 signal pathway-related proteins was evaluated by Western blot.
MSCs and SAB individually or combination treatment have protective effect in CI/RI rats. More importantly, the rats with the combination treatment showed a better behavioral recovery, neurogenesis and smaller infarct size compared with the rats administered with MSCs or SAB individually. Further research showed that the combination treatment decreased CI/RI induced inflammatory cytokines and oxidative stress, including inhibiting the production of IL-1β, IL-6, TNF-α, decreasing the levels of malondialdehyde (MDA), and increased the activity of superoxide dismutase (SOD). In addition, the neuroprotection effect of SAB and MSCs combination was achieved through the regulation of TLR4/NF-κB/MyD88 signaling pathway related proteins, including inhibition the protein levels of TLR4, MYD88, p-NF-κB p65, TRAF6-and action of SIRT1 in brain tissues.
The present study indicated that the MSCs and SAB combination treatment had better protective effect against rat ischemic brain injury. The combination of SAB and MSCs may provide a potent and promising strategy for the treatment of ischemic stroke and is worthy for further development.
间充质干细胞（MSCs）和丹酚酸 B（SAB）已被证实具有强大的抗炎和抗氧化作用。但是，SAB 和 MSCs 联合治疗对脑缺血/再灌注损伤（CI/RI）的影响尚不清楚。
建立 CI/RI 动物模型后，单独或联合给予大鼠 MSCs 和 SAB 治疗。通过行为学测试、TTC 染色、苏木精-伊红（HE）染色和免疫荧光分析来评估神经保护和内源性神经发生；通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）染色和酶联免疫吸附试验（ELISA）评估抗凋亡和抗炎作用；同时通过 Western blot 评估 TLR4/NF-κB/MYD88 信号通路相关蛋白的表达。
MSCs 和 SAB 单独或联合治疗对 CI/RI 大鼠均具有保护作用。更重要的是，与单独给予 MSCs 或 SAB 的大鼠相比，联合治疗的大鼠表现出更好的行为学恢复、神经发生和更小的梗死面积。进一步的研究表明，联合治疗可降低 CI/RI 诱导的炎症细胞因子和氧化应激，包括抑制白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）的产生，降低丙二醛（MDA）水平，增加超氧化物歧化酶（SOD）活性。此外，SAB 和 MSCs 联合的神经保护作用是通过调节 TLR4/NF-κB/MyD88 信号通路相关蛋白实现的，包括抑制 TLR4、MYD88、p-NF-κB p65、TRAF6 的蛋白水平以及 SIRT1 在脑组织中的作用。
本研究表明，MSCs 和 SAB 联合治疗对大鼠缺血性脑损伤具有更好的保护作用。SAB 和 MSCs 的联合应用可能为缺血性脑卒中的治疗提供一种强大而有前景的策略，值得进一步开发。
