Abstract
We investigated the effects of Shenfu Injection (SFI) on HCN4 activity in bone marrow mesenchymal stem cells (BMSCs). The sample of BMSCs was divided into six groups: a control group, a high-dose SFI group (0.25 ml/ml), a middle-dose SFI group (0.1 ml/ml), a low-dose SFI group (0.05 ml/ml), an adenovirus-encoded control vector group, and an adenovirus-encoded HCN4 group. Cell ultrastructure was observed using a transmission electron microscope. Quantitative reverse transcription PCR (RT-qPCR) was performed to detect HCN4 expression, and HCN4 activity was detected using the whole-cell patch clamp technique. An enzyme-linked immunosorbent assay was performed to detect cAMP content. Application of flow cytometry confirmed that the isolated cells showed BMSC-like phenotypes. Differentiation of BMSCs in both the SFI and the adenovirus-encoding HCN4 groups occurred according to the cellular ultrastructure. Application of the whole-cell patch clamp technique revealed that SFI could activate the inward pacing current of BMSCs in a concentration-dependent manner. The RT-qPCR results showed that HCN4 expression was significantly higher in the high-dose SFI group than in the medium- and low-dose groups, whereas the cAMP content in the overexpressed HCN4 group decreased significantly; this content in the high-dose SFI group increased significantly. In conclusion, SFI promotes HCN4 activity in BMSCs, which could explain its treatment effect when administered to patients with cardiovascular diseases.
Highlights
Sick sinus syndrome (SSS) refers to a group of heart rhythm disorders caused by problems relating to the sinus node
We investigated the effects of Shenfu Injection (SFI) on HCN4 activity in bone marrow mesenchymal stem cells (BMSCs). e findings of this study have important implications for the use of SFI in the treatment of heart diseases
To assess the effect of SMI on HCN4 activity, we transfected the BMSCs with AAV-encoding HCN4 gene treated as a positive control
Summary
Sick sinus syndrome (SSS) refers to a group of heart rhythm disorders caused by problems relating to the sinus node. E HCN4 gene is the molecular basis of pacemaker current, which, through depolarization, provides a large proportion of inward current and plays an important role in the generation and autonomous regulation of the heart rate [8,9,10]. This gene is linked to the diastolic depolarization process of the sinoatrial node [10, 11] and is thought to be a determining channel for cardiocytes [12]. We investigated the effects of SFI on HCN4 activity in BMSCs. e findings of this study have important implications for the use of SFI in the treatment of heart diseases
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