The findings of this study demonstrated that AuNPs inhibited the expression of pro-inflammation factors and AD-related protein (T-Tau, P-Tau, and Aβ), moreover, AuNPs could ameliorate the inhibition of neural stem cell differentiation due to Aβ accumulation. Then, AuNPs increased mitochondrial membrane potential, activated expression of Drp1, NRF1, and TFAM, and decreased ROS production in hNSCs. Thirdly, apoptosis of hNSCs induced by Aβ was inhibited with AuNPs co-treatment, and expression of proteins associated with apoptosis was also reduced in AuNPs co-treatment group. Fourthly, The miR-21-5p was unregulated in AuNPs treated cells, while the effect of AuNPs treatment on hNSCs was reversed after transfection with miR-21-5p inhibitor. Finally, inhibited miR-21-5p expression in hNSCs also reduced mitochondrial function induced by Aβ accumulation.
AD is the most common form of dementia and causes problems with memory, thinking, and behavior. It is generally accepted that Aβ is the main toxic form, and there are multiple strategies for drug development against this hypothesis, including BACE1 inhibitors that inhibit Aβ production, antibodies that promote Aβ clearance, and then none of them have been successfully marketed. The reason for this may be that the therapeutic modality is intervened too late, Aβ appears much earlier than the occurrence of clinical symptoms of AD, and toxic forms of Aβ have caused irreversible neurological damage when cognitive impairment appears (Wang et al. 2021). Therefore, the development of Aβ-labeled small molecules is important for the early diagnosis and treatment of AD.
The AuNPs preparation process is simple and controllable, allows a variety of functionalized modifications to the surface, and has the characteristics of good biocompatibility, high specific surface area, and low toxicity (Gao et al. 2015). AuNPs, as a new method, may have some advantages in eliminating Aβ-induced cytotoxicity in neurodegeneration studies. Kim et al. (Kim et al. 2017) suggested that AuNPs could act as an effective gene regulator and decrease the aggregation of Aβ. In addition, AuNPs have a protective effect on Aβ1-42-induced apoptosis, and the process is related to NF-KB-induced neuroinflammation (Ali et al. 2016). Muller et al. (Muller et al. 2017) also found that AuNPs have an important role in improving mitochondrial function, neuroinflammation as well as oxidative stress in diabetic rats. The results of this study revealed that AuNPs could ameliorate the cytotoxicity of hNSCs induced by Aβ enrichment, including decreasing apoptosis, promoting cell proliferation and differentiation, increasing mitochondria-related gene expression, and decreasing ROS production. This was consistent with previously reported results and AuNPs can improve the cytotoxicity induced by Aβ accumulation.
AuNPs are promising agents and materials for therapeutic applications such as drug delivery. In recent years, more and more attention has been paid to the role of miRNAs in the disease's progression. There have been several studies on AuNPs in AD drug therapy, but the mechanism of action of AuNPs on miRNA regulation during treatment is not yet clear. AuNPs nanomaterials, designed by Song et al. (Song et al. 2022), can accurately diagnose miRNA-125b and miR-361 in exosomes of AD patients and have great potential for AD clinical diagnosis as well as future dementia research and treatment fields. Gao et al. (Gao et al. 2021) found DNAzyme machines combined with AuNPs to detect specific miRNAs in living cells and this technology can accurately distinguish diseased cells from healthy cells and indicate a promising tool for cancer diagnosis and prognosis. Ven et al. (van der Ven et al. 2021) coupled AuNP and miRNAs mimics and improved drug delivery efficiency. Ng et al. (Ng et al. 2016) used AuNPs intravenously to inject AD rats and found that miR-327 expression in rat cells showed differential regulation, and the process may be related to pulmonary inflammation. The present results showed that miR-21-5p was unregulated in AuNPs treated cells, while the effect of AuNPs treatment on hNSCs was reversed after transfection with miR-21-5p inhibitor. Differential miRNA expression may be the result of AuNPs playing a role in cells and in promoting cells against Aβ.
In addition to Aβ and tau protein being involved in the initiation and progression of AD, impaired mitochondrial bioenergetics and dynamics are likely major etiological factors in AD pathogenesis (Onyango et al. 2021). Several reports had demonstrated the neuroprotection effects of AuNPs in different aspects. Chiang et al. (Chiang et al. 2020) had proved that AuNPs protect mitochondrial function and morphology from Aβ in hNCSs. This is consistent with our findings that in this study, we found that AuNPs protect mitochondrial function, consistent with up-regulation of miR-21-5p action, therefore, AuNPs in inhibiting Aβ aggregation and reducing Aβ cytotoxicity, may be related by regulating miR-21-5p.
This study has shortcomings, only hNSCs cells are used for analytical studies, and subsequent studies will be conducted using animal tests to further improve the rigor. Besides, the research process regulated miR-21-5p expression by AuNPs and improved Aβ-induced hNSCs toxicity, especially on mitochondrial function regulation, while the miR-21-5p downstream targeted genes have not been deeply explored, which will be analyzed in subsequent studies. Our study is the first to show AuNPs can ameliorate Aβ-induced cytotoxicity in hNSCs, and the process is associated with miR-21-5p up-regulation and further acts through mitochondrial function regulation.