J. Mater. Sci. Technol. ›› 2020, Vol. 43: 197-207.DOI: 10.1016/j.jmst.2019.10.013
• Research Article • Previous Articles Next Articles
Yongyong Xuea, Na Wanga, Zhi Zengbc, Jinpeng Huangbc, Zhiming Xiangcd, Yan-Qing Guanabc*()
Received:
2019-08-01
Revised:
2019-09-29
Accepted:
2019-10-09
Published:
2020-04-15
Online:
2020-04-26
Contact:
Guan Yan-Qing
Yongyong Xue, Na Wang, Zhi Zeng, Jinpeng Huang, Zhiming Xiang, Yan-Qing Guan. Neuroprotective effect of chitosan nanoparticle gene delivery system grafted with acteoside (ACT) in Parkinson’s disease models[J]. J. Mater. Sci. Technol., 2020, 43: 197-207.
Fig. 2. (A) FITC spectra of mPEG-PLA, ACT-PLA-mPEG, ACT and ACT-PLA-mPEG-CTS-pDNA; (B) UV-vis spect of mPEG-PLA, ACT and ACT-PLA-mPEG; (C) The Grafting rate and Loading capacities of pDNA, NGF and ACT.
Fig. 3. (A) Size distribution histograms and TEM image of mPEG-PLA, ACT-PLA-mPEG, ACT-PLA-mPEG-CTS-pDNA and ACT-PLA-mPEG-CTS-pDNA-NGF; (B) AFM observation of ACT-PLA-mPEG-CTS-pDNA and ACT-PLA-mPEG-CTS-pDNA-NGF; (C) Zeta potential of mPEG-PLA, ACT-PLA-mPEG, ACT-PLA-mPEG-CTS-pDNA and ACT-PLA-mPEG-CTS-pDNA-NGF; (D) Variation of ACT-PLA-mPEG-CTS-pDNA-NGF particle size with time gradient in cell culture medium.
Fig. 4. (A) Viabilities of PC12 cells 24 h after MPP+ treatment in the absence or presence of pre-incubated nano-drug with different groups were measured with MTT assays; (B) Bright field and confocal fluorescence images of PC12 cells treated with different groups. The nuclei of cells werestained with DAPI (blue); (C) Apoptosis rate of different groups were assessed by flow cytometry methods; (D) The PC12 cells apoptosis quantified by the flow cytometry at 24 h after various treated with different groups (PBS + PBS; MPP++PBS; MPP++ mPEG-PLA; MPP++mPEG-PLA-ACT; MPP++mPEG-PLA-ACT-CTS-pDNA; MPP++mPEG-PLA-ACT-pDNA-NGF) (Q1: dead cells; Q2: late apoptotic cells; Q3: live cells; Q4: early apoptotic cells)).
Fig. 5. (A, B) Confocal images of PC12 cells at 24 h after various treatments. Immunofluorescence staining was used to detect the nucleus (blue fluorescence) of DAPI, alpha synuclein (SNCA) and nerve growth factor receptor (NGFR) (green fluorescence) staining; (C, D) Expression of NGFR and SNCA after various treatment with different groups. The relative levels are plotted at a significance of p<0.05 indicated by *, 0.001<p<0.01 indicated by **, and p<0.001 indicated by ***, in comparison to the MPP++PBS group.
Fig. 6. (A) Schematic representation of the open field system; (B) The photos of mice after various treatments (Saline + Saline; MPTP + Saline; MPTP + APPDN); (C) The body weight changes of mice during various treatments; (D-H) The behavior detection in mice after various treatments; (D, G) Open field test; (E) Pole-jump test; (H) Gait analysis Morris water maze (n = 5 biologically independent samples; two-way ANOVA with Tukey HSD test). The relative levels are plotted at a significance of p<0.05 indicated by *, 0.001<p<0.01 indicated by **, and p<0.001 indicated by ***, in comparison to the MPTP + saline group.
Fig. 7. (A1, A2) Representative images and head semi-quantitative analysis of APPDN/CY5 fluorescence imaging; (B) A representative picture of the retention of fluorescent labeled drugs in the substantia nigra at different times after intraperitoneal injection of APPDN/CY5 in mice. The nucleus is marked blue with DAPI and the APPDN is marked pink with CY5; (C, D) Immunofluorescence and alpha-syn immunohistochemical images of TH in substantia nigra of three groups (Saline + Saline; MPTP + Saline; MPTP + APPDN). The red circle indicates abnormal accumulation of alpha -syn cells, TH uses green fluorescent labeling; (E1, E2) Expression levels of TH and α-syn in the substantia nigra pars compacta of C57bl/6 mice and the quantitative results of ratio; (F) The images of H&E stained brain, lung, liver, spleen, kidney and heart after various treatments; (G-I) Serological and toxicology test of C57 mice, including the density of platelet, the density of white blood cell, and the density of red blood cell in different groups (Saline + Saline; MPTP + Saline; MPTP + APPDN). The relative levels are plotted at a significance of p<0.05 indicated by *, 0.001<p<0.01 indicated by **, and p<0.001 indicated by ***, in comparison to the MPTP + saline group.
Fig. 8. Schematic illustration of the APPDN treatment process. The APPDNs nano- micelle complex is internalized by PC12 cells, and PDNA degrades mRNA through a series of actions, thus inhibiting SNCA expression. ACT inhibits cell senescence by increasing the activity of telomerase and reducing the production of malondialdehyde in the cytoplasm.
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