Oxidative Medicine and Cellular Longevity
·
2021
·
IF 6.543
Xiao-lu Wang,Liang Wang,Fo-lan Lin,Si-si Li,Ting-xuan Lin,Ren-wang Jiang
Copper/zinc superoxide dismutase (SOD1) can clear cisplatin- (CP-) induced excessive reactive oxygen species (ROS), but exogenous SOD1 cannot enter cells because of its low biomembrane permeability. Cell-penetrating peptides (CPPs) can rapidly cross plasma membranes. This study is aimed at identifying an efficient and stable CPP-SOD1 and investigating its effects on CP-induced nephrotoxicity. We recombined SOD1 with 14 different CPPs and purified them using an NTA-Ni2+ column. In in vitro experiments, CPPs-SOD1 cell membrane penetration ability and JNK/p38 MAPK signaling pathway were evaluated using Western blotting. ROS production, mitochondrial membrane potential (MMP), and cell apoptosis were determined using flow cytometry and immunofluorescence staining in VERO and HK-2 cells. For in vivo experiments, mice were administered PSF-SOD1 for 2 h before cotreatment with a single CP injection for an additional 4 days. Blood and kidney samples were collected for renal function assessment (creatinine, urea nitrogen, histopathology, TUNEL assay, and JNK/p38 MAPK signaling pathway). Compared with TAT-SOD1, we found that PSF-SOD1 is more efficient at crossing the cell membrane and is stable after transduction into cells. Pretreatment with PSF-SOD1 inhibited CP-induced apoptosis, ROS generation, and JNK/p38 MAPK activation and restored CP-induced MMP loss in VERO and HK-2 kidney cells. Treatment of mice with PSF-SOD1 inhibited CP-induced serum creatinine, blood urea nitrogen elevation, and JNK/p38 MAPK activation. H&E staining and TUNEL assay indicated that kidney tissue damage was alleviated following PSF-SOD1 pretreatment. Overall, PSF-SOD1 ameliorated CP-induced renal damage by partially reducing oxidative stress and cell apoptosis by regulating JNK/p38 MAPK signaling pathway and might be a better cytoprotective agent than TAT-SOD1.
研究领域:
药理学
·
肾病学
·
分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2025
·
IF 3.8
Zhang Jin,Su Ning,Liu Wei,Li Mengran,Zheng Haoyang,Li Bing,Jin Xue,Gao Mingxia,Zhang Xiangmin
Extracellular vesicles (EVs) have been demonstrated to own the advantages in evading phagocytosis, crossing biological barriers, and possessing excellent biocompatibility and intrinsic stability. Based on these characteristics, EVs have been used as effective therapeutic carriers for drug delivery, but the low drug loading capacity greatly limits further applications. Herein, we developed a drug loading method based on cell-penetrating peptide (CPP) to enhance the encapsulation of therapeutic reagents in EVs, and EVs-based drug delivery system achieved higher killing efficacy to tumor cells. Urinary EVs and chemotherapy reagent doxorubicin (DOX) were used as model. It is easy to conjugate CPP with DOX (CPP-DOX) through the linker N-succinimidyl 3-maleimidopropionate (SMP). CPP-DOX was incubated with EVs under a mild condition, promoting the encapsulation of DOX into EV cavities. CPP-DOX-EVs showed strong anticancer ability since EVs delivery facilitated the uptake by cancer cells. EVs loading of CPP-DOX exhibited higher drug loading efficiency at 37.18%, presenting about 2.5 times increase in efficiency over EVs loading of DOX through passive incubation. Easy operation and controllable condition further reinforce the advantages compared with other loading methods. CPP-based drug loading method provides an effective strategy for EVs-based drug delivery system. Graphical
研究领域:
药理学
·
生物医学工程
·
纳米医学
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产品关联
DOI
PubMed
高影响因子
ACS Applied Nano Materials
·
2024
·
IF 5.3
Peng Xu,Simei Zhong,Yan Ma,Xinxin Duan,Yunfei Wei,Meng Zhang,Jing Xu,Yu-Hui Zhang
Vesicle entrapment remains a major bottleneck for the application of fluorescent nanoprobes (FNPs) with excellent optical properties in the field of super-resolution imaging of subcellular structures in living cells. Here, we developed a cell-penetrating peptide, fR8, which enables efficient cytoplasmic delivery of 3.5 nm FNPs instead of being trapped in vesicles, with a delivery efficiency of up to 75.6%. Further, we have successfully combined fusion protein technology with FNPs-fR8 to achieve high specificity and stable labeling of FNPs to various subcellular structures within live cells. Utilizing the advantages of FNPs, such as high brightness and resistance to photobleaching, FNPs modified with fR8 and the HaloTag ligand can be used for long-term super-resolution imaging of subcellular structures in living cells, allowing us to study the dynamic interactions between them. This work provides a practical tool for applying FNPs to live-cell super-resolution imaging and expands the application of super-resolution imaging technology in live-cell studies.
研究领域:
生物化学
·
分子生物学
·
细胞生物学
✓ 已验证
产品关联
DOI
高影响因子
PeerJ
·
2017
·
IF 2.177
Jia Yao,Yinyun Ma,Wei Zhang,Li Li,Yun Zhang,Li Zhang,Hui Liu,Jingman Ni,Rui Wang
TH(AGYLLGHINLHHLAHL(Aib)HHIL-NH 2 ), a histidine-rich, cell-penetrating peptide with acid-activated pH response, designed and synthesized by our group, can effectively target tumor tissues with an acidic extracellular environment. Since the protonating effect of histidine plays a critical role in the acid-activated, cell-penetrating ability of TH, we designed a series of new histidine substituents by introducing electron donating groups (Ethyl, Isopropyl, Butyl) to the C-2 position of histidine. This resulted in an enhanced pH-response and improved the application of TH in tumor-targeted delivery systems. The substituents were further utilized to form the corresponding TH analogs (Ethyl-TH, Isopropyl-TH and Butyl-TH), making them easier to protonate for positive charge in acidic tumor microenvironments. The pH-dependent cellular uptake efficiencies of new TH analogs were further evaluated using flow cytometry and confocal laser scanning microscopy, demonstrating that ethyl-TH and butyl-TH had an optimal pH-response in an acidic environment. Importantly, the new TH analogs exhibited relatively lower toxicity than TH. In addition, these new TH analogs were linked to the antitumor drug camptothecin (CPT), while butyl-TH modified conjugate presented a remarkably stronger pH-dependent cytotoxicity to cancer cells than TH and the other conjugates. In short, our work opens a new avenue for the development of improved acid-activated, cell-penetrating peptides as efficient anticancer drug delivery vectors.
研究领域:
生物化学
·
肿瘤学
·
药物递送系统
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 104
·期 24
·10503-10513
·
IF 3.53
Hu Gaowei,Miao Yingjie,Luo Xi,Chu Wenhui,Fu Yongqian
Abstract Cell membranes are a great obstacle for entrance of gene therapeutic agents. Cell-penetrating peptides (CPPs) have been proven as a promising gene delivery tool. However, the early TAT peptide derived from the HIV transcription activator protein has been proven that the sequence contains Furin protease cleaved motifs which limited the TAT application in delivery of exogenous active molecules. In the present study, through the bioinformatics and experimental approach, we have identified a novel CPP derived from the N terminus of VP1 protein of chicken anemia virus (CAV), designated as CVP1-N2, which is rich in arginine residues and contains α-helical structure. Then, the ability of CVP1-N2 cell penetrating was detected using confocal imaging and flow cytometry. FITC-labeled CVP1-N2 peptide could rapidly internalize into different types of live cells with dose dependence and without cytotoxic effects by MTT assay. Surprisingly, CVP1-N2 with a pattern of nuclear sub-location has shown the higher uptake efficiency than TAT. At 10, 1, and 0.1 μM, the mean relative internalization of CVP1-N2 was respectively 1.08-, 12-, and 75-fold higher than that of CVP1, as well as 1.6-, 56-, and 75-fold higher than that of TAT. Moreover, using endocytic inhibitors along with low-temperature stress validated that the CVP1-N2 internalization route is direct translocation pathway. Finally, the capacity of CVP1-N2 for delivery of gene into cells was determined, where it was able to carry red fluorescent protein (RFP) and apoptin genes into cells respectively and induce the apoptosis. All these data indicate that CVP1-N2 could be used as a novel gene delivery vehicle for gene therapy in the future. Key points • 1CVP1-N2 was identified as a novel more efficient cell-penetrating peptide.
• 2. CVP1-N2 localized to the nucleus through the direct transduction pathway.
• 3. CVP1-N2 was able to deliver the apoptin gene into HCT116 cells and induce apoptosis.
研究领域:
分子生物学
·
基因治疗
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病毒学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2019
·
IF 4.959
Hongshuai Wu,Chaoqun You,Fanghui Chen,Jia Jiao,Zhiguo Gao,Peijing An,Baiwang Sun,Renjie Chen
Recently, the emergence of cell-penetrating peptides (CPPs) like TAT has greatly improved the efficiency of cancer therapy by enhancing cellular uptake of nanomaterials. Here, we designed a near-infrared (NIR) triggered TAT-based targeted nanoplatform ( [email protected] ), which co-delivered anticancer drug doxorubicin (DOX) and biocompatible dye indocyanine green (ICG) to realize combined chemo/photothermal/photodynamic therapy of cancer in vitro. The resulting nanoparticles showed favorable monodispersity and colloidal stability. Impressively, the DOX could be released in a promoted manner once the nanoparticles were exposed to NIR light. Confocal laser scanning microscopy (CLSM) and flow cytometry analysis demonstrated an immensely enhanced cellular accumulation of DOX after the simultaneous introduction of targeted ligand cRGD and CPP TAT. In addition, the obtained nanoparticles exhibited explosive temperature elevation and reactive oxygen species (ROS) generation mediated by encapsulated ICG under NIR irradiation, and in vitro cytotoxicity assay confirmed the [email protected] had an increasing cytotoxicity and excellent synergistic inhibition capacity. Thus, TAT-based nanosystems provide a high-efficient drug delivery strategy for optimizing combined therapy efficiency of cancer.
研究领域:
肿瘤学
·
纳米医学
·
药理学
✓ 已验证
DOI
PubMed
高影响因子
PLoS One
·
2017
·Vol. 12
·期 11
·e0186184
·
IF 2.806
Tingyu Yu,Jiao Lin,Jin Zhao,Wei Huang,Lingwen Zeng,Zhiyuan Fang,Ning Xu
The analysis of intracellular ATP can reveal the response of cells to different treatments and is important for individualized medicine. In the present study, we developed a cell penetrating peptides (CPPs) tagged luciferase (TAT-LUC) for tumor chemosensitivity assay. The activity of recombinant TAT-LUC was evaluated using ATP standard solution and tumor cells. This recombinant TAT-LUC was then used for the analysis of sensitivity index (SI) of four strains of tumor cells. The results showed that TAT-LUC could detect less than 10 nM extracellular ATP with a strong correlation between the luminescence intensity and the ATP content (R 2 = 0.994). Without cell lysis, the detection limit for intracellular ATP analysis was 40 tumor cells. Furthermore, chemosensitivity of four strains of tumor cells (Skov-3/DDP, A549/DDP, MDA-MB-231, Huh-7) was determined by this assay successfully. The cell penetration ability of TAT-LUC enables the assay not only to reflect drug resistance of tumor cells real-timely but also to minimize the test time, which can be a valuable aid for personalized cancer chemotherapy.
研究领域:
肿瘤学
·
分子生物学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Jove-Journal of Visualized Experiments
·
2022
·
IF 1.424
Dan Fang,Rui Wang,Xinyi Yu,Yuan Tian
Cancer has been a grand challenge in global health. However, the complex tumor microenvironment generally limits the access of therapeutics to deeper tumor cells, leading to tumor recurrence. To conquer the limited penetration of biological barriers, cell-penetrating peptides (CPPs) have been discovered with excellent membrane translocation ability and have emerged as useful molecular transporters for delivering various cargoes into cells. However, conventional linear CPPs generally show compromised proteolytic stability, which limits their permeability across biological barriers. Thus, the development of novel molecular transporters that can penetrate biological barriers and exhibit enhanced proteolytic stability is highly desired to promote drug delivery efficiency in biomedical applications. We have previously synthesized a panel of short cyclic CPPs with aromatic crosslinks, which exhibited superior permeability in cancer cells and tissues compared to their linear counterparts. Here, a concise protocol is described for the synthesis of the fluorescently labeled cyclic polyarginine R8 peptide and its linear counterpart, as well as key steps for investigating their cell permeability.
研究领域:
肿瘤学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY
·
2019
·
IF 2.606
Yuling Mao,Shuang Feng,Xiaojing Zhang,Qinfu Zhao,Yu Fang,Siling Wang
For orally administered nanocarriers , being efficiently absorbed by the epithelium and penetrating through the mucus that covers the epithelium are twin challengeable barriers on the way to make drugs work. However, mucus penetration and epithelial absorption require quite different surface properties that nanocarriers possess. Thereby, we designed a dual-surficial-functionalized mesoporous silica nanoparticles (MSN) with thiolation Polymer (PSH) and Cell-Penetrating Peptide (CPP), which promote the mucoadhesion and cellular interactions, respectively, to boost the intestinal permeability of MSN-CPP-PSH, for efficient delivery of oral drugs. As a result, MSN-CPP-PSH showed excellent penetration across the mucus layer and high epithelial absorption using a mucus-secreting epithelial cell model. After investigating the behavior and integrity of MSN-CPP-PSH in the process of mucus penetrating and epithelium uptaking, the PSH molecules were observed gradually dissociated from the surface of MSN as it penetrated across mucus, and the CPP-covering core was revealed in time for the subsequent transepithelial transport. Biodistribution studies demonstrated this newly thiolated-polymers had quite mucoadhesive strength and greatly increased the intestinal residence time. Finally, MSN-CPP-PSH exhibits 6.76-fold in the relative bioavailability of lopinavir (LNV), as compared to oral free LNV. Our study demonstrated this newly synthesized PSH can play as a dissociable “mucus-adherent” agent to facilitate mucoadhesion and mucopenetration of nanocarriers, and also provided a strategy to overcome the diffusion barrier of mucus and absorption barrier of epithelium simultaneously.
研究领域:
药剂学
·
生物材料学
✓ 已验证
DOI
高影响因子
Biomedical Materials
·
2022
·Vol. 17
·期 4
·045006
·
IF 4.103
Li Li,Anqi Chen,Bingmi Liu,Hao Pan,Yanjie Yu,Yu Liu
The article presents a hepatocellular carcinoma cell surface-specific ligand glycyrrhetinic acid (GA) and cell-penetrating peptide (TAT) with good cell membrane penetration to modify the anti-tumor drug pingyangmycin (PYM) liver delivery system, which achieve targeted delivery of drugs and improve anti-tumor efficiency. In this study, we synthesized the pingyangmycin liposome modified by glycyrrhetinic acid and cell penetrating peptide(GA-TAT-PYM-L) and evaluated the anti-tumor effect of GA-TAT-PYM-L in vitro. Using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenylte-trazolium bromidecell proliferation method, GA-TAT-PYM-L had a stronger inhibitory effect on HepG2 cells than the free drug PYM at the same concentration. Acridine orange-ethidium bromide staining assays showed that GA-TAT-PYM-L had stronger apoptosis promotion effects on HepG2 cells in comparison to PYM. Pharmacokinetic studies indicated that, compared with PYM, GA-TAT-PYM-L enhanced mean residence time (MRT0–∞) and area under curve (AUC0–∞) by about 2.79-fold and 2.45-fold. The T1/2 was prolonged to 140.23 ± 14.13 min. Tissue distribution results showed that the PYM concentrations in livers from the GA-TAT-PYM-L group were always higher than other tissues at each monitoring period after 5 min, indicating that GA-TAT-PYM-L can achieve liver targeting.
研究领域:
药学
·
肿瘤学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Pharmaceutics
·
2023
·Vol. 15
·期 2
·670
·
IF 5.4
Wen-Juan Shen,Duo-Mei Tian,Le Fu,Biao Jin,Yu Liu,Yun-Sheng Xu,Yong-Bin Ye,Xiao-Bo Wang,Xiao-Jun Xu,Chun Tang,Fang-Ping Li,Chun-Fei Wang,Gang Wu,Le-Ping Yan
Cell-penetrating peptides (CPPs) are attractive non-viral gene delivery vectors due to their high transfection capacity and safety. Previously, we have shown that cell-penetrating peptide RALA can be a promising gene delivery vector for chronic wound regeneration application. In this study, we engineered a novel peptide called RALA-E by introducing elastin-derived VGVAPG fragment into RALA, in order to target the elastin-binding protein on the cell surface and thus improve delivery efficacy of RALA. The transfection efficiency of RALA-E was evaluated by transfecting the HEK-293T and HeLa cell lines cells with RALA-E/pDNA complexes and the flow-cytometry results showed that RALA-E significantly increased the transfection efficiency by nearly 20% in both cell lines compared to RALA. Inhibition of pDNA transfection on HEK-293T cells via chlorpromazine, genistein and mβCD showed that the inhibition extent in transfection efficiency was much less for RALA-E group compared to RALA group. In addition, RALA-E/miR-146a complexes showed up to 90% uptake efficiency in macrophages, and can escape from the endosome and enter the nucleus to inhibit the expression of inflammation genes. Therefore, the developed RALA-E peptide has high potential as a safe and efficient vector for gene therapy application.Keywords:cell-penetrating peptide;gene delivery;elastin-derived peptide;elastin-binding protein;immune modulation
研究领域:
生物技术
·
药学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Drug Delivery and Translational Research
·
2020
·Vol. 11
·期 5
·1969-1982
·
IF 2.664
Zhao Haijun,Chen Mengyuan,Zhao Zhujiang,Zhu Linjia,Yuan Shaofei
Balancing the antitumor activity and systemic toxicity of tripterine still faces a big challenge due to the narrow therapeutic window. To address this issue, we report a microemulsion system based on tripterine, brucea oil, and glycyrrhizin, and dual modified with both transferrin and cell-penetrating peptide SA-R 6 H 4 (Tf/SA-R 6 H 4 -TBG-MEs) for combinational and tumor-targeted cancer therapy. Such a microemulsion exhibited a spherical shape with a size of ~50 nm and a mildly-negative charge. The half-maximal inhibitory concentration (IC 50 ) of Tf/SA-R 6 H 4 -TBG-MEs against ovarian cancer SKOV3 cells was 0.27 ± 0.43 μg tripterine/mL, which was 5.85 times lower than that of free tripterine. The cellular uptake of tripterine after treatment with Tf/SA-R 6 H 4 -TBG-MEs was 1.56 times higher than that of TBG-MEs (non-modified microemulsion). In pharmacokinetics studies, the area under the curve of Tf/SA-R 6 H 4 -TBG-MEs increased by 1.97 times compared with that of the physical mixture group. The tumoral accumulation of tripterine was significantly improved in Tf/SA-R 6 H 4 -TBG-MEs group than TBG-MEs-treated group. In antitumor efficacy in vivo, Tf/SA-R 6 H 4 -TBG-MEs exhibited the strongest inhibition of tumor growth and the longest survival period among all the groups, which is associated with the rational combination, microemulsion system, and dual modification with tumor-targeted ligands. Importantly, Tf/SA-R 6 H 4 -TBG-MEs significantly reduced the toxicity of tripterine against the liver and kidney. Our design provides a new approach for efficient and safe ovarian cancer therapy based on a multicomponent combination.
研究领域:
药学
·
肿瘤学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）