Cholesterol lowering effects of mono-lactose-appended β-cyclodextrin in Niemann–Pick type C disease-like HepG2 cells

The Niemann–Pick type C disease (NPC) is one of inherited lysosomal storage disorders, emerges the accumulation of unesterified cholesterol in endolysosomes. Currently, 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) has been applied for the treatment of NPC. HP-β-CyD improved hepatosplenomegaly in NPC patients, however, a high dose of HP-β-CyD was necessary. Therefore, the decrease in dose by actively targeted-β-CyD to hepatocytes is expected. In the present study, to deliver β-CyD selectively to hepatocytes, we newly fabricated mono-lactose-appended β-CyD (Lac-β-CyD) and evaluated its cholesterol lowering effects in NPC-like HepG2 cells, cholesterol accumulated HepG2 cells induced by treatment with U18666A. Lac-β-CyD (degree of substitution of lactose (DSL) 1) significantly decreased the intracellular cholesterol content in a concentration-dependent manner. TRITC-Lac-β-CyD was associated with NPC-like HepG2 cells higher than TRITC-β-CyD. In addition, TRITC-Lac-β-CyD was partially localized with endolysosomes after endocytosis. Thus, Lac-β-CyD entered NPC-like HepG2 cells via asialoglycoprotein receptor (ASGPR)-mediated endocytosis and decreased the accumulation of intracellular cholesterol in NPC-like HepG2 cells. These results suggest that Lac-β-CyD may have the potential as a drug for the treatment of hepatosplenomegaly in NPC disease.

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Synthesis of multi-lactose-appended β-cyclodextrin and its cholesterol-lowering effects in Niemann–Pick type C disease-like HepG2 cells

Beilstein Journal of Organic Chemistry ◽

10.3762/bjoc.13.2 ◽

2017 ◽

Vol 13 ◽

pp. 10-18 ◽

Cited By ~ 9

Author(s):

Keiichi Motoyama ◽

Rena Nishiyama ◽

Yuki Maeda ◽

Taishi Higashi ◽

Yoichi Ishitsuka ◽

...

Keyword(s):

Hepg2 Cells ◽

High Dose ◽

Unesterified Cholesterol ◽

Peanut Lectin ◽

Cholesterol Lowering ◽

Cholesterol Levels ◽

Type C ◽

Intracellular Cholesterol ◽

Niemann Pick ◽

Very High

Niemann–Pick type C (NPC) disease, characterized by intracellular accumulation of unesterified cholesterol and other lipids owing to defects in two proteins NPC1 and NPC2, causes neurodegeneration and other fatal neurovisceral symptoms. Currently, treatment of NPC involves the use of 2-hydroxypropyl-β-cyclodextrin (HP-β-CD). HP-β-CD is effective in the treatment of hepatosplenomegaly in NPC disease, albeit at a very high dose. One of the methods to reduce the required dose of HP-β-CD for treatment of NPC is to actively targeting hepatocytes with β-cyclodextrin (β-CD). The aim of the present study was to synthesize a novel multi-lactose-appended β-CD (multi-Lac-β-CD) and to evaluate its cholesterol-lowering effect in U18666A-treated HepG2 (NPC-like HepG2) cells. Further, the study aimed at delivering β-CD to hepatocytes via cholesterol-accumulated HepG2 cells, and indicated that the newly synthesized multi-Lac-β-CD had an average degree of substitution of lactose (DSL) of 5.6. This newly synthesized multi-Lac-β-CD was found to significantly decrease the concentration of intracellular cholesterol with negligible cytotoxicity as compared to HP-β-CD. An increased internalization of TRITC-multi-Lac-β-CD (DSL 5.6) as compared to TRITC-HP-β-CD was observed in NPC-like HepG2 cells. Further, the dissociation constant of peanut lectin with multi-Lac-β-CD (DSL5.6) was found to be extremely low (2.5 × 10−8 M). These results indicate that multi-Lac-β-CD (DSL5.6) diminished intracellular cholesterol levels in NPC-like HepG2 cells via asialoglycoprotein receptor (ASGPR)-mediated endocytosis.

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Improved Method for Obtaining of Arctigenin from Arctium Lappa L. and its Antiproliferative Effect on Human Hepatocarcinoma HepG2 Cells

Current Bioactive Compounds ◽

10.2174/1573407214666181115124223 ◽

2020 ◽

Vol 16 (3) ◽

pp. 358-362

Author(s):

Renan S. Teixeira ◽

Paulo H.D. Carvalho ◽

Jair A.K. Aguiar ◽

Valquíria P. Medeiros ◽

Ademar A. Da Silva Filho ◽

...

Keyword(s):

Antitumor Activity ◽

Crude Extract ◽

Hepg2 Cells ◽

Liver Carcinoma ◽

Adhesion Assay ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Arctium Lappa ◽

Nih 3T3

Background: Arctigenin is a lignan found in Arctium lappa L. (Asteraceae) that displays anti-inflammatory activities. Previous studies showed that the crude extract of A. Lappa has antitumor activity in human liver carcinoma, lung and stomach cancer cells. The aim of this study was to obtain arctigenin from A. lappa L., as well as to evaluate its antiproliferative effects in cells of liver carcinoma (HepG2) and fibroblasts (NIH/3T3). Methods: Arctigenin was obtained from the hydrolysis of arctiin, which was isolated from the crude extract of A. lappa. The effects of arctigenin and arctiin on HepG2 cell viability and cell adhesion were analyzed by MTT method. Adhesion assay was also carried out to evaluate the antitumor activity. Results: Our results showed that the analytical process to obtain arctigenin was fast and easy. In vitro experiments showed that arctigenin (107-269 μM) decreased HepG2 cells viability and did not cause cytotoxicity on NIH/3T3 cells. Arctigenin (27-269 μM) demonstrated anti-adhesion in HepG2 cells in a concentration-dependent manner, when compared with control. Conclusion: These results suggest a promising pharmacological activity for arctigenin as an antiproliferative compound.

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Invariant natural killer T cells are not affected by lysosomal storage in patients with Niemann-Pick disease type C

European Journal of Immunology ◽

10.1002/eji.201141821 ◽

2012 ◽

Vol 42 (7) ◽

pp. 1886-1892 ◽

Cited By ~ 12

Author(s):

Anneliese O. Speak ◽

Nicholas Platt ◽

Mariolina Salio ◽

Danielle te Vruchte ◽

David A. Smith ◽

...

Keyword(s):

T Cells ◽

Natural Killer T Cells ◽

Disease Type ◽

Lysosomal Storage ◽

Niemann Pick Disease ◽

Type C ◽

Niemann Pick ◽

Killer T Cells ◽

Invariant Natural Killer ◽

Pick Disease

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In vivo Efficacy and Safety Evaluation of Lactosyl-β-cyclodextrin as a Therapeutic Agent for Hepatomegaly in Niemann-Pick Type C Disease

Nanomaterials ◽

10.3390/nano9050802 ◽

2019 ◽

Vol 9 (5) ◽

pp. 802 ◽

Cited By ~ 4

Author(s):

Yuki Maeda ◽

Keiichi Motoyama ◽

Rena Nishiyama ◽

Taishi Higashi ◽

Risako Onodera ◽

...

Keyword(s):

Free Cholesterol ◽

Subcutaneous Administration ◽

Safety Evaluation ◽

Autosomal Recessive Disorder ◽

Liver Cells ◽

High Dose ◽

Efficacy And Safety ◽

Type C ◽

Niemann Pick

Niemann-Pick type C disease (NPC) is a fatal, autosomal recessive disorder, which causes excessive accumulation of free cholesterol in endolysosomes, resulting in progressive hepatomegaly and neurodegeneration. Currently, 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) is used at a high dose for the treatment of NPC, risking lung toxicity and hearing loss during treatment. One method to reduce the required dose of HP-β-CyD for the treatment of hepatomegaly is to actively deliver β-cyclodextrin (β-CyD) to hepatocytes. Previously, we synthesized lactosyl-β-CyD (Lac-β-CyD) and demonstrated that it lowers cholesterol in NPC model liver cells. In the present study, we studied the efficacy and safety of Lac-β-CyD treatment of hepatomegaly in Npc1−/− mice. After subcutaneous administration, Lac-β-CyD accumulated in the liver and reduced hepatomegaly with greater efficacy than HP-β-CyD. In addition, subcutaneous administration of a very high dose of Lac-β-CyD was less toxic to the lungs than HP-β-CyD. Notably, the accumulation of intracellular free cholesterol in endolysosomes of NPC-like liver cells was significantly lower after administration of Lac-β-CyD than after treatment with HP-β-CyD. In conclusion, these results suggest that Lac-β-CyD is a candidate for the effective treatment of hepatomegaly in NPC.

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Human iNSC-derived brain organoid model of lysosomal storage disorder in Niemann–Pick disease type C

Cell Death and Disease ◽

10.1038/s41419-020-03262-7 ◽

2020 ◽

Vol 11 (12) ◽

Author(s):

Seung-Eun Lee ◽

Nari Shin ◽

Myung Geun Kook ◽

Dasom Kong ◽

Nam Gyo Kim ◽

...

Keyword(s):

Stem Cells ◽

Lysosomal Storage Disorder ◽

Disease Type ◽

Lysosomal Storage ◽

Niemann Pick Disease ◽

Type C ◽

Storage Disorder ◽

Niemann Pick ◽

Pick Disease

AbstractRecent studies on developing three-dimensional (3D) brain organoids from stem cells have allowed the generation of in vitro models of neural disease and have enabled the screening of drugs because these organoids mimic the complexity of neural tissue. Niemann-Pick disease, type C (NPC) is a neurodegenerative lysosomal storage disorder caused by mutations in the NPC1 or NPC2. The pathological features underlying NPC are characterized by the abnormal accumulation of cholesterol in acidic compartments, including late endosomes and lysosomes. Due to the inaccessibility of brain tissues from human NPC patients, we developed NPC brain organoids with induced neural stem cells from NPC patient-derived fibroblasts. NPC organoids exhibit significantly reduced size and proliferative ability, which are accompanied by accumulation of cholesterol, impairment in neuronal differentiation, and autophagic flux and dysfunction of lysosomes; therefore, NPC organoids can recapitulate the main phenotypes of NPC patients. Furthermore, these pathological phenotypes observed in NPC organoids were reversed by treatment with valproic acid and HPBCD, which are known to be an effective treatment for several neurodegenerative diseases. Our data present patient-specific phenotypes in 3D organoid-based models of NPC and highlight the application of this model to drug screening in vitro.

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Niemann-Pick Type C Disease and Intracellular Cholesterol Trafficking

Journal of Biological Chemistry ◽

10.1074/jbc.r400040200 ◽

2005 ◽

Vol 280 (22) ◽

pp. 20917-20920 ◽

Cited By ~ 112

Author(s):

Ta-Yuan Chang ◽

Patrick C. Reid ◽

Shigeki Sugii ◽

Nobutaka Ohgami ◽

Jonathan C. Cruz ◽

...

Keyword(s):

Cholesterol Trafficking ◽

Type C ◽

Intracellular Cholesterol ◽

Niemann Pick

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Evaluation of Anti-Inflammatory Activities of Qingre-Qushi Recipe (QRQS) against Atopic Dermatitis: Potential Mechanism of Inhibition of IL-33/ST2 Signal Transduction

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2017/2489842 ◽

2017 ◽

Vol 2017 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Mengjiao Chen ◽

Peijun Ding ◽

Lili Yang ◽

Xufeng He ◽

Chunjie Gao ◽

...

Keyword(s):

Signal Transduction ◽

Molecular Mechanisms ◽

Histological Study ◽

High Dose ◽

Dependent Manner ◽

Hacat Cells ◽

Concentration Dependent Manner ◽

Dermal Thickness ◽

Significant Difference ◽

Anti Inflammatory

To evaluate the anti-inflammatory activities of QRQS against AD and the inhibitory molecular mechanisms of IL-33/ST2 signal transduction, BALB/c mice were divided into six groups (normal control, OVA control, low-dose of QRQS, middle-dose of QRQS, high-dose of QRQS, and cetirizine) and epicutaneously exposed to ovalbumin or PBS for 3 weeks and treated with QRQS for 2 weeks. Skin biopsies and blood samples were obtained for histological study, antibody analysis, and RNA isolation. HaCaT cells, stimulated by TNF-α and IFN-γ, were treated with QRQS to evaluate mRNA and protein expression by RT-PCR and ELISA. QRQS decreased both epidermal and dermal thickness, alleviated dermatitis, and reduced IL-33 and ST2 positive cell numbers. The concentration of specific IgE, IgG, IgG1, and IgG2a antibodies in serum and the expression of IL-33, ST2, IL-1RAcP, IL-4, and IL-13 mRNA in the skin were suppressed. No significant difference exists in TNF-α or IFN-γ. QRQS decreased IL-33 mRNA and protein secretion in HaCaT cells exposed to TNF-α and IFN-γ in a time- and concentration-dependent manner. QRQS regulates related molecule expression of ovalbumin-induced dermatitis involved in the IL-33/ST2 signaling axis in the treatment of acute AD.

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