Connect with us

Heartland

CLG from Hemp Seed Inhibits LPS-Stimulated Neuroinflammation in BV2 Microglia by Regulating NF-κB and Nrf-2 Pathways

The healthy benefits of hemp (Cannabis sativa L.) seed have often been attributed to its oils and proteins. Recent studies reveal that hemp seed phenylpropionamides could also show various bioactivities. Continuation of our study on hemp seed provided a phenylpropionamide, coumaroylaminobutanol glucopyranoside (CLG). This work investigated the neuroprotective effect of CLG and its underlying mechanism using lipopolysaccharide-induced BV2 microglia. Our study demonstrated that CLG increased…

Republished by Plato

Published

on


. 2019 Sep 26;4(15):16517-16523.

doi: 10.1021/acsomega.9b02168. eCollection 2019 Oct 8.

Affiliations

Free PMC article

Item in Clipboard

Shanshan Wang et al. ACS Omega. .

Free PMC article

Abstract

The healthy benefits of hemp (Cannabis sativa L.) seed have often been attributed to its oils and proteins. Recent studies reveal that hemp seed phenylpropionamides could also show various bioactivities. Continuation of our study on hemp seed provided a phenylpropionamide, coumaroylaminobutanol glucopyranoside (CLG). This work investigated the neuroprotective effect of CLG and its underlying mechanism using lipopolysaccharide-induced BV2 microglia. Our study demonstrated that CLG increased adenosine monophosphate-activated protein kinase (AMPK) expression, suppressed the nuclear factor-kappa B (NF-κB) signaling pathway by inhibiting the phosphorylation of IκBα and NF-κB p65 and decreased proinflammatory cytokine levels in a concentration-dependent manner. Furthermore, CLG reduced the production of cellular reactive oxygen species and stimulated the nuclear factor erythroid 2-related factor 2 (Nrf-2) signaling pathway. Collectively, these results suggested that CLG effectively and simultaneously inhibited inflammatory responses and oxidative stress through the NF-κB and Nrf-2 signaling pathways. AMPK was also involved in the anti-inflammatory effect of CLG. This study provides new insights into the diverse bioactive constituents of hemp seed.

Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1

Figure 1

Effect of CLG on the cell viability of
BV2 microglia with or without
LPS stimulation. (a) Structure of CLG. (b) BV2 cells were cotreated
with various concentrations of CLG (5, 10, 15 μM) or RES (10
μM) with or without LPS (100 ng/mL) for 24 h. Cell viability
was determined by the MTT assay. (The data are expressed as the mean
± SD of three experiments.)

Figure 2

Figure 2

CLG reduced proinflammatory cytokine production
in LPS-stimulated
BV2 microglia. BV2 cells were pretreated with 5, 10, and 15 μM
CLG for 1 h and then stimulated with LPS (100 ng/mL). After coincubation
for 24 h, the supernatants were collected for the measurement of IL-1β
(a) and IL-6 (b) by ELISA. After coincubation for 6 h, total RNA was
isolated, and relative IL-1β (c) and IL-6 (d) mRNA expression
was measured by RT-PCR. (Data are presented as the mean ± SD
from at least three independent experiments. *p <
0.05, **p < 0.01, and ***p <
0.001 compared with the LPS-treated group; ###p < 0.001 compared with the control group.)

Figure 3

Figure 3

CLG inhibited
TLR4/NF-κB protein expression in LPS-induced
BV2 microglia. BV2 cells were pretreated with CLG for 1 h and stimulated
with LPS. After stimulation for 24 h, cell extracts were prepared
and subjected to western blotting with TLR4 and MyD88 antibodies.
After stimulation for 1 h, cell extracts were prepared and subjected
to western blotting with IκBα, phospho-IκBα,
NF-κB p65, and phospho-NF-κB p65 antibodies. β-Actin
was used as the internal control for normalization. (a) Western blot
bands of TLR4 and MyD88. (b) The density of TLR4 bands was measured,
and their ratio was calculated. (c) Density ratio of MyD88 bands.
(d) Western blot bands of IκBα, phospho-IκBα,
NF-κB p65, and phospho-NF-κB p65. (e–h) Density
ratio of phospho-NF-κB p65, p65, phospho-IκBα, and
IκBα. (The results are presented as the mean ± SD
from at least three independent experiments. *p <
0.05, **p < 0.01, and ***p <
0.001 compared with cells treated with LPS; #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the control.)

Figure 4

Figure 4

CLG reduced cellular ROS generation and promoted Nrf-2/HO-1 expression
in LPS-induced BV2 microglia. BV2 cells were treated with CLG for
1 h prior to LPS stimulation for 24 h. (a) Cellular ROS
levels were measured using DCFH-DA by flow cytometry. (b) Quantification
of the relative % of cells with ROS production. (c) Cell lysates were
collected and analyzed using western blotting for Nrf-2 and HO-1.
(d,e) The density of Nrf-2 and HO-1 bands was measured, and their
ratio was calculated. β-Actin was used as the internal control
for normalization. (The results are presented as the mean ± SD
from at least three independent experiments. **p <
0.01 compared with cells treated with LPS; ##p < 0.01 and ###p < 0.001 compared
with the control.)

Figure 5

Figure 5

CLG increased AMPK activation in LPS-induced BV2 microglia. BV2
cells were treated with CLG for 1 h prior to LPS stimulation for 24
h. Cell lysates were collected and analyzed using western blotting
for phospho-AMPK and AMPK (a). The density of phospho-AMPK and AMPK
bands was measured, and their ratio was calculated (b). Cells were
pretreated with or without CLG and C–C (1 μM) for 1 h
and then coincubated with LPS for 6 h, total RNA was isolated, and
relative IL-1β (c) and IL-6 (d) mRNA expression was measured
by RT-PCR. β-Actin was used as the internal control. (The results
are presented as the mean ± SD from at least three independent
experiments. *p < 0.05 and ***p < 0.001 compared with cells treated with LPS; ###p < 0.001 compared with the control.)

Similar articles

References

    1. Xu L.; He D.; Bai Y. Microglia-mediated inflammation and neurodegenerative disease. Mol. Neurobiol. 2016, 53, 6709–6715. 10.1007/s12035-015-9593-4. – DOI PubMed
    1. Smith J. A.; Das A.; Ray S. K.; Banik N. L. Role of pro-inflammatory cytokines released from microglia in neurodegenerative diseases. Brain Res. Bull. 2012, 87, 10–20. 10.1016/j.brainresbull.2011.10.004. – DOI PubMed
    1. Callaway J. C. Hempseed as a nutritional resource: An overview. Euphytica 2004, 140, 65–72. 10.1007/s10681-004-4811-6. – DOI
    1. Zhou Y.; Wang S.; Lou H.; Fan P. Chemical constituents of hemp (Cannabis sativa L.) seed with potential anti-neuroinflammatory activity. Phytochem. Lett. 2018, 23, 57–61. 10.1016/j.phytol.2017.11.013. – DOI
    1. Luo Q.; Yan X.; Bobrovskaya L.; Ji M.; Yuan H.; Lou H.; Fan P. Anti-neuroinflammatory effects of grossamide from hemp seed via suppression of TLR-4-mediated NF-κB signaling pathways in lipopolysaccharide-stimulated BV2 microglia cells. Mol. Cell. Biochem. 2017, 428, 129–137. 10.1007/s11010-016-2923-7. – DOI PubMed

LinkOut – more resources

  • Full Text Sources

Source: https://pubmed.ncbi.nlm.nih.gov/31616830/?utm_source=no_user_agent&utm_medium=rss&utm_campaign=pubmed-2&utm_content=1lg0RU4SkRVNqPFA7e6OZattw-Uzcw0dBEajJBQqmhHlNZ8MdX&fc=20200804212534&ff=20200812123210&v=2.11.5

Heartland

My Mom Is In A Lot Of Pain. So Much Nerve Damage They Had To Remove 3 Disks In Her Neck. Assistance On CBD And Med Interactions, Please!

Republished by Plato

Published

on

My mom is taking Januvia, Glipizide, Lantus Injections, and Trulicity Injections. I really think gummies with CBD:THC 100:1 would really help her. Unfortunately, I have heard that diabetes medications might interfere with CBD and vice versa. Does anyone have any experience with interactions with these medications? None of her prescriptions have the grapefruit warning on the labels.

Edited to make corrections.

Source: https://www.reddit.com/r/CBD/comments/l23ooi/my_mom_is_in_a_lot_of_pain_so_much_nerve_damage/

Continue Reading

Heartland

MP Trae el Flow del Año 3000: Su Papel en Red Bull, Su Anhelo de FMS y su Tema con Kódigo

ElPlanteo.com entrevista al freestyler y músico MP, también conocido como “El Juvenil”. Nacido en 1999 su nombre viene, según dice, de que se ve muy joven en escena.

Republished by Plato

Published

on


MP Trae el Flow del Año 3000: Su Papel en Red Bull, Su Anhelo de FMS y su Tema con Kódigo | High Times





























Source: https://hightimes.com/espanol/mp-trae-el-flow-del-ano-3000-su-papel-en-red-bull-su-anhelo-de-fms-y-su-tema-con-kodigo/

Continue Reading

Heartland

Bert Kreischer on Comedy Shows During COVID And Hanging Out With Snoop Dogg

Comedian Bert Kreisher is a “go big or go home” type of man.

Republished by Plato

Published

on


Bert Kreischer on Comedy Shows During COVID And Hanging Out With Snoop Dogg | High Times






























Source: https://hightimes.com/celebrities/bert-kreischer-comedy-shows-during-covid-hanging-out-with-snoop-dogg/

Continue Reading
News4 days ago

What are Some Major Factors to Keep in Mind When Buying CBD Vape Oil?

Heartland1 day ago

MagicMed: Bringing the Drug Candidate Library Model to Psychedelics

Heartland4 days ago

Investors in This Cannabis Stock Are Leaving $800 Million on the Table

Heartland4 days ago

What’s New With Cannabis Stocks for the Week Ending 01/15/21

Heartland4 days ago

What is Delta-8 THC? Everything You Need to Know

Heartland5 days ago

Optical Coherence Tomography Findings in Cannabis Users

Heartland3 days ago

Alcanna to Spin Out Cannabis Business and Pursue Value Segment Strategy

Heartland14 hours ago

10 Herbs That Can Help With Sleep

News3 days ago

CBD News: NIHC chairman appointed to board advising U.S. on trade policy

Heartland3 days ago

Fire & Flower Expands Free Same-Day Delivery Across Its Ontario Network

Heartland2 days ago

Nordic Oil FAQ: The CBD questions you’ve always wanted answers to

News2 days ago

CBD News: Restrictions on shipment of vaping products could impact hemp industry

News3 days ago

Illinois Collects $62 Million in Cannabis Revenue to Support Neighborhoods

Heartland2 days ago

Hemp & CBD Blog | Northeast Kingdom Hemp

Heartland2 days ago

CBD Blog: News, Info & Tips

Heartland3 days ago

Heavy metals in cbd

Heartland3 days ago

Blog – Global Cannabinoids | CBD Wholesale & Bulk | White Label | Private Label

Heartland2 days ago

Informational blogs about CBD and Hemp

Heartland3 days ago

A Beginners Guide to Pests

Heartland3 days ago

Clinical and biochemical heterogeneity of Parkinson’s disease

Heartland2 days ago

Easy Day Hemp Blog

Heartland2 days ago

CBD Blogs | Ergo Hemp Co.

News3 days ago

BC Craft Supply Co Announces Letter of Intent with Psilocybin Research and Development Company Ava Pathways

News3 days ago

CBD News: Circle K wants to block hemp trademark that resembles its own

Uncategorized3 days ago

10xPURE-GOLD CBDa Muscle & Joint Relief Cream

Heartland2 days ago

Heartland2 days ago

Does CBD Affect Men and Women Differently?

Heartland3 days ago

How Red White & Bloom Established A Base In Michigan’s Cannabis Scene—And Keeps Expanding

Heartland17 hours ago

19 in texas I know it is technically legal to possess hemp flower I’m wondering about the penalties of possession under 21

Heartland2 days ago

Straight Hemp CBD Blog

Trending

A Cloud Nine Capital Entity Copyright © 2020 – All Rights Reserved Proudly Made in America