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The anti-inflammatory effects of luminal Lactobacillus rhamnosus GG perfusion via regulation of JNK related pathway

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TitleInfo
Title
The anti-inflammatory effects of luminal Lactobacillus rhamnosus GG perfusion via regulation of JNK related pathway
Name (type = personal)
NamePart (type = family)
Yang
NamePart (type = given)
Bowen
NamePart (type = date)
1991-
DisplayForm
Bowen Yang
Role
RoleTerm (authority = RULIB)
author
Name (type = personal)
NamePart (type = family)
Ferraris
NamePart (type = given)
Ronaldo P
DisplayForm
Ronaldo P Ferraris
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
chair
Name (type = personal)
NamePart (type = family)
Bonder
NamePart (type = given)
Edward M
DisplayForm
Edward M Bonder
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = personal)
NamePart (type = family)
Gao
NamePart (type = given)
Nan
DisplayForm
Nan Gao
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (type = corporate)
NamePart
Graduate School - Newark
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (qualifier = exact)
2017
DateOther (qualifier = exact); (type = degree)
2017-10
CopyrightDate (encoding = w3cdtf); (qualifier = exact)
2017
Place
PlaceTerm (type = code)
xx
Language
LanguageTerm (authority = ISO639-2b); (type = code)
eng
Abstract (type = abstract)
Lactobacillus rhamnosus GG (LGG) is a well studied and commercially available probiotic, known to be able to alleviate intestinal disorders. From previous research in Dr. Ferraris’s lab, we discovered that by direct perfusion of active LGG to the jejunum of the small intestine, we were able to promote the production of anti-inflammatory cytokines IFN-γ, IL10, IL2, IL4, IL5, and IL6 (P<0.05) in four hours. To further study the properties of the LGG perfusate, we conducted a feeding study, where we gavage fed Mus Musculus (30 mice, 22.3±2 g Body Weight) with either Saline or LGG (6.6 x 108 CFU/mL in Saline) two times a day for days, then perfused with bacterial LPS or Poly I:C to the small intestine to determine whether prior consumption of LGG prevented bacterial or viral induced inflammation (n=5). Unfortunately, the results generated were statistically insignificant. Suggesting that gavage feeding LGG may have failed to stimulate an intestinal reaction, probably because of stress and dilution with endogenous bacterial. We performed another perfusion experiment followed by transcriptional analysis of the small intestinal flora. LGG and Saline perfused mice results of the RNAseq were analyzed by IPA (Ingenutiy Pathway Analysis), a software that is able to predict the activities of molecular pathways. Among pathways predicted by IPA to be affected by LGG perfusion, “LPS/IL-1 Mediated Inhibition of RXR (Retinoid X receptor) Function” has the most significant P value. Hence, using IPA’s result as a map, I determined to focused my attention towards LGG’s effect on this pathway. The “LPS/IL-1 Mediated Inhibition of RXR Function” pathway is a Mitogen-activated protein kinase (MAPK) pathway, with JNK as the MAPK. Bacterial LPS would activate JNK by phosphorylation. The pJNK would in turn inhibits RXR activation, thus inhibiting the production of its’ downstream associate enzymes and transporter. According to our RNAseq data, IPA predicted the pathway inhibited with LGG perfusion. Meaning that LGG would decrease pJNK, resulting in an increase in downstream transcription. There are a total of 25 effectors (enzymes and transporters) associated with xenobiotic metabolism downstream of RXRα. Among the effectors, the mRNA levels of 9 (CYP2A6, CYP2C8, CYP2C9, CYP3A5, CYP3A7, ALDH, PAPSS2, SULT, GST) were measured in previous RNAseq experiment, 12 (MGMT, SOD3, FMO2, UGT, MRP2, MRP3, MRP4, OATP2, CES, MAOA, CAT, MDR1) were measured by qPCR. Two receptors within the RXR complex (CAR, PXR), a known downstream substrate of JNK (c-JUN), and an upstream receptor of the pathway were also measured by qPCR. The phosphorylation of JNK and EGFR were detected and analyzed with western bloting. From RNAseq, the mRNA expression of 9 effectors (CYP2A6, CYP2C8, CYP2C9, CYP3A5, CYP3A7, ALDH, PAPSS2, SULT, GST) showed an increase of >1.5 fold with LGG perfusion compared to Saline Perfusion (P<0.05). From our qPCR results, downstream of RXR, when compared with saline perfused samples, the mRNA expression of 10 effectors (SOD3, FMO2, UGT, MRP2, MRP3, MRP4, OATP2, CES, MAOA, MDR1) increased more than 1.2 fold with LGG perfusion (P<0.05). The phosphorylation of JNK decreased in LGG perfused samples when compared with the control. In conclusion, we have discovered strong evidence showing that LGG has the ability to increase xenobiotic metabolism in the small intestine through the inhibition of the “LPS/IL-1 Mediated Inhibition of RXR Function” pathway. More experiments are still needed to further prove this theory. However, EGFR (Epidermal growth factor receptor), increased in phosphorylation in LGG perfused samples when compared with the control. EGFR is known to be associated with the production of cytokines, hence this can be used for future experiments in explaining the reaction observed with LGG perfusion.
Subject (authority = RUETD)
Topic
Biology
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_8280
PhysicalDescription
Form (authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
1 online resource (x, 63 p. : ill.)
Note (type = degree)
M.S.
Note (type = bibliography)
Includes bibliographical references
Subject (authority = ETD-LCSH)
Topic
Intestine, Small
Subject (authority = ETD-LCSH)
Topic
Inflammation
Subject (authority = ETD-LCSH)
Topic
Probiotics
Note (type = statement of responsibility)
by Bowen Yang
RelatedItem (type = host)
TitleInfo
Title
Graduate School - Newark Electronic Theses and Dissertations
Identifier (type = local)
rucore10002600001
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier (type = doi)
doi:10.7282/T39K4F9P
Genre (authority = ExL-Esploro)
ETD graduate
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RightsDeclaration (ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder (type = personal)
Name
FamilyName
Yang
GivenName
Bowen
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2017-08-08 00:48:11
AssociatedEntity
Name
Bowen Yang
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - Newark
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Copyright
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Copyright protected
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Open
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ETD
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windows xp
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DateCreated (point = end); (encoding = w3cdtf); (qualifier = exact)
2017-09-21T23:20:10
DateCreated (point = end); (encoding = w3cdtf); (qualifier = exact)
2017-09-21T23:20:10
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