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Article ; Online: Lipopolysaccharide regulation of antiinflammatory tristetraprolin family and proinflammatory gene expression in mouse macrophages.

Cao, Heping

2024 Volume 17, Issue 1, Page(s) 82

Abstract: Objective: Tristetraprolin (TTP/ZFP36) family proteins exhibit antiinflammatory effects by destabilizing proinflammatory mRNAs. Previous studies showed that bacterial endotoxin lipopolysaccharides (LPS) stimulated TTP and tumor necrosis factor (TNF) ... ...

Abstract	Objective: Tristetraprolin (TTP/ZFP36) family proteins exhibit antiinflammatory effects by destabilizing proinflammatory mRNAs. Previous studies showed that bacterial endotoxin lipopolysaccharides (LPS) stimulated TTP and tumor necrosis factor (TNF) gene expression, but less was known about LPS effects on TTP homologues and other proinflammatory gene expression in macrophages. The objective was to investigate LPS regulation of TTP family gene and TTP-targeted gene expression in mouse RAW264.7 macrophages using much higher concentrations of LPS and much longer treatment time than previous studies. Results: MTT assay showed that LPS was not toxic to the cells under LPS treatment up to 1000 ng/mL for 2-24 h. LPS mildly affected the soluble protein content in the cells. qPCR assay showed that LPS stimulated TTP mRNA rapidly but not sustainably with 40, 10, and 3 fold of the DMSO control after 2, 8 and 24 h treatment, respectively. Immunoblotting confirmed qPCR results on LPS stimulation of TTP gene expression in the mouse macrophages. LPS exhibited minimal effects on ZFP36L1, ZFP36L2 and ZFP36L3 mRNA levels. LPS increased mRNA levels of TNF, COX2, GM-CSF, INFγ and IL12b up to 311, 418, 11, 9 and 4 fold, respectively. This study demonstrated that LPS did not affect macrophage viability, dramatically increased antiinflammatory TTP gene expression as well as proinflammatory TNF and COX2 gene expression but had only mild effects on TTP homologues and other proinflammatory cytokine gene expression in the mouse macrophages.
MeSH term(s)	Mice ; Animals ; Lipopolysaccharides/pharmacology ; Lipopolysaccharides/metabolism ; Tristetraprolin/genetics ; Tristetraprolin/metabolism ; Cyclooxygenase 2/genetics ; Cyclooxygenase 2/metabolism ; Macrophages/metabolism ; Anti-Inflammatory Agents/pharmacology ; Gene Expression ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Tumor Necrosis Factor-alpha/genetics ; Tumor Necrosis Factor-alpha/metabolism ; Gene Expression Regulation
Chemical Substances	Lipopolysaccharides ; Tristetraprolin ; Cyclooxygenase 2 (EC 1.14.99.1) ; Anti-Inflammatory Agents ; RNA, Messenger ; Tumor Necrosis Factor-alpha
Language	English
Publishing date	2024-03-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2413336-X
ISSN	1756-0500 ; 1756-0500
ISSN (online)	1756-0500
ISSN	1756-0500
DOI	10.1186/s13104-024-06743-6
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Editorial for the Special Issue on Plant Polyphenols in the Immune and Inflammatory Responses.

Cao, Heping

Biomolecules

2023 Volume 13, Issue 5

Abstract: Inflammation and associated immune diseases have placed a heavy burden on health care systems [ ... ]. ...

Abstract	Inflammation and associated immune diseases have placed a heavy burden on health care systems [...].
MeSH term(s)	Humans ; Polyphenols/pharmacology ; Inflammation ; Plants
Chemical Substances	Polyphenols
Language	English
Publishing date	2023-05-11
Publishing country	Switzerland
Document type	Editorial
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom13050814
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Article ; Online: Bacterial endotoxin lipopolysaccharides regulate gene expression in human colon cancer cells.

Cao, Heping

BMC research notes

2023 Volume 16, Issue 1, Page(s) 216

Abstract: Objective: Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria. Colon bacteria contribute to LPS which promotes colon cancer metastasis. The objective of this study was to survey the effect of LPS on cell viability and gene ...

Abstract	Objective: Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria. Colon bacteria contribute to LPS which promotes colon cancer metastasis. The objective of this study was to survey the effect of LPS on cell viability and gene expression of 55 molecular targets in human colon cancer cells. Results: LPS did not affect the viability of COLO 225 cells under the culture conditions but affected the expression of a number of genes important in inflammatory responses and cancer development. LPS increased TTP family, GLUT family and DGAT1 mRNA levels but decreased DGAT2a and DGAT2b expression in the human colon cancer cells. LPS also increased COX2, CXCL1, ELK1, ICAM1, TNFSF10 and ZFAND5 but decreased BCL2L1, CYP19A1 and E2F1 mRNA levels in the colon cancer cells. These data suggest that LPS has profound effects on gene expression in human colon cancer cells.
MeSH term(s)	Humans ; Lipopolysaccharides/pharmacology ; Endotoxins ; Colonic Neoplasms/genetics ; Gene Expression
Chemical Substances	Lipopolysaccharides ; Endotoxins
Language	English
Publishing date	2023-09-13
Publishing country	England
Document type	Journal Article
ZDB-ID	2413336-X
ISSN	1756-0500 ; 1756-0500
ISSN (online)	1756-0500
ISSN	1756-0500
DOI	10.1186/s13104-023-06506-9
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Article ; Online: Bacterial endotoxin lipopolysaccharides regulate gene expression in human colon cancer cells

Cao, Heping

BMC Research Notes. 2023 Sept. 13, v. 16, no. 1 p.216-

2023

Abstract	Objective: Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria. Colon bacteria contribute to LPS which promotes colon cancer metastasis. The objective of this study was to survey the effect of LPS on cell viability and gene expression of 55 molecular targets in human colon cancer cells. Results: LPS did not affect the viability of COLO 225 cells under the culture conditions but affected the expression of a number of genes important in inflammatory responses and cancer development. LPS increased TTP family, GLUT family and DGAT1 mRNA levels but decreased DGAT2a and DGAT2b expression in the human colon cancer cells. LPS also increased COX2, CXCL1, ELK1, ICAM1, TNFSF10 and ZFAND5 but decreased BCL2L1, CYP19A1 and E2F1 mRNA levels in the colon cancer cells. These data suggest that LPS has profound effects on gene expression in human colon cancer cells.
Keywords	carcinogenesis ; cell viability ; cell wall components ; colon ; colorectal neoplasms ; endotoxins ; gene expression ; humans ; intercellular adhesion molecule-1 ; lipopolysaccharides ; metastasis
Language	English
Dates of publication	2023-0913
Publishing place	Springer Science and Business Media LLC
Document type	Article ; Online
Note	Resource is Open Access
ZDB-ID	2413336-X
ISSN	1756-0500
ISSN	1756-0500
DOI	10.1186/s13104-023-06506-9
Database	NAL-Catalogue (AGRICOLA)

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Article: Editorial: Woody oil crops: key trait formation and regulation.

Cao, Heping / Gong, Wenfang / Rong, Jun / Yuan, Deyi

Frontiers in plant science

2023 Volume 14, Page(s) 1328990

Language	English
Publishing date	2023-11-10
Publishing country	Switzerland
Document type	Editorial
ZDB-ID	2613694-6
ISSN	1664-462X
ISSN	1664-462X
DOI	10.3389/fpls.2023.1328990
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Article ; Online: Plant Polyphenol Gossypol Induced Cell Death and Its Association with Gene Expression in Mouse Macrophages.

Cao, Heping / Sethumadhavan, Kandan

Biomolecules

2023 Volume 13, Issue 4

Abstract: Gossypol is a complex plant polyphenol reported to be cytotoxic and anti-inflammatory, but little is known about its effect on gene expression in macrophages. The objective of this study was to explore gossypol's toxicity and its effect on gene ... ...

Abstract	Gossypol is a complex plant polyphenol reported to be cytotoxic and anti-inflammatory, but little is known about its effect on gene expression in macrophages. The objective of this study was to explore gossypol's toxicity and its effect on gene expression involved in the inflammatory response, glucose transport and insulin signaling pathways in mouse macrophages. Mouse RAW264.7 macrophages were treated with multiple concentrations of gossypol for 2-24 h. Gossypol toxicity was estimated by MTT assay and soluble protein content. qPCR analyzed the expression of anti-inflammatory tristetraprolin family (TTP/ZFP36), proinflammatory cytokine, glucose transporter (GLUT) and insulin signaling genes. Cell viability was greatly reduced by gossypol, accompanied with a dramatic reduction in soluble protein content in the cells. Gossypol treatment resulted in an increase in TTP mRNA level by 6-20-fold and increased ZFP36L1, ZFP36L2 and ZFP36L3 mRNA levels by 26-69-fold. Gossypol increased proinflammatory cytokine TNF, COX2, GM-CSF, INFγ and IL12b mRNA levels up to 39-458-fold. Gossypol treatment upregulated mRNA levels of GLUT1, GLUT3 and GLUT4 genes as well as INSR, AKT1, PIK3R1 and LEPR, but not APP genes. This study demonstrated that gossypol induced macrophage death and reduced soluble protein content, which was accompanied with the massive stimulation of anti-inflammatory TTP family and proinflammatory cytokine gene expression, as well as the elevation of gene expression involved in glucose transport and the insulin signaling pathway in mouse macrophages.
MeSH term(s)	Mice ; Animals ; Polyphenols/pharmacology ; Polyphenols/metabolism ; Gossypol/pharmacology ; Macrophages/metabolism ; Insulin/metabolism ; Anti-Inflammatory Agents/pharmacology ; Cytokines/metabolism ; Gene Expression ; RNA, Messenger/metabolism ; Cell Death ; Glucose/metabolism ; Tristetraprolin/genetics ; Tristetraprolin/metabolism ; Tristetraprolin/pharmacology
Chemical Substances	Polyphenols ; Gossypol (KAV15B369O) ; Insulin ; Anti-Inflammatory Agents ; Cytokines ; RNA, Messenger ; Glucose (IY9XDZ35W2) ; Tristetraprolin
Language	English
Publishing date	2023-03-30
Publishing country	Switzerland
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom13040624
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Article ; Online: Identification of

Cao, Heping / Sethumadhavan, Kandan

Molecules (Basel, Switzerland)

2022 Volume 27, Issue 21

Abstract: Cottonseed contains many bioactive molecules including plant polyphenols. Cottonseed value might be increased by providing high-value bioactive polyphenols for improving nutrition and health. However, there was a lack of molecular evidence for cottonseed ...

Abstract	Cottonseed contains many bioactive molecules including plant polyphenols. Cottonseed value might be increased by providing high-value bioactive polyphenols for improving nutrition and health. However, there was a lack of molecular evidence for cottonseed bioactivity in mammalian cells. One widely used method for evaluating the bioactivity of natural products is quantitative real-time-PCR (qPCR). The selection of stably expressed internal reference genes is a crucial task of qPCR assay for data analysis. The rationale for reference gene selection is that a lower standard deviation of the cycle of threshold (Cq) among the treatments indicates a more stable expression of the gene. The objective of this study was to select reference genes in human colon cancer cells (COLO 205) treated with cottonseed-derived gossypol and bioactive extracts along with bacterial endotoxin lipopolysaccharides (LPS). SYBR Green qPCR was used to analyze the mRNA levels of a wide range of biomarkers involved in glucose transport, lipid biosynthesis, inflammatory response, and cancer development. qPCR data (10,560 Cq values) were generated from 55 genes analyzed from 64 treatments with triplicate per treatment for each gene. The data showed that B-cell lymphoma 2 (
MeSH term(s)	Animals ; Humans ; Cottonseed Oil/analysis ; Gossypol/pharmacology ; Gossypol/analysis ; Lipopolysaccharides/pharmacology ; Colonic Neoplasms/drug therapy ; Colonic Neoplasms/genetics ; Bacteria ; RNA, Messenger ; Mammals ; Proto-Oncogene Proteins c-bcl-2
Chemical Substances	Cottonseed Oil ; Gossypol (KAV15B369O) ; Lipopolysaccharides ; RNA, Messenger ; BCL2 protein, human ; Proto-Oncogene Proteins c-bcl-2
Language	English
Publishing date	2022-11-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1413402-0
ISSN	1420-3049 ; 1431-5165 ; 1420-3049
ISSN (online)	1420-3049
ISSN	1431-5165 ; 1420-3049
DOI	10.3390/molecules27217560
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Article ; Online: Identification of Bcl2 as a Stably Expressed qPCR Reference Gene for Human Colon Cancer Cells Treated with Cottonseed-Derived Gossypol and Bioactive Extracts and Bacteria-Derived Lipopolysaccharides

Heping Cao / Kandan Sethumadhavan

Molecules, Vol 27, Iss 7560, p

2022 Volume 7560

Abstract	Cottonseed contains many bioactive molecules including plant polyphenols. Cottonseed value might be increased by providing high-value bioactive polyphenols for improving nutrition and health. However, there was a lack of molecular evidence for cottonseed bioactivity in mammalian cells. One widely used method for evaluating the bioactivity of natural products is quantitative real-time-PCR (qPCR). The selection of stably expressed internal reference genes is a crucial task of qPCR assay for data analysis. The rationale for reference gene selection is that a lower standard deviation of the cycle of threshold (Cq) among the treatments indicates a more stable expression of the gene. The objective of this study was to select reference genes in human colon cancer cells (COLO 205) treated with cottonseed-derived gossypol and bioactive extracts along with bacterial endotoxin lipopolysaccharides (LPS). SYBR Green qPCR was used to analyze the mRNA levels of a wide range of biomarkers involved in glucose transport, lipid biosynthesis, inflammatory response, and cancer development. qPCR data (10,560 Cq values) were generated from 55 genes analyzed from 64 treatments with triplicate per treatment for each gene. The data showed that B-cell lymphoma 2 ( Bcl2 ) mRNA was the most stable among the 55 mRNAs analyzed in the human colon cancer cells. Glyceraldehyde 3 phosphate dehydrogenase ( Gapdh ) and ribosome protein L32 ( Rpl32 ) mRNAs were not good qPCR references for the colon cancer cells. These observations were consistent regardless of the treatment comparison between gossypol and LPS, glanded and glandless seed extracts, seed coat and kernel extracts, or treatment for 8 and 24 h. These results suggest that Bcl2 is a preferable reference gene for qPCR assays in human colon cancer cells treated with cottonseed-derived gossypol and bioactive extracts as well as LPS. The extensive qPCR results firmly support the conclusion that the Bcl2 gene is stably expressed at the mRNA level in the human colon cancer cells regardless of ...
Keywords	bioactivity ; colon cancer cell ; cottonseed extract ; gene expression ; gossypol ; lipopolysaccharides ; Organic chemistry ; QD241-441
Subject code	610
Language	English
Publishing date	2022-11-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Identification of the major diacylglycerol acyltransferase mRNA in mouse adipocytes and macrophages.

Cao, Heping

BMC biochemistry

2018 Volume 19, Issue 1, Page(s) 11

Abstract: Background: Triacylglycerols (TAGs) are the major form of energy storage in eukaryotes. Diacylglycerol acyltransferases (DGATs) catalyze the final and rate-limiting step of TAG biosynthesis. Mammalian DGATs are classified into DGAT1 and DGAT2 ... ...

Abstract	Background: Triacylglycerols (TAGs) are the major form of energy storage in eukaryotes. Diacylglycerol acyltransferases (DGATs) catalyze the final and rate-limiting step of TAG biosynthesis. Mammalian DGATs are classified into DGAT1 and DGAT2 subfamilies. It was unclear which DGAT was the major isoform expressed in animal cells. The objective was to identify the major DGAT mRNA expressed in cultured mouse adipocytes and macrophages and compared it to that expressed in tung tree seeds. Methods: qPCR evaluated DGAT mRNA levels in mouse 3 T3-L1 adipocytes and RAW264.7 macrophages and tung tree seeds. Results: TaqMan qPCR showed that DGAT2 mRNA levels were 10-30 fold higher than DGAT1 in adipocytes and macrophages, and DGAT mRNA levels in adipocytes were 50-100-fold higher than those in macrophages. In contrast, the anti-inflammatory tristetraprolin/zinc finger protein 36 (TTP/ZFP36) mRNA levels were 2-4-fold higher in macrophages than those in adipocytes and similar to DGAT1 in adipocytes but 100-fold higher than DGAT1 in macrophages. SYBR Green qPCR analyses confirmed TaqMan qPCR results. DGAT2 mRNA as the major DGAT mRNA in the mouse cells was similar to that in tung tree seeds where DGAT2 mRNA levels were 10-20-fold higher than DGAT1 or DGAT3. Conclusion: The results demonstrated that DGAT2 mRNA was the major form of DGAT mRNA expressed in mouse adipocytes and macrophages and tung tree seeds.
MeSH term(s)	3T3-L1 Cells ; Adipocytes/cytology ; Adipocytes/drug effects ; Adipocytes/enzymology ; Aleurites/enzymology ; Animals ; Diacylglycerol O-Acyltransferase/genetics ; Diacylglycerol O-Acyltransferase/metabolism ; Dimethyl Sulfoxide/pharmacology ; Macrophages/cytology ; Macrophages/drug effects ; Macrophages/enzymology ; Mice ; Plant Proteins/genetics ; Plant Proteins/metabolism ; Protein Isoforms/genetics ; Protein Isoforms/metabolism ; RAW 264.7 Cells ; RNA Stability ; RNA, Messenger/metabolism ; Real-Time Polymerase Chain Reaction ; Seeds/enzymology
Chemical Substances	Plant Proteins ; Protein Isoforms ; RNA, Messenger ; Diacylglycerol O-Acyltransferase (EC 2.3.1.20) ; Dimethyl Sulfoxide (YOW8V9698H)
Language	English
Publishing date	2018-12-14
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2041216-2
ISSN	1471-2091 ; 1471-2091
ISSN (online)	1471-2091
ISSN	1471-2091
DOI	10.1186/s12858-018-0103-y
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Article ; Online: Regulation of Cell Viability and Anti-inflammatory Tristetraprolin Family Gene Expression in Mouse Macrophages by Cottonseed Extracts

Heping Cao / Kandan Sethumadhavan

Scientific Reports, Vol 10, Iss 1, Pp 1-

2020 Volume 11

Abstract: Abstract Bioactive plant extracts have been used for the prevention and treatment of various diseases. One of the major classes of bioactive compounds is plant polyphenols. Cottonseed ethanol extracts were determined by HPLC-MS analysis to be essentially ...

Abstract	Abstract Bioactive plant extracts have been used for the prevention and treatment of various diseases. One of the major classes of bioactive compounds is plant polyphenols. Cottonseed ethanol extracts were determined by HPLC-MS analysis to be essentially free of toxic gossypol. The objective of this study was to investigate the effect of cottonseed ethanol extracts on the cytotoxicity and regulation of anti-inflammatory tristrataprolin (TTP) family gene expression in mouse cells. MTT, qPCR and immunoblotting assays tested the effects of cottonseed extracts in mouse RAW264.7 macrophages and 3T3-L1 adipocytes. No cytotoxicity effect was observed in macrophages treated with extracts from the coat or kernel of glanded and glandless cottonseed. Similarly, the viability of mouse adipocytes was not affected by cottonseed extracts. In contrast, gossypol and lipopolysaccharides were toxic to macrophages but not adipocytes under high concentration or long time treatment. Cottonseed extracts exhibited modest effect on TTP family gene expression in macrophages but glandless cottonseed coat extract significantly increased TTP mRNA and protein levels with a magnitude similar to cinnamon and green tea polyphenol extract and insulin. These results demonstrated that cottonseed extracts are harmless towards the mouse cells and that glandless cottonseed coat extract stimulates TTP gene expression. We propose that glandless cottonseed is a safe source of plant polyphenols with anti-inflammatory property.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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