Human genome screen to identify the genetic basis of the anti-inflammatory effects of Boswellia in microvascular endothelial cells.
Inflammatory disorders represent a substantial health problem. Medicinal plants belonging to the Burseraceae family, including Boswellia, are especially known for their anti-inflammatory properties. The gum resin of Boswellia serrata contains boswellic acids, which inhibit leukotriene biosynthesis. A series of chronic inflammatory diseases are perpetuated by leukotrienes. Although Boswellia extract has proven to be anti-inflammatory in clinical trials, the underlying mechanisms remain to be characterized. TNF alpha represents one of the most widely recognized mediators of inflammation. One mechanism by which TNFalpha causes inflammation is by potently inducing the expression of adhesion molecules such as VCAM-1. We sought to test the genetic basis of the antiinflammatory effects of BE (standardized Boswellia extract, 5-Loxin) in a system of TNF alpha-induced gene expression in human microvascular endothelial cells. We conducted the first whole genome screen for TNF alpha- inducible genes in human microvascular cells (HMEC). Acutely, TNF alpha induced 522 genes and downregulated 141 genes in nine out of nine pairwise comparisons. Of the 522 genes induced by TNF alpha in HMEC, 113 genes were clearly sensitive to BE treatment. Such genes directly related to inflammation, cell adhesion, and proteolysis. The robust BE-sensitive candidate genes were then subjected to further processing for the identification of BE-sensitive signaling pathways. The use of resources such as GenMAPP, KEGG, and gene ontology led to the recognition of the primary BE-sensitive TNF alpha-inducible pathways. BE prevented the TNF alpha-induced expression of matrix metalloproteinases. BE also prevented the inducible expression of mediators of apoptosis. Most strikingly, however, TNF alpha-inducible expression of VCAM-1 and ICAM-1 were observed to be sensitive to BE. Realtime PCR studies showed that while TNF alpha potently induced VCAM-1 gene expression, BE completely prevented it. This result confirmed our microarray findings and built a compelling case for the anti-inflammatory property of BE. In an in vivo model of carrageenan-induced rat paw inflammation, we observed a significant antiinflammatory property of BE consistent with our in vitro findings. These findings warrant further research aimed at identifying the signaling mechanisms by which BE exerts its anti-inflammatory effects.
Roy
Sashwati
S
Laboratory of Molecular Medicine, Department of Surgery, The Ohio State University Medical Center, Columbus, Ohio 43210, USA.
Khanna
Savita
S
Shah
Hiral
H
Rink
Cameron
C
Phillips
Christina
C
Preuss
Harry
H
Subbaraju
Gottumukkala V
GV
Trimurtulu
Golakoti
G
Krishnaraju
Alluri V
AV
Bagchi
Manashi
M
Bagchi
Debasis
D
Sen
Chandan K
CK
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D029743:Boswellia / Q000737:chemistry*; D002351:Carrageenan / Q000633:toxicity; D017931:DNA Primers; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy / Q000473:pathology; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005528:Foot / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D005796:Genes / Q000187:drug effects* / Q000235:genetics; D015894:Genome, Human; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D015289:Leukotrienes / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010936:Plant Extracts / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use; D010991:Plethysmography; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D014315:Triterpenes / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D017931:DNA Primers; D015289:Leukotrienes; D010936:Plant Extracts; D014315:Triterpenes; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; C054625:boswellic acid; D002351:Carrageenan; D020782:Matrix Metalloproteinases