Effects of 13-alkyl-substituted berberine alkaloids on the expression of COX-II, TNF-alpha, iNOS, and IL-12 production in LPS-stimulated macrophages.
Berberine, a major alkaloidal component of Coptidis Rhizoma, has antibacterial activity, anti-inflammatory effect, antitumor and antimotility actions. We suggested that one of possible mechanisms of anti-bacterial activity of berberine may be based on the production of interleukin (IL)-12. Recently 13-alkyl-substituted berberines were shown to be better activity than berberine against certain bacteria species and human cancer cell lines. In the present study, therefore, the effects of 13-methylberberine (13-MB) and 13-ethylberberine (13-EB) on the production of IL-12 and expression of iNOS, TNF-alpha and COX-II were investigated using macrophages in culture. In LPS-stimulated RAW 264.7 cells, these alkaloids decreased the nitrites, concentration-dependently. The concentration of 50% inhibition of NO production (IC50) by 13-MB and 13-EB was 11.64 and 9.32 microM, respectively. The suppressed expression of iNOS protein was responsible for the reduction of NO production. Neither the expression of mRNA of iNOS, COX-II and TNF- alpha nor protein of COX-II and TNF-alpha was affected by both 13-MB and 13-EB, but production of PGE2 in LPS-stimulated RAW 264.7 cells was significantly reduced. Another striking finding of the present study is that 13-MB and 13-EB increased production of IL-12 in LPS-treated splenic macrophages. These results indicate that posttranscriptional regulatory mechanism of iNOS gene expression by 13-MB and 13-EB is involved, and COX-II activity is inhibited by 13-MB and 13-EB, respectively. In conclusion, the present study demonstrates that 13-methyl- and 13-ethylberberine alkaloids can be useful as an immunotherapeutic compound for induction of IL-12, which is potentially applicable for tumors, infectious disease, and airway inflammation.
Lee
Dong-Ung
DU
Department of Biochemistry, College of Natural Science, Dongguk University, Gyeongju 780-714, South Korea.
Kang
Young Jin
YJ
Park
Min Kyu
MK
Lee
Young Soo
YS
Seo
Han Geuk
HG
Kim
Tae Sung
TS
Kim
Cheorl-Ho
CH
Chang
Ki Churl
KC
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001600:Berberine Alkaloids / Q000494:pharmacology*; D002460:Cell Line; D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000096:biosynthesis; D004926:Escherichia coli; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D018664:Interleukin-12 / Q000096:biosynthesis*; D007527:Isoenzymes / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008811:Mice, Inbred DBA; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D013154:Spleen / Q000166:cytology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
D000975:Antioxidants; D001600:Berberine Alkaloids; D007527:Isoenzymes; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone