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Effects of HB-13 and HP-13 on NF-κB and p38MAPK Signaling Pathways

Author: JiangHui
Tutor: LiXinYu；ChangBaoZhu
School: Peking Union Medical College , China
Course: Dermatology and Venereology
Keywords: Hydrochloric acid 13 - hexyl berberine Hydrochloric acid, 13 - have Chiba A Martin Inflammatory dermatosis T lymphocytes Keratinocytes IL-8 IL-1β Tumor necrosis factor α IFN-γ Nuclear factor -kappa B p38 mitogen-activated protein kinase
CLC: R285
Type: Master's thesis
Year: 2009
Downloads: 130
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Abstract

Inflammation-related skin diseases account for more than half the incidence in dermatology to psoriasis, atopic dermatitis, as the representative. Of such diseases is a T lymphocyte-mediated immune disorders skin diseases, activated T lymphocytes, Th1-type cytokines (TNF-α, IFN-γ as the representative) are closely related. T cells and keratinocyte cells to secrete cytokines in the inflammatory skin disease complex cytokine network is formed, resulting in its unique immune pathological damage. Nuclear factor-kappa B (NF-κB) signaling pathway, and p38 mitogen original activated protein kinase (p38MAPK) abnormal inflammatory response of the signal transduction pathways are closely related, inflammatory skin disease pathogenesis and drug treatment targets hot spots. TNF-α and IFN-γ is important pathological factor in skin inflammation, keratinocytes can cause a chain reaction of intracellular protein kinase, activation of these two pathways involved in the occurrence and development of the regulation of the inflammatory response. Berberine series Ranunculaceae Coptis the plant Coptis rhizome extract active ingredients, the traditional antimicrobial, anti-inflammatory bioactive. Hydrochloric acid, 13 - hexyl of berberine (HB-13) is hydrochloric berberine (BBR) derivatives; palmatine (palmatine) from the yellow vines and other plants, hydrochloric acid, 13 - have Chiba A Martin (HP -13) palmatine hydrochloride derivatives. HB-13 and HP-13 the structure is very similar. Early studies have shown that the two compounds with the primary compound (berberine and Palmatine) are not available, or the strength exceeds the primary compound, the biological activity, such as antibacterial, anti-herpes virus and anti-inflammatory activity. In this study, HB-13 and HP-13 as the objective of the study compound, derived from normal human skin keratinocytes immortalized cell line HaCaT cells is formed as a research system discussed HB-13 and HP-13, TNF-α and / or the HaCaT cells NF-κB signaling pathway and p38MAPK signaling pathway activation caused by the impact of IFN-γ, and two compounds of T lymphocyte activation and cytokine expression function. The study results are divided into three aspects: first, the activation of T lymphocytes, HB-13 and HP-13 on keratinocyte cell IL-8 expression and expression of IL-1β mouse peritoneal macrophages vitro cultured murine spleen cells with concanavalin A (ConA) as a stimulant, HB-13 and HP-13 mouse T lymphocyte activation using the MTT assay; rhTNF-α-induced HaCaT cells, with ELISA, HB-13 and HP-13 rhTNF-α cells up-regulated expression of IL-8 level; mouse peritoneal macrophages by LPS stimulation, ELISA test both LPS-stimulated mouse peritoneal macrophages IL -1β expression. The results showed that (1) HB-13 from in 0.04875μg/ml to 0.39μg/ml, HP-13 within the concentration range of 0.78μg/ml ~ 3.12μg/ml inhibit T lymphocyte proliferation inhibition rates were 12.7% -78.2% 14.3% -92.5% inhibition is dose-dependent trend to the IC ₅₀ values ??to approximately 0.123μg/ml and 1.563μg/ml; (2) HB-13 and HP-13 in 0.39μg/ml ~ 3.12μg/ml experimental concentration range can significantly reduce by rhTNF-α raised the level of IL-8, IL-8 expression inhibition rate of about 85.7% -99.5% 68.4% -95.1%; (3) HB-13 expression in LPS-stimulated mouse peritoneal macrophage IL-1β inhibition rate of over 50% in the concentration range of 0.01μg/ml ~ 1μg/ml, inhibition dose - trend, IC ₅₀ value of approximately 0.013μg/ml. 1μg/ml HP-13 significantly inhibited LPS-stimulated mouse peritoneal macrophage IL-1β expression, inhibition rate of over 90%. These results suggest that HB-13 and HP-13, T lymphocyte activation and inflammatory cytokines IL-8 and IL-1β inhibit the expression. The second chapter of HB-13 and HP-13 pairs of human epidermal keratinocytes cells NF-κB signaling transduction pathways HaCaT cells cultured in vitro, exogenous rhTNF-α or rhIFN-γ-induced cells IκB-α protein phosphorylation, Western blot detection HB-13 and HP-13, TNF-α or IFN-γ-induced the HaCaT cells NF-κB activation. The results showed that (1) HB-13 and HP-13 in the concentration range of 0.39μg/ml ~ 1.56μg/ml inhibited rhTNF-α-induced expression of p-IκB-α, inhibition rates were 47.4% -67.5% and 32% -67.2% inhibition with a dose-dependent trend IC ₅₀ the value approximately 0.441μg/ml and 0.832μg/ml. (2) HB-13 and HP-13 rhIFN-γ stimulation caused by the expression of p-IκB-α also inhibited in 0.39μg/ml to 1.56μg/ml concentration range, the inhibition rates were 37.8% -57.4%, and 37.5% -43.6% inhibition was dose-dependent trend IC ₅₀ values ??were 0.923μg/ml and 4.556μg/ml. These results suggest that HaCaT keratinocytes HB-13 and HP-13 rhTNF-α rhlFN-γ, caused NF-κB activation was inhibited. The third chapter HB-13 and HP-13 in human epidermal keratinocytes cells p38MAPK signal transduction pathway in HaCaT cells cultured in vitro, exogenous rhTNF-α induced cell p38MAPK protein phosphorylation as a stimulant, the use of the Western blot detection of HB-13 and HP-13 the p38MAPK pathway activation of TNF-α-stimulated HaCaT cells. The results show that the expression of p-p38-HB-13 and HP-13 at 0.39μg/ml ~ 1.56μg/ml experimental concentration range of rhTNF-α stimulation caused no significant inhibition. The above results show that the HaCaT keratinocytes cells p38MAPK pathway activation-HB-13 and HP-13 of rhTNF-α caused no significant inhibition. The anti-inflammatory effects of the two compounds may be independent of p38MAPK signal transduction pathway.

Effects of HB-13 and HP-13 on NF-κB and p38MAPK Signaling Pathways

Abstract

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