Objective: To investigate the effects of Chinese herbal monomer shikonin which has the function of cooling blood and detoxification on abnormally activated T lymphocyte and its related signal pathways and to elucidate...Objective: To investigate the effects of Chinese herbal monomer shikonin which has the function of cooling blood and detoxification on abnormally activated T lymphocyte and its related signal pathways and to elucidate the role of shikonin in the pathological state of psoriasis and its mechanism of treating psoriasis. Materials and Methods: Jurkat E6?1 T lymphocytes were activated with phorbol ester and ionomycin. The shikonin concentration of 0.5–2 μg/mL was applied to the cells, and the proliferation of T lymphocytes was detected by Cell Counting Kit?8 assay. Flow cytometry was used to measure CD69 expression on the cell membrane and intracellular free calcium ion concentration([Ca^(2+)]i); enzyme?linked immunosorbent assay was used to detect the levels of interleukin(IL)?2, interferon?γ(IFN?γ), and(TNF?α) released by activated T lymphocytes; quantitative polymerase chain reaction and Western blot were used to observe the expression of nuclear transcription factor m RNA and protein, respectively. Results: Different concentrations of shikonin could significantly inhibit cell proliferation, CD69 expression, and secretion of Th1 cytokines. In addition, shikonin could effectively reduce the [Ca^(2+)]i and protein kinase C phosphorylation proteins. Besides that, shikonin could significantly reduce the nuclear transcription factor nuclear factor of activated T lymphocytes mRNA expression, downregulate c?Jun mRNA and protein expression, and inhibit NF?κB protein expression. All the above indicators show a certain dose–effect relationship. Conclusions: Shikonin can exert immune regulation by inhibiting the function of overactivated T lymphocytes. Hence, this study provides experimental basis for the mechanism and application prospect of shikonin in the treatment of psoriasis.展开更多
基金supported by the Natural Science Foundation of China for“Study on the Molecular Mechanism of Stat3 MediatedγδT‑KC Interaction by Yangxue Jiedu Decoction Based on Psoriasis of Blood Dryness Syndrome with‘Mutual Transformation of Dryness and Dampness,and Internal Generation of Dampness Toxin’”(No.81573974)“Study on the Effect of Yangxue Jiedu Decoction on the Mechanism of Microvascular Regression in Psoriasis of Blood Dryness Syndrome Based on Survivin/PI3K/βcatenin Pathway”(No.81673974)+1 种基金and Beijing Municipal Natural Science Foundation for“Study on the Effect of Nourishing Blood and Detoxification on the Blood Vessel of the Psoriasis of Blood Dryness Syndrome Based on Survivin/Tcf/Lef Pathway”(No.7172097)the Project of Beijing Hospital of TCM for“Study on mechanism of vascular inflammation attenuated by Linagxue Jiedu Fang in psoriasis via S‑nitrosoglutathione NF‑kB(No.YJ‑201821).
文摘Objective: To investigate the effects of Chinese herbal monomer shikonin which has the function of cooling blood and detoxification on abnormally activated T lymphocyte and its related signal pathways and to elucidate the role of shikonin in the pathological state of psoriasis and its mechanism of treating psoriasis. Materials and Methods: Jurkat E6?1 T lymphocytes were activated with phorbol ester and ionomycin. The shikonin concentration of 0.5–2 μg/mL was applied to the cells, and the proliferation of T lymphocytes was detected by Cell Counting Kit?8 assay. Flow cytometry was used to measure CD69 expression on the cell membrane and intracellular free calcium ion concentration([Ca^(2+)]i); enzyme?linked immunosorbent assay was used to detect the levels of interleukin(IL)?2, interferon?γ(IFN?γ), and(TNF?α) released by activated T lymphocytes; quantitative polymerase chain reaction and Western blot were used to observe the expression of nuclear transcription factor m RNA and protein, respectively. Results: Different concentrations of shikonin could significantly inhibit cell proliferation, CD69 expression, and secretion of Th1 cytokines. In addition, shikonin could effectively reduce the [Ca^(2+)]i and protein kinase C phosphorylation proteins. Besides that, shikonin could significantly reduce the nuclear transcription factor nuclear factor of activated T lymphocytes mRNA expression, downregulate c?Jun mRNA and protein expression, and inhibit NF?κB protein expression. All the above indicators show a certain dose–effect relationship. Conclusions: Shikonin can exert immune regulation by inhibiting the function of overactivated T lymphocytes. Hence, this study provides experimental basis for the mechanism and application prospect of shikonin in the treatment of psoriasis.
