穿心莲内酯调节cGAS-STING信号通路对银屑病小鼠的治疗作用

doi:10.11958/20230914

天津医药 ›› 2024, Vol. 52 ›› Issue (4): 379-386.doi: 10.11958/20230914

穿心莲内酯调节cGAS-STING信号通路对银屑病小鼠的治疗作用

何亚男(), 蔡翔^△(), 邱百怡, 孙邦梅, 李伶华

武汉市中医医院皮肤科（邮编430050）

收稿日期:2023-06-16 修回日期:2023-11-01 出版日期:2024-04-15 发布日期:2024-04-19
通讯作者: ^△E-mail：caixiang1983c@163.com
作者简介:何亚男（1992），女，住院医师，主要从事皮肤病的中医药治疗方面研究。E-mail：rfmi47q6@163.com
基金资助:
武汉市中医药科研项目(WZ22C75);武汉市中医药科研项目(WZ22C72)

Study on the therapeutic effect of andrographolide on psoriasis mouse model by regulating cGAS-STING signal pathway

HE Yanan(), CAI Xiang^△(), QIU Baiyi, SUN Bangmei, LI Linghua

Department of Dermatology, Wuhan Traditional Chinese Medicine Hospital, Wuhan 430050, China

Received:2023-06-16 Revised:2023-11-01 Published:2024-04-15 Online:2024-04-19
Contact: ^△E-mail：caixiang1983c@163.com

何亚男, 蔡翔, 邱百怡, 孙邦梅, 李伶华. 穿心莲内酯调节cGAS-STING信号通路对银屑病小鼠的治疗作用[J]. 天津医药, 2024, 52(4): 379-386.

HE Yanan, CAI Xiang, QIU Baiyi, SUN Bangmei, LI Linghua. Study on the therapeutic effect of andrographolide on psoriasis mouse model by regulating cGAS-STING signal pathway[J]. Tianjin Medical Journal, 2024, 52(4): 379-386.

摘要/Abstract

摘要：

目的探究穿心莲内酯（Andro）调节GMP-AMP合酶-干扰素基因刺激物（cGAS-STING）信号通路对银屑病小鼠的治疗作用及机制。方法 90只BALB/c小鼠分为对照组（Control组），模型组（Model组），穿心莲内酯低、中、高剂量组（Andro-L、M、H组，10、30、50 mg•kg^-1•d^-1 Andro）和穿心莲内酯高剂量+STING激活剂DMXAA组（Andro-H+DMXAA组，50 mg•kg^-1•d^-1 Andro+5 mg•kg^-1•d^-1 DMXAA）。除Control组外，其余组小鼠背部施用咪喹莫特（IMQ）建立银屑病小鼠模型。观察小鼠银屑病面积并进行严重程度指数（PASI）评分，酶联免疫吸附试验（ELISA）检测血清白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α、IL-1β、IL-23、IL-17A、干扰素（IFN）-γ和IFN-β水平，苏木素-伊红（HE）染色和甲苯胺蓝染色测定表皮厚度和肥大细胞数，免疫荧光染色检测Ki-67、CD4阳性细胞表达率，RT-qPCR检测cGAS和STING mRNA表达水平，免疫印迹实验检测cGAS、STING、干扰素调节因子3（IRF3）、p-IRF3蛋白水平。结果与Control组相比，Model组小鼠背部皮肤出现严重的红斑、鳞屑，有大量的炎性细胞浸润；抓挠次数、PASI评分、表皮厚度、肥大细胞数量、IL-6、IL-1β、TNF-α、IL-23、IL-17A、IFN-γ和IFN-β水平、Ki-67和CD4阳性表达率、cGAS和STING mRNA和蛋白表达水平及IRF3磷酸化水平升高（P<0.05）。与Model组相比，Andro-L组、Andro-M组和Andro-H组小鼠皮肤红斑、鳞屑、炎性细胞浸润现象减轻；抓挠次数、PASI评分、表皮厚度、肥大细胞数量、IL-6、IL-1β、TNF-α、IL-23、IL-17A、IFN-γ和IFN-β水平、Ki-67和CD4阳性表达率、cGAS和STING mRNA和蛋白表达水平及IRF3磷酸化水平呈剂量依赖性降低（P<0.05）；STING激活剂DMXAA逆转了穿心莲内酯对银屑病小鼠的保护作用（P<0.05）。结论穿心莲内酯可通过抑制cGAS-STING信号通路减轻炎症反应，改善小鼠银屑病症状。

关键词: 穿心莲内酯, 银屑病, 炎症, GMP-AMP合酶-干扰素基因刺激物信号通路

Abstract:

Objective To explore the therapeutic mechanism of andrographolide (Andro) on psoriasis mice by regulating GMP-AMP synthase-stimulator of interferon genes signal pathway (cGAS STING signal pathway). Methods Ninety BALB/c mice were divided into the control group, the model group, the Andro low dose group, the Andro medium dose group, the Andro high dose groups (Andro-L, M, H groups, 10, 30 and 50 mg•kg^-1•d^-1Andro) and the high dose Andro+STING activator DMXAA group (Andro-H+DMXAA group, 50 mg•kg^-1•d^-1 Andro+5 mg•kg^-1•d^-1 DMXAA). Except for the control group, mice of the other groups were administered imiquimod (IMQ) on back to establish the psoriasis mouse model. The psoriasis area of mice was observed, and severity index (PASI) scoring was performed. Serum levels of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-23, IL-17A, interferon (IFN)-γ and IFN-β were detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin eosin (HE) staining and toluidine blue staining were used to detect the epidermal thickness and mast cell count. Immunofluorescence staining was applied to detect the expression rates of Ki-67 and CD4 positive cells. RT-qPCR was applied to detect the expression level of cGAS and STING mRNA, and Western blot assay was applied to detect levels of cGAS, STING, interferon regulatory factor-3 (IRF3) and p-IRF3 proteins. Results Compared with the control group, the back skin of mice in the model group showed severe erythema, scales and a large number of inflammatory cell infiltration. The scratch times, PASI score, epidermal thickness, number of mast cells, levels of IL-6, IL-1β, TNF-α, IL-23, IL-17A, IFN-γ and IFN-β, the expression rates of Ki-67 and CD4 positive cell, the expression level of cGAS and STING mRNA and protein, and IRF3 phosphorylation level were increased obviously (P<0.05). Compared with the model group, skin erythema, scales and inflammatory cell infiltration were reduced in the Andro-L, Andro-M and Andro-H groups. The scratch times, PASI score, epidermal thickness, number of mast cells, levels of IL-6, IL-1β, TNF-α, IL-23, IL-17A, IFN-γ and IFN-β, the expression rates of Ki-67 and CD4 positive cells, the expression levels of cGAS and STING mRNA and protein, and IRF3 phosphorylation level were decreased in a dose-dependent manner (P<0.05). STING activator DMXAA reversed the protective effect of Andro on psoriasis mice (P<0.05). Conclusion Andrographolide can improve psoriasis symptoms in mice by inhibiting the cGAS-STING signaling pathway.

Key words: Andrographolide, psoriasis, inflammation, GMP-AMP synthase-stimulator of interferon genes signaling pathway

中图分类号:

R758.63

图/表 12

参考文献 27

[1]	YIN Q, SUN L, CAI X, et al. Lidocaine ameliorates psoriasis by obstructing pathogenic CGRP signaling-mediated sensory neuron-dendritic cell communication[J]. J Invest Dermatol, 2022, 142(8):2173-2183.e6. doi:10.1016/j.jid.2022.01.002.
[2]	BURGOS R A, ALARCÓN P, QUIROGA J, et al. Andrographolide, an anti-inflammatory multitarget drug:all roads lead to cellular metabolism[J]. Molecules, 2020, 26(1):5. doi:10.3390/molecules26010005.
[3]	MUSSARD E, JOUSSELIN S, CESARO A, et al. Andrographis paniculata and its bioactive diterpenoids protect dermal fibroblasts against inflammation and oxidative stress[J]. Antioxidants, 2020, 9(5):432. doi:10.3390/antiox9050432.
[4]	LI C X, LI H G, ZHANG H, et al. Andrographolide suppresses thymic stromal lymphopoietin in phorbol myristate acetate/calcium ionophore A23187-activated mast cells and 2,4-dinitrofluorobenzene-induced atopic dermatitis-like mice model[J]. Drug Des Devel Ther, 2016, 10:781-791. doi:10.2147/dddt.s94056.
[5]	DECOUT A, KATZ J D, VENKATRAMAN S, et al. The cGAS-STING pathway as a therapeutic target in inflammatory diseases[J]. Nat Rev Immunol, 2021, 21(9):548-569. doi:10.1038/s41577-021-00524-z.
[6]	HOPFNER K P, HORNUNG V. Molecular mechanisms and cellular functions of cGAS-STING signalling[J]. Nat Rev Mol Cell Biol, 2020, 21(9):501-521. doi:10.1038/s41580-020-0244-x.
[7]	HUANG C, LI W, REN X, et al. The crucial roles and research advances of cGAS-STING pathway in cutaneous disorders[J]. Inflammation, 2023, 46(4):1161-1176. doi:10.1007/s10753-023-01812-7.
[8]	杨梅, 唐彩红, 桂雨晴, 等. 咪喹莫特诱导银屑病模型小鼠T细胞免疫功能改变与性别的关系[J]. 中国药理学通报, 2022, 38(4):589-597.
	YANG M, TANG C H, GUI Y Q, et al. The relationship between T cell immune function changes and gender in psoriasis model mice induced by imiquimod[J]. Chinese Pharmacological Bulletin, 2022, 38(4):589-597. doi:10.12360/CPB202108021.
[9]	YU Y, XUE X, TANG W, et al. Cytosolic DNA-mediated STING-dependent inflammation contributes to the progression of psoriasis[J]. J Invest Dermatol, 2022, 142(3 Pt B):898-906.e4. doi:10.1016/j.jid.2021.08.430.
[10]	CHEN Y, YAN Y, LIU H, et al. Dihydroartemisinin ameliorates psoriatic skin inflammation and its relapse by diminishing CD8⁺ T-cell memory in wild-type and humanized mice[J]. Theranostics, 2020, 10(23):10466-10482. doi:10.7150/thno.45211.
[11]	GANGWAR R S, GUDJONSSON J E, WARD N L. Mouse models of psoriasis:a comprehensive review[J]. J Invest Dermatol, 2022, 142(3PtB):884-897. doi:10.1016/j.jid.2021.06.019.
[12]	KAMIYA K, KISHIMOTO M, SUGAI J, et al. Risk factors for the development of psoriasis[J]. Int J Mol Sci, 2019, 20(18):4347. doi:10.3390/ijms20184347.
[13]	XU H, LIU J, NIU M, et al. Soluble IL-6R-mediated IL-6 trans-signaling activation contributes to the pathological development of psoriasis[J]. J Mol Med (Berl), 2021, 99(7):1009-1020. doi:10.1007/s00109-021-02073-3.
[14]	ZHANG L J. Type1 interferons potential initiating factors linking skin wounds with psoriasis pathogenesis[J]. Front Immunol, 2019, 10:1440. doi:10.3389/fimmu.2019.01440.
[15]	INDIRAPRIYADARSHINI R, KANIMOZHI G, NATARAJAN D, et al. Andrographolide protects acute ultraviolet-B radiation-induced photodamages in the mouse skin[J]. Arch Dermatol Res, 2023, 315(5):1197-1205. doi:10.1007/s00403-022-02504-2.
[16]	朱玉婷, 晏文, 应理晟, 等. 川芎嗪对银屑病小鼠皮损内TNF-α,IL-17,VEGF表达的影响[J]. 天津医药, 2023, 51(6):590-595.
	ZHU Y T, YAN W, YING L S, et al. Effects of tetramethylpyrazine on expression levels of TNF-α,IL-17 and VEGF in skin lesions of psoriatic mice[J]. Tianjin Med J, 2019, 51(6):590-595. doi:10.11958/20221433.
[17]	陈若玺, 郑胜, 郭春燕, 等. 黄芪多糖对咪喹莫特诱导小鼠银屑病样皮炎的干预作用及其机制[J]. 中国应用生理学杂志, 2022, 38(2):154-159.
	CHEN R X, ZHENG S, GUO C Y, et al. The intervention effect and mechanism of Astragalus polysaccharides on imiquimod induced psoriasis like dermatitis in mice[J]. Chinese Journal of Applied Physiology, 2022, 38(2):154-159. doi:10.12047/j.cjap.6214.2022.022.
[18]	RAMEZANI M, SHAMSHIRI A, ZAVATTARO E, et al. Immunohistochemical expression of P53, Ki-67,and CD34 in psoriasis and psoriasiform dermatitis[J]. Biomedicine(Taipei), 2019, 9(4):26. doi:10.1051/bmdcn/2019090426.
[19]	CHIRICOZZI A, ROMANELLI P, VOLPE E, et al. Scanning the immunopathogenesis of psoriasis[J]. Int J Mol Sci, 2018, 19(1):179-209. doi:10.3390/ijms19010179.
[20]	JIANG B W, ZHANG W J, WANG Y, et al. Convallatoxin induces HaCaT cell necroptosis and ameliorates skin lesions in psoriasis-like mouse models[J]. Biomed Pharmacother, 2020, 121:109615. doi:10.1016/j.biopha.2019.109615.
[21]	ÖZCAN A, SAHIN D, IMPELLIZZIERI D, et al. Nanoparticle-coupled topical methotrexate can normalize immune responses and induce tissue remodeling in psoriasis[J]. J Invest Dermatol, 2020, 140(5):1003-1014.e8. doi:10.1016/j.jid.2019.09.018.
[22]	HUANG Z Z, XU Y, XU M, et al. Artesunate alleviates imiquimod-induced psoriasis-like dermatitis in BALB/c mice[J]. Int Immunopharmacol, 2019, 75:105817. doi:10.1016/j.intimp.2019.105817.
[23]	BECK M A, FISCHER H, GRABNER L M, et al. DNA hypomethylation leads to cGAS-induced autoinflammation in the epidermis[J]. EMBO J, 2021, 40(22):e108234. doi:10.15252/embj.2021108234.
[24]	ERGUN S L, LI L. Structural insights into STING signaling[J]. Trends Cell Biol, 2020, 30(5):399-407. doi:10.1016/j.tcb.2020.01.010.
[25]	PAN Y, YOU Y, SUN L, et al. The STING antagonist H-151 ameliorates psoriasis via suppression of STING/NFκB-mediated inflammation[J]. Br J Pharmacol, 2021, 178(24):4907-4922. doi:10.1111/bph.15673.
[26]	LI N, ZHOU H, WU H, et al. STING-IRF3 contributes to lipopolysaccharide-induced cardiac dysfunction, inflammation, apoptosis and pyroptosis by activating NLRP3[J]. Redox Biol, 2019, 24:101215. doi:10.1016/j.redox.2019.101215.
[27]	LI X H, ZHANG Z T, YU Y J, et al. Activation of the STING-IRF3 pathway involved in psoriasis with diabetes mellitus[J]. J Cell Mol Med, 2022, 26(8):2139-2151. doi:10.1111/jcmm.17236.

组别	抓挠次数/次	PASI评分/分
Control组	5.16±0.61	0
Model组	29.43±4.06^a	2.93±0.46
Andro-L组	23.74±3.81^b	2.47±0.50^b
Andro-M组	16.52±2.04^bc	1.93±0.26^bc
Andro-H组	10.76±1.61^bcd	0.73±0.46^bcd
Andro-H+DMXAA组	13.42±1.74^e	1.40±0.50^e
F	170.859^＊＊	56.872^＊＊

组别	抓挠次数/次	PASI评分/分
Control组	5.16±0.61	0
Model组	29.43±4.06^a	2.93±0.46
Andro-L组	23.74±3.81^b	2.47±0.50^b
Andro-M组	16.52±2.04^bc	1.93±0.26^bc
Andro-H组	10.76±1.61^bcd	0.73±0.46^bcd
Andro-H+DMXAA组	13.42±1.74^e	1.40±0.50^e
F	170.859^＊＊	56.872^＊＊

组别	IL-6	IL-1β	TNF-α	IL-23	IL-17A
Control组	86.36±7.03	54.16±6.89	80.43±8.16	33.86±3.05	152.23±16.08
Model组	159.76±12.46^a	117.24±12.03^a	246.48±21.31^a	101.62±9.11^a	326.06±28.57^a
Andro-L组	143.86±10.35^b	103.66±7.43^b	183.35±16.25^b	87.38±7.12^b	275.24±24.46^b
Andro-M组	127.71±9.82^bc	94.26±8.51^bc	129.52±11.07^bc	62.10±6.08^bc	244.18±20.31^bc
Andro-H组	103.52±9.21^bcd	72.31±6.56^bcd	98.34±9.76^bcd	43.91±5.07^bcd	187.09±19.17^bcd
Andro-H+DMXAA组	115.38±10.94^e	83.19±8.01^e	113.71±10.92^e	51.24±5.10^e	208.85±19.22^e
F	105.412^＊＊	107.583^＊＊	309.341^＊＊	267.107^＊＊	126.150^＊＊

组别	IL-6	IL-1β	TNF-α	IL-23	IL-17A
Control组	86.36±7.03	54.16±6.89	80.43±8.16	33.86±3.05	152.23±16.08
Model组	159.76±12.46^a	117.24±12.03^a	246.48±21.31^a	101.62±9.11^a	326.06±28.57^a
Andro-L组	143.86±10.35^b	103.66±7.43^b	183.35±16.25^b	87.38±7.12^b	275.24±24.46^b
Andro-M组	127.71±9.82^bc	94.26±8.51^bc	129.52±11.07^bc	62.10±6.08^bc	244.18±20.31^bc
Andro-H组	103.52±9.21^bcd	72.31±6.56^bcd	98.34±9.76^bcd	43.91±5.07^bcd	187.09±19.17^bcd
Andro-H+DMXAA组	115.38±10.94^e	83.19±8.01^e	113.71±10.92^e	51.24±5.10^e	208.85±19.22^e
F	105.412^＊＊	107.583^＊＊	309.341^＊＊	267.107^＊＊	126.150^＊＊

组别	IFN-γ	IFN-β
Control组	50.43±6.07	16.46±1.53
Model组	174.58±18.03^a	89.14±9.76^a
Andro-L组	137.16±13.54^b	64.53±6.01^b
Andro-M组	102.15±9.43^bc	43.68±4.96^bc
Andro-H组	67.28±7.11^bcd	23.72±2.11^bcd
Andro-H+DMXAA组	83.49±7.07^e	34.68±2.94^e
F	262.388^＊＊	389.008^＊＊

穿心莲内酯调节cGAS-STING信号通路对银屑病小鼠的治疗作用

Study on the therapeutic effect of andrographolide on psoriasis mouse model by regulating cGAS-STING signal pathway

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 12

参考文献 27

相关文章 15

编辑推荐

Metrics

本文评价

组别	表皮厚度/μm	肥大细胞数量/个
Control组	6.25±0.23	4.31±0.37
Model组	15.12±1.04^a	57.64±4.89^a
Andro-L组	13.48±0.83^b	42.17±4.31^b
Andro-M组	11.62±0.86^bc	28.39±3.04^bc
Andro-H组	8.31±0.32^bcd	11.75±1.67^bcd
Andro-H+DMXAA组	10.83±0.61^e	17.96±2.06^e
F	105.306^＊＊	203.736^＊＊

组别	Ki-67阳性表达率	CD4阳性表达率
Control组	9.43±1.03	5.23±0.74
Model组	48.67±3.84^a	27.96±3.01^a
Andro-L组	36.86±4.03^b	20.52±2.14^b
Andro-M组	27.74±2.42^bc	16.83±1.36^bc
Andro-H组	16.32±1.67^bcd	9.45±1.01^bcd
Andro-H+DMXAA组	22.87±2.15^e	12.89±1.21^e
F	133.356^＊＊	107.263^＊＊

组别	cGAS	STING
Control组	1.00±0.06	1.00±0.04
Model组	3.07±0.26^a	4.75±0.31^a
Andro-L组	2.34±0.21^b	3.81±0.24^b
Andro-M组	1.52±0.13^bc	3.06±0.25^bc
Andro-H组	1.18±0.10^bcd	1.43±0.12^bcd
Andro-H+DMXAA组	1.87±0.15^e	2.72±0.18^e
F	109.537^＊＊	227.060^＊＊

组别	cGAS	STING	p-IRF3/IRF3
Control组	0.41±0.04	0.21±0.01	0.12±0.01
Model组	1.34±0.15^a	0.91±0.08^a	0.67±0.06^a
Andro-L组	1.16±0.11^b	0.72±0.04^b	0.52±0.04^b
Andro-M组	0.95±0.08^bc	0.57±0.05^bc	0.38±0.03^bc
Andro-H组	0.55±0.05^bcd	0.38±0.02^bcd	0.20±0.02^bcd
Andro-H+DMXAA组	0.70±0.04^e	0.47±0.04^e	0.27±0.02^e
F	83.805^＊＊	147.397^＊＊	182.914^＊＊

[1]	陈晶晶, 农章嵩, 谭良源, 杨培培, 梁英业, 唐宏亮, 王开龙. 小胶质细胞极化在神经病理性疼痛中作用研究进展[J]. 天津医药, 2024, 52(9): 1000-1003.
[2]	范慧慧, 任玉梅, 田新磊, 张凯, 李晓丽. 止咳平喘方对支气管哮喘小鼠气道炎症及TLR4/TRAF6/NF-κB通路的影响[J]. 天津医药, 2024, 52(9): 924-929.
[3]	贾维宁, 鲍亚玲, 雷慧, 殷晓宁. 夏枯草提取物对脓毒症小鼠炎症反应和腹腔巨噬细胞的影响[J]. 天津医药, 2024, 52(9): 930-935.
[4]	吴波, 朱卓农, 郑丽娟, 陈俊如. 苦参碱对特应性皮炎炎症、氧化应激和伤口愈合的影响[J]. 天津医药, 2024, 52(6): 566-571.
[5]	袁满, 冯子瀚, 谢敏, 王柏军. 大黄素对骨关节炎模型小鼠痛觉行为的调节机制[J]. 天津医药, 2024, 52(6): 572-577.
[6]	蒋韬, 程红艳, 吴琼. 棕矢车菊素调节SDF-1α/CXCR4信号通路对妊娠糖尿病大鼠炎症反应的影响[J]. 天津医药, 2024, 52(6): 594-598.
[7]	霍晶辰, 王悦, 李华, 邱嵘, 苏景伟, 王卓凡, 杨洁. 系统免疫炎症指数对根治性放疗Ⅲ期肺鳞癌患者长期生存的预测价值[J]. 天津医药, 2024, 52(6): 634-638.
[8]	慕静然, 骆延, 梁璇, 徐陶, 曾俊伟, 刘晓红. 补体系统激活参与阿尔茨海默病的研究进展[J]. 天津医药, 2024, 52(6): 663-668.
[9]	韩正怡, 李锐, 陈齐, 王家友, 盛奎, 宋洁, 张野. 收肌管阻滞联合全麻对老年全膝关节置换术患者术后疼痛和认知功能的影响[J]. 天津医药, 2024, 52(5): 523-527.
[10]	吕梦娜, 李建斌, 吴锐. 自身炎症性疾病患者合并COVID-19严重程度的早期预测指标探讨[J]. 天津医药, 2024, 52(5): 528-531.
[11]	黎敏, 龚坚, 吴伟伟, 刘巧. Nrf2/HO-1通路在银屑病中作用的研究进展[J]. 天津医药, 2024, 52(5): 552-556.
[12]	贾西瑞, 刘莉洁. 小胶质细胞在脓毒症相关性脑病中的作用及研究进展[J]. 天津医药, 2024, 52(5): 557-560.
[13]	吴倩, 王意, 陈念, 周凯, 田昕, 徐晖, 苟小霞. 诱导化疗对鼻咽癌患者免疫功能及炎症指标的影响[J]. 天津医药, 2024, 52(4): 397-402.
[14]	肖雨倩, 孙可心, 万俊, 陈淑颖, 陈丽敏, 王岩, 白艳杰. RNA m6A甲基化在卒中后认知障碍中的研究进展[J]. 天津医药, 2024, 52(3): 331-336.
[15]	易娜, 肖雯, 田源, 袁李礼. BMAL1减轻H₂O₂诱导的心肌细胞损伤机制研究[J]. 天津医药, 2024, 52(2): 119-123.