基于PI3K/Akt/NF-κB信号通路探讨黄连素对特应性皮炎大鼠皮肤病理变化的治疗作用

doi:10.11958/20242370

摘要/Abstract

摘要：

目的基于磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）/核因子κB（NF-κB）信号通路探讨黄连素对特应性皮炎（AD）大鼠的治疗作用及其机制。方法将60只成年雄性Wistar大鼠随机均分为空白组（正常大鼠）、对照组（构建AD模型，50 mg/kg黄连素治疗）以及实验组（构建AD模型，200 mg/kg黄连素治疗）。干预1 d、7 d、14 d时，采用酶联免疫吸附试验检测白细胞介素（IL）-4、IL-13、肿瘤坏死因子（TNF）-α水平；Western blot检测PI3K、p-PI3K、Akt、p-Akt、NF-κB p65蛋白表达；HE染色观察大鼠皮肤组织病理变化。结果干预1 d、7 d、14 d时，对照组血清IL-4、IL-13、TNF-α水平及PI3K、p-PI3K、Akt、p-Akt、NF-κB p65蛋白表达水平均高于空白组（P<0.05）；干预7 d、14 d时，实验组上述指标水平均低于对照组（P<0.05）。干预14 d时，与空白组相比，对照组和实验组大鼠皮肤组织出现明显的病理改变，表皮层增厚，角质层过度角化，棘层肥厚，真皮层有大量炎性细胞浸润；实验组大鼠皮肤组织的病理损伤明显减轻。结论黄连素可通过抑制PI3K/Akt/NF-κB信号通路的激活降低AD大鼠血清炎性因子水平，减轻皮肤组织病理损伤。

关键词: 皮炎，特应性, 小檗碱, 磷酸肌醇3-激酶类, 原癌基因蛋白质c-akt, NF-κB, 细胞因子类, 皮肤组织

Abstract:

Objective To explore the therapeutic mechanism of berberine in atopic dermatitis (AD) rats based on PI3K/Akt/NF-kappa B signaling pathway. Methods Sixty adult male Wistar rats were randomly divided into the blank group (normal rats), the control group (AD model, 50 mg/kg berberine treatment) and the experimental group (AD model, 200 mg/kg berberine treatment), with 20 rats in each group. The levels of interleukin-4 (IL-4), interleukin-13 (IL-13) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA) at 1 d, 7 d and 14 d of intervention. The protein levels of PI3K, p-PI3K, Akt, p-Akt and NF-κB p65 were detected by Western blot assay. Pathological changes of rat skin tissue were analyzed by HE staining. Results After intervention for 1 d, 7 d and 14 d, serum levels of IL-4, IL-13, TNF-α and PI3K, p-PI3K, Akt, p-Akt and NF-kappa B p65 were higher in the control group than those in the blank group (P<0.05). After intervention for 7 d and 14 d, the levels of the above indicators were lower in the experimental group than those in the control group (P<0.05). After 14 days of intervention, compared with the blank group, the skin tissue of rats in the control group and the experimental group showed obvious pathological changes, including thickening of epidermis layer, excessive keratinization of the stratum corneum, thickening of spinous layer and a large infiltration of inflammatory cells in dermis. The pathological damage of rat skin tissue was significantly alleviated in the experimental group. Conclusion Berberine can inhibit the activation of PI3K/Akt/NF-kappa B signaling pathway, reduce serum level of inflammatory factors and reduce pathological damage of skin tissue in AD rats.

Key words: dermatitis, atopic, berberine, phosphatidylinositol 3-kinases, proto-oncogene proteins c-akt, NF-kappa B, cytokines, skin tissue

中图分类号:

R751

图/表 4

参考文献 20

[1]	中华医学会, 中华医学会杂志社, 中华医学会皮肤性病学分会, 等. 特应性皮炎基层诊疗指南(2022年)[J]. 中华全科医师杂志, 2022, 21(7):609-619.
	Chinese Medical Association, Chinese Medical Association Magazine, Chinese Medical Association Dermatology Branch, et al. Guidelines for primary diagnosis and treatment of atopic dermatitis (2022)[J]. Chinese Journal of General Practitioners, 2022, 21(7):609-619. doi:10.3760/cma.j.cn114798-20220215-00102.
[2]	马婉婷, 王清滢, 崔晗, 等. 基于"心为五脏六腑之大主"理论辨证论治特应性皮炎[J]. 世界中西医结合杂志, 2024, 19(4):835-839.
	MA W T, WANG Q Y, CUI H, et al. Syndrome differentiation and treatment of atopic dermatitis based on the theory that "the Heart is the Chief of the Five Zang - Organs and Six Fu - Organs"[J]. World Journal of Integrated Traditional and Western Medicine, 2024, 19(4):835-839. doi:10.13935/j.cnki.sjzx.240435.
[3]	牛蔚露, 陈恋恋, 冯敏, 等. 三种中药单体体外对特应性皮炎优势菌群活性的影响[J]. 时珍国医国药, 2022, 33(2):345-348.
	NIU W L, CHEN L L, FENG M, et al. Effects of three traditional Chinese medicine monomers on the activity of dominant flora in atopic dermatitis in vitro[J]. LiShiZhen Medicine and Materia Medica Research, 2022, 33(2):345-348. doi:10.3969/j.issn.1008-0805.2022.02.23.
[4]	LI L, JIANG W, YU B, et al. Quercetin improves cerebral ischemia/reperfusion injury by promoting microglia/macrophages M2 polarization via regulating PI3K/Akt/NF-κB signaling pathway[J]. Biomed Pharmacother, 2023,168:115653. doi:10.1016/j.biopha.2023.115653.
[5]	秦宗碧, 徐爱琴, 蔡翔, 等. 黄连素调节PI3K/AKT/NF-κB信号通路对慢性湿疹大鼠皮肤损伤的影响[J]. 天津医药, 2023, 51(8):834-840.
	QIN Z B, XU A Q, CAI X, et al. Effects of berberine on skin lesions in rats with chronic eczema by regulating PI3K/AKT/NF-kappa B signaling pathway[J]. Tianjin Med J, 2023, 51(8):834-840. doi:10.11958/20221627.
[6]	汤舒玲, 黎晓红, 段亚菊, 等. 葫芦巴碱调节PI3K/Akt/NF-κB信号通路对变应性接触性皮炎大鼠免疫反应的影响[J]. 河北医药, 2023, 45(21):3211-3216.
	TANG S L, LI X H, DUAN Y J, et al. Effects of cucurbitine on immune response in rats with allergic contact dermatitis by regulating PI3K/Akt/NF-kappa B signaling pathway[J]. Hebei Medical Journal, 2023, 45(21):3211-3216. doi:10.3969/j.issn.1002-7386.2023.21.002.
[7]	陈娟平, 彭圆, 洪学敏, 等. 基于PI3K/Akt/Nrf2信号通路探讨天王补心丹加减对睡眠剥夺小鼠皮肤影响的作用机制[J]. 中国实验方剂学杂志, 2024, 30(11):120-128.
	CHEN J P, PENG Y, HONG X M, et al. To explore the mechanism of the effect of Tianwang Buxin Dan on the skin of sleep-deprived mice based on PI3K/Akt/Nrf2 signaling pathway[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2024, 30(11):120-128. doi:10.13422/j.cnki.syfjx.20240505.
[8]	崔梦迪, 高加巍, 朱吕群, 等. 基于SIRT1和PI3K/Akt信号通路探讨黄芪甲苷对慢性难愈性创面大鼠模型修复愈合的影响及作用机制[J]. 中国实验方剂学杂志, 2024, 30(6):101-108.
	CUI M D, GAO J W, ZHU L Q, et al. To explore the effect and mechanism of astragaloside A on the repair and healing of chronic refractory wounds in a rat model based on SIRT1 and PI3K/Akt signaling pathways[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2024, 30(6):101-108. doi:10.13422/j.cnki.syfjx.20240116.
[9]	LIU J, JIA S, YANG Y, et al. Exercise induced meteorin-like protects chondrocytes against inflammation and pyroptosis in osteoarthritis by inhibiting PI3K/Akt/NF-κB and NLRP3/caspase-1/GSDMD signaling[J]. Biomed Pharmacother, 2023,158:114118. doi:10.1016/j.biopha.2022.114118.
[10]	杨雅, 包海燕, 康莹莹, 等. 基于网络药理学及实验验证探讨薰衣草对皮肤光损伤的防护作用机制[J]. 中国实验方剂学杂志, 2022, 28(8):167-174.
	YANG Y, BAO H Y, KANG Y Y, et al. To explore the protective mechanism of lavender on skin photodamage based on network pharmacology and experimental verification[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2022, 28(8):167-174. doi:10.13422/j.cnki.syfjx.20220514.
[11]	TIAN Y, LIU B, LI Y, et al. Activation of RARα receptor attenuates neuroinflammation after SAH via promoting M1-to-M2 phenotypic polarization of microglia and regulating Mafb/Msr1/PI3K-Akt/NF-κB pathway[J]. Front Immunol, 2022,13:839796. doi:10.3389/fimmu.2022.839796.
[12]	程可, 刘晓, 林尽染, 等. 小檗碱在皮肤疾病应用中的研究进展[J]. 中国临床药学杂志, 2023, 32(4):316-320.
	CHENG K, LIU X, LIN J R, et al. Research progress of berberine in the application of skin diseases[J]. Chinese Journal of Clinical Pharmacy, 2023, 32(4):316-320. doi:10.19577/j.1007-4406.2023.04.016.
[13]	QU J, LI J, ZHANG Y, et al. AKR1B10 promotes breast cancer cell proliferation and migration via the PI3K/AKT/NF-κB signaling pathway[J]. Cell Biosci, 2021, 11(1):163. doi:10.1186/s13578-021-00677-3.
[14]	徐进, 隆康健, 钱英明, 等. 黄连素液对产ESBLs大肠埃希菌感染创面的愈合作用及抗耐药机制[J]. 中华医院感染学杂志, 2023, 33(8):1216-1220.
	XU J, LONG K J, QIAN Y M, et al. Healing effect and anti-drug resistance mechanism of berberine solution on ESBLs-producing Escherichia coli infection wounds[J]. Chinese Journal of Hospital Infectious Diseases, 2023, 33(8):1216-1220. doi:10.11816/cn.ni.2023-221354.
[15]	GONG Y, QIU J, JIANG T, et al. Maltol ameliorates intervertebral disc degeneration through inhibiting PI3K/AKT/NF-κB pathway and regulating NLRP3 inflammasome-mediated pyroptosis[J]. Inflammopharmacology, 2023, 31(1):369-384. doi:10.1007/s10787-022-01098-5.
[16]	杨宇琦, 吴丽娜, 孙克, 等. 黄连素通过重塑肠道菌群和调节PI3K/AKT信号通路改善多囊卵巢综合征小鼠的作用机制研究[J]. 疑难病杂志, 2024, 23(3):352-359.
	YANG Y Q, WU L N, SUN K, et al. Study on the mechanism of berberine improving polycystic ovary syndrome mice by remodeling intestinal flora and regulating PI3K/AKT signaling pathway[J]. Chinese Journal of Difficult and Complicated Cases, 2024, 23(3):352-359. doi:10.3969/j.issn.1671-6450.2024.03.018.
[17]	崔玉娟, 赵辉, 苏东霞, 等. 黄连素下调NRAV和PI3K/AKT通路减轻RSV感染致HEp-2细胞的损伤[J]. 中国药理学通报, 2024, 40(4):747-755.
	CUI Y J, ZHAO H, SU D X, et al. Berberine downregulates NRAV and PI3K/AKT pathway to attenuate the damage of HEp-2 cells induced by RSV infection[J]. Chinese Pharmacological Bulletin, 2024, 40(4):747-755. doi:10.12360/CPB202312017.
[18]	李春霖, 李时超, 贾英田, 等. 基于JAK/STAT信号通路探究黄连素对溃疡性结肠炎小鼠结肠皮细胞凋亡作用[J]. 中国药理学通报, 2023, 39(5):938-945.
	LI C L, LI S C, JIA Y T, et al. Exploring the effect of berberine on apoptosis of colonic epithelial cells in mice with ulcerative colitis based on JAK/STAT signaling pathway[J]. Chinese Pharmacological Bulletin, 2023, 39(5):938-945. doi:10.12360/CPB202210070.
[19]	SUN Y, ZHANG H, WU Z, et al. Quercitrin rapidly alleviated depression-like behaviors in lipopolysaccharide-treated mice:The involvement of PI3K/AKT/NF-κB signaling suppression and CREB/BDNF signaling restoration in the hippocampus[J]. ACS Chem Neurosci, 2021, 12(18):3387-3396. doi:10.1021/acschemneuro.1c00371.
[20]	LI C, GONG L, JIANG Y, et al. Sanguisorba officinalis ethyl acetate extract attenuates ulcerative colitis through inhibiting PI3K-AKT/NF-κB/STAT3 pathway uncovered by single-cell RNA sequencing[J]. Phytomedicine, 2023,120:155052. doi:10.1016/j.phymed.2023.155052.

组别	IL-4			IL-13			TNF-α
组别	1 d	7 d	14 d	1 d	7 d	14 d	1 d	7 d	14 d
空白组	15.36±4.31	14.39±5.22	15.68±4.07	18.35±4.96	18.64±5.01	19.31±6.32	38.42±5.62	37.45±6.06	38.25±7.21
对照组	73.82±6.35^a	56.31±8.45^a	50.28±7.24^a	69.72±10.45^a	45.36±9.72^a	39.71±4.46^a	115.35±13.45^a	78.74±9.37^a	70.46±7.42^a
实验组	74.21±7.06^a	35.44±9.27^ab	33.25±5.71^ab	70.01±9.83^a	38.45±8.31^ab	31.45±4.38^ab	116.82±14.01^a	69.42±8.31^ab	48.31±6.45^ab

组别	IL-4			IL-13			TNF-α
组别	1 d	7 d	14 d	1 d	7 d	14 d	1 d	7 d	14 d
空白组	15.36±4.31	14.39±5.22	15.68±4.07	18.35±4.96	18.64±5.01	19.31±6.32	38.42±5.62	37.45±6.06	38.25±7.21
对照组	73.82±6.35^a	56.31±8.45^a	50.28±7.24^a	69.72±10.45^a	45.36±9.72^a	39.71±4.46^a	115.35±13.45^a	78.74±9.37^a	70.46±7.42^a
实验组	74.21±7.06^a	35.44±9.27^ab	33.25±5.71^ab	70.01±9.83^a	38.45±8.31^ab	31.45±4.38^ab	116.82±14.01^a	69.42±8.31^ab	48.31±6.45^ab

组别	PI3K			p-PI3K
组别	1 d	7 d	14 d	1 d	7 d	14 d
空白组	0.83±0.12	0.80±0.25	0.79±0.13	0.45±0.09	0.44±0.10	0.46±0.08
对照组	1.73±0.20^a	1.45±0.31^a	1.31±0.20^a	0.96±0.21^a	0.73±0.09^a	0.69±0.07^a
实验组	1.75±0.18^a	0.98±0.15^ab	0.82±0.05^b	0.98±0.25^a	0.67±0.03^ab	0.50±0.25^b
组别	Akt			p-Akt			NF-κB p65
组别	1 d	7 d	14 d	1 d	7 d	14 d	1 d	7 d	14 d
空白组	0.72±0.11	0.73±0.09	0.71±0.12	0.25±0.07	0.28±0.03	0.19±0.05	0.62±0.13	0.67±0.08	0.65±0.09
对照组	1.41±0.23^a	1.02±0.17^a	0.98±0.20^a	0.91±0.25^a	0.76±0.20^a	0.68±0.14^a	1.62±0.13^a	1.23±0.10^a	1.02±0.07^a
实验组	1.43±0.25^a	0.76±0.45^b	0.81±0.10^ab	0.93±0.26^a	0.54±0.10^ab	0.31±0.06^ab	1.61±0.15^a	0.79±0.21^ab	0.71±0.14^b

组别	PI3K			p-PI3K
组别	1 d	7 d	14 d	1 d	7 d	14 d
空白组	0.83±0.12	0.80±0.25	0.79±0.13	0.45±0.09	0.44±0.10	0.46±0.08
对照组	1.73±0.20^a	1.45±0.31^a	1.31±0.20^a	0.96±0.21^a	0.73±0.09^a	0.69±0.07^a
实验组	1.75±0.18^a	0.98±0.15^ab	0.82±0.05^b	0.98±0.25^a	0.67±0.03^ab	0.50±0.25^b
组别	Akt			p-Akt			NF-κB p65
组别	1 d	7 d	14 d	1 d	7 d	14 d	1 d	7 d	14 d
空白组	0.72±0.11	0.73±0.09	0.71±0.12	0.25±0.07	0.28±0.03	0.19±0.05	0.62±0.13	0.67±0.08	0.65±0.09
对照组	1.41±0.23^a	1.02±0.17^a	0.98±0.20^a	0.91±0.25^a	0.76±0.20^a	0.68±0.14^a	1.62±0.13^a	1.23±0.10^a	1.02±0.07^a
实验组	1.43±0.25^a	0.76±0.45^b	0.81±0.10^ab	0.93±0.26^a	0.54±0.10^ab	0.31±0.06^ab	1.61±0.15^a	0.79±0.21^ab	0.71±0.14^b