Effect of baicalin regulating cAMP/PKA/CREB signaling pathway on skin barrier function in eczema rats

doi:10.11958/20230416

Abstract

Abstract:

Objective To investigate the effect of baicalin (BA) regulating cyclic adenosine phosphate (cAMP)/protein kinase A(PKA)/cAMP response elemen-binding protein (CREB) pathway on skin barrier function in eczema rats. Methods SD rats were randomly divided into the control group (NC group), the model group, the low-dose BA group (BA-L group, 25 mg/kg), the medium-dose BA group (BA-M group, 50 mg/kg), the high-dose BA group (BA-H group, 100 mg/kg), the prednisone group (PNS group, 25 mg/kg), the BA-H+cAMP inhibitor (SQ22536) group (100 mg/kg+2.13 mg/kg) and the BA-H+PKA inhibitor (H-89) group (100 mg/kg+5 mg/kg), 12 animals in each group. Except for the NC group, eczema rat model was constructed in the other groups. Two days after successful modeling, drug administration was performed in groups. Changes of eczema area and severity index (EASI) score, transcutaneous water loss (TEWL) and cuticle water content (WCSC) were detected. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of immunoglobulin E (IgE), interferon-γ (IFN-γ) and interleukin-4 (IL-4) in rat serum and the expression of cAMP protein in rat back lesions. HE staining was used to detect pathological changes of skin lesions on the back of rats. Western blot assay was used to detect aquaporin 3 (AQP3), cathelicidin related antimicrobial peptide (CRAMP), p-PKA, p-CREB protein expression in rat back lesions. Results Compared with the NC group, rats had serious pathological lesions on the back of the tested area, increased EASI score, TEWL, IgE and IL-4 levels, and decreased WCSC, IFN-γ, AQP3, CRAMP, cAMP, p-PKA and p-CREB protein levels in the model group (P < 0.05). Compared with the model group, pathological lesion of the tested area in the back of rats was relieved, and EASI score, TEWL, IgE and IL-4 levels were decreased, WCSC, IFN-γ levels, AQP3, CRAMP, cAMP, p-PKA and p-CREB protein were increased in the BA-L group, the BA-M group, the BA-H group and the PNS group (P<0.05). Changes of above indexes in the BA-L group, the BA-M group, the BA-H group were dose-dependent. SQ22536 or H-89 attenuated the improvement effect of high dose BA on skin barrier function in eczema rats. Conclusion BA may improve skin barrier function in eczema rats by activating cAMP/PKA/CREB signaling pathway.

Key words: eczema, baicalin, cyclic adenosine monophosphate/protein kinase A/cyclic-AMP response binding protein pathway, skin barrier function

CLC Number:

R285.5

Figures/Tables 5

References 22

[1]	BOYLE R J, SHAMJI M H. Aetiology and prevention of eczema[J]. Clin Exper Allergy, 2021, 51(3):380-381. doi:10.1111/cea.13626.
[2]	GALL C. Causes and symptoms of atopic eczema[J]. J Allergy Ther, 2021, 12(3):1.
[3]	陈兴, 黄丽霞, 李忻红, 等. 中药外治湿疹用药规律的数据挖掘研究[J]. 辽宁中医杂志, 2021, 48(9):86-90.
	CHEN X, HUANG L X, LI X H, et al. Data mining research on the law of external treatment of eczema with traditional Chinese medicine[J]. Liaoning J Trad Chin Med, 2021, 48(9):86-90. doi:10.13192/j.issn.1000-1719.2021.09.023.
[4]	BOHANNON M, LIU M, NADEAU P, et al. Topical doxycycline monohydrate hydrogel 1% targeting proteases/PAR2 pathway is a novel therapeutic for atopic dermatitis[J]. Exp Dermatol, 2020, 29(12):1171-1175. doi:10.1111/exd.14201.
[5]	彭清, 孙柳, 苗雪圆, 等. 天然产物化合物作为肿瘤免疫调节剂的研究进展[J]. 中国中西医结合外科杂志, 2021, 27(1):136-140.
	PENG Q, SUN L, MIAO X Y, et al. Research progress of natural product compounds as tumor immunomodulators[J]. Chinese Journal of Surgery of Integrated Traditional and Western Medicine, 2021, 27(1):136-140. doi:10.3969/j.issn.1007-6948.2021.01.027.
[6]	WANG L, XIAN Y F, LOO S K F, et al. Baicalin ameliorates 2,4-dinitrochlorobenzene-induced atopic dermatitis-like skin lesions in mice through modulating skin barrier function,gut microbiota and JAK/STAT pathway[J]. Bioorg Chem, 2022, 119:105538. doi:10.1016/j.bioorg.2021.105538.
[7]	TSAI Y F, CHEN C Y, LIN I W, et al. Imperatorin alleviates psoriasiform dermatitis by blocking neutrophil respiratory burst,adhesion,and chemotaxis through selective phosphodiesterase 4 inhibition[J]. Antioxid Redox Signal, 2021, 35(11):885-903. doi:10.1089/ars.2019.7835.
[8]	王永慧, 王思农, 肖静秋, 等. 藏药二十五味儿茶凝胶对大鼠慢性湿疹模型LTB-4、LTC-4因子的影响[J]. 中国皮肤性病学杂志, 2018, 32(5):568-570.
	WANG Y H, WANG S N, XIAO J Q, et al. Effects of Tibetan Drug Ershiwuweiercha Gel on factors ltb-4 and ltc-4 in model rats of chronic eczema[J]. Chin J Dermatology and Venereology, 2018, 32(5):568-570. doi:10.13735/j.cjdv.1001-7089.201711042.
[9]	FANG F, XIE Z, QUAN J, et al. Baicalin suppresses Propionibacterium acnes-induced skin inflammation by downregulating the NF-κB/MAPK signaling pathway and inhibiting activation of NLRP3 inflammasome[J]. Braz J Med Biol Res, 2020, 53(12):e9949. doi:10.1590/1414-431X20209949.
[10]	汪园园, 金明华, 黄颖娟, 等. 苓桂术甘汤联合热量限摄对湿疹模型大鼠皮损的影响及其机制分析[J]. 中国中西医结合杂志, 2020, 40(4):465-469.
	WANG Y Y, JIN M H, HUANG Y J, et al. Effect of calorie-restriction therapy combined Zhugan Decoction on eczema model rats[J]. Chinese Journal of Integrated Traditional and Western Medicine, 2020, 40(4):465-469. doi:10.7661/j.cjim.20190928.286.
[11]	汪雪峰, 陈锋, 宋顺德, 等. 腺苷酸环化酶抑制剂和激动剂在脂多糖诱导的急性肺损伤中的作用[J]. 中国药理学通报, 2017, 33(10):1410-1414.
	WANG X F, CHEN F, SONG S D, et al. Effect of adenylate cyclase antagonists andagonist in acute lung injury induced by lipopolysaccharide[J]. Chinese Pharmacological Bulletin, 2017, 33(10):1410-1414. doi:10.3969/j.issn.1001-1978.2017.10.016.
[12]	崇显瑾, 杨历新. 利拉鲁肽通过cAMP/PKA/CREB通路干预2型糖尿病大鼠骨质疏松的机制研究[J]. 中国病理生理杂志, 2021, 37(11):1949-1956.
	CHONG X J, YANG L X. Mechanism of liraglutide intervening osteoporosis in type 2 diabetic rats through cAMP/PKA/CREB signaling pathways[J]. Chinese Journal of Pathophysiology, 2021, 37(11):1949-1956. doi:10.3969/j.issn.1000-4718.2021.11.004.
[13]	AIERKEN K, LUO Y, MAIWULANJIANG M, et al. The suppressive effect of mamiran cream on atopic dermatitis-like skin lesions in vivo[J]. Evid Based Complement Alternat Med, 2021, 2021(1):2854238-2854249. doi:10.1155/2021/2854238.
[14]	汪秀梅, 武亦阁, 贺乙, 等. 针灸对湿热蕴肤型湿疹豚鼠皮肤屏障功能的影响[J]. 中国针灸, 2022, 42(7):779-784,810.
	WANG X M, WU Y G, HE Y, et al. Effect of acupuncture and moxibustion on skin barrier function in guinea pigs with eczema of skin damp-heat accumulation[J]. Chinese Journal of Acupuncture and Moxibustion, 2022, 42(7):779-784,810. doi:10.13703/j.0255-2930.20210619-k0001.
[15]	BOLLAG W B, AITKENS L, WHITE J, et al. Aquaporin-3 in the epidermis:more than skin deep[J]. Am J Physiol Cell Physiol, 2020, 318(6):C1144-C1153. doi:10.1152/ajpcell.00075.2020.
[16]	吴百威, 羊小华, 占润之. 甘草酸对特应性皮炎小鼠皮肤屏障功能的影响[J]. 中国临床药理学杂志, 2020, 36(10):1324-1327.
	WU B W, YANG X H, ZHAN R Z. Effects of glycyrrhizic acid on skin barrier function in mice with atopic dermatitis[J]. Chinese Journal of Clinical Pharmacology, 2020, 36(10):1324-1327. doi:10.13699/j.cnki.1001-6821.2020.10.042.
[17]	WANG X, XIE L, LONG J, et al. Therapeutic effect of baicalin on inflammatory bowel disease:A review[J]. J Ethnopharmacol, 2022, 283:114749. doi:10.1016/j.jep.2021.114749.
[18]	SHERWANI M A, YANG K, JANI A, et al. Protective effect of baicalin against TLR4-mediated UVA-induced skin inflammation[J]. Photochem Photobiol, 2019, 95(2):605-611. doi:10.1111/php.13021.
[19]	HUNG C H, WANG C N, CHENG H H, et al. Baicalin ameliorates imiquimod-induced psoriasis-like inflammation in mice[J]. Planta Med, 2018, 84(15):1110-1117. doi:10.1055/a-0622-8242.
[20]	MA X, HAO M, ZHANG M H, et al. Integrated strategy of network analysis prediction and experimental validation to elucidate the possible mechanism of compound Turkish gall ointment in treating eczema[J]. Chin Med, 2022, 17:89-109. doi:10.1186/s13020-022-00643-2.
[21]	WU J, GAN Y, LUO H, et al. β-Patchoulene ameliorates water transport and the mucus barrier in 5-fluorouracil-induced intestinal mucositis rats via the cAMP/PKA/CREB signaling pathway[J]. Front Pharmacol, 2021, 12:689491-689503. doi:10.3389/fphar.2021.689491.
[22]	YUAN X H, YAO C, OH J H, et al. Vasoactive intestinal peptide stimulates melanogenesis in B16F10 mouse melanoma cells via CREB/MITF/tyrosinase signaling[J]. Biochem Biophys Res Commun, 2016, 477(3):336-342. doi:10.1016/j.bbrc.2016.06.105.

组别	EASI评分/分	TEWL/ （g·m^-2·h^-1）	WCSC/%
NC组	0	8.68±0.37	63.57±3.11
Model组	7.33±0.32^a	46.52±1.89^a	18.86±0.82^a
BA-L组	6.84±0.15^b	40.14±1.57^b	25.57±1.13^b
BA-M组	6.11±0.10^bc	32.28±1.23^bc	34.43±1.66^bc
BA-H组	3.32±0.09^bcd	17.73±0.82^bcd	46.68±2.23^bcd
PNS组	3.30±0.10^bcd	17.69±0.75^bcd	46.73±2.19^bcd
BA-H+SQ22536组	6.58±0.12^e	35.52±1.63^e	30.36±1.47^e
BA-H+H-89组	6.54±0.11^e	35.48±1.57^e	30.45±1.45^e
F	3 384.000^＊	1 139.000^＊	698.800^＊

组别	EASI评分/分	TEWL/ （g·m^-2·h^-1）	WCSC/%
NC组	0	8.68±0.37	63.57±3.11
Model组	7.33±0.32^a	46.52±1.89^a	18.86±0.82^a
BA-L组	6.84±0.15^b	40.14±1.57^b	25.57±1.13^b
BA-M组	6.11±0.10^bc	32.28±1.23^bc	34.43±1.66^bc
BA-H组	3.32±0.09^bcd	17.73±0.82^bcd	46.68±2.23^bcd
PNS组	3.30±0.10^bcd	17.69±0.75^bcd	46.73±2.19^bcd
BA-H+SQ22536组	6.58±0.12^e	35.52±1.63^e	30.36±1.47^e
BA-H+H-89组	6.54±0.11^e	35.48±1.57^e	30.45±1.45^e
F	3 384.000^＊	1 139.000^＊	698.800^＊

组别	IgE/（mg/L）	IFN-γ/（ng/L）	IL-4/（ng/L）
NC组	112.28±5.44	63.32±2.79	38.67±1.73
Model组	283.36±12.25^a	24.42±1.13^a	84.45±3.83^a
BA-L组	243.43±10.20^b	33.58±1.42^b	72.29±2.97^b
BA-M组	200.12±8.45^bc	42.28±1.87^bc	61.33±2.41^bc
BA-H组	134.66±5.45^bcd	54.47±2.05^bcd	46.67±1.72^bcd
PNS组	132.28±5.61^bcd	54.36±1.98^bcd	47.72±1.66^bcd
BA-H+SQ22536组	223.37±10.54^e	35.57±1.62^e	68.87±2.69^e
BA-H+H-89组	220.69±10.31^e	35.63±1.59^e	69.03±2.58^e
F	563.800^＊	599.500^＊	445.000^＊

组别	IgE/（mg/L）	IFN-γ/（ng/L）	IL-4/（ng/L）
NC组	112.28±5.44	63.32±2.79	38.67±1.73
Model组	283.36±12.25^a	24.42±1.13^a	84.45±3.83^a
BA-L组	243.43±10.20^b	33.58±1.42^b	72.29±2.97^b
BA-M组	200.12±8.45^bc	42.28±1.87^bc	61.33±2.41^bc
BA-H组	134.66±5.45^bcd	54.47±2.05^bcd	46.67±1.72^bcd
PNS组	132.28±5.61^bcd	54.36±1.98^bcd	47.72±1.66^bcd
BA-H+SQ22536组	223.37±10.54^e	35.57±1.62^e	68.87±2.69^e
BA-H+H-89组	220.69±10.31^e	35.63±1.59^e	69.03±2.58^e
F	563.800^＊	599.500^＊	445.000^＊

组别	cAMP/（nmol/L）	AQP3/β-actin	CRAMP/β-actin	p-PKA/PKA	p-CREB/CREB
NC组	13.36±0.54	1.56±0.12	1.68±0.13	0.94±0.03	0.92±0.08
Model组	4.67±0.23^a	0.42±0.03^a	0.72±0.05^a	0.23±0.02^a	0.41±0.03^a
BA-L组	5.89±0.26^b	0.59±0.05^b	0.93±0.07^b	0.36±0.02^b	0.52±0.04^b
BA-M组	7.34±0.28^bc	0.82±0.07^bc	1.14±0.11^bc	0.49±0.05^bc	0.60±0.05^bc
BA-H组	11.32±0.43^bcd	1.28±0.11^bcd	1.36±0.12^bcd	0.82±0.07^bcd	0.84±0.08^bcd
PNS组	11.36±0.45^bcd	1.30±0.12^bcd	1.38±0.13^bcd	0.84±0.07^bcd	0.86±0.07^bcd
BA-H+SQ22536组	6.64±0.37^e	0.73±0.06^e	1.03±0.09^e	0.42±0.03^e	0.54±0.04^e
BA-H+H-89组	11.40±0.41	0.75±0.06^e	1.06±0.11^e	0.44±0.03^e	0.56±0.05^e
F	420.100^＊	135.700^＊	49.350^＊	207.300^＊	63.960^＊