Effects of Xiebai San on the Morphological Structures of Lung and Intestinal Tissues and Expression Levels of PI3K and Akt in Rats with Allergic Asthma

doi:10.12300/j.issn.1674-5817.2025.084

Abstract

Abstract:

Objective To investigate the mechanism by which Xiebai San regulates respiratory tract and intestinal mucosal immunity in rats with allergic asthma. Methods Forty male SD rats were randomly divided into four groups based on body weight: control group, model group, positive control group, and Xiebai San group. The model group, positive control group, and Xiebai San group were sensitized with ovalbumin to establish a rat model of allergic asthma. From day 21 (the aerosol challenge phase), each group received daily gavage interventions simultaneously: the positive control group was administered dexamethasone (0.068 mg/kg), the Xiebai San group received Xiebai San solution (2 g/mL, 11.3 mL/kg), while the control and model groups were given an equal volume of normal saline, once daily for 14 consecutive days. After euthanasia, lung and intestinal tissues were collected. Hematoxylin and eosin staining was used to observe histopathological changes. Transmission electron microscopy was employed to examine tissue ultrastructure. Immunohistochemistry was applied to detect the positive reaction areas of phosphatidyl-inositol 3-kinase (PI3K) and protein kinase B (Akt) proteins. Total protein and total RNA were extracted from lung and intestinal tissues, then the protein and mRNA expression levels of PI3K and Akt genes were detected by Western blotting and real-time quantitative PCR, respectively. Results Histopathological results showed alveolar emphysema accompanied by inflammatory cell infiltration, and intestinal mucosal injury with inflammatory cell infiltration in the model group as compared with the control group; the cellular structure of lung tissues was disrupted in the model group, with reduced organelles, while the ultrastructural lesions in the intestine were relatively mild. Compared with the model group, Xiebai San group exhibited milder pathological changes in lung tissues, with occasional alveolar wall damage and a small amount of inflammatory cell infiltration; the intestinal mucosal structure was improved, glands were arranged regularly, and pathological changes such as tissue loosening and inflammatory infiltration were alleviated; the cellular structure of lung tissues was relatively intact with reduced severity of lesions, and no ultrastructural pathological changes were observed in intestinal tissues. Immunohistochemistry and Western blotting results showed that compared with the control group, the specific positive reaction areas of PI3K and Akt in lung and intestinal tissues were significantly increased in the model group (all P<0.001); meanwhile, the protein expression levels of PI3K and Akt were significantly upregulated (all P<0.05). Compared with the model group, the positive area of Akt protein in lung tissue was significantly reduced in the Xiebai San group (P<0.001), and the positive area of PI3K in intestinal tissue was also significantly decreased (P<0.000 1). Additionally, the protein expression levels of PI3K and Akt in lung and intestinal tissues were significantly downregulated (all P<0.01). Real-time quantitative PCR results showed that compared with the control group, the mRNA expression levels of PI3K and Akt genes in lung and intestinal tissues were significantly elevated in the model group (all P<0.05). Compared with the model group, the mRNA expression levels of PI3K and Akt genes in lung and intestinal tissues were significantly reduced in the Xiebai San group (all P<0.05). Conclusion Xiebai San exerts protective effects on rats with allergic asthma by inhibiting the expression of key nucleic acids and proteins in the PI3K-Akt signaling pathway in lung and intestinal tissues, improving the morphological structure of lung tissue, and maintaining intestinal mucosal integrity, and regulating intestinal mucosal immune function.

Key words: Xiebai San, Asthma, Mucosal immunity, PI3K-Akt signaling pathway, Rats

CLC Number:

Q95-33

SONG Jing,YANG Zongtong,LI Xiaojing,et al. Effects of Xiebai San on the Morphological Structures of Lung and Intestinal Tissues and Expression Levels of PI3K and Akt in Rats with Allergic Asthma[J]. Laboratory Animal and Comparative Medicine, 2026, 46(2): 191-204. DOI: 10.12300/j.issn.1674-5817.2025.084.

Figures/Tables 9

References 30

[1]	任雪交, 黄巍, 邱菲菲, 等. 支气管哮喘动物模型研究的文献分析与评价[J]. 实验动物与比较医学, 2022, 42(1): 74-80. DOI:10.12300/j.issn.1674-5817.2021.044 .
	REN X J, HUANG W, QIU F F, et al. Progress in animal models for bronchial asthma[J]. Lab Anim Comp Med, 2022, 42(1): 74-80. DOI:10.12300/j.issn.1674-5817.2021.044 .
[2]	罗世雄, 张赛, 陈慧. 常见哮喘动物模型的建立方法与评价研究进展[J]. 实验动物与比较医学, 2025, 45(2): 167-175. DOI:10.12300/j.issn.1674-5817.2024.120 .
	LUO S X, ZHANG S, CHEN H. Research progress in establishment and evaluation of common asthma animal models[J]. Lab Anim Comp Med, 2025, 45(2): 167-175. DOI:10.12300/j.issn.1674-5817.2024.120 .
[3]	兰露莎, 赵兵兵, 杨红宇, 等. 氧化苦参碱对小鼠哮喘模型保护作用的初步研究[J]. 实验动物与比较医学, 2017, 37(4): 320-323. DOI:10.3969/j.issn.1674-5817.2017.04.013 .
	LAN L S, ZHAO B B, YANG H Y, et al. The protective effect of oxymatrine on asthma mice[J]. Lab Anim Comp Med, 2017, 37(4): 320-323. DOI:10.3969/j.issn.1674-5817.2017.04.013 .
[4]	AN J, LIU Y Q, WANG Y Q, et al. The role of intestinal mucosal barrier in autoimmune disease: a potential target[J]. Front Immunol, 2022, 13:871713. DOI:10.3389/fimmu.2022.871713 .
[5]	CERVANTES-GARCÍA D, JIMÉNEZ M, RIVAS-SANTIAGO C E, et al. Lactococcus lactis NZ9000 prevents asthmatic airway inflammation and remodelling in rats through the improvement of intestinal barrier function and systemic TGF-β production[J]. Int Arch Allergy Immunol, 2021, 182(4):277-291. DOI:10.1159/000511146 .
[6]	洪天一. 蝎黄解痉治哮颗粒影响NF-κB和HIF-1α信号传导通路治疗哮喘的调控机制[D]. 长春: 长春中医药大学, 2020. DOI: 10.26980/d.cnki.gcczc.2020.000459 .
	HONG T Y. Regulatory mechanism of Xiehuang Jiezhuan Zhixiao granules on the NF-κB and HIF-1α signaling pathways in the treatment of asthma[D]. Changchun: Changchun University of Chinese Medicine, 2020. DOI: 10.26980/d.cnki.gcczc.2020.000459 .
[7]	马兰兰, 陈玲, 王琴, 等. 肺通气功能正常的儿童支气管哮喘控制情况及急性发作随访的研究[J]. 中国当代儿科杂志, 2024, 26(5): 476-480. DOI:10.7499/j.issn.1008-8830.2311149 .
	MA L L, CHEN L, WANG Q, et al. Control status and follow-up of acute attacks in children with bronchial asthma with normal pulmonary ventilation function[J]. Chin J Contemp Pediatr, 2024, 26(5): 476-480. DOI:10.7499/j.issn.1008-8830.2311149 .
[8]	丁云录, 郑明昱, 南敏伦, 等. 基于ERK信号通路探讨鹿茸大补汤颗粒对哮喘缓解期豚鼠的调控及作用机制[J]. 中国老年学杂志, 2023, 43(21):5309-5313. DOI:10.3969/j.issn.1005-9202.2023.21.054 .
	DING Y L, ZHENG M Y, NAN M L, et al. Based on ERK signal pathway, this paper discusses the regulation and mechanism of Lulong Dabutang Granule on guinea pigs with asthma in remission stage[J]. Chin J Gerontol, 2023, 43(21):5309-5313. DOI:10.3969/j.issn.1005-9202.2023.21.054 .
[9]	李玉丽, 易腾达, 谭志强, 等. 经典名方泻白散的源流及古今应用考究[J]. 中国实验方剂学杂志, 2021, 27(4):168-174. DOI:10.13422/j.cnki.syfjx.20202328 .
	LI Y L, YI T D, TAN Z Q, et al. Literature research on origin and application of classical prescription xiebaisan[J]. Chin J Exp Tradit Med Formulae, 2021, 27(4):168-174. DOI:10.13422/j.cnki.syfjx.20202328 .
[10]	刘凌志, 梁敏兰, 郭栋伟. 黄芩泻白散对支气管哮喘小鼠的治疗作用及Wnt/β-catenin信号轴的影响[J]. 湖南中医杂志, 2025, 41(6):140-146. DOI:10.16808/j.cnki.issn1003-7705.2025.06.028 .
	LIU L Z, LIANG M L, GUO D W. Therapeutic effect of Huangqin Xiebai powder on rats with bronchial asthma and its effect on the Wnt/β-catenin signaling axis[J]. Hunan J Tradit Chin Med, 2025, 41(6):140-146. DOI:10.16808/j.cnki.issn1003-7705.2025.06.028 .
[11]	张天柱, 张景龙, 樊湘泽, 等. 泻白散对小鼠过敏性哮喘气道炎症的作用及机制[J]. 中国实验方剂学杂志, 2014, 20(20):173-177. DOI:10.13422/j.cnki.syfjx.2014200173 .
	ZHANG T Z, ZHANG J L, FAN X Z, et al. Effects and its mechanism of Xiebai San on allergic airway inflammation in asthma mouse[J]. Chin J Exp Tradit Med Formulae, 2014, 20(20):173-177. DOI:10.13422/j.cnki.syfjx.2014200173 .
[12]	JUTEL M, MOSNAIM G S, BERNSTEIN J A, et al. The One Health approach for allergic diseases and asthma[J]. Allergy, 2023, 78(7):1777-1793. DOI:10.1111/all.15755 .
[13]	JOHNSON C C, OWNBY D R. The infant gut bacterial microbiota and risk of pediatric asthma and allergic diseases[J]. Transl Res, 2017, 179:60-70. DOI:10.1016/j.trsl.2016.06.010 .
[14]	杨阳, 张桂信, 杨琦, 等. 从肠-肺轴相关微环境角度诠释中医"肺与大肠相表里"的新内涵[J]. 中国中西医结合外科杂志, 2025, 31(02): 295-299. DOI:10.3969/j.issn.1007-6948.2025.02.027 .
	YANG Y, ZHANG G X, YANG Q, et al. Interpretation of the new connotation of "lung and large intestine are exterior and interior" in traditional Chinese medicine from the perspective of microenvironment related to intestine-lung axis[J]. Chin J Surg Integr Tradit West Med, 2025, 31(2): 295-299. DOI:10.3969/j.issn.1007-6948.2025.02.027 .
[15]	刘寻, 周婷婷, 朱紫冰, 等. 槟榔碱通过上调PI3K/Akt/mTOR信号通路促进口腔黏膜下纤维化的体内外实验研究[J]. 中国临床药理学与治疗学, 2025, 30(07): 865-875. DOI:10.12092/j.issn.1009-2501.2025.07.001 .
	LIU X, ZHOU T T, ZHU Z B, et al. Arecoline promotes oral submucous fibrosis by upregulating PI3K/Akt/mTOR signaling pathway in vivo and in vitro [J]. Chin J Clin Pharmacol Ther, 2025, 30(07): 865-875. DOI:10.12092/j.issn.1009-2501.2025.07.001 .
[16]	YAO T, WU Y H, FU L Y, et al. Christensenellaceae minuta modulates epithelial healing via PI3K-AKT pathway and macrophage differentiation in the colitis[J]. Microbiol Res, 2024, 289:127927. DOI:10.1016/j.micres.2024.127927 .
[17]	ACOSTA-MARTINEZ M, CABAIL M Z. The PI3K/Akt pathway in meta-inflammation[J]. Int J Mol Sci, 2022, 23(23):15330. DOI:10.3390/ijms232315330 .
[18]	徐东川. 基于网络药理学方法研究泻白散治疗哮喘的作用机制[D]. 济南: 山东中医药大学, 2022. DOI:10.27282/d.cnki.gsdzu.2022.000841 .
	XU D C. Investigating the mechanism of Xie Bai San in the treatment of asthma using network pharmacology methods[D]. Jinan: Shandong University of Traditional Chinese Medicine, 2022. DOI:10.27282/d.cnki.gsdzu.2022.000841 .
[19]	MCGUIGAN R M, MULLENIX P, NORLUND L L, et al. Acute lung injury using oleic acid in the laboratory rat: establishment of a working model and evidence against free radicals in the acute phase[J]. Curr Surg, 2003, 60(4):412-417. DOI:10.1016/S0149-7944(02)00775-4 .
[20]	覃芳芳, 丘琴, 韦红杏, 等. 泻白散的研究进展及质量标志物(Q-Marker)预测分析[J]. 中华中医药学刊, 2025, 43(11): 155-161. DOI:10.13193/j.issn.1673-7717.2025.11.031 .
	QIN F F, QIU Q, WEI H X, et al. Research progress of Xiebai Powder and prediction analysis of quality markers (Q-markers)[J]. Chin Arch Tradit Chin Med, 2025, 43(11): 155-161. DOI:10.13193/j.issn.1673-7717.2025.11.031 .
[21]	LAMPIASI N. Macrophage polarization: learning to manage it 2.0[J]. Int J Mol Sci, 2023, 24(24):17409. DOI: 10.3390/ijms242417409 .
[22]	张会敏, 杨宗统, 苏本正, 等. 高效液相色谱指纹图谱评价泻白散及其3种单味药内在质量[J]. 化学分析计量, 2021, 30(9): 48-53. DOI:10.3969/j.issn.1008 –6145.2021.09.011.
	ZHANG H M, YANG Z T, SU B Z, et al. Evaluation of internal quality of Xiebai powder and its three single herbs by HPLC fingerprint[J]. Chem Anal Meterage, 2021, 30(9): 48-53. DOI:10.3969/j.issn.1008 –6145.2021.09.011.
[23]	徐东川, 刘瑾, 李晓晶, 等. 泻白散大鼠体内入血成分研究[J]. 中国药房, 2022, 33(1):38-45. DOI:10.6039/j.issn.1001-0408.2022.01.07 .
	XU D C, LIU J, LI X J, et al. Study on absorbed components of Xiebai powder in rat blood[J]. China Pharm, 2022, 33(1):38-45. DOI:10.6039/j.issn.1001-0408.2022.01.07 .
[24]	杨宗统, 徐东川, 刘瑾, 等. 基于非靶向代谢组学和肠道菌群探究经典名方泻白散对过敏性哮喘大鼠的保护作用[J]. 中国实验动物学报, 2024, 32(2): 177-189. DOI:10.3969/j.issn.1005-4847.2024.02.005 .
	YANG Z T, XU D C, LIU J, et al. Protective effect of Xiebaisan on allergic asthma in rats based on non-targeted metabolomics and intestinal bacterial flora[J]. Acta Lab Anim Sci Sin, 2024, 32(2): 177-189. DOI:10.3969/j.issn.1005-4847.2024.02.005 .
[25]	WANG J, HE M, YANG M, et al. Gut microbiota as a key regulator of intestinal mucosal immunity[J]. Life Sci, 2024, 345:122612. DOI: 10.1016/j.lfs.2024.122612 .
[26]	ZHENG S H, XUE T Y, WANG B, et al. Chinese medicine in the treatment of ulcerative colitis: the mechanisms of signaling pathway regulations[J]. Am J Chin Med, 2022, 50(7): 1781-1798. DOI: 10.1142/S0192415X22500756 .
[27]	YAO T, WU Y H, FU L Y, et al. Christensenellaceae minuta modulates epithelial healing via PI3K-AKT pathway and macrophage differentiation in the colitis[J]. Microbiol Res, 2024, 289:127927. DOI: 10.1016/j.micres.2024.127927 .
[28]	ACOSTA-MARTINEZ M, CABAIL M Z. The PI3K/Akt pathway in meta-inflammation[J]. Int J Mol Sci, 2022, 23(23): 15330. DOI: 10.3390/ijms232315330 .
[29]	ZHANG M, SONG X, LIU S B, et al. Magnolin inhibits intestinal epithelial cell apoptosis alleviating Crohn's disease-like colitis by suppressing the PI3K/AKT signaling pathway[J]. Int Immunopharmacol, 2024, 134: 112181. DOI: 10.1016/j.intimp. 2024.112181
[30]	王百乔, 林小茹, 韩敏, 等. 二甲双胍对阿尔茨海默症模型大鼠认知功能障碍及PI3K/Akt通路的影响[J]. 实验动物与比较医学, 2021, 41(4): 313-320. DOI:10.12300/j.issn.1674-5817.2020.185 .
	WANG B Q, LIN X R, HAN M, et al. Effects of metformin on cognitive dysfunction and PI3K/Akt pathway in Alzheimer's disease rats[J]. Lab Anim Comp Med, 2021, 41(4): 313-320. DOI:10.12300/j.issn.1674-5817.2020.185 .

基因名称 Gene name	引物序列（5'→3'） Primer sequence (5'→3')	产物大小/bp Product size/bp
PI3K	F: GCGTGACATGTAGGCTCTCG	349
PI3K	R: GGGCAGTGCTGGTGGAT	349
Akt	F: CCGCCTGATCAAGTTCTCCT	118
Akt	R: TTCAGATGATCCATGCGGGG	118
β-actin	F: CGCAGCCACTGTCGAGTC	96
β-actin	R: GTCATCCATGGCGAACTGGT	96

基因名称 Gene name	引物序列（5'→3'） Primer sequence (5'→3')	产物大小/bp Product size/bp
PI3K	F: GCGTGACATGTAGGCTCTCG	349
PI3K	R: GGGCAGTGCTGGTGGAT	349
Akt	F: CCGCCTGATCAAGTTCTCCT	118
Akt	R: TTCAGATGATCCATGCGGGG	118
β-actin	F: CGCAGCCACTGTCGAGTC	96
β-actin	R: GTCATCCATGGCGAACTGGT	96

程序 Program	温度/℃ Temperature/℃	时间 Time
预变性 Initial denaturation	95	3 min
扩增 Amplification	95	20 s
45个循环 45 cycles	61	15 s
熔解曲线 Melting curve	95	5 s
	65	1 min
	97	10 s

程序 Program	温度/℃ Temperature/℃	时间 Time
预变性 Initial denaturation	95	3 min
扩增 Amplification	95	20 s
45个循环 45 cycles	61	15 s
熔解曲线 Melting curve	95	5 s
	65	1 min
	97	10 s

组别 Group	肺气肿积分 Pulmonary emphysema score	肺充/出血积分 Pulmonary congestion/bleeding score	肺间质增厚积分 Pulmonary interstitial thickening score	炎细胞浸润积分 Inflammatory cell infiltration score	总积分 Total score
空白组 Control group	0.00±0.00	0.67±0.58	0.67±0.58	0.67±0.58	2.00±1.00
模型组 Model group	2.56±0.53^∗∗	1.56±0.53^∗	1.89±0.60^∗	1.89±0.78^∗	7.89±1.05^∗∗
阳性药组 Positive control group	1.86±0.69^##	1.43±0.53	2.14±0.38	1.57±0.53	7.00±1.15
泻白散组 Xiebai San group	1.46±0.88^##	1.54±0.52	1.46±0.52	1.46±0.52	5.92±1.26^#