茯苓多糖通过调节肠道微生物群减轻环磷酰胺引起的肠道屏障和免疫损伤

doi:10.12122/j.issn.1673-4254.2026.01.04

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (1): 34-46.doi: 10.12122/j.issn.1673-4254.2026.01.04

茯苓多糖通过调节肠道微生物群减轻环磷酰胺引起的肠道屏障和免疫损伤

张越¹^,²(), 段雨婷¹(), 张晨¹, 喻璐喆¹, 刘颖颖¹, 邢丽花¹^,³, 王雷¹^,³^,⁵, 俞年军¹, 彭代银¹^,³^,⁴^,⁵, 陈卫东¹^,³^,⁴^,⁵(), 王妍妍¹^,³^,⁴()

^1.安徽中医药大学药学院，安徽合肥 230012
^2.安徽协和成药业饮片有限公司，安徽亳州 236000
^3.省部共建安徽道地中药材品质提升协同创新中心，安徽合肥 230012
^4.中药资源保护与开发研究所，安徽合肥 230012
^5.安徽省中药复方重点实验室，安徽合肥 230012

收稿日期:2025-07-11 出版日期:2026-01-20 发布日期:2026-01-16
通讯作者: 陈卫东,王妍妍 E-mail:zhangyue@ahtcm.edu.cn;wdchen@ahtcm.edu.cn;wangyanyan@ahtcm.edu.cn
作者简介:张越，博士后，E-mail: zhangyue@ahtcm.edu.cn
张越，博士后，E-mail: zhangyue@ahtcm.edu.cn
第一联系人：段雨婷，硕士，E-mail: 1651240443@qq.com
共同第一作者
基金资助:
国家自然科学基金青年项目(82505171);国家自然科学基金青年项目(82204748);国家自然科学基金区域创新发展联合基金(U25A20173);安徽省科技创新攻坚计划项目(202423l10050002);安徽省科技特派员示范项目(皖科生秘〔2024〕407号);高水平中医药重点学科建设项目(zyyzdxk-2023095);安徽省高校科研计划项目团队项目(2022AH010036)

Poria cocos polysaccharide alleviates cyclophosphamide-induced intestinal barrier dysfunction and inflammation in mice by modulating gut flora

Yue ZHANG¹^,²(), Yuting DUAN¹(), Chen ZHANG¹, Luzhe YU¹, Yingying LIU¹, Lihua XING¹^,³, Lei WANG¹^,³^,⁵, Nianjun YU¹, Daiyin PENG¹^,³^,⁴^,⁵, Weidong CHEN¹^,³^,⁴^,⁵(), Yanyan WANG¹^,³^,⁴()

^1.School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China
^2.Anhui Xiehecheng Pharmacy Traditional Chinese Medicine Co. , Ltd, Bozhou 236000, China
^3.MOE-Anhui Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials, Hefei 230012, China
^4.Institute of Traditional Chinese Medicine Resources Protection and Development, Hefei 230012, China
^5.Anhui Provincial Key Laboratory of Chinese Medicinal Formula, Hefei 230012, China

Received:2025-07-11 Online:2026-01-20 Published:2026-01-16
Contact: Weidong CHEN, Yanyan WANG E-mail:zhangyue@ahtcm.edu.cn;wdchen@ahtcm.edu.cn;wangyanyan@ahtcm.edu.cn
Supported by:
Natural Science Foundation for the Youth (NSFY) of China(82505171)

摘要/Abstract

摘要：

目的研究茯苓多糖（PCP）对环磷酰胺（CTX）诱导的肠道粘膜损伤的修复作用，探讨其对肠道微生物群和代谢产物的影响，以及对肠粘膜损伤小鼠的潜在保护机制。方法建立CTX诱导的小鼠肠道黏膜损伤模型，观察PCP对肠道屏障和免疫功能的保护作用。小鼠随机分为6组：正常对照组（NC）、CTX模型组（MC）、CTX+谷氨酰胺阳性药物组（PC）、CTX+PCP低剂量组（LD）、CTX+PCP中剂量组（MD）、CTX+PCP高剂量组（HD）。除NC组外，其余各组小鼠连续3 d腹腔注射CTX（80 mg/kg），NC组注射生理盐水。PC组给予谷氨酰胺（300 mg/kg），LD、MD、HD组分别给予PCP（75、150、300 mg/kg），连续灌胃7 d。检测各组小鼠紧密连接蛋白（occludin、ZO-1）表达水平、血清内毒素、D-乳酸盐、二胺氧化酶（DAO）水平、肠道通透性、结肠损伤情况以及细胞因子（IL-4、IL-22、IL-17A、IFN-γ等）水平变化。16S rRNA测序分析肠道微生物群组成变化，气相色谱-质谱法检测短链脂肪酸（SCFAs，如乙酸、丙酸）水平，Western blotting检测GPR41表达。粪菌移植（FMT）实验验证肠道微生物在PCP修复肠道损伤中的作用。结果与模型组相比，PCP处理组小鼠结肠组织中紧密连接蛋白Occludin和ZO-1表达显著升高（P<0.001），血清内毒素、D-乳酸盐、DAO水平及肠道通透性均显著降低（P<0.05），结肠组织中IL-4、IL-22、IL-17A和IFN-γ的mRNA表达水平显著升高（P<0.001）。PCP处理组小鼠肠道中Muribaculaceae相对丰度增加（P<0.01），Lactobacillus和Bacteroides相对丰度减少（P<0.05），结肠内容物中乙酸和丙酸含量升高（P<0.05），GPR41蛋白表达水平显著升高（P<0.001）。FMT实验证实了肠道微生物在PCP修复肠粘膜损伤中的关键作用。结论 PCP能够保护CTX引起的肠粘膜损伤，其机制可能通过调节肠道微生物群和短链脂肪酸的代谢，从而增强肠道的防御能力。

关键词: 茯苓多糖, 肠粘膜损伤, 肠道菌群, 短链脂肪酸, GPR41

Abstract:

Objective To investigate the protective effects of Poria cocos polysaccharide (PCP) against cyclophosphamide (CTX)-induced intestinal mucosal injury and its impact on gut flora and their metabolites in mice. Methods Adult BALB/C mice were randomized into normal control group, CTX model group, glutamine (positive control) group, and low-, medium- and high-dose PCP treatment groups. In all but the normal control group, the mice were subjected to modeling of CTX-induced intestinal mucosal injury by intraperitoneal CTX injections for 3 days, followed by treatment with gavage of normal saline, glutamine (300 mg/kg), or PCP at 75, 150, or 300 mg/kg for 7 consecutive days. The colonic expressions of tight junction proteins (occludin and ZO-1), serum endotoxin, D-lactate, and DAO levels, intestinal permeability, colon injury, and colonic cytokine levels (IL-4, IL-22, IL-17A, and IFN-γ mRNA) were assessed. Gut microbiota, short-chain fatty acids (SCFAs; mainly acetates and propionates) and colonic GPR41 expression were analyzed using 16S rRNA sequencing, GC-MS, and Western blotting, respectively. Fecal microbiota transplantation (FMT) experiment was conducted to validate the role of gut microbes in PCP-mediated repair of intestinal injuries. Results Compared with those in the model group, the mice treated with PCP showed significantly increased colonic occludin and ZO-1 expressions, reduced serum endotoxin, D-lactate and DAO levels, and lowered intestinal permeability with increased colonic expressions of IL-4, IL-22, IL-17A, and IFN-γ mRNA. PCP treatment obviously increased the abundance of Muribaculaceae, decreased Lactobacillus and Bacteroides, increased the contents of acetate and propionate in the colon, and upregulated colonic GPR41 expression. The results of FMT experiment confirmed the crucial role of gut microbes in PCP-mediated repair of CTX-induced intestinal injuries in mice. Conclusion PCP can protect against CTX-induced intestinal mucosal injury in mice possibly by modulating gut flora and SCFAs metabolism to enhance intestinal defense capacity.

Key words: Poria cocos polysaccharide, intestinal mucosal injury, gut flora, short-chain fatty acids, GPR41

张越, 段雨婷, 张晨, 喻璐喆, 刘颖颖, 邢丽花, 王雷, 俞年军, 彭代银, 陈卫东, 王妍妍. 茯苓多糖通过调节肠道微生物群减轻环磷酰胺引起的肠道屏障和免疫损伤[J]. 南方医科大学学报, 2026, 46(1): 34-46.

Yue ZHANG, Yuting DUAN, Chen ZHANG, Luzhe YU, Yingying LIU, Lihua XING, Lei WANG, Nianjun YU, Daiyin PENG, Weidong CHEN, Yanyan WANG. Poria cocos polysaccharide alleviates cyclophosphamide-induced intestinal barrier dysfunction and inflammation in mice by modulating gut flora[J]. Journal of Southern Medical University, 2026, 46(1): 34-46.

图/表 9

图1 PCP对CTX 治疗小鼠结肠生理状态的影响

Fig.1 Effect of PCP on physiological state of the colon in CTX-treated mice. A: Illustration of the experiment design. i.p.:Intraperitoneal injection,i.g.: Irrigation. B: Spleen index (spleen weight/body weight). C: Thymus index (thymus weight/body weight) of the mice. D: Colon length of the mice. E: Colon index (colon weight/body weight). F: HE staining of the colon tissue (Original magnification: ×7.5). NC: Control group; MC: CTX model group; PC: CTX+Glutamine (positive drug) group; LD: CTX+PCP low dose group; MD: CTX+PCP medium dose group; HD: CTX+PCP high dose group. Data are presented as Mean±SD (n=6). *P<0.05, ***P<0.001 vs NC group; #P<0.05, ##P<0.01, ###P<0.001 vs MC group.

图2 PCP对CTX处理小鼠肠道屏障功能的影响

Fig.2 Effect of PCP on intestinal barrier function in CTX-treated mice. A-D: Immunofluorescence staining of occludin and ZO-1 (green: occludin and ZO-1; blue: nucleus; ×7.5). E: PAS staining showing mucus-secreting epithelial cells (×7.5). F: MUC2 contents in colonic tissues. G: Endotoxin levels in serum. H: DAO levels in serum. I: D-Lactate levels in serum. Data are presented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs NC group; #P<0.05, ##P<0.01, ###P<0.001 vs MC group.

图3 PCP对小鼠结肠黏膜免疫细胞因子的影响

Fig.3 Effect of PCP on immune cytokines in the colonic mucosa of the mice. A-H: Relative mRNA expression levels of IL-4, IL-5, IL-13, IL-17A, IL-17F, IL-22, IFN-γ, and TGF-β detected with RT-qPCR, using β-actin as the reference control. Data are represented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs NC group; ##P<0.01, ###P<0.001 vs MC group.

表1 NC组、MC组与HD组肠道菌群多样性指数比较

Tab.1 Diversity indexes of gut microbiota in NC, MC, and HD groups (Mean±SD, n=6)

Group	Chao1	ACE	Shannon	Simpson	Goods_coverage
NC	195.22±50.78	195.79±51.02	5.81±0.21	0.97±0.00	1.00
MC	182.98±17.05	183.05±16.94	5.51±0.23	0.96±0.01	1.00
HD	275.21±52.61	274.09±51.86	6.32±0.44	0.98±0.01	1.00

图4 PCP对CTX处理小鼠肠道菌群组成的影响

Fig.4 Effect of PCP on gut microbiota composition in CTX-treated mice. A: Species abundance distribution curve. B: Principal coordinates analysis. C: Venn graph showing species overlap. D: Phylum-level microbial composition. E: Genus-level microbial composition. F: Firmicutes abundance. G: Bacteroidetes abundance. H: Firmicutes-to-Bacteroidetes ratio. I: Muribaculaceae abundance. J: Lactobacillus abundance. K: Bacteroides abundance. L: LDA score. M: Phylogenetic distribution of discriminant taxa. Data are represented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs NC group; #P<0.05, ##P<0.01, ###P<0.001 vs MC group.

图5 PCP对CTX处理小鼠SCFAs含量的影响

Fig.5 Effect of PCP on SCFAs content in CTX-treated mice. A: Standard curve of each component of SCFAs to be measured. B: GC chromatogram of mixed control solution of SCFAs (1: acetic acid; 2: propionic acid; 3: butyric acid; 4: isobutyric acid; 5: valeric acid; 6: isovaleric acid; 7: 2-ethylbutyric acid). C: Acetate levels. D: Propionate levels. E: Butyrate levels. F: Valerate levels. G: Correlation analysis between gut microbiota and short-chain fatty acids. H: Representative bands of GPR41. I: Protein expression of GPR41. Data are presented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs NC group; #P<0.05, ##P<0.01, ###P<0.001 vs MC group.

图6 FMT对CTX处理小鼠结肠生理状态的影响

Fig.6 Effect of FMT on colon injury in CTX-treated mice. A: Illustration of the design of fecal microbiota transplantation (FMT) experiment. B: Splenic somatic index (organ-to-body weight ratio). C: Thymus index (thymus weight/body weight). D: Colon length. E: Colon index (colon weight/body weight). F: HE staining (×100). FMTNC: FMT normal group; FMTCTX: FMT CTX model group; FMTPCP: FMT-Poria cocos polysaccharide. G: FMT modulates immune function in CTX-treated mice (Relative mRNA expression levels of cytokines: IL-4, IL-5, IL-13, IL-17A, IL-17F, IL-22, IFN-γ, and TGF-β). Data are presented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs FMTNC group; #P<0.05, ##P<0.01, ###P<0.001 vs FMTMC group.

图7 FMT对小鼠结肠通透性的影响

Fig.7 Effect of FMT on intestinal permeability of the mice. A-D: Immunofluorescence staining of occludin and ZO-1. (green: Occludin and ZO-1; blue: nucleus; ×100). E: PAS staining showing mucus-secreting epithelial cells (×100). F: MUC2 content in colonic tissue. G: Endotoxin levels in serum. H: DAO levels in serum. I: D-Lactate levels in serum. Data are presented as Mean±SD (n=6). *P<0.05, ***P<0.001 vs FMTNC group; #P<0.05, ##P<0.01, ###P<0.001 vs FMTMC group.

图8 FMT对小鼠结肠SCFAs含量和GPR41的影响

Fig. 8 Effect of FMT on SCFAs content and GPR41 in the colon of the mice. A: Acetate. B: Propionate. C: Butyrate. D: Valerate concentrations. E: Representative bands of GPR41. F: Quantitative analysis of GPR41 protein expression normalized to β-actin. Data are presented as Mean±SD (n=6). **P<0.01, ***P<0.001 vs FMTNC group; #P<0.05, ###P<0.001 vs FMTMC group.

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茯苓多糖通过调节肠道微生物群减轻环磷酰胺引起的肠道屏障和免疫损伤

Poria cocos polysaccharide alleviates cyclophosphamide-induced intestinal barrier dysfunction and inflammation in mice by modulating gut flora

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