芪黄健脾滋肾颗粒通过抑制MyD88/NF-κB通路减轻MRL/lpr小鼠肾损害

doi:10.12122/j.issn.1673-4254.2025.08.07

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (8): 1625-1632.doi: 10.12122/j.issn.1673-4254.2025.08.07

• • 上一篇

芪黄健脾滋肾颗粒通过抑制MyD88/NF-κB通路减轻MRL/lpr小鼠肾损害

汤忠富¹(), 黄传兵¹^,²(), 李明¹^,², 程丽丽¹, 陈君洁¹, 尚双双¹, 刘思娣¹

^1.安徽中医药大学第一附属医院
^2.新安医学与中医药现代化研究所，安徽合肥 230031

收稿日期:2025-03-06 出版日期:2025-08-20 发布日期:2025-09-05
通讯作者: 黄传兵 E-mail:2835192721@qq.com;chuanbinh@163.com
作者简介:汤忠富，在读博士研究生，医师，E-mail: 2835192721@qq.com
基金资助:
国家自然科学基金(82104782);安徽省临床医学研究转化专项(202304295107020114);安徽省临床医学研究转化专项(202304295107020115);大健康研究院新安医学与中医药现代化研究所专项(2023CXMMTCM004);大健康研究院新安医学与中医药现代化研究所专项(2023CXMMTCM015)

Qihuang Jianpi Zishen Granules ameliorate renal damage in MRL/lpr mice by inhibiting the MyD88/NF-κB pathway

Zhongfu TANG¹(), Chuanbing HUANG¹^,²(), Ming LI¹^,², Lili CHENG¹, Junjie CHEN¹, Shuangshuang SHANG¹, Sidi LIU¹

^1.First Affiliated Hospital of Anhui University of Chinese Medicine
^2.Center for Xin'an Medicine and Modernization of Traditional Chinese Medicine of IHM, Hefei 230031, China

Received:2025-03-06 Online:2025-08-20 Published:2025-09-05
Contact: Chuanbing HUANG E-mail:2835192721@qq.com;chuanbinh@163.com
Supported by:
National Natural Science Foundation of China(82104782)

摘要/Abstract

摘要：

目的基于MyD88/NF-κB通路探讨芪黄健脾滋肾颗粒（QJZ）改善MRL/lpr小鼠肾损害的作用机制。方法 6只雌性C57BL/6小鼠为正常对照组（Control组），将30只雌性MRL/lpr小鼠随机分为模型组（Model组）、芪黄健脾滋肾颗粒低剂量组（QJZ-L组）、芪黄健脾滋肾颗粒中剂量组（QJZ-M组）、芪黄健脾滋肾颗粒高剂量组（QJZ-H组）、泼尼松组（Pred组），6只/组。干预8周后取各组小鼠的尿液、全血及肾脏组织，生化试剂盒检测肾损害指标：24 h尿蛋白定量（24 h PRO）、尿总蛋白肌酐比值（UTPCR）、尿蛋白肌酐比（UACR）。ELISA法检测血清中免疫球蛋白G（IgG）、补体3（C3）、补体4（C4）、抗双链DNA抗体（anti-dsDNA）、干扰素γ（IFN-γ）、白细胞介素17（IL-17）。肾脏组织进行HE染色和透射电镜观察肾小球超微结构。采用RT-qPCR、Western blotting和免疫组化法检测肾组织中MyD88/NF-κB通路相关分子的表达。结果与Model组相比，QJZ-L、QJZ-M、QJZ-H和Pred组24 h PRO、UTPCR、UACR、IgG、anti-dsDNA、IFN-γ、IL-17降低（P<0.01），C3、C4水平升高（P<0.01）。HE染色结果显示，与Model组比较，各组肾小球内皮细胞增生、系膜增厚减轻。透射电镜显示，各组肾小球电子致密物沉积、炎性细胞浸润较Model组减轻。RT-qPCR和免疫组化结果显示，与Model组相比，QJZ-L、QJZ-M、QJZ-H和Pred组肾脏中MyD88、NF-κB表达下降（P<0.05）。Western blotting结果显示，各组肾脏中p65、p52蛋白水平低于Model组（P<0.01）。结论芪黄健脾滋肾颗粒能够改善MRL/lpr小鼠肾损害，其机制可能与抑制MyD88/NF-κB通路过度激活有关。

关键词: MRL/lpr小鼠, MyD88/NF-κB通路, 系统性红斑狼疮, 肾损害, 芪黄健脾滋肾颗粒

Abstract:

Objective To investigate the mechanism of Qihuang Jianpi Zishen Granules (QJZ) for ameliorating renal damage in MRL/lpr mice. Methods With 6 female C57BL/6 mice as the normal control group, 30 female MRL/lpr mice were randomized into model group, QJZ treatment groups at low, moderate and high doses, and prednisone treatment group (n=6). After 8 weeks of treatment, the mice were examined for 24-h urine protein, creatinine and albumin levels, serum levels of IgG, complement 3 (C3), C4, anti-dsDNA, interferon γ (IFN‑γ) and interleukin 17 (IL-17). Kidney tissues were sampled for histopathological examination with HE staining and observation of glomerular ultrastructure changes using transmission electron microscopy (TEM). The expressions of MyD88/NF-κB pathway-related molecules in the kidney tissue were detected using RT-qPCR, Western blotting and immunohistochemistry. Results Compared with those in the model group, the mice treated with QJZ at the 3 doses and prednisone showed significant reductions in the renal injury biomarkers and serum IgG, anti-dsDNA, IFN‑γ and IL-17 levels and elevation of serum C3 and C4 levels. HE staining revealed lessened glomerular endothelial cell proliferation and mesangial thickening in all the treatment groups. TEM observation further demonstrated reduced electron-dense deposits and diminished inflammatory cell infiltration in the glomeruli in the intervention groups. QJZ at the 3 doses and prednisone treatment all significantly lowered renal expression levels of MyD88, NF-κB, p65 and p52 in the mouse models. Conclusion QJZ can improve renal damage in MRL/lpr mice possibly by inhibiting overactivation of the MyD88/NF-κB pathway.

Key words: MRL/lpr mice, MyD88/NF?κB pathway, systemic lupus erythematosus, renal damage, Qihuang Jianpi Zishen Granules

汤忠富, 黄传兵, 李明, 程丽丽, 陈君洁, 尚双双, 刘思娣. 芪黄健脾滋肾颗粒通过抑制MyD88/NF-κB通路减轻MRL/lpr小鼠肾损害[J]. 南方医科大学学报, 2025, 45(8): 1625-1632.

Zhongfu TANG, Chuanbing HUANG, Ming LI, Lili CHENG, Junjie CHEN, Shuangshuang SHANG, Sidi LIU. Qihuang Jianpi Zishen Granules ameliorate renal damage in MRL/lpr mice by inhibiting the MyD88/NF-κB pathway[J]. Journal of Southern Medical University, 2025, 45(8): 1625-1632.

图/表 7

表1 引物序列

Tab.1 Primer sequences for RT-qPCR

Gene

Primer sequence (5'→3')

Length (bp)

GAPDH

Forward:GCAGTGGCAAAGTGGAGATTG

Reverse:CGCTCCTGGAAGATGGTGAT

169

MyD88

Forward:TCATGTTCTCCATACCCTTGGT

Reverse:AAACTGCGAGTTGGGGTCAG

175

NF-κB

Forward:GCTCCTGTTCGAGTCTCCAT

Reverse:TTGCGCTTCTCTTCAATCCG

119

图1 QJZ对各组小鼠24 h PRO、UTPCR、UACR的影响

Fig.1 Effect of Qihuang Jianpi Zishen Granules (QJZ) at low, moderate and high doses (QJZ-L, QJZ-M and QJZ-H, respectively) and prednisone (Pred) on 24-h PRO, UTPCR, and UACR of MRL/lpr mice. **P<0.01 vs Control group (C57BL/6 mice); ##P<0.01 vs Model group. UPCR: Urine protein creatinine ratio; UACR: Urinary albumin creatinine ratio.

图2 QJZ对各组小鼠免疫炎症的影响

Fig.2 Effects of QJZ on immune inflammation in MRL/lpr mice.**P<0.01 vs Control group; ##P<0.01 vs Model group.

图3 QJZ对各组小鼠肾脏病理学的影响

Fig.3 Effect of QJZ on renal pathology in MRL/lpr mice (Left, original magnification: ×40; Right,×100).**P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group. A: HE staining; B: Glomerular score.

图4 各组小鼠肾小球超微结构

Fig.4 Transmission electron microscopy for observation of glomerular ultrastructure of the mice in each group (Original magnification: ×25 000).

图5 QJZ对各组小鼠肾组织中MyD88、NF-κB表达的影响

Fig.5 Effect of QJZ on expressions of MyD88 and NF-κB in renal tissues of the mice in each group (Scale bar=50 μm). **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group. A,B:Relative expression of MyD88 and NF‑κB mRNA; C-E: immunohistochemistry results.

图6 QJZ对各组小鼠肾组织中p65、p52蛋白水平的影响

Fig.6 Effect of QJZ on p65 and p52 protein levels in the renal tissues of the mice in each group detected by Western blotting. **P<0.01 vs Control group; ##P<0.01 vs Model group.

参考文献 39

[1]	Zhao G, Li X, Zhang Y, et al. Intricating connections: the role of ferroptosis in systemic lupus erythematosus[J]. Front Immunol, 2025, 16: 1534926. doi：10.3389/fimmu.2025.1534926
[2]	Halfon M, Tankeu AT, Ribi C. Mitochondrial dysfunction in systemic lupus erythematosus with a focus on lupus nephritis[J]. Int J Mol Sci, 2024, 25(11): 6162. doi：10.3390/ijms25116162
[3]	Accapezzato D, Caccavale R, Paroli MP, et al. Advances in the pathogenesis and treatment of systemic lupus erythematosus[J]. Int J Mol Sci, 2023, 24(7): 6578. doi：10.3390/ijms24076578
[4]	Zucchi D, Silvagni E, Elefante E, et al. Systemic lupus erythematosus: one year in review 2023[J]. Clin Exp Rheumatol, 2023, 41(5): 997-1008.
[5]	Crow MK. Pathogenesis of systemic lupus erythematosus: risks, mechanisms and therapeutic targets[J]. Ann Rheum Dis, 2023, 82(8): 999-1014. doi：10.1136/ard-2022-223741
[6]	陈少敏, 李红毅, 刘炽, 等. 基于CiteSpace的中医药治疗系统性红斑狼疮可视化分析[J].新中医, 2024, 56(24): 184-91. doi：10.13457/j.cnki.jncm.2024.24.037
[7]	朱雅文, 黄传兵, 李明, 等. 芪黄健脾滋肾颗粒治疗系统性红斑狼疮患者疗效观察及对外周血Toll样受体表达水平的影响[J]. 中医药学报, 2025, 53(2): 67-71.
[8]	钱爱, 程园园, 朱雅文, 等.芪黄健脾滋肾颗粒治疗系统性红斑狼疮的疗效观察[J]. 安徽中医药大学学报, 2023, 42(6): 7-12. doi：10.3969/j.issn.2095-7246.2023.06.003
[9]	李明, 尚双双, 李云飞, 等. 芪黄健脾滋肾颗粒治疗脾肾亏虚型系统性红斑狼疮的临床疗效观察[J]. 时珍国医国药, 2023, 34(2): 370-3. doi：10.3969/j.issn.1008-0805.2023.02.31
[10]	陈君洁, 黄传兵, 程丽丽, 等. 芪黄健脾滋肾颗粒调控JAK1/STAT1信号通路对MRL/lpr狼疮小鼠足细胞凋亡的影响[J]. 中成药, 2024,46(12): 4145-50.
[11]	Baek WY, Choi YS, Lee SW, et al. Toll-like receptor signaling inhibitory peptide improves inflammation in animal model and human systemic lupus erythematosus[J]. Int J Mol Sci, 2021, 22(23): 12764. doi：10.3390/ijms222312764
[12]	Tao B, Xiang W, Li XL, et al. Regulation of Toll-like receptor-mediated inflammatory response by microRNA-152-3p-mediated demethylation of MyD88 in systemic lupus erythematosus[J]. Inflamm Res, 2021, 70(3): 285-96. doi：10.1007/s00011-020-01433-y
[13]	Yang Z, Ji S, Liu L, et al. Promotion of TLR7-MyD88-dependent inflammation and autoimmunity in mice through stem-loop changes in Lnc-Atg16l1[J]. Nat Commun, 2024, 15(1): 10224. doi：10.1038/s41467-024-54674-4
[14]	Tilstra JS, Kim M, Gordon RA, et al. B cell-intrinsic Myd88 regulates disease progression in murine lupus[J]. J Exp Med, 2023, 220(12): e20230263. doi：10.1084/jem.20230263
[15]	吕冬菊, 黄东明, 段伟静. 黄芪甲苷调控HMGB1/NF-κB通路对脓毒症大鼠巨噬细胞极化和急性肾损伤的机制研究[J]. 中国煤炭工业医学杂志, 2025, 28(1): 1-7.
[16]	Reynolds JA, Li Y, Herlitz L, et al. Novel biomarker discovery through comprehensive proteomic analysis of lupus mouse serum[J]. J Autoimmun, 2024, 142: 103134. doi：10.1016/j.jaut.2023.103134
[17]	Chalmers SA, Ayilam Ramachandran R, Garcia SJ, et al. The CD6/ALCAM pathway promotes lupus nephritis via T cell-mediated responses[J]. J Clin Invest, 2022, 132(1): e147334. doi：10.1172/jci147334
[18]	Desai SB, Ahdoot R, Malik F, et al. New guidelines and therapeutic updates for the management of lupus nephritis[J]. Curr Opin Nephrol Hypertens, 2024, 33(3): 344-53. doi：10.1097/mnh.0000000000000969
[19]	尚双双, 李明, 黄传兵.健脾滋肾法治疗系统性红斑狼疮的理论探源[J].中国中医基础医学杂志,2023,29(04):663-5.
[20]	钱爱, 黄传兵, 朱雅文, 等. 基于“肾-脾-肠轴” 探讨健脾滋肾法调节肠道菌群治疗系统性红斑狼疮的可行性[J]. 上海中医药杂志, 2024, 58(12): 52-5, 70.
[21]	Tian HY, An LZ, Wang PW, et al. Review of Astragalus membranaceus polysaccharides: extraction process, structural features, bioactivities and applications[J]. Chin Herb Med, 2025, 17(1): 56-69. doi：10.1016/j.chmed.2024.09.004
[22]	张慧, 罗进辉, 杨柳, 等.黄芪多糖调节TGF-β1/Smad/AP-1信号通路对慢性肾衰竭大鼠肾纤维化的影响[J].中国老年学杂志, 2024, 44(16): 4040-4.
[23]	陈鲁宁, 胡扬, 辛国松, 等. 菟丝子化学成分、药理作用研究进展及其质量标志物(Q-Marker)预测[J]. 中草药, 2024, 55(15): 5298-314. doi：10.7501/j.issn.0253-2670.2024.15.029
[24]	Liu ZH, Xu QY, Wang Y, et al. Catalpol from Rehmannia glutinosa targets Nrf2/NF‑κB signaling pathway to improve renal Anemia and fibrosis[J]. Am J Chin Med, 2024, 52(5): 1451-85. doi：10.1142/s0192415x24500575
[25]	Bailly C. Atractylenolides, essential components of Atractylodes-based traditional herbal medicines: Antioxidant, anti-inflammatory and anticancer properties[J]. Eur J Pharmacol, 2021, 891: 173735. doi：10.1016/j.ejphar.2020.173735
[26]	Ding X, Li S, Huang H, et al. Bioactive triterpenoid compounds of Poria cocos (Schw.) Wolf in the treatment of diabetic ulcers via regulating the PI3K-AKT signaling pathway[J]. J Ethnopharmacol, 2024, 325: 117812. doi：10.1016/j.jep.2024.117812
[27]	施江南, 温乐乐, 江丽洁, 等. 基于文献计量学的覆盆子研究热点与趋势分析[J].中草药, 2025, 56(2): 598-616. doi：10.7501/j.issn.0253-2670.2025.02.022
[28]	王平, 孙文娟, 潘保朝, 等. 基于NLRP3通路研究金樱子抑制肾小管上皮细胞间质化治疗糖尿病肾病的作用机制[J].天津中医药,2024, 41(11): 1468-74.
[29]	Chen Y, Tian B. IFN-γ promotes the development of systemic lupus erythematosus through the IFNGR1/2-PSTAT1-TBX21 signaling axis[J]. Am J Transl Res, 2022, 14(10): 6874-88.
[30]	Richter P, Macovei LA, Mihai IR, et al. Cytokines in systemic lupus erythematosus-focus on TNF-α and IL-17[J]. Int J Mol Sci, 2023, 24(19): 14413. doi：10.3390/ijms241914413
[31]	Bayer AL, Alcaide P. MyD88: At the heart of inflammatory signaling and cardiovascular disease[J]. J Mol Cell Cardiol, 2021, 161: 75-85. doi：10.1016/j.yjmcc.2021.08.001
[32]	Owen AM, Luan L, Burelbach KR, et al. MyD88-dependent signaling drives toll-like receptor-induced trained immunity in macrophages[J]. Front Immunol, 2022, 13: 1044662. doi：10.3389/fimmu.2022.1044662
[33]	Yu T, Xiaojuan F, Jinxi L, et al. Extracellular HMGB1 induced glomerular endothelial cell injury via TLR4/MyD88 signaling pathway in lupus nephritis[J]. Mediators Inflamm, 2021, 2021: 9993971. doi：10.1155/2021/9993971
[34]	Feng X, Yang R, Tian Y, et al. HMGB1 protein promotes glomerular mesangial matrix deposition via TLR2 in lupus nephritis[J]. J Cell Physiol, 2020, 235(6): 5111-9. doi：10.1002/jcp.29379
[35]	Hövelmeyer N, Schmidt-Supprian M, Ohnmacht C. NF‑κB in control of regulatory T cell development, identity, and function[J]. J Mol Med, 2022, 100(7): 985-95. doi：10.1007/s00109-022-02215-1
[36]	Caster DJ, Powell DW. Precision targeting of NF-κB signaling in lupus nephritis[J]. Lupus: Los Angel, 2020, 6(1): 160.
[37]	Chen JS, Chen PH, Song YJ, et al. STING upregulation mediates ferroptosis and inflammatory response in lupus nephritis by upregulating TBK1 and activating NF‑κB signal pathway[J]. J Biosci, 2024, 49(1): 9. doi：10.1007/s12038-023-00381-z
[38]	He LY, Niu SQ, Yang CX, et al. Cordyceps proteins alleviate lupus nephritis through modulation of the STAT3/mTOR/NF-кB signaling pathway[J]. J Ethnopharmacol, 2023, 309: 116284. doi：10.1016/j.jep.2023.116284
[39]	Su B, Ye H, You X, et al. Icariin alleviates murine lupus nephritis via inhibiting NF-κB activation pathway and NLRP3 inflammasome[J]. Life Sci, 2018, 208: 26-32. doi：10.1016/j.lfs.2018.07.009

芪黄健脾滋肾颗粒通过抑制MyD88/NF-κB通路减轻MRL/lpr小鼠肾损害

Qihuang Jianpi Zishen Granules ameliorate renal damage in MRL/lpr mice by inhibiting the MyD88/NF-κB pathway

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