The Study on the Action Mechanism of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) Couplet Herbs on Membranous Nephropathy Based on Network Pharmacology

Abstract

Objective Our objective was to explore the action mechanism of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs in the treatment of membranous nephropathy (MN) based on network pharmacology.

Methods The active ingredients and targets of Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) were screened by systematic pharmacology database and analysis platform. Online Human Mendelian Genetic database and GeneCards database were used to retrieve MN-related targets. The active ingredient-related targets and MN disease targets were introduced into Venny 2.1, and Wayne diagram was drawn. The intersection targets were the potential targets of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs in the treatment of MN. The protein interaction network of potential targets was constructed, and the core targets were screened with String platform. Metascape platform was used for functional enrichment analysis of gene ontology (GO) and pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG). The “herb-active ingredient-target-pathway” networks were drawn by using Cytoscape software, and the key components, targets, and signaling pathways were screened.

Results A total of 8 active ingredients and 193 related targets in Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) were screened out; a total of 1,621 targets of MN disease and 105 potential targets for the treatment of MN were obtained in the treatment with Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs; 40 core targets were screened by protein–protein interaction network topology analysis; a total of 1,978 results were obtained by GO function enrichment analysis, and 206 signal pathways were obtained by KEGG pathway enrichment analysis and screening. The network topology analysis of “herb-active ingredient-target-pathway” showed that the key components included quercetin, kaempferol, β-sitosterol, etc.; the key targets included protein kinase Bα (AKT), mitogen-activated protein kinase 1 (MAPK1), B-cell lymphoma-2 (BCL2), prostaglandin-endoperoxide synthase 2 (PTGS2), etc.; the key pathways included advanced glycation end product/receptor of AGE signaling pathway, phosphatidyl inositol 3-kinase (PI3K)/AKT signaling pathway, MAPK signaling pathway, hypoxia-inducible factor-1 signaling pathway, Ras signaling pathway, nuclear factor kappa-B (NF-κB) signaling pathway, Toll-like receptors signaling pathway, p53 signaling pathway and vascular endothelial growth factor signaling pathway in the late stage of diabetic complications.

Conclusion The Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs can regulate PI3K/AKT, MAPK, NF-κB signaling pathways in MN by targeting proteins of AKT1, MAPK8, PTGS2 through key components of quercetin, β-sitosterol, and kaempferol, so as to inhibit the overexpression of inflammatory factors in renal tissues, regulate inflammatory response, and improve renal function.

Keywords

membranous nephropathy - MN - Jinyingzi (Rosae Laevigatae Fructus) - Qianshi (Semen Euryales) - network pharmacology - action mechanism

Introduction

Membranous nephropathy (MN) is a common glomerular disease in China, with a large number of clinical manifestations of proteinuria, hypoalbuminemia, edema, and hyperlipidemia. It is one of the most common pathological types of nephrotic syndrome, with uniform thickening of glomerular capillary matrix and diffuse deposition of subcutaneous immune complexes as its pathological characteristics. Studies have shown that the detection rate of MN in all renal biopsy samples is about 23.4%, which is the second most common glomerular disease after immunoglobulin A (IgA) nephropathy.[1] It has been found that in idiopathic MN, about 30% of patients can completely spontaneously resolve, 30% would develop lifelong proteinuria while maintaining normal renal function, and the rest would progress to end-stage renal failure.[2] At present, the Western medicine treatments for MN are mainly glucocorticoids combined with immunosuppressive agents.[3] However, these treatments, with expensive treatment costs, often produce serious adverse reactions and their side effects can further aggravate the renal burden. For the limitations of Western medicine, more and more patients and clinicians choose to combine it with Chinese herbs for the treatment of MN.

There is no discussion on the name, etiology, pathogenesis and related treatment of MN in traditional Chinese medicine (TCM). According to the symptoms and signs, modern doctors classify MN into the categories of edema, low back pain, fatigue, kidney wind, etc., and summarize that the pathogenesis lies in deficient foundation with excess external manifestations, and the deficiency is mainly about the spleen and kidney, and the excess is mainly about wind pathogen and blood stasis. If spleen qi becomes deficient and fails to transport, transform and ascend the lucid, it will lead to disordered distribution of the body fluid which will accumulate locally or overflow to the skin and result in edema; failure of lucid qi to ascend and leakage of essence will bring about proteinuria. Deficient kidney qi, dysfunction of the bladder to transpire and fluid immersion in the skin will cause edema; insufficient kidney qi to consolidate and store essence may lead to proteinuria. When Chinese herbs are applied to treat MN, it is based on tonifying the spleen and kidney, supplemented by promoting diuresis and detumescence, and eliminating proteins.[4] Protein belongs to the category of “essence” and “essential qi” in the theory of TCM. Therefore, the application of essence-consolidating and urine-astringing herbs in the treatment of MN has significant clinical efficacy in eliminating urinary protein. Professor Pingdong Zheng, in the treatment of proteinuria, advocated consolidating essence to achieve the purpose of “plugging the flow” while consolidating the foundation. He chose Jinyingzi (Rosae Laevigatae Fructus), Qianshi (Euryales Semen), Fupenzi (Rubi Fructus), Wuweizi (Schisandrae Chinensis Fructus) to consolidate the kidney and astringe essence.[5] In the treatment of MN, Professor Liqun Song applied the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs to achieve the efficacy of tonifying the kidney, strengthening the spleen, astringing, and consolidating the essence of the lower energizer.[6] Professor Hongtao Yang often applied the method of tonifying the kidney and spleen with Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Semen Euryales) to consolidate the kidney and essence.[7] It is obvious that when many doctors treat MN, they choose kidney-nourishing and essence-consolidating herbs represented by Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen). Jinyingzi (Rosae Laevigatae Fructus) is sour in taste and mild in nature and pertains to the lung, spleen, kidney, and bladder meridians. It has the effects of consolidating essence and astringing urine, checking metrorrhagia and leukorrhagia, astringing the large intestine, and relieving diarrhea. It is often used in clinical treatment of spermatorrhea, metrorrhagia, leukorrhagia and diarrhea, etc.[8] Modern pharmacological studies have confirmed that the chemical components contained in Jinyingzi (Rosae Laevigatae Fructus) are phenolic acid, steroids, triterpenes, phenylpropanoids, etc., which have antioxidant, anti-inflammatory, and bacteriostatic effects and can improve renal function and immunity, reduce blood sugar and fight against tumor, etc.[9] Qianshi (Euryales Semen) is sweet and astringent in taste, mild in nature, and pertains to the spleen and kidney meridians. It has the functions of invigorating the spleen and stopping diarrhea, tonifying the kidney and consolidating essence, removing dampness, and relieving leukorrhagia.[10] Studies have shown that the superfine powder of Qianshi (Euryales Semen) can reduce the inflammatory response and inhibit the phosphatidylinositol 3-kinase (PI3K) /protein kinase B (AKT)/mammal target of rapamycin (mTOR) signaling pathway in type 2 diabetic mice, thus protecting the renal structure and function in diabetic nephropathy.[11]

Network pharmacology takes the concept of TCM as the basic research method and draws disease-related biomolecules and herb action targets to the biomolecular network to understand complex diseases and systemic diagnosis and treatment. It has incomparable advantages in clarifying the pharmacodynamic components and action mechanisms of Chinese herbs and compound preparations.[12] Therefore, the network pharmacology was used to explore the potential biological mechanism of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs in the treatment of MN, hoping to provide theoretical evidence for its clinical development and application.

Materials and Methods

The Active Ingredients of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) Couplet Herbs and Screening of Their Related Action Targets

The chemical components were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, http://tcmspw.com/tcmsp.php) with the keywords of Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen). The active ingredients of the above herbs were obtained according to the screening criteria of oral bioavailability ≥ 30% and herb-like properties (drug likeness) ≥ 0.18. Target proteins associated with the active ingredients were obtained on the TCMSP, and the target protein names were standardized through the Unitprot database (http://www.Unitprot.org).

Acquisition of Targets for Membranous Nephropathy and Potential Targets for the Treatment of Membranous Nephropathy by Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) Couplet Herbs

With the keyword of “membraous nephropathy,” the Online Human Mendelian Inheritance in Man (OMIM, https://www.omim.org) and GeneCards database (https://www.genecards.org) were used to search the targets related to MN, integrate the collected disease targets, remove the duplicate values, and finally obtain MN disease targets. The selected herb active ingredient-related targets and MN disease targets were introduced into Venny 2.1 (https://bioinfogp. cnb.csic.es/tools/venny/), Wayne diagram was drawn, and the intersection targets of the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs for active ingredient treatment of MN were obtained, which were the potential targets of these couplet herbs for the treatment of MN.

Establishment of Protein–Protein Interaction Network

The potential targets were input into the online String 11.5 database (https://string-db.org/), the specie was selected as “Homo sapiens”, the “highest configuration” was set to 0.9, the single node was deleted, the protein–protein interaction (PPI) network was drawn, and the results in TSV format were exported. The obtained data were input into Cytoscape 3.9.0, topology analysis was visualized and performed with a network analyzer, and the core targets were selected according to the network node degree value (degree).

Functional Enrichment Analysis of Gene Ontology and Pathway Enrichment Analysis of Kyoto Encyclopedia of Genes and Genes)

The screened core targets were uploaded to the Metascape database (https://metascape.org/) for gene ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genes (KEGG) pathway enrichment analysis. p < 0.01 was set as the screening criteria, and the enrichment results were visualized through the bioinformatics cloud platform (http://www.bioinformations.com.cn/).

Construction of Herb-Active Ingredient-Target-Pathway Network

The active ingredients, core targets, and KEGG pathways of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) were uploaded to Cytoscape 3.9.0 software to construct a “herb-active ingredient-target-pathway” visualization network. A network analyzer was used to analyze the topology of the network, and key components, key targets, and key pathways were screened according to the network node degree.

Results

Active Ingredients and Related Targets of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) Couplet Herbs

Seven active ingredients of Jinyingzi (Rosae Laevigatae Fructus) and two active ingredients of Qianshi (Euryales Semen) were retrieved in TCMSP, and nine compounds in total. The active ingredients for which target information could not be obtained were eliminated, and eight active ingredients of Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) were finally obtained. The relevant targets of active ingredients were obtained from the TCMSP database, and the targets were standardized using the UniProt database. One hundred and ninety-three targets were obtained after deduplication. See [Table 1].

Table 1 Active ingredients of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs

Chinese herbs	MOLID	Marks	Active ingredients	OB/%	DL
Jinyingzi (Rosae Laevigatae Fructus)	MOL001494	JYZ1	Mannitol	42.00	0.19
	MOL000358	JYZ2	β-sitosterol	36.91	0.75
	MOL000422	JYZ3	Kaempferol	41.88	0.24
	MOL005030	JYZ4	Gondor acid	30.70	0.20
	MOL008628	JYZ5	4'-Methyl-N-Methyl Chlorophyll	53.43	0.26
	MOL000098	JYZ6	Quercetin	46.43	0.28
Qianshi (Euryales Semen)	MOL002773	QS	β-carotene	37.18	0.58
	MOL007180	QS1	Vitamin E	32.29	0.70

Abbreviation: OB, oral bioavailability.

Membranous Nephropathy-Related Targets and Potential Targets of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) Couplet Herbs for the Treatment of Membranous Nephropathy

GeneCards database was used to retrieve the targets of MN, and the score was set to be greater than the average for screening, and 1,186 targets related to MN were obtained. The OMIM database was used to retrieve 476 targets related to MN. The retrieved disease targets were combined, and the duplicate values were removed. Finally, 1,621 targets related to MN were obtained. The active-ingredient targets and the MN targets were passed through a Venny platform to draw a Wayne diagram, and the intersection targets were taken to obtain 105 potential targets for the treatment of MN by Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs. See [Fig. 1].

Fig. 1 Wayne plot of herb-disease intersection targets.

Protein–Protein Interaction Network and Core Targets

The obtained intersection targets were uploaded to the String platform, the single node was deleted, the PPI network relationship of the intersection targets was obtained, and the TSV file was exported. The TSV file was imported into Cytoscape 3.9.0 for visualization, and the network consisted of 105 nodes and 378 edges. A total of 40 core targets were obtained, including AKT1, interleukin-2 (IL-2), tumor protein p53, caspase 3 (CASP3), caveolin-1, signal transducer and activator of transcription 1, androgen receptor, MYC, epidermal growth factor receptor, CASP8, mitogen-activated protein kinase 8 (MAPK8), MAPK1, tumor necrosis factor (TNF), IL-10 and IL-6, etc. See [Fig. 2].

Fig. 2 Intersection targets PPI network. PPI, protein–protein Interaction.

Gene Ontology Function Enrichment Analysis and Kyoto Encyclopedia of Genes and Genes Pathway Enrichment Analysis

Metascap Platform was used for GO function enrichment analysis and KEGG pathway enrichment analysis of the core targets for the treatment of MN with Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs, and the analysis results were visualized by using the Bioinformatics Cloud Platform, as shown in [Fig. 3]. A total of 1 978 results were obtained by GO function enrichment analysis (p < 0.01), in which the biological process involved response to an inorganic substance, regulation of cell motility, response to cytokine, response to bacterium, response to hormone, etc.; cell composition mainly included membrane raft, membrane microdomain, side of membrane, etc.; molecular function mainly involved cytokine receptor binding, receptor ligand activity, cytokine activity, DNA-transcription factor binding, etc. A total of 206 signaling pathways were obtained through KEG pathway enrichment analysis and screening (p < 0.01), mainly including HIF-1 signaling pathway, MAPK signaling pathway, PI3K/AKT signaling pathway, JAK kinase/STAT signaling pathway, advanced glycation end (AGE)/receptor of AGE (RAGE) signaling pathway, TNF signaling pathway, IL-17 signaling pathway, Toll-like receptor signaling pathway, p53 signaling pathway, NF-κB signaling pathway, VEGF signaling pathway, etc. The top 20 enrichment analysis items were selected to draw the bubble diagram. The results are shown in [Figs. 3] [4] [5] to [6].

Fig. 3 BP bubble diagram of GO function enrichment analysis. BP, biological process; GO, gene ontology.

Fig. 4 CC Bubble diagram of GO function enrichment analysis. CC, cell composition; GO, gene ontology.

Fig. 5 MF Bubble diagram for GO function enrichment analysis. MF, molecular function; GO, gene ontology.

Fig. 6 Bubble diagram of KEGG pathway enrichment analysis. KEGG, Kyoto Encyclopedia of Genes and Genomes.

Herb-Active Ingredient-Target-Pathway Network

The herbs, active ingredients, core targets, and pathways were introduced into Cytoscape 3.9.0 software to construct the “herb-active ingredient-target-pathway” network diagram, as shown in [Fig. 7]. The analysis by network analyzer showed that quercetin was the main ingredient in the treatment of MN with Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs; the other ingredients, respectively, included kaempferol, β-sitosterol, 4'-methyl-N-methyl chlorophyll, and β-carotene, as shown in [Table 2]. The top key targets of connectivity, media and compactness in the network were AKT1, MAPK1, JUN, B-cell lymphoma-2 (BCL2), prostaglandin-endoperoxide synthase 2 (PTGS2), RELA, CASP3, TNF, protein kinase C alpha (PRKCA), VEGFA, MAPK8 and IL-6, as shown in [Table 3]. Key pathways mainly include AGE/RAGE signaling pathway, PI3K/AKT signaling pathway, MAPK signaling pathway, TNF signaling pathway, JAK/STAT signaling pathway, HIF-1 signaling pathway, IL-17 signaling pathway, Ras signaling pathway, NF-κB signaling pathway, Toll-like receptor signaling pathway, p53 signaling pathway, VEGF signaling pathway, and mTOR signaling pathway in diabetic complications, as shown in [Table 4].

Table 2 Characteristic parameters of network nodes of main active ingredients of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs

MOLID	Active ingredients	Degree of connectivity	Betweenness	Compactness
MOL000098	Quercetin	90	0.250160989	0.678217822
MOL000422	Kaempferol	47	0.067931795	0.479020979
MOL000358	β-sitosterol	27	0.050802488	0.420245399
MOL008628	4'-Methyl-N-Methyl Chlorophyll	16	0.012957271	0.362433862
MOL002773	β-carotene	14	0.005805591	0.38700565

Table 3 Characteristic parameters of core target network nodes of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs

Core targets	Degree of connectivity	Betweenness	Compactness	Core targets	Degree of connectivity	Betweenness	Tightness
AKT1	20	0.021786549	0.485815603	CASP3	14	0.014190824	0.489285714
MAPK1	18	0.015607588	0.462837838	TNF	13	0.010916685	0.459731544
JUN	17	0.022550500	0.500000000	PRKCA	13	0.013550801	0.482394366
BCL2	17	0.021823919	0.500000000	VEGFA	12	0.005565254	0.444805195
PTGS2	17	0.041232989	0.500000000	MAPK8	12	0.005614411	0.439102564
RELA	17	0.013437304	0.475694444	IL-6	12	0.005743961	0.444805195

Table 4 Characteristic parameters of core pathway network nodes of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs

Core pathway	Degree of connectivity	Betweenness	Tightness
AGE/RAGE signaling pathway in diabetic complications	31	0.022937366	0.417682927
PI3K-Akt signaling pathway	26	0.013528230	0.405325444
MAPK signaling pathway	25	0.013089274	0.402941176
TNF signaling pathway	20	0.009066171	0.400584795
HIF-1 signaling pathway	20	0.008853526	0.391428571
IL-17 signaling pathway	19	0.008134330	0.395953757
Ras signaling pathway	16	0.004216075	0.382681564
NF-κB signaling pathway	15	0.006124239	0.389204545
Toll-like receptor signaling pathway	15	0.004166944	0.380555556
p53 signaling pathway	13	0.003692008	0.368279570
VEGF signaling pathway	11	0.002746478	0.380555556
mTOR signaling pathway	8	0.000979000	0.366310160

Fig. 7 Herb-active ingredient-target-pathway network diagram. Notes: JYZ: Jinyingzi (Rosae Laevigatae Fructus); QS: Qianshi (Euryales Semen); diamond: the target; round: the active ingredient; inverted triangle: the pathway; larger node area and darker color indicate that the node is more important.

Discussion

In TCM, it believes that the basic therapeutic principle of MN is to tonify the kidney and consolidate essence, and the herbs represented by Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) have been widely used in clinical practice and have achieved remarkable clinical efficacy. For example, Professor Chuanhui Ye 's self-formulated Ye's Xiaobai prescription was used to eliminate proteinuria with Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) to tonify the kidney and consolidate essence.[13] Professor Yueyi Deng used Jinyingzi (Rosae Laevigatae Fructus) to nourish the kidney and fill the essence.[14] Professor Daning Zhang also likes to use Jinyingzi (Rosae Laevigatae Fructus) to consolidate essence and astringe urine, Qianshi (Euryales Semen), and Shayuanzi (Astragali Complanati Semen) to tonify the kidney and consolidate essence in the treatment of MN.[15] In the treatment of MN in the nonedema stage, Professor Qinguo Hong also chose the herbs Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen) to strengthen the kidney and consolidate essence, which means that sufficient kidney qi ensures sealed and consolidated essence, and the essence will be controlled rather than being leaked out.[16] Professor Shili Cao also treated MN by benefiting the kidney and consolidating essence with Jinyingzi (Rosae Laevigatae Fructus) and Qianshi (Euryales Semen).[17]

According to this study and related studies, the main active ingredient quercetin in Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs has anti-inflammatory, antioxidant, antidiabetic and immunity-regulating effects, and it has certain renal protection efficacy.[18] [19] Quercetin can effectively inhibit the expression of TNF, IL-1β, IL-17, IL-6, and also inhibit the phosphorylation of inflammatory signaling pathways such as NF-κB, TLR4, and STAT3, and reduce the expression of inflammatory factors and renal inflammatory response by inhibiting the activation of PI3K/AKT signaling pathway, thereby improving renal function.[20] [21] [22] [23] Kaempferol has antioxidation, anticancer, antiviral and immunity-regulating effects,[24] and it has been proved by experiments that kaempferol can significantly reduce the expression of IL-6 and TNF-α inflammatory factors in kidney tissues and inhibit inflammatory response.[25] β-sitosterol has antioxidant, anti-inflammatory, and antitumor effects and plays an important role in kidney diseases and immune diseases.[26] [27] β-carotene has antioxidation, inflammatory response-regulating effects, which can enhance body immunity and improve the anticancer effect.[28] Experiments have proved that β-carotene could inhibit the expression of NF-κB and IL-6.[29]

Through KEGG enrichment analysis, it was found that Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs involved multiple signaling pathways in the treatment of MN and was closely related to the main targets. It was found that PI3K/AKT, as an upstream signaling pathway, could regulate and control the downstream mTOR signaling pathway,[30] and excessive activation of PI3K/AKT/mTOR pathway could accelerate the occurrence and development of renal fibrosis in rats with MN.[31] Inhibition of PI3K/AKT/mTOR pathway can slow down podocyte injury in rats with MN, thus achieving the purpose of protecting renal function in rats with MN.[32] MAPK-related signaling pathways have also been confirmed to be involved in the process of renal fibrosis, and c-Jun amino-terminal kinase is a key mediator. MAPK signaling pathways reduce the thickening of glomerular basement membrane by inhibiting the phosphorylation of c-Jun.[33] [34] Studies have shown that overexpression of IL-17 can be used as one of the indicators to determine the progression of renal fibrosis.[35] IL-17 signaling pathway, as an inflammatory pathway, can promote the secretion of inflammatory factors such as IL-6, TNF-α, thus playing a proinflammatory role.[36] Inhibiting this pathway can effectively reduce the production of inflammatory factors in patients with MN. HIF-1 signaling pathway is directly related to hypoxia in the body and can induce increased apoptosis of glomeruli and renal tubules through oxidative stress. However, there is an obvious oxidative stress response in rats with MN. Down-regulation of HIF-1 signaling pathway can antagonize oxidative stress response and achieve renal protection.[37] [38] Activation of NF-κB signaling pathway is critical in the occurrence and development of kidney disease.[39] Studies have found that up-regulation of NF-κB expression in renal tissues of rats with MN can promote glomerular mesangial cell proliferation.[40] [41] Therefore, inhibiting the excessive activation of NF-κB signaling pathway, reducing the release of inflammatory factors, and reducing glomerular mesangial cell proliferation can delay the progression of MN.

Through analysis, it was found that the Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs could act on key targets such as AKT1, MAPK1, JUN, CASP3, TNF, VEGFA, MAPK8, IL-6, so as to inhibit signaling pathways of PI3K/AKT, MAPK, TNF, HIF-1, IL-17, NF-κB, VEGF, mTOR, etc., which plays a role in down-regulating inflammatory response, reducing urinary protein excretion and edema, and thus achieving the purpose of treating MN. This study has preliminarily confirmed the multicomponent, multitarget, multipathway synergistic effect of Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs in the treatment of MN, which provides the theoretical evidence for the clinical application of this couplet herbs and provides directions and references for future experimental studies.

Conclusion

The Jinyingzi (Rosae Laevigatae Fructus)–Qianshi (Euryales Semen) couplet herbs can regulate PI3K/AKT, MAPK, NF-κB signaling pathways in MN by targeting proteins of AKT1, MAPK8, PTGS2 through key components of quercetin,β-sitosterol and kaempferol, so as to inhibit the overexpression of inflammatory factors in renal tissues, regulate inflammatory response, and improve renal function.

Conflict of Interest

The authors declare no conflict of interest.

CRediT Authorship Contribution Statement

Haoyi Tian: Conceptualization, data curation, formal analysis, visualization, and writing -original draft. Tian Yun: Funding acquisition, supervision, writing-review & editing.

• References

• 1 Xu X, Wang GB, Chen N. et al. Long-term exposure to air pollution and increased risk of MN in China. J Am Soc Nephrol 2016; 27 (12) 3739-3746
• 2 Ren S, Wang Y, Xian L. et al. Comparative effectiveness and tolerance of immunosuppressive treatments for idiopathic membranous nephropathy: a network meta-analysis. PLoS One 2017; 12 (09) e0184398
• 3 Chen Z. Analysis of reactivity factors after the treatment of idiopathic membranous nephropathy. Electron J Clin Med Lit 2019; 6 (64) 50
• 4 He BY. Experience and experience of TCM treatment for idiopathic membranous nephropathy. Clin J Tradit Chin Med 2019; 31 (02) 273-276
• 5 Zhu TT, Wang C. Essence of clinical experience of Zheng Pingdong in treating chronic nephritis proteinuria. Liaoning J Tradit Chin Med 2016; 43 (01) 34-35
• 6 Chen HL, Song LQ. Medication analysis of Song Liqun's treatment of membranous nephropathy. Acta Chin Med 2021; 36 (04) 787-792
• 7 Wang L, Yang ZH, Yang K. et al. Medication rule analysis of Professor Yang Hongtao in the treatment of MN based on data mining. China Med Her 2021; 18 (07) 122-125
• 8 Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China: the 2020 edition. Beijing: China Medical Science and Technology Press; 2020: 232
• 9 Chen Q, Li N, Zhang YL. et al. Research progress on Jinyingzi (Rosa laevigata michx). Guid J Tradit Chin Med Pharm 2018; 24 (19) 106-110
• 10 Liu XS, Li XL, Fu XQ. et al. Pharmaceutical research progress of Qianshi (Euryales Semen). Mod Heal Pres 2019; 19 (14) 61-62
• 11 Zhu YD. Hypoglycemic effect and renal function regulation of ultrafine powder of gorgon Euryale seed on type 2 diabetic mice. Hefei: Hefei University of Technology; 2020
• 12 Niu M, Zhang SQ, Zhang B. et al. Interpretation of network pharmacology evaluation method guidance. Chin Tradit Herb Herbs 2021; 52 (14) 4119-4129
• 13 Yang YC, Liu XC, Ma BM. et al. Professor Chuanhui Ye's experience in treating idiopathic membranous nephropathy. Shaanxi J Tradit Chin Med 2015; 36 (01) 78-80
• 14 Lu ZZ, Chen WJ, Liu WY. et al. Summary of Professor Yueyi Deng's experience in treating idiopathic membranous nephropathy. Chin J Integr Tradit West Nephrol 2018; 19 (10) 909-910
• 15 Li YD. Daning Zhang's experience in treating membranous nephropathy. Guangming J Chin Med 2018; 33 (13) 1868-1869
• 16 Wang C, Yang C, Tang SF. Professor Qinguo Hong's clinical experience in differentiating and treating idiopathic membranous nephropathy. Hebei J Tradit Chin Med 2018; 40 (03) 325-328
• 17 Zhi Y, Zhao W, Lin Y. et al. Shili Cao's experience in treating MN from collateral theory. J Basic Clin Med 2020; 26 (08) 1170-1171 , 1174
• 18 Ma N, Li YJ, Fan JP. Research progress on pharmacological action of quercetin. Liaoning Zhongyiyao Daxue Xuebao 2018; 20 (08) 221-224
• 19 Albatany M, Meakin S, Bartha R. The monocarboxylate transporter inhibitor quercetin induces intracellular acidification in a mouse model of glioblastoma multiforme: in-vivo detection using magnetic resonance imaging. Invest New Drugs 2019; 37 (04) 595-601
• 20 Wang YY, Li GT, Wang MS. et al. Quercetin attenuates spinal cord injury by inhibiting TLR4/ NF-κB-mediated inflammatory response. Orthop J China 2020; 28 (14) 1311-1316
• 21 Li LM, Lin SS, Zhang DH. et al. The anti-inflammatory effects of quercetin on acute laryngitis and its action mechanism. Hebei Med J 2020; 42 (18) 2730-2734
• 22 Cai JH, Ren YH, Li JL. et al. Quercetin regulates IL-6/STAT3 signaling pathway in mouse models of inflammation-related colon cancer: a mechanistic study. Zhongguo Yaowu Yu Linchuang 2020; 20 (06) 896-899
• 23 Wu HH, Yang H, Wu P. Regulatory mechanism of quercetin on A549 cell PI3K/Akt signaling pathway and malignant characteristics of tumor. Zhongchengyao 2021; 43 (12) 3476-3479
• 24 Chen YH, Zhou KY, Yuan HY. Research progress of the pharmaceutical effect of kaempferol. Guangdong Yixue 2010; 31 (08) 1064-1066
• 25 Wu QM, Ni HX, Lu X. et al. The study of protective effects of kaempferol on renal tissues in db/db mice. Prev Treat Cardio Cereb Vasc Dis 2017; 17 (01) 19-22
• 26 Lu CC, Wang GH, Lu J. et al. Role of podocyte injury in glomerulosclerosis. Adv Exp Med Biol 2019; 1165: 195-232
• 27 Chen YK, Zeng A, Luo ZH. et al. Advances on pharmacology of β-sitosterol. Guangdong Yaoxueyuan Xuebao 2021; 37 (01) 148-153
• 28 Wu CS, Huang JJ. (AI/YI) XL, et al. Research progress of β-carotene. Zhongguo Yiyuan Yaoxue Zazhi 2008; 28 (16) 1381-1383
• 29 Yan CM, Tian H, Huo MX. et al. Effects of β-carotene on expressions of NF-kB, TGF-β1 and IL-6 in rats with acute pancreatitis. Jiangsu Med J 2018; 44 (04) 357-360
• 30 Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell 2017; 168 (06) 960-976
• 31 Choi ME. Autophagy in kidney disease. Annu Rev Physiol 2020; 82: 297-322
• 32 Lin J, OuYang H, Liang CL. et al. Effect of total glucosides of Paeonia on renal protection and autophagy in rats with membranous nephropathy. Tradit Chin Herb Res. Clin Pharmacol 2019; 30 (09) 1025-1031
• 33 Yao L, Li J. Research progress of renal fibrosis related MAPK signaling pathway and Chinese herbs intervention. Zhonghua Zhongyiyao Zazhi 2015; 30 (07) 2431-2433
• 34 Hao CX, Su YJ, Wang QJ. et al. Inhibition of curcumin on up regulation c-fos and c-Jun mRNA in kidney of diabetic rats. J Med Pest Control 2010; 26 (12) 1089-1091
• 35 Yu WL, Wu XY, Zhang LL. et al. Relationship between the expression of interleukin-17 in the kidney of diabetic rats and diabetic nephropathy. Chin J Integr Tradit West Nephrol 2010; 11 (11) 958-960 , 1037
• 36 Nguyen H, Chiasson VL, Chatterjee P, Kopriva SE, Young KJ, Mitchell BM. Interleukin-17 causes Rho-kinase-mediated endothelial dysfunction and hypertension. Cardiovasc Res 2013; 97 (04) 696-704
• 37 Yang LL, Wang YJ, Liu G. et al. Renal protection of N-acetylcysteine in rats with membranous nephropathy. J Pract Med 2019; 35 (06) 848-854 , 859
• 38 Li B, Zhou J, Zhang GQ. et al. Relationship between the level of hypoxia-inducible factor-1α and the risk factors for progression of IgA nephropathy. Ningxia Med J 2019; 41 (04) 289-291
• 39 Zhou Y, Hong Y, Huang H. Triptolide attenuates inflammatory response in membranous glomerulo-nephritis rat via downregulation of NF-κB signaling pathway. Kidney Blood Press Res 2016; 41 (06) 901-910
• 40 Liu CM, Sun YZ, Sun JM, Ma JQ, Cheng C. Protective role of quercetin against lead-induced inflammatory response in rat kidney through the ROS-mediated MAPKs and NF-κB pathway. Biochim Biophys Acta 2012; 1820 (10) 1693-1703
• 41 Fan HW, Xue HY, Yao JH. et al. Ligustrazine reduces inflammatory response and kidney injury in rats with MNby inhibiting NF-κB signaling pathway. Herb Eval Res 2021; 44 (08) 1666-1673

Publikationsverlauf

Eingereicht: 18. April 2022

Angenommen: 13. Juni 2022

Artikel online veröffentlicht:
27. Oktober 2022