Abstract

Introduction

   Chronic glomerulonephritis (CGN) is a common glomerular disease with
   multifactorial pathogenesis. Huangkui capsule (HKC), a traditional
   Chinese herbal formulation, demonstrates therapeutic potential in CGN;
   however, its molecular mechanisms remain insufficiently characterized.
   This study aimed to clarify the therapeutic mechanisms of HKC in CGN by
   integrating proteomic analysis with network pharmacology.

Methods

   We employed liquid chromatography–mass spectrometry (LC-MS) to identify
   the active components of HKC. A CGN rat model was established and
   treated with HKC. Renal function parameters and serum inflammatory
   cytokines were assessed. Histopathological alterations and IgG
   deposition in kidney tissues were examined using hematoxylin–eosin (HE)
   staining and immunofluorescence, respectively. Proteomic profiling of
   renal tissue was conducted, and network pharmacology analysis was
   applied to identify potential therapeutic targets of HKC.

Results

   A total of 39 active compounds were identified in HKC. HKC
   administration significantly improved renal function and mitigated
   glomerular injury in CGN rats. Proteomic analysis revealed 2,079
   differentially expressed proteins, predominantly associated with
   oxidoreductase activity. Network pharmacology identified 462 targets
   related to HKC and 1,835 targets associated with CGN, with 13
   overlapping targets, including STAT3, PIK3R1, AKT1, HIF-1α, and VEGF,
   which were downregulated following HKC treatment.

Conclusion

   HKC exerts renoprotective effects in CGN by regulating multiple
   signaling pathways, notably HIF-1, VEGF, PI3K-Akt, MAPK, and PPAR.
   Through attenuation of inflammatory and oxidative responses, HKC
   alleviates renal pathological damage and supports kidney function,
   offering mechanistic insight into its multi-target therapeutic
   potential.

   Keywords: Huangkui capsule, proteomics, network pharmacology, chronic
   glomerulonephritis, mechanism

1 Introduction

   CGN is a common immune-mediated glomerular disorder characterized by
   proteinuria, hematuria, hypertension, and edema, often leading to renal
   insufficiency and, ultimately, end-stage renal disease (ESRD)
   ([34]Chebotareva et al., 2015; [35]Satirapoj et al., 2015). The global
   burden of CGN is rising, with mortality increasing annually from 2015
   to 2020, reaching 52,260 deaths in 2020 ([36]Foti et al., 2021). In
   China alone, chronic kidney disease (CKD) affects approximately 132.3
   million individuals, contributing nearly one-fifth of the global burden
   ([37]Kriz et al., 1998). Research suggests that cytokine- and
   immunokine-mediated microcirculatory disturbances, cellular
   proliferation, and immune complex deposition are key pathological
   mechanisms ([38]Ahmad and Bomback, 2020). Abelmoschus manihot (L.)
   Medic (AM), a traditional Chinese medicine, has been used in China for
   centuries. HKC, a single-herb traditional Chinese medicine preparation
   derived from AM corolla extract. It was approved by the China Food and
   Drug Administration (CFDA) in 1999 (Z19990040).The primary active
   components of AM are flavonoids, including rutin, hyperoside,
   hibifolin, isoquercitrin, myricetin, quercetin, and
   quercetin-3-O-robinobioside ([39]Guo et al., 2015; [40]Li et al.,
   2021). The chemical spectrum and structural formulas of these compounds
   are shown in [41]Figure 1. In vivo, these flavonoids can be metabolized
   into sulfate-glucuronide conjugates, which are considered key
   contributors to the nephroprotective effects of HKC ([42]Guo et al.,
   2016). Multiple multicenter randomized controlled trials (RCTs) have
   confirmed that HKC improves renal function in patients with primary
   glomerular diseases, IgA nephropathy, and diabetic nephropathy (DN)
   ([43]Li et al., 2017; [44]Li et al., 2020; [45]Zhao et al., 2022).
   Compared with conventional medications such as angiotensin-converting
   enzyme ([46]Regev et al., 2022) inhibitors and angiotensin receptor
   blockers (ARBs), HKC demonstrates distinct advantages in treating
   chronic kidney disease (CKD), as it improves renal function and reduces
   proteinuria without adverse effects on blood pressure. Both preclinical
   and clinical studies consistently indicate that HKC alleviates renal
   injury, reduces urinary protein excretion, and improves renal function
   ([47]Cai et al., 2017). The key active ingredients, including
   quercetin, myricetin, rutin, and hyperoside, exert therapeutic effects
   via multitarget mechanisms, such as anti-inflammatory, antioxidant, and
   immunomodulatory pathways ([48]Wan et al., 2022). Therefore, with its
   remarkable renoprotective effects, HKC has gradually become the
   clinical treatment of choice for CGN ([49]Pei and Li, 2021). However,
   studies on its direct targets and signaling pathways are still limited,
   and the specific mechanisms remain to be elucidated ([50]Liu et al.,
   2012).

FIGURE 1.

   [51]FIGURE 1
   [52]Open in a new tab

   Determination of 39 compounds in Huangkui capsule by LC-MS/MS.18.
   Rutin; 20. Hyperoside; 21. Isoquercetin.; 25. Hibifolin; 33. Myricetin;
   36. Quercetin. (A) Structural formulae of Rutin, Hyperoside,
   Isoquercetin, Hibifolin, Myricetin and Quercetin (B).

   This study will combine network pharmacology and proteomics to
   investigate the effectiveness and potential molecular mechanism of CGN
   treatment with HKC, which will provide new perspectives and scientific
   references for further revealing its potential pharmacological effects,