Huangkui Capsule in Combination with Leflunomide Improves Immunoglobulin A Nephropathy by Inhibiting the TGF-β1/Smad3 Signaling Pathway

Abstract

To investigate the efficacy and potential molecular mechanism of Huangkui capsule in combination with leflunomide (HKL) for the treatment of immunoglobulin A nephropathy (IgAN) METHODS: IgAN rat models were constructed by treating rats with bovine serum albumin, lipopolysaccharide, and tetrachloromethane. Th22 cells were isolated from the blood samples of patients with IgAN using a CD4+ T cell isolation kit. The expression levels of the components of the TGF-b1/Smad3 signaling pathway, namely, TGF-b1, Smad2, Smad3, Smad4, and Smad7, were detected using quantitative reverse transcription polymerase chain reaction. Cell proliferation was determined using the MTT assay, cell viability was determined using the WST 1 method, and the chemotaxis of Th22 cells was observed using the wound healing assay. Changes in the histology of the kidney tissues were analyzed using hematoxylin and eosin staining. RESULTS: Compared with IgAN rats, the rats subjected to HKL treatment showed good improvement in kidney injuries, and the combined drug treatment performed much better than the single-drug treatment. In addition, following HKL treatment, the viability, proliferation, and chemotaxis of Th22 cells dramatically decreased (*po0.05, **po0.01, and ***po0.001). In addition, CCL20, CCL22, and CCL27 levels decreased and the expression of the key components of the TGF-b1/Smad3 signaling pathway was downregulated in IgAN rats and Th22 cells (*po0.05, ***po0.001). CONCLUSIONS: By targeting the TGF-b1/Smad3 signaling pathway, HKL treatment can improve kidney injury in IgAN rats as well as the excessive proliferation and metastasis of Th22 cells.

Spectrum-Effect Relationship and Component Knock-Out or Knock-In in Total Flavones of Abelmoschus manihot

“…The traditional Chinese medicine preparations have also achieved a certain effect on the treatment of CKD, and Huangkui capsules is one of them [14][15][16] .The main components of Huangkui capsules are the total avonoids extracted from Abelmoschus manihot. They have been widely used to treat CKD, DKD and IgA nephropathy [17][18][19][20][21][22][23] . However, it is rarely reported whether the proportion relationship among seven avonoid monomers has signi cant effect in the treatment of DKD.…”

“…MTT assay was used to determine cell viability by the manufacturer's instruction. MPC-5 cells were plated in 96-well plates at a density of 5×10 3 cells/well and incubated with normal glucose (NG, 5.5 mM glucose+ 24.5 mM mannitol), hign glucose (HG, 30 mM glucose), osmotic control (24.5 mM Mannitol was added to the NG group) and HG with TFA at different concentrations (5,10,20, 40 μg/mL) for 48 h. Then, an MTT solution (5 mg/mL) was added to each well, and the MPC-5 cells were incubated for 2-3 h at 37 °C. The supernatants were then aspirated, and 100 μL of dimethyl sulfoxide (DMSO) was added to solubilize formazan crystals with shaking for 10 min.…”

Efficacy of Huangkui Capsules in the Treatment of Diabetic Kidney Disease: A Systematic Review and Using Network Pharmacology

“…[34] Additionally, accumulated evidence has demonstrated that HKC could ameliorate proteinuria induced by primary glomerular diseases, such as primary immunoglobulin A nephropathy and membranous nephropathy. [35] Proteinuria, a symptom of various diseases, is usually caused by metabolic and immune-mediated disturbances of glomerular filtration. Generally speaking, the pathogenesis of metabolic-mediated proteinuria is complicated, including but not limited to glomerular hyperfiltration, highly increased trans-glomerular pressure, and the consequent glomerular and tubulointerstitial damage.…”

Traditional Chinese medicine for the treatment of immune-related nephropathy: A review

“…ChiCTR1900025273), and leflunomide is a clinical drug commonly used in IgAN treatment. A combination of the above two drugs inhibits the expression of the TGF- β 1/small mothers against decapentaplegic 3 (SMAD3) signaling pathway, and the therapeutic effect is superior to that of single drug therapy 80 , 81 .…”