RESUMEN
BACKGROUND: Diabetic nephropathy (DN) is one of the main causes of end-stage renal disease with scantly effective treatment. Numerous evidences indicated that macrophages play an important role in the occurrence and pathogenesis of DN by secreting inflammatory cytokines. Mincle is mainly expressed in macrophages and promotes kidney inflammation and damage of acute kidney injury. However, the role of Mincle in DN is unclear. In this study, we aim to investigate the effect of Mincle-related macrophage inflammation on DN, and whether it can be identified as the therapeutic target for Astragalus mongholicus Bunge and Panax notoginseng Formula (A&P), a widely used Chinese herbal decoction for DN treatment. METHODS: In vivo experiments high-fat and high-sugar diet and streptozotocin was used to establish a diabetic nephropathy model, while in vitro experiments inflammation model was induced by high-glucose in mouse Bone Marrow-Derived Macrophages (BMDM) cells and mouse mesangial (MES) cells. Kidney pathological staining is used to detect kidney tissue damage and inflammation, Western blotting, Real-time PCR and ELISA are performed to detect Mincle signaling pathway related proteins and inflammatory cytokines. RESULTS: Mincle was mainly expressed in infiltrated macrophage of DN kidney, and was significant decreased after A&P administration. The in vitro experiments also proved that A&P effectively down-regulated the expression of Mincle in macrophage stimulated by high glucose. Meanwhile, the data demonstrated that A&P can reduce the activation of NFκB, and the expression and secretion of inflammatory cytokines in DN kidney or BMDM cells. Notably, we set up a co-culture system to conform that BMDM cells can aggravate the inflammatory response of mesangial (MES) cells under high glucose stimulation. Furthermore, we found that the anti-injury role of A&P in MES cells was dependent on inhibition of the Mincle in macrophage. CONCLUSION: In summary, our study found that A&P is effective in reducing renal pathological damage and improving renal function and inflammation in diabetic nephropathy by a mechanism mainly related to the inhibition of the Mincle/Card9/NFκB signaling pathway.
Asunto(s)
Astragalus propinquus , Nefropatías Diabéticas/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Lectinas Tipo C/metabolismo , Macrófagos/efectos de los fármacos , Proteínas de la Membrana/metabolismo , Células Mesangiales/efectos de los fármacos , Panax notoginseng , Animales , Proteínas Adaptadoras de Señalización CARD/metabolismo , Riñón/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Animales , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
PURPOSE: Chronic kidney disease (CKD) is a global nephrotic syndrome characterized by chronic inflammation, oxidative stress and fibrosis in the kidney. Isoliquiritigenin (ISL), a flavonoid from licorice, has historically been reported to inhibit innate immune responses to inflammation and fibrosis in vivo. However, the effect of ISL on CKD progression is largely unknown. MATERIALS AND METHODS: In this study, we employed the inflammatory and fibrotic models of LPS/TGF-ß-induced bone marrow-derived macrophages (BMDM) in vitro and unilateral ureteral obstruction (UUO) model in vivo to explore the potential effects and mechanism of ISL on renal inflammation and fibrosis. RESULTS: Our results manifest that ISL improved UUO-induced renal dysfunction and reduced tubular damage with a significantly downregulated mRNA expression and secretion of IL-1ß, IL-6, TNF-α and MCP-1 in vitro and in vivo. It is worth noting that ISL can strongly inhibit the mRNA and protein expression of Mincle (macrophage-induced c-type lectin) in BMDM and UUO. ISL inhibited the phosphorylation of Syk and NF-kappa B and simultaneously reduced the expression of α-SMA and Col III in vivo and in vitro. More interestingly, when dealing with TDB, a ligand of Mincle, it revealed significant reversal of protein expression levels as that observed with ISL. The expressions of IL-1ß, IL-6, TNF-α, iNOS, p-Syk, p-NF-kappa B, α-SMA and FN in BMDM inflammatory model were significantly upregulated with TDB treatment. This confirms that ISL inhibits inflammation and fibrosis of macrophage by suppressing Mincle/Syk/NF-kappa B signaling pathway. CONCLUSION: To conclude, ISL protects UUO-induced CKD by inhibiting Mincle-induced inflammation and suppressing renal fibrosis, which might be a specific renal protective mechanism of ISL, making it a novel drug to ameliorate CKD.
Asunto(s)
Chalconas/farmacología , Fibrosis/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Riñón/efectos de los fármacos , Sustancias Protectoras/farmacología , Transducción de Señal/efectos de los fármacos , Obstrucción Ureteral/tratamiento farmacológico , Administración Oral , Animales , Células Cultivadas , Chalconas/administración & dosificación , Chalconas/química , Relación Dosis-Respuesta a Droga , Fibrosis/metabolismo , Fibrosis/patología , Inflamación/metabolismo , Inflamación/patología , Riñón/metabolismo , Riñón/patología , Lectinas Tipo C/antagonistas & inhibidores , Lectinas Tipo C/metabolismo , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Proteínas de la Membrana/antagonistas & inhibidores , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Estructura Molecular , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Sustancias Protectoras/administración & dosificación , Sustancias Protectoras/química , Relación Estructura-Actividad , Quinasa Syk/antagonistas & inhibidores , Quinasa Syk/metabolismo , Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Factor de Crecimiento Transformador beta/farmacología , Obstrucción Ureteral/metabolismo , Obstrucción Ureteral/patologíaRESUMEN
To introduce the origin, development and theoretical source of the bloodletting therapy overseas. The bloodletting therapy, which was based on Hippocrates's four fluid theories, had the therapeutical effect and therefor spread abroad. Many people not only thought this therapy was benefit, but also let out blood regularly. During the medieval Europ, the bloodletting therapy reached its top, which was caused by the part-time job of barber. It was questioned when spreading in the America. After that, as the development of the other medical method, the disadvantage was exposed. With several clinical mistherapies, it faded away from the stage of history. The recent research shows that the normative bloodletting therapy could treat some diseases.