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With the improvement of people's living standard and the change of dietary structure, the prevalence of gout has increased gradually with the increased intake of protein, sugar and fat. There has been a positive correlation between gout and age, and the age of onset decreased gradually. The inflammation induced by sodium urate crystal is the pathological basis of gout, which activates innate immunity, releases many kinds of inflammatory mediators, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α, and then causes inflammatory cascade reaction and acute attacks, such as joint redness, swelling and heat pain. There is a spontaneous remission mechanism in gout. For one thing, macrophages reduce the stimulation of monosodium urate(MSU) through phagocytosis of MSU crystals as foreign bodies, for another, differentiated and mature macrophages secrete anti-inflammatory factor transforming growth factor(TGF)-β1, inhibit the expression of inflammatory factors and promote spontaneous relief of acute gout attack. In addition to the activation mechanism of intracellular signaling molecules associated with inflammatory response, the inflammatory mechanism of gout also involves complement activation, cell activation and other pathways. The complications caused by gout, such as cardiovascular system damage and joint destruction, are seriously harmful to human health. At present, western drugs, such as allopurinol and febuxostat, exert an effect in inhibiting xanthine oxidase. Benzimarone has effect in reducing renal absorption of uric acid and promoting uric acid excretion by inhibiting uric acid transporter 1(URAT1) and glucose transporter 9(GLUT9). Even Lesinurad and other medicines in current studies are based on the inhibition of uric acid re-absorption, but with adverse reactions that limit the clinical application. The treatment of gout with traditional Chinese medicine(TCM) has multi-target characteristics, with advantages in reducing uric acid, resisting inflammation and improving joint function and a high safety. It has been gradually popularized and applied in clinical treatment of gout. Therefore, it is a promising research direction to treat gout with TCM and western medicine based on the pathomechanism of gout.
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Objective: To explore the effect of hirudin on hyperuricemia rats and its mechanism. Methods: Male Wistar rats were randomly divided into control group, model group, allopurinol (30 mg/kg) group, hirudin low-, middle- and high-dose (0.2, 0.4, 0.8 g/kg) group. Rats were ig potassium oxonate (0.75 g/kg) to induce hyperglycemia model, once a day for five weeks. And all administration groups were respectively ig corresponding doses of drugs. The level of uric acid in serum and urine of rats were measured by biochemical method; The level of organic anion transporter 1 (OAT1) in kidney was measured by immunohistochemistry; The protein expressions of glucose transporter 9 (GLUT9), OAT1 and urate transporter 1 (URAT1) in kidney were measured by western blotting; The expression levels of GLUT9, OAT1 and URAT1 mRNA in kidney were detected by qRT-PCR. Results: Compared with control group, the level of uric acid in serum and urine of rats in model group was significantly increased (P < 0.01), the expressions of GLUT9, URAT1 mRNA and protein were significantly increased (P < 0.01), the expressions of OAT1 mRNA and protein were significantly decreased (P < 0.01). Compared with model group, the level of uric acid in serum and urine of rats in hirudin group were significantly decreased (P < 0.01), the expressions of GLUT9, URAT1 mRNA and protein were significantly reduced (P < 0.01), the expressions of OAT1 mRNA and protein were significantly increased (P < 0.01). Conclusion Hirudin can reduce the uric acid by regulating the expressions of renal urate transporters OAT1, URAT1 and GLUT9 in hyperuricemia rats.
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Urate transporter 1 (URAT1) is a validated target for the treatment of hyperuricemia. Based on the structure of URC-102, which is currently under a phase Ⅱ clinical trial, fourteen novel analogs were designed and synthesized. Among them, four compounds (9b, 9c, 10e and 10g) exhibited substantial inhibitory effect against URAT1. The most active compound 9b showed an IC50 value of 0.061 μmol·L-1, which is significantly more potent than Lesinurad and Benzbromarone. Preliminary SAR was drawn, providing clues for further structural optimization.
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Objective To investigate the mechanism of total flavonoids of Mori Cortex (TFMC) in improving hyperlipidemia and hyperuricemia related nephropathy. Methods The molecular structure of URAT1 protein and structure of Mori Cortex total flavonoids extract were docked by selecting the effective components of total flavonoids extract of Mori Cortex and related genes of uric acid. Forty SD rats were randomly divided into four groups: normal group, model group, TFMC group (1.0 g/kg) and benzbromarone group (6.25 mg/kg). The hyperlipidemia and hyperuricemia model was established by feeding with high fat diet plus adenine and ethylamine butanol. After 16 weeks, the levels of blood glucose and blood lipids in serum, uric acid (UA), creatinine (CRE), and urea nitrogen (BUN) of rats in each group were compared. The renal pathological changes were observed by hematoxylin eosin staining. The expressions of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), uric acid anion exchange transporter 1 (URAT1) and organic anion transporter 1 (OAT1) were detected by qRT-PCR. Results The main component of moracin of total flavonoids of Mori Cortex was the high target of the genes related to uric acid, suggesting that the moracin might be the main active component in the improvement of hyperlipidemia and hyperuricemia related nephropathy. After 16 weeks of drug intervention, the serum levels of Glu, TC, TG, LDL-C, UA, CRE and BUN in the model group were significantly higher than those in the normal group, and the level of HDL-C in the model group was significantly higher than that in the normal group (P < 0.05, 0.01). Compared with the model group, the above biochemical indexes in the TFMC group and the benzbromarone group were significantly decreased and HDL-C was significantly increased (P < 0.05, 0.01). The results of HE staining showed that the epithelial cells of renal tubules in the model group were swollen and necrotized, and the protein tubules could be seen in the renal tubules. In the TFMC group, some renal tubular epithelial cells were slightly swollen, and no inflammatory cells infiltrated around them. The structures of renal cortex and medullary were clear, and no hyperplasia or atrophy in glomerular were found, no tubular type and inflammatory cell infiltration in renal interstitial tissue of rats in benzbrommarone group were observed. The results of qRT-PCR showed that, compared with the normal group, the content of IL-6, TNF-α, and URAT1 mRNA was significantly increased, the content of OAT1 mRNA was significantly decreased in the model group; The content of above indicators was decreased and OAT1 was increased after drug intervention, (P < 0.05, 0.01). Conclusion The improvement of hyperlipidemia and hyperuricemia associated nephropathy may be related to the regulation of IL-6, TNF-α, URAT1, and OAT1 mRNA. Mori Cortex has obvious influence on the key factor of hyperlipidemia and hyperuricemia with URAT1 as its potential target, and the results of molecular virtual docking and animal experiments are similar. It provides a theoretical basis for further study on the improvement of hyperlipidemia and hyperuricemia related nephropathy and provides a reference for the further molecular mechanism.
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Objective: To observe the effect of phlegm and blood stasis on the expressions of sirtuin 3(SIRT3)protein and urate transporter 1(URAT1) mRNA in skeletal muscle of diabetic rats with gout. Method: The 40 healthy rats, excepting the normal group, the remaining groups were fed with high-fat diet combined with low-dose streptozotocin solution (40 mg·kg-1) once a day, with blood glucose "16.7 mmol·L-1" as the criterion for the diabetes model. After 4 days, the 5% sodium urate solution was injected into the joint cavity once to induce the gout model. After the successful modeling, the Biling group (10 g·kg-1), the indomethacin group (5 mg·kg-1) and the pioglitazone group (10 mg·kg-1) continued to be administered for 21 days. The normal group and the model group were given the same amount of normal saline. The expression of SIRT3 protein in skeletal muscle tissue was determined by Western blot, URAT1 mRNA expression in bone tissue was detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR),and blood was collected to measure blood glucose (GLU), blood uric acid (UA) and C-reactive protein (CRP). Result: Compared with the normal group, GLU, UA and CRP in the model group were significantly increased (PPPPPPPPConclusion: Biling Qutong prescription with effects in purging turbidity, detoxifying and dredging collaterals can significantly reduce the content of serum inflammatory factor CRP, significantly increase the protein expression of SIRT3 in skeletal muscle tissue of model rats, lower the content of URAT1 mRNA, reduce the blood glucose and blood uric acid levels in diabetic gout rats, and protect joints.
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Lesinurad (RDEA 594), discovered by Ardea Biosciences, is a new drug used for the treatment of gout. RDEA594 could increase the excretion of uric acid to reduce uric acid levels by inhibiting the uric acid salt transport protein 1 (URAT1). In this essay, we summarize synthetic methods of lesinurad reported in recent years, evaluate and analyze them briefly, aiming to lay a foundation for lesinurad synthesis process to achieve industrialization.
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Aim To investigate the effects of 3 ,5 ,2 ’ , 4’-tetrahydroxychalcone (P40) on urate excretion, as well as mRNA and protein expressions of renal URAT1 and GLUT9 in hyperuricemic mice. Methods Sixty Kunming mice were randomly divided into six groups:normal control group, hyperuricemic group ( model group), P40 2. 0, 4. 0, 8. 0 mg·kg-1 groups and positive control group. All drugs were administered in-tragastrically to mice for 5 doses. Hyperuricemic mice were induced by intraperitoneal injection of uric acid (0. 15 g·kg-1 body weight) for 3 times. The urate levels were assayed with the phosphotungstic acid method. The mRNA and protein expressions of GLUT9 and URAT1 were determined by RT-PCR and Western blot. Results P40 at a dose of 4. 0 and 8. 0 mg · kg-1 significantly reduced the serum urate levels in a dose-dependent manner, when compared with untreat-ed hyperuricemic mice ( P<0. 05 or 0. 01 ) . The he-patic urate contents decreased in untreated-and treated-hyperuricemic mice as compared with normal mice ( P<0. 01 ) . Furthermore, P40 had no influence on the renal URAT1 mRNA and protein expression levels, while it could down-regulate renal GLUT9 protein ex-pression but not mRNA expression in hyperuricemic mice. Conclusion P40 possesses potent uricosuric effects associated with urate reabsorption by down-regu-lating the protein expression of GLUT9 in kidney.
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Objective To investigate the metabolic profile of uric acid and the significance of the altered renal expression of urate trans-porter 1(URAT1) in patients with uric acid nephrolithiasis. Methods The data of 24 patients in our hospital from January 2012 to October 2013 were analyzed retrospectively. Participants in the research were divided into three groups:patients with uric acid nephrolithiasis,other patients with nephrolithiasis and normal participants. The basic clinical data and the related data of uric acid metabolition of participants were collected,URAT1 gene expression in renal tissures of three groups was detected by Real-time PCR technique. All data were statistically ana-lyzed and compared between these groups. Results Uric acid levels in plasma,body mass index and age were significantly higher in patients with uric acid nephrolithiasis than other two groups (P0. 05). The result of Real-time PCR suggested that the URAT1 renal expression was significantly higher in patients with uric acid nephrolithiasis than other two groups (P<0. 05). Conculusion Patients with uric acid nephrolithiasis are closely related with hyperuricemia,but unrelated with renal over-ex-cretion of uric acid. The upregulated URAT1 expression in the kidney may be an important molecular mechanism of the clinical features.
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A 24-year-old male visited our hospital because of pain in both flanks. His biochemistry profile showed an elevated serum creatinine level and low serum uric acid level. History taking revealed that he had undertaken exercise prior to the acute kidney injury (AKI) event, and he stated that family members had a history of urolithiasis. His renal profile improved after hydration and supportive care during hospitalization. Although the patient was subsequently admitted again due to AKI, his status recovered with similar treatment. Since the diagnosis of the patient was familial renal hypouricemia with exercise-induced AKI, we performed genotyping of SLC22A12, which encodes human urate transporter 1. The diagnosis was confirmed by the detection of a homozygous mutation of W258X. We herein, report a case of familial renal hypouricemia confirmed by genotyping of SLC22A12, and review the relevant literature.
Subject(s)
Humans , Male , Young Adult , Acute Kidney Injury , Biochemistry , Creatinine , Diagnosis , Hospitalization , Uric Acid , UrolithiasisABSTRACT
Objective To investigate a new single nucleotid polymorphism (SNP) intron5(+4668C/T) in SLC22A12 in primary gout patients and the association between clinical characteristics and genotypes. Methods One hundred and one primary gout patients and 186 healthy subjects were recruited into this study. Blood pressure, body mass index (BMI) was recorded. Serum uric acid, glucose, lipid and creatinine were detected. DNA was extracted from peripheral blood to amplify the fragment located in intron 5. The genotypes of SLC22A12 can be detected with high-resolution melting (HRM) assay, followed by sequencing analysis. Chi-square test was used for statistical analysis. Results ① A new SNP in intron 5 of SLC22A12 was identi-fied successfully by HRM, which was defined as intron 5 (+4668C/T). CC, CT and TT genotypes were unam-biguously distinguished with HRM technology, which was fully concordant with sequencing. ②The genotypes of CC, CT and TT in male and female groups were 28.1%, 33.7%, 38.2% and 20.0%, 47.1%, 32.9%, respectively.③ However, no significant differences of genotype distribution were found concerning BMI, blood pressure, creatinine, total cholesterol and triglyceride in both male group and female group. But the serum uric acid levels in the CC genotype were significantly higher than those with the CT+TT genotypes. ④ The genotype frequencies of CC and CT+TT in high uric acid group were remarkably different from those in low uric acid group (21.2%, 78.8%,; 35.0%, 65.0%; P<0.05). Conclusion A new SNP has been successfully discovered with HRM technology with simplicity, rapidity and accuracy. T allele of intron 5 (+4668C/T) may be a genetic protective factor for hyperuricemia among Chinese population.
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Objective To study the association between hURAT1 gene single nucleotide polymorphism(SNP) and primary hyperuricemia(HUA). Methods A total of 215 patients with HUA and 323 healthy subjects were chosen to investigate SNP of hURAT1. Exon 2 to 4 and flanking introns of the hURAT1 gene in patients and control individuals were screened with PCR. The relationship between SNP of hURAT1 gene with HUA was studied with statistical analysis. Results The frequency of AA/AG genotype was significantly increased in HUA patients as compared with that in healthy controls( 11.6% vs 3.7% ,P =3.81 × 10-4). Allele A of hURAT1 intron 3, 11 G ＞A was found significantly higher in the group of HUA patients, being detected in 6.0% of the HUA patients alleles and in 1.9% of the healthy control alleles (P =2.66 × 10-5 ). Those carrying the low frequency AA/AG genotype had a risk effect on the morbidity of HUA and the odds ratio for the HUA patients versus controls was 3.41 with AA/AG genotype versus GG genotype( OR = 3.41,95% CI = 1.67 - 6.95 ). The HT4 haplotype, which carried the intron 3,11A allele, was associated with a significantly increased risk of HUA(69.44% vs 30.56% ,P ＜ 0.001). Conclusion The SNP of 11G ＞A in the intron 3 of hURAT1 gene was apparently associated with HUA, thus suggesting the genetic effect of hURAT1 gene in the pathogenesis of HUA.
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Idiopathic renal hypouricemia is a disorder characterized by impaired urate handling in the renal tubules. Most patients with hypouricemia are asymptomatic and are found incidentally, but the condition is known to be at high risk for exercise-induced acute renal failure or urolithiasis. URAT1 protein encoded by SLC22A12 gene has been identified recently as a urate/anion exchanger in the human kidney. Inactivation mutations in SLC22A12 gene have been shown to cause renal idiopathic hypouricemia. We experienced a 3-year-old boy who presented with persistent orange-colored urine since infancy. His urine contained many uric acid crystals, while the serum showed hypouricemia(0.7 mg/dL). The fractional excretion of uric acid was increased to 41.7%. SLC22a12 gene analysis revealed W258X homozygote alleles. Renal hypouricemia must be included in the differential diagnosis of red-urine and SLC22A12 gene analysis is recommended in idiopathic renal hypouricemia.