Beneficial effects of Bifidobacteria in a gastroresistant seamless capsule on hyperhomocysteinemia in hemodialysis patients.

Intestinal microflora is deranged in hemodialysis (HD) patients as an increase in aerobic bacteria such as Escherichia coli and a decrease in anaerobic bacteria such as Bifidobacterium . Bifidobacteria ferment carbohydrates to produce acetic acid and lactic acid, which inhibit the intestinal putrefaction. Thus, intake of Bifidobacteria effectively restores the disturbed microflora to normal. However, Bifidobacteria in most medical products and healthy foods cannot usually survive because of exposure to gastric juices before it reaches the intestines. A gastroresistant seamless capsule prevents Bifidobacteria from inactivation by acidic gastric juice and allows it to be active in the intestines. We showed that the oral administration of Bifidobacterium longum in a gastroresistant seamless capsule to HD patients is effective in decreasing the pre-HD serum levels of homocysteine, indoxyl sulfate, and triglyceride. The reduction in the serum level of homocysteine is mainly attributable to the supply of folate produced by Bifidobacterium longum in the human intestines.

Interactions of human organic anion as well as cation transporters with indoxyl sulfate.

Various uremic toxicants including indoxyl sulfate exert a number of biological effects on uremic patients. In order to elucidate the molecular mechanisms for the pharmacokinetics of indoxyl sulfate in human, we examined the interactions of human organic anion transporters (human-OATs) and human organic cation transporters (human-OCTs) with indoxyl sulfate using stable transfectants. Indoxyl sulfate inhibited human-OAT1, human-OAT3 and human-OAT4, but not human-OAT2, human-OCT1 and human-OCT2. Kinetic analysis revealed that the K(i) values for human-OAT1, human-OAT3 and human-OAT4 were 22.7, 168.7 and 181.3 microM, respectively. Human-OAT1 and human-OAT3 mediated the uptake of indoxyl sulfate and human-OAT4 mediated not only the uptake but also the efflux of indoxyl sulfate. In conclusion, by comparing the K(i) values with the plasma concentration of unbound indoxyl sulfate, it was predicted that human-OAT1 and human-OAT3 mediate the transport of indoxyl sulfate in vivo. In addition, it was suggested that human-OAT1 and human-OAT3 are involved in the urinary excretion of indoxyl sulfate, the exacerbation of renal dysfunction and the induction of uremic encephalopathy by indoxyl sulfate.

An inhibitor of advanced glycation end product formation reduces N epsilon-(carboxymethyl)lysine accumulation in glomeruli of diabetic rats.

BACKGROUND: An inhibitor of advanced glycation, OPB-9195, retards the progression of nephropathy in Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model of non-insulin-dependent diabetes mellitus. The aim of this study is to evaluate histologically the role of N(epsilon)-(carboxymethyl)lysine (CML) in the development of diabetic nephropathy and investigate whether inhibition of CML accumulation by OPB-9195 is associated directly with the prevention of glomerular lesions in OLETF rats. METHODS: Kidneys of OLETF and Long-Evans Tokushima Otsuka rats were obtained at ages 7, 20, 50, and 68 weeks after collecting their blood and urine samples. OPB-9195 had been administered to the rats from age 24 weeks to the end of the experiments. CML in kidneys was detected by using a monoclonal antibody against CML according to an indirect immunofluorescence technique. CML-positive glomerular area was measured using NIH Image software (Research Services Branch of NIMH, Bethesda, MD). Hyalinized and/or sclerotic areas in glomeruli and mesangial and glomerular volume were measured using a point-counting technique. RESULTS: CML-positive area in glomeruli correlated closely not only with urinary albumin excretion (r = 0.912; P = 0.001), but also with volumes of mesangium and hyalinized and/or sclerotic lesions (r = 0.859; P = 0.0019 and r = 0.833; P = 0.0027, respectively). Treatment with OPB-9195 reduced CML-positive area and prevented the increase in mesangial volume, with no significant change in glomerular volume at age 68 weeks. The volume of hyalinized and/or sclerotic lesions also decreased by treatment with OPB-9195 in three of four rats at age 68 weeks. CONCLUSION: CML is a major advanced glycation end product contributing to the development of diabetic nephropathy, and inhibition of its accumulation by OPB-9195 results in amelioration of glomerular lesions in OLETF rats.

Bifidobacterium in gastro-resistant seamless capsule reduces serum levels of indoxyl sulfate in patients on hemodialysis.

BACKGROUND: Intestinal microflora is deranged in hemodialysis (HD) patients as increased aerobacteria such as Escherichia coli, and decreased anaerobacteria such as bifidobacteria. Indole, a precursor of indoxyl sulfate, is produced by E coli but not by bifidobacteria. The serum levels of indoxyl sulfate are increased markedly in HD patients and cannot be reduced efficiently by HD because of its albumin binding. METHODS: To compare the effect of oral administration of Bifidobacterium longum in gastro-resistant seamless capsule (Bifina) on indoxyl sulfate levels with that of Bifidobacteria in powder formulation (Lac B), Bifina was administered to 11 HD patients for 5 weeks, and Lac B to another group of 11 HD patients for 5 weeks. The authors measured the serum level of indoxyl sulfate by using high-performance liquid chromatography. RESULTS: The pre-HD serum levels of indoxyl sulfate significantly decreased in Bifina-treated patients (before, 4.9 +/- 1.7 mg/dL, 4.5 mg/dL, mean +/- SD, median, after 5 weeks, 3.5 +/- 1.3 mg/dL, 3.8 mg/dL; P < 0.005). However, they did not decrease in the Lac B-treated patients (before, 4.8 +/- 1.4 mg/dL, 4.5 mg/dL, after 5 weeks, 5.2 +/- 2.0 mg/dL, 5.1 mg/dL). CONCLUSION: Oral administration of Bifina to HD patients is effective in reducing the serum levels of indoxyl sulfate by correcting the intestinal microflora. Gastro-resistant seamless capsule prevents bifidobacteria from its inactivation in acidic gastric juice, and allows it to be actived in the intestine.

Role of organic anion transporters in the tubular transport of indoxyl sulfate and the induction of its nephrotoxicity.

In uremic patients, various uremic toxins are accumulated and exert various biologic effects on uremia. Indoxyl sulfate (IS) is one of uremic toxins that is derived from dietary protein, and serum levels of IS are markedly increased in both uremic rats and patients. It has been previously reported that the accumulation of IS promotes the progression of chronic renal failure (CRF). This study demonstrates the role of rat organic anion transporters (rOATs) in the transport of IS and the induction of its nephrotoxicity. The administration of IS to 5/6-nephrectomized rats caused a faster progression of CRF, and immunohistochemistry revealed that IS was detected in the proximal and distal tubules where rOAT1 (proximal tubules) and/or rOAT3 (proximal and distal tubules) were also shown to be localized. In in vitro study, the proximal tubular cells derived from mouse that stably express rOAT1 (S2 rOAT1) and rOAT3 (S2 rOAT3) were established. IS inhibited organic anion uptake by S2 rOAT1 and S2 rOAT3, and the Ki values were 34.2 and 74.4 microM, respectively. Compared with mock, S2 rOAT1 and S2 rOAT3 exhibited higher levels of IS uptake, which was inhibited by probenecid and cilastatin, organic anion transport inhibitors. The addition of IS induced a decrease in the viability of S2 rOAT1 and S2 rOAT3 as compared with the mock, which was rescued by probenecid. These results suggest that rOAT1 and rOAT3 play an important role in the transcellular transport of IS and the induction of its nephrotoxicity.