Studies On The Blood Concentration Of Indoxyl Sulfate And Its Degradation By Gene Engineering L.Lactis In Chronic Renal Failure

Part I The level of serum indoxyl sulfate in patients with chronic kidney disease and its relationship with renal functionObjectiveTo explore the serum level of indoxyl sulfate in chronic kidney diseases patients with different renal function and different therapies.Methods1. Experimental subjects:75patients with chronic kidney diseases in our department and20healthy participants were enrolled in this study (the eGFR was measured by the modified MDRD equation).then the subjects were divided into different group.2. Experiment with sub-groups:1) According to CKD stage and their treatment as follows:CKD3(7); CKD4(15);CKD5(53); the controlled(20);And2)gastrointestinal treatment (56); hemodialysis (19); the controlled (20).2. Experimental procedure:1) Collection of general information, clinical characteristics and biochemical indicators of the subjects.2) Detection of the serum and urine level of indoxyl sulfate by using high performance liquid chromatography-fluorescence.Results:χ±S1.The serum and urine level of indoxyl sulfate in healthy participants were0.085±0.064ug/ml and18.761±12.248ug/ml respectively;2. The plasma and urine level of indoxyl sulfate in CKD patients with different renal function:1) Compared with healthy participants: Serum IS:CKD3:0.127+0.122ug/ml, P=0.246; CKD4:0.482±0.533ug/ml, P＜0.0001; CKD5:2.745±2.493ug/ml, P＜0.0001. Urine IS:CKD3:3.616±2.885ug/ml, P<0.0001; CKD4:3.184±3.045ug/ml, P＜0.0001; CKD5:2.540±1.684ug/ml, P＜0.0001.2) Compared with CKD3stage patients: Serum IS:CKD4:0.482±0.533ug/ml, P=0.026; CKD5:2.745±2.493ug/ml, P＜0.0001. Urine IS:CKD4:3.184+3.045ug/ml, P=0.771; CKD5:2.540±1.684ug/ml, P=0.199.3) Compared with CKD4stage patients: Serum IS:CKD5:2.745±2.493ug/ml, P＜0.0001. Urine IS:CKD5:2.540±1.684ug/ml, P=0.465.3. The plasma and urine level of indoxyl sulfate in CKD patients with different therapies:1) Compared with healthy participants: Serum IS:gastrointestinal therapy:1.484±1.315, P＜0.0001; hemodialysis:5.002±2.532, P＜0.0001. Urine IS:gastrointestinal therapy:2.422±1.680,P＜0.0001; hemodialysis:3.879±2.249,P＜0.0001.2) Compared with gastrointestinal therapy: Serum IS:hemodialysis:5.002±2.532, P＜0.0001. Urine IS:hemodialysis:3.879±2.249, P＜0.01.4.There are a positive relationship between with serum indoxyl sulfate and the blood urea nitrogen (r=0.665, P＜0.0001),the serum creatinine (r=0.789, P＜0.0001) and the triglycerides (r=0.222, P=0.032); while, a negative relationship of serum indoxyl sulfate and hemoglobin (r=-0.548, P＜0.0001),eGFR(r=-0.807, P＜0.0001),low-density lipoprotein Cholesterol (r=-0.35, P=0.002)Conlusion1. The levels of serum indoxyl sulfate in CKD patients with different stages were significantly different, and its serum concentration of indoxyl sulfate was negatively related with eGFR, but its urine concentration were positively correlated with eGFR. 2. The serum levels of indoxyl sulfate in Hemodialysis patients was significantly higher than the gastrointestinal treatment patients.3.There are a positive relationship between serum indoxyl sulfate and TG, a negative relationship between with serum indoxyl sulfate and LDL-C and Hb, which suggest that indoxyl sulfate may play a very important role in the development of CKD complication of cardiovascular disease and renal anemia. Part II The decomposition role of gene engineering of The L.lactis on indoxyl sulfateObjectiveTo observer the decomposition role of gene engineering L.lactis on indoxyl sulfate both In vitro and in vivo.Methods1. In vitro:19of patients with regular dialysis choose from blood purification center, gene engineering L.lactis and serum with the saturation1:4,after24h reaction in anaerobic environment at37℃, detecting plasma indoxyl sulfate by using HPLC-GLU before and after interfere.2. In vitro:6-week-old male SD rats were randomly divided into2groups:1.controlled group (n=14):NS2mL, i.g Qd; experimental group (n=21):gene engineering L.lactis1.5×108cfu/ml×2ml,i.g Qd; the rats of two groups were underwent5/6nephrectomy. for renal failure model successed, Blood、urine and stool were collected to detect concentrations of indoxyl sulfate before and4week after administration. Two groups of rats were executed after4weeks of administration, to observe the degradation ability of gene engineering L.lactis to indoxyl sulfate.Results:χ±S1.19of uremic patients completed the trial, the baseline values of indoxyl sulfate were5.6282±2.4637ug/ml,the level of indoxyl sulfate after gene engineering L.lactis processed for24h trial were4.2478±2.0347ug/ml,which had significantly difference between two groups (P <0.001).2. There had no significant difference between each group in the body weight of rats before administration. The body weight of rats in the experimental group significantly reduced compared with the controlled group at4th week(P<0.07).3. At4th week, compared with the controlled, the:the concentration of plasma and urine indoxyl sulfate of the experimental were decreased by1.5240±1.1665ug/ml (P＜0.0001),3.3542±2.5924ug/ml (P＜0.0001), respectively.Conclusion1. gene engineering L.lactis can decrease the level of serum indoxyl sulfate in patients of chronic renal failure.2. gene engineering L.lactis can decrease the level of serum indoxyl sulfate in rats of chronic renal failure.