| Objective:1.to investigate the renal protective function and mechanism of active vitamin D3 through regulating NF-kB signaling pathway in type 2 diabetic kidney disease rats;2.to find the best concentration of vitamin D in the treatment of diabetic kidney disease.Methods:60 SD rats were randomly divided into normal control group(10 rats)and DKD model group(50 rats,and were divided into peanut oil group,irbesartan group,vitamin D low-dose group,vitamin D medium-dose group and vitamin D high-dose group with 10 rats in each group).The normal group was fed with normal diet,and the model group was administered by gavage with the different drugs.After 6 weeks,the rats were killed and weighed.The serum was separated to detect fasting blood glucose(FBG),triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-c),insulin(INS),C peptide(CP).The 24 hour--urine was collected to note urine volume and measure 24 hour-urine protein quantitative(uPro).The renal tissue was retained and HE staining was used to observe the pathological changes of renal tissue;Real-time fluorescent quantitative PCR was used to detect the expression of NF-kB mRNA and its downstream monocyte chemotactic protein(MCP-1)mRNA in renal tissue;Western booting was used to detect the expression of NF-kB and MCP-1 protein in renal tissue.The t test was performed for pair-wise comparison and wilcoxon rank sum test was used for ranked data.Results:(1)when the levels of FBG,24h urine protein quantitative,TG,LDL-c were compared in each group,vitamin D low-dose group(18.20±5.95?32.24±14.28?0.89±0.18?0.30±0.10)were significantly decreased than that of peanut oil group(23.55±2.08?72.52±140.76?1.12±0.25?0.39±0.15)(P<0.05);(2)When the levels of TC was compared,irbesaftan group(1.76±0.60)?vitamin D low-dose group(1.52±1.17)?medium-dose group(1.78± 1.83)?high-dose group(2.03±0.60)were significantly decreased than that of peanut oil group(3.50± 1.49)(P<0.05);(3)when the levels of Insulin were compared in each group,vitamin D low-dose group(148.00± 166.25)were significantly increased than that of peanut oil group(74.00±82.00)(P<0.05).(4)In normal group,HE staining showed that the glomerular morphology was intact,the mesangial area had no obvious hyperplasia,the capillary cavity loope regularly shaped.While in peanut oil group,the glomerular volume got narrowed,the mesangial area was broadened obviously,the glomerular mesangial cells proliferated and capillary wall thickened.The irbesartan group was less prolific than peanut oil group.The vitamin D high-dose group,middle-dose group and low-dose group all had a broaden mesangial area,but low-dose group had the lightest degree.(5)the mRNA of NF-kB and MCP-1 in vitamin D low-dose group(1.72±0.002,0.67±0.01)were significantly decreased compared with the normal group(1.00±0.00,1.00±0.00)and peanut oil group(3.57±0.40,3.57±0.37)(P<0.05),but there was no statistical difference between irbesartan group and vitamin D low-dose group;(6)The expression of the NF-kB protein in each group was not statistically significant.The expression of MCP-1 protein in vitamin D low-dose group(0.62±0.30)was obvious decreased than that of peanut oil group(1.19±0.09)and vitamin D middle-dose group dose group(1.12±0.23)(P<0.05).Conclusion:1.vitamin D could effectively alleviate renal injury in diabetic kidney disease by down-regulation of NF-kB inflammatory signaling pathway;2.vitamin D low-dose group(0.03ug/kg/d)is the best concentration in the protective effect of kidney disease.Objective:to investigate the renal protective function and mechanism of active vitamin D3 through regulating JNK signaling pathway in type 2 diabetic kidney disease rats.Methods:Take the renal tissue of diabetic kidney disease rats and normal rats from the first part of study.ELISA was used to detect the content of interleukin-1(IL-1)and interleukin-6(IL-6)in renal tissues,Western booting was used to detect the expression of inflammatory factors p-MEK-4,p-JNK1/2/3,p-AP-1 and p-ATF-2 protein in renal tissue.Results:1.Compared with peanut oil group(17±0.16),irbesartan group(10.26±0.24),vitamin D medium-dose group(13.89± 1.07)and vitamin D high-dose group(12.17±0.70),the level of IL-1 in vitamin D low-dose group(7.53 ±0.91)was significantly decreased(P<0.05);2.Compared with peanut oil group(5.07±0.47),irbesartan group(3.66±0.12),vitamin D medium-dose group(4.49 ±0.20)and vitamin D high-dose group(3.87 ±0.14),the level of IL-6 in vitamin D low-dose group(3.13 ±0.13)was significantly decreased(P<0.05);3.The protein levels of p-MEK4,p-JNKl/2/3,p-ATF-2 in irbesartan group(0.22±0.007,0.24±0.11,0.46±0.03)and vitamin D low dose group(0.20±0.17,0.41 ±0.17,0.38±0.09)were significantly decreased compared with peanut oil group(0.46±0.05,0.98±0.12,1.50±0.10),vitamin D medium-dose group(0.33±0.03,0.72±0.06,0.74±0.19)and the high-dose vitamin D group(0.35±0.01,1.32±0.19,1.46±0.32)(P<0.05),but there was no statistical difference between irbesartan group and the vitamin D low-dose group.4.Compared with peanut oil group(1.51±0.11),irbesartan group(0.80 ± 0.04),vitamin D medium-dose group(1.44±0.12)and vitamin D high-dose group(1.46±0.32),the level of p-AP-1 in vitamin D low-dose group(0.57±0.10)was significantly decreased(P<0.05).Conclusion:Vitamin D low-dose of(0.03ug/kg/d)can reduce the release of inflammatory factors and improve renal injury by inhibiting the activation of JNK signaling pathway. |
PDF Full Text Request