| Objectives To investigate the effects of lanthanum carbonate on hyperphosphatemia in rats with chronic renal failure and the preventive effect of lanthanum carbonate on vascular calcification,which may involve the inhibiting effects of TLR4/NF-κB signaling pathway,the expression of inflammatory factors such as IL-1β,IL-6,TNF-α and osteogenic transcription factor MSX2,CBFA1,ALP.Methods First of all,50 rats were randomly divided into 3 groups(blank group 10,sham group 10,building model group 30)after adaptive feeding for 1 week: rats in blank group would not be treated;rats in control group rats were anesthetized by 10% chloral hydrate,and only removed renal capsule,not renal tissue;rats in building model group were achieved renal failure by surgical 5/6 nephrectomy(Platt method).2ml blood was drawn from tail vein of 50 rats after 4 weeks of normal diet,measured the level of BUN,Scr,Ca and P by biochemical analyzer,confirmed that the rats in the building model group was successfully established the model of chronic renal failure.In addition,the diet of the original sham operation group was unchanged,and 30 rats in building model group were randomly divided into model group,lanthanum carbonate group and calcium carbonate group,with 10 rats in each group.Model group was only given high phosphorus diet,calcium carbonate group was given high phosphorus diet and calcium carbonate,lanthanum carbonate group was given high phosphorus diet and lanthanum carbonate.Then,a total of 40 rats in the sham operation group and the building model group were anesthetized,their whole thoracic aorta blood and their whole thoracic aorta and kidney samples were collected.The kidney samples were stained with HE to evaluate the renal failure model again.Von Kossa staining was used to detect calcium deposition in the upper half of the aorta to establish a vascular calcification model.The concentration of Serum P,Ca,ALP were measured by biochemical analyzer in half of blood to evaluate the effect of lanthanum carbonate on calcium and phosphorus metabolism.The expression of TLR4,NF-κB,MSX2,CBFA1,ALP was detected by immunohistochemical staining in the remaining half of the upper segment of the aorta.And TLR4,NF-κB,MSX2,CBFA1genes’ expression quantity was detected by real-time PCR in the lower segment of the aorta.Il-1β,IL-6 and TNF-α’s concentration was measured in the other half of the blood by elisa.Calcium content was detected in the middle aorta.Final data analysis.Results 1 After 4 weeks of general feeding,there was no significant difference in serum Scr,BUN,P,Ca index between the blank group and the sham group(P>0.05);The concentration of Scr,BUN,P in modeling group was higher,and Ca was significantly lower than that in blank group and sham group(P<0.05).This indicated successful modeling of renal failure.2 After 10 weeks of high phosphorus and drug feeding,HE staining of renal pathology in 4 groups showed that there were no significant abnormalities in glomerular tubules and surrounding interstitium in sham operation group;however,the other three groups showed varying degrees of glomerular morphological changes,with obvious thickening of vascular loops,proliferation of mesangial matrix,widening of basement membrane,increase of inflammatory cells,and dilation of renal tubules,which further indicated that the renal failure model was successfully created except for the sham operation group.3 VK staining was performed on the upper half of thoracic aorta in the 4groups.No obvious black particle calcium salt deposition was found in the sham operation group.In the model group,large area of calcium salt deposits were observed,which were connected into sheets.The area of calcium carbonate group was smaller than that of model group.The deposition of calcium salt of the lanthanum carbonate group was the lowest.This indicated that vascular calcification was modeled successfully except in the sham operation group.4 After 10 weeks of high phosphorus and drug feeding,compared with sham operation group,the serum P and ALP concentrations in other three groups were higher,Ca was lower than it(P<0.05);The levels of P and ALP in model group were higher than,while the levels of Ca were lower than those in calcium and lanthanum group(P<0.05);The P and ALP levels of calcium group were higher than,and Ca levels were higher than those in lanthanum group(P<0.05).P and ALP were least expression in lanthanum group.5 TLR4,NF-κB,MSX2,CBFA1 genes’ expression in the lower thoracic aorta were detected by PCR.Other groups’ expression levels were higher than those of sham operation group(P<0.05);The other three groups’ expression levels from high to low were model group,calcium group,lanthanum group(P<0.05).6 The expression of TLR4,NF-κB,MSX2,CBFA1 in the upper segment of thoracic aorta was detected by immunohistochemistry.The dark brown-yellow staining was the least in the sham group and the most in the model group,and lanthanum group’s gene expression and AOD were lower than calcium group.7 Compared with sham group,other part of thoracic aorta’s serum IL-1β,IL-6,TNF-α of the other 3 group was higher than that it(P<0.05);Model group was higher than calcium group and lanthanum group(P<0.05);Calcium group was higher than lanthanum group(P<0.05).8 The expression of ALP in the upper aorta of the rats in the four groups was detected by immunohistochemistry.The dark brown-yellow staining was the least in the sham operation group and the most in the model group.Lanthanum carbonate group’s expression and AOD were lower than calcium group.9After10 weeks of high phosphorus and drug feeding,compared with sham operation group,the middle thoracic aorta’s calcium content of other 3 groups was higher than it(P<0.05);Model group was higher than calcium group,which higher than lanthanum group(P<0.05).Conclusions 1 The phosphorus reducing effect of lanthanum carbonate on chronic renal failure rats is better than that of calcium carbonate.2 The expression of TLR4/NF-κB was down-regulated by lanthanum carbonate through phosphorus reduction,and the expressions of MSX2,CBFA1 and their induced products ALP are decrease,and the production of IL-1β,IL-6 and TNF-α are decrease.3 Phosphorus reducing drug lanthanum carbonate can significantly delay VC,and the effect is more obvious than calcium carbonate.Figure 4;Table 10;Reference 67... |
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