| Obejective:In this study, by constructing rat model of renal calcium oxalate stones, observation CaSR andClaudin-14expression in renal tissue of rats in both groups, while observing CaSR and Claudin-14with ratserum BUN, Cr, Ca2+levels,24h urine volume,24h urine calcium and urinary pH correlation.Methods:30adult male S-D rats were randomly divided into two groups (n=15), control group and stone group.1%ethylene glycol containing free drinking water+2%ammonium chloride(2ml) gavaged per day, in order toestablish rat model of renal calcium oxalate stones;Experiment28days collecting24h urine of rats, thecalcium content determination for24h urine. Chloral hydrate anesthesia puncture of the inferior vena cavablood make4ml,3000r/min4℃centrifuged for10min, take the supernatant, detection of serum BUN,Cr and Ca2+content; taking the left kidney surgery after the above steps, formaldehyde solution fixed,paraffin embedding slices after routine HE staining and observe the CaSR immunohistochemicalstaining;RT-PCR method to detect Claudin-14mRNA expression in kidney tissue; Western Blotting methodto detect renal tissue CaSR and Claudin-14protein expression.Results:1.Compared with control group, stone group, serum Cr, BUN increased significantly (P<0.01), stone groupof24h urine calcium excretion,24h urine volume increased significantly (P<0.01), the blood calcium levelsimilar between the two groups have no statistical significance (P>0.05),24h urine pH differences betweenthe two groups has no statistical significance (P>0.05).2.Visible to the naked eye stone group in the rat kidney tissue swelling, surface dotted with yellowish-whitecrystalline or calcification. HE staining, ordinary optical microscope found most kidney tissue in rat renalcalculi groups small tube cavity diffuse spreading colorless, transparent, flaky crysta.3.Were observed under light see CaSR renal tubular epithelial cells in rats in two groups all have differentdegrees of positive expression, and stone group rats CaSR in diffuse renal tubular epithelial cells andstrength increased obviously.4.Compared with control group, Claudin-14mRNA of stone group had a significantly higher, differencebetween groups was statistically significant (P<0.01).5. Compared with control group, stone group CaSR protein content increased significantly, the differencewas statistically significant (P<0.05); Claudin-14protein content increased significantly, in the controlgroup did not express, the difference was statistically significant (P<0.01). Meanwhile, CaSR andClaudin-14protein expression is also positively correlation (r=0.896, P<0.01).6.Expression of CaSR and Claudin-14showed a positively correlation with the24h urine calcium excretion,respectively, r=0.878, P <0.01; r=0.934, P <0.01.Conclusion: 1.Using1%glycol free drinking water+2%ammonium chloride daily2ml lavage can establish stable renalcalcium oxalate calculus model in rats.2.The high expression CaSR and Claudin-14in kidney tissue of kidney stone rats, and showed a positivelycorrelation with the calcium24h urine calcium excretion, respectively, suggesting that the expression CaSRin kidney stone rats increased by upregulating the expression of Claudin-14inhibit tubular calciumreabsorption, thus contributing to increased urinary calcium excretion, and thus participate in the formationof tubular crystal stones. |
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