Study On The Effect And Mechanism Of Matrix-Gla Protein On The Retention Of Calcium Oxalate Crystal

Objective:Establish animal model of calcium oxalate crystals by using ethylene glycol in SD rat, Warfarin intervene the protein like MGP, Investigate the effect and mechanism of MGP on calcium oxalate crystal retentionMethods:50 male Sprague-Dawley rats are divided into the control group:the blank control group(n=11), the Warfarin control group(n=12) and the model group:the Glycol model group(n=12), the Warfarin & EG group(n=15).Calcium oxalate calculus in male Sprague-Dawley rats is induced by 0.75% EG in drinking water for up to 1 week and the control group is given plain water, Intraperitoneal inject Warfarin to inactivate y-carboxylation of MGP. At day 5 post treatment,24 hour urine samples from rats is assayed for oxalate, calcium and others. A week later, the rats are killed to collect the blood serum, and analyzed of serum calcium, phosphorus, creatinine, uric acid and other biochemical indicators; Microscopic CT is used to observe renal papillary surface and internal crystal distribution characteristics; Kidney is respectively stained by hematoxylin-eosin (HE) staining and Von-Kossa’s staining; Polarizing microscope is used to observe the crystal retention around kidney calcification and artificial analysis; The distributio of MGP expression in renal tissue of rats is detected by immunohistochemistry.Results:1. The general condition of the rats:during the experiment,3 rats died; the weight of administration group was less than that of blank control group, the Warfarin & EG group, Warfarin control group decrease significantly compared with blank control group.2. Serum biochemical results:There are significant differences between the Glycol model group, the Warfarin control group, the Warfarin & EG group and the blank control group in Urea. There are no significant differences between the warfarin administration group and other group. The groups of serum P, Mg2+ are no significant differences.3.24h urine biochemical result:24h urine creatinine of administration group is slightly higher than that in control group; The experimental groups in 24h UA, P, Ca2+are significantly lower than the control group, and higher in urine oxalic acid. The, The inter-group comparison show no significant difference in experimental groups and control groups. Compared with the blank control group, the Warfarin & EG group has a statistically significant difference in 24h urine Ca2+、 UA、P、Ox. Compared with the Warfarin control group, the Warfarin & EG group has a statistically significant difference in 24h urine Ca2+、UA、Ox. Compared with the blank control group, The Glycol model group has a statistically significant difference in 24h urine UA、P. Compared with the Warfarin control group, the Glycol model group has a statistically significant difference in 24h urine Ca2+、P. Urine Mg2+ in each group has no significant statistical difference. Urinary citrate, the blank control group, the Glycol model group are significantly higher than that of warfarin administration group, Compared with the Glycol model group and the blank control group, the Warfarin & EG group has significant difference, The Warfarin control group has significant difference compared with the Warfarin control group, The Glycol model group compared with the blank control group, has no significant difference.4. Micro CT results on renal papillary:The control groups have no obvious calcification shadow, The experimental groups are observed with calcification shadows, and compared with the Glycol group, the Warfarin & EG group appeare high ratio of calcium plaque, there are significant differences, and the calcium spots more obvious and widely.5. Comparison of pathological staining and crystal count:The control group show no calcium salt staining, The experimental group is found in the renal tubules of calcium oxalate crystals in HE staining and Von-Kossa’s staining, the experimental group is found in the renal tubules of calcium oxalate crystals in HE staining and Von-Kossa’s staining, the tubular lumen expansion; artificial statistics for each section of the number of crystals, the Warfarin & EG group is significantly higher than the number of crystal retention of the Glycol model group, there was significant difference in count.6、Immunohistochemistry:Staining is extensively seen in renal cortex and medulla tubular epithelial cells both in the normal control rats and hyperoxaluria group.Conclusion:1.Calcium oxalate crystal in renal tubule and hyperoxaluria and hypocitraturia is induced successfully within 7 days in Sprague-Dawley male rats2. The inactivate MGP protein has obvious promoting the formation and retention of calcium oxalate crystal in SD rats.3. MGP is widely expressed in the renal cortex and medulla in the normal group and the crystallization of calcium oxalate group.