草酸钙结石形成对肾组织bikunin和IαI表达的影响

目的研究肾草酸钙结石形成对肾组织bikunin基因和间α胰蛋白酶抑制物(IαI)蛋白表达的影响,探讨bikunin在尿结石形成中的意义。方法诱导实验性大鼠肾草酸钙结石模型,收集结石大鼠、正常大鼠、临床肾结石和非结石患者每组各8例的肾组织标本。采用免疫组化、逆转录聚合酶链反应(RT-PCR)和计算机图像分析技术分别检测所有大鼠和人肾组织中bikuninmRNA和IαI蛋白的表达水平。结果正常大鼠和非结石患者肾组织均存在bikuninmRNA和IαI蛋白的表达。肾草酸钙结石形成后,结石大鼠肾组织IαI蛋白的灰度值和bikuninmRNA的相对表达水平分别为198.43±15.17、0.70±0.14;肾结石患者肾组织IαI蛋白的灰度值和bikuninmRNA的相对表达水平分别为263.25±17.41、1.27±0.13,分别和对照组相比,均显著增加(P<0.05)。结论Bikunin作为构成IαI的轻链结构,在肾草酸钙结石形成后,bikuninmRNA的表达迅速增强,提示机体通过肾脏合成更多的bikunin来抑制肾草酸钙结石的形成。     

草酸钙结石形成对肾组织bikunin和ⅠαⅠ表达的影响

目的研究肾草酸钙结石形成对肾组织bikunin基因和间α胰蛋白酶抑制物(ⅠαⅠ)蛋白表达的影响,探讨bikunin在尿结石形成中的意义.方珐诱导实验性大鼠肾草酸钙结石模型,收集结石大鼠、正常大鼠、临床肾结石和非结石患者每组各8例的肾组织标本.采用免疫组化、逆转录聚合酶链反应(RT-PCR)和计算机图像分析技术分别检测所有大鼠和人肾组织中bikuninmRNA和ⅠαⅠ蛋白的表达水平.结果正常大鼠和非结石患者肾组织均存在bikunin mRNA和IαI蛋白的表达.肾草酸钙结石形成后,结石大鼠肾组织ⅠαⅠ蛋白的灰度值和bikunin mRNA的相对表达水平分别为198.43士15.17、0.70士0.14;肾结石患者肾组织IαI蛋白的灰度值和bikunin mRNA的相对表达水平分别为263.25士17.41、1.27±0.13,分别和对照组相比,均显著增加(P＜0.05).结论Bikunin作为构成IαI的轻链结构,在肾草酸钙结石形成后,bikunin mRNA的表达迅速增强,提示机体通过肾脏合成更多的bikunin来抑制肾草酸钙结石的形成.     

草酸钙结石形成对肾组织bikunin和IαⅠ表达的影响

目的研究肾草酸钙结石形成对肾组织bikunin基因和间α胰蛋白酶抑制物(ⅠαⅠ)蛋白表达的影响,探讨bikunin在尿结石形成中的意义.方珐诱导实验性大鼠肾草酸钙结石模型,收集结石大鼠、正常大鼠、临床肾结石和非结石患者每组各8例的肾组织标本.采用免疫组化、逆转录聚合酶链反应(RT-PCR)和计算机图像分析技术分别检测所有大鼠和人肾组织中bikuninmRNA和ⅠαⅠ蛋白的表达水平.结果正常大鼠和非结石患者肾组织均存在bikunin mRNA和IαI蛋白的表达.肾草酸钙结石形成后,结石大鼠肾组织ⅠαⅠ蛋白的灰度值和bikunin mRNA的相对表达水平分别为198.43士15.17、0.70士0.14;肾结石患者肾组织IαI蛋白的灰度值和bikunin mRNA的相对表达水平分别为263.25士17.41、1.27±0.13,分别和对照组相比,均显著增加(P＜0.05).结论Bikunin作为构成IαI的轻链结构,在肾草酸钙结石形成后,bikunin mRNA的表达迅速增强,提示机体通过肾脏合成更多的bikunin来抑制肾草酸钙结石的形成.     

钙敏感受体活性改变对大鼠泌尿系草酸钙结石形成的影响

目的探讨钙敏感受体（CaSR）活性改变对草酸钙结石形成的影响。方法在实验期间给予乙二醇和氯化铵诱导雄性SD大鼠产生泌尿系草酸钙结石。在造模期间给予不同剂量的CaSR抑制剂（NPS-2390）。实验结束时检测各组大鼠血尿素氮（BUN）、肌酐（Cr）、血磷、血钙、血镁、PTH的含量、24h尿量、尿pH值、尿钙、镁、尿草酸的分泌量,显微镜下观察肾组织切片中草酸钙结晶沉积及病理变化情况及肾脏中CaSR表达情况。从而评价CaSR活性的改变对泌尿系结石形成的影响。结果成石对照组大鼠血BUN、Cr、尿草酸、尿钙较空白组明显升高并且有大量结晶形成,表明建模成功。CaSR抑制剂组较成石对照组甲状旁腺激素（PTH）的分泌增加并且血Ca2＋升高尿钙升高。肾组织病理学检查显示CaSR抑制组的肾脏组织中的草酸钙结晶较成石对照组明显增加,组织病理损伤也较重。结论肾脏中及甲状旁腺中CaSR的表达下降可以导致草酸钙结晶的形成增加。     

苄丙酮香豆素对实验性大鼠肾草酸钙结石形成的影响

目的：探讨Vit．K拮抗剂苄丙酮香豆素(商品名华法令)对大鼠肾草酸钙结石形成的影响。方法：采用乙二醇饮水和氯化铵灌胃作成石剂，30只Wistar大鼠随机分为对照组(A组)、成石组(B组)、华法令组(C组)。饲养4周后，检测大鼠肾组织钙含量和草酸钙晶体形成、24h尿钙、尿草酸含量及血生化指标。结果：成石组和华法令组肾组织中钙、镁含量，24h尿草酸及尿钙、镁排泄量差异无显著性意义；镜下观察发现：华法令组大鼠肾脏草酸钙结晶形成多于成石组，但组间比较差异无显著性意义。结论：苄丙酮香豆素对大鼠肾草酸钙结石的形成无显著影响。     

钙拮抗剂抑制大鼠肾草酸钙结石生成的实验研究

目的观察不同剂量硝苯地平灌喂对大鼠肾草酸钙结石生成的影响。方法选用60只雄性SD大鼠,体重200~250g,随机分为6组,分别为空白对照组、单纯硝苯地平组、单纯诱石组、诱石 3、6、10mg·kg-1·d-1硝苯地平干预组,每组各10只。应用乙二醇诱导大鼠产生肾草酸钙结石,4周后观察大鼠肾小管结晶沉积情况、肾组织自由基水平、细胞凋亡情况和大鼠血和尿多项生化指标的变化。结果与单纯诱石组相比,各硝苯地平干预组的肾小管草酸钙结晶评分分别降低37.0%、55.6%、66.7%(P均<0.05),肾小管上皮细胞凋亡数分别减少30.2%、44.6%、48.7%(P均<0.05),两者有显著相关性(r=0.8251);肾组织中MDA含量也分别减少14.5%、20.4%、21.8%,而SOD活性增加3.5%、8.7%、12.6%(P均<0.05);量效关系呈正相关。结论硝苯地平通过减少高尿草酸所致的肾小管上皮细胞凋亡,能有效抑制饮用诱石剂大鼠的肾小管结石生成。     

大鼠肾草酸钙结石模型制备效果的比较研究

目的 验证大鼠肾草酸钙结石模型制备方法,为癸壬化石丸治疗肾结石的药理作用及机制研究选择有效的大鼠肾草酸钙结石模型.方法 按文献报道的四种大鼠肾草酸钙结石模型制备方法,将SD大鼠分为A、B、C、D四组,A组采用1％乙二醇+2％氯化铵溶液进行灌胃,B组采用1％乙二醇+2％氯化铵+10％葡萄糖进行灌胃,C组采用1％乙二醇+10％葡萄糖酸钙进行灌胃,D组采用1％乙二醇+2％氯化铵+10％葡萄糖酸钙进行灌胃,28 d后检查尿常规、尿生化、血生化和肾脏标本的病理切片后比较肾草酸钙结石模型制备效果.结果 四种方法制备大鼠肾草酸钙结石模型,造模效果D组＞A组＞B组＞C组.结论 采用1％乙二醇+2％氯化铵+10％葡萄糖酸钙所形成大鼠肾草酸钙结石模型效果较好.     

泽泻提取物对大鼠尿结石形成和间α胰蛋白酶抑制物表达的影响

目的　观察中药泽泻的有效部位提取物对实验性大鼠尿草酸钙结石形成和间α胰蛋白酶抑制物 (IαI)表达的影响。方法　采用乙二醇和氯化铵诱导形成大鼠肾草酸钙结石模型 ,将健康成年雄性Wistar大白鼠随机分成对照组 (A组 )、成石组 (B组 )、泽泻组 (C组 )。用免疫组织化学和计算机图像分析技术检测IαI在肾组织的表达情况 ,并测定血、尿生化和草酸钙晶体在肾组织的分布。结果　泽泻组大鼠肾组织草酸钙晶体分布、血生化等指标均明显低于结石模型组 ;泽泻组大鼠肾组织IαI的灰度值为 2 18.3 2± 7.5 8、肾钙含量 (7.3 2± 1.5 9)mg/g、2 4h尿钙分泌量 (5 .3 4±1.10 ) μmol/2 4h ,模型组大鼠IαI的灰度值为 2 0 3 .40± 14 .69、肾钙含量 (12 .63± 2 .2 9)mg/g、2 4h尿钙分泌量 (8.5 3± 1.73 ) μmol/2 4h ,两组间差异均有显著性 (P <0 .0 5 )。 结论　泽泻的乙酸乙酯浸膏的乙酸乙酯洗脱液提取物可能通过抑制肾组织内草酸钙晶体的形成和减少肾IαI的表达 ,从而能抑制尿结石的形成。     

药用醋膏预防大鼠泌尿系结石形成及其机制的初步研究

目的:通过应用药用醋膏喂养泌尿系结石模型大鼠,观察大鼠肾组织病理变化及测定肾组织草酸和钙离子含量,观察醋酸预防泌尿系结石的效果,进一步探讨其机制。方法:将适宜体重的健康Wister大鼠随机分为空白组、1%乙二醇+2%氯化铵诱石液造模对照组、30g/100ml高剂量醋膏+1%乙二醇+2%氯化铵诱石液组、10g/100ml低剂量醋膏+1%乙二醇+2%氯化铵诱石液组,喂养28天后处死,取肾组织HE染色,观察草酸钙结晶面积分布情况,测定各组大鼠肾组织中草酸与钙离子含量。结果:空白组与其它三组比较,肾组织草酸钙结晶面积分布及草酸和钙离子含量差异均有统计学意义(均P0.05);造模对照组与高剂量醋膏组比较,肾组织草酸钙结晶面积分布及草酸和钙离子含量差异均有统计学意义(P0.05),而与低剂量醋膏组比较,肾组织结晶面积分布及草酸和钙离子含量差异均无统计学意义(P0.05);高剂量醋膏组与低剂量醋膏组比较,肾组织草酸和钙离子含量差异有统计学意义(P0.05),肾组织草酸钙结晶面积分布差异无统计学意义(P0.05)。结论:在正常食用剂量范围内,高剂量醋膏对大鼠泌尿系结石的形成有预防作用,可降低肾组织中草酸及钙离子含量。     

尿凝血酶原片断1在肾结石模型大鼠肾组织的表达及其意义

目的　研究尿液中尿凝血酶原片段 1(UPTF1)的来源和UPTF1在肾结石模型大鼠肾组织的表达 ,探讨尿结石形成对肾组织UPTF1表达的影响及其在尿结石形成中的意义。方法用乙二醇和 1α 羟基维生素D3 灌胃制作大鼠肾草酸钙结石模型。采用半定量逆转录聚合酶链反应(RT PCR)检测UPTF1mRNA在结石模型大鼠和正常对照大鼠肾组织中的表达及水平变化。结果　偏光显微镜下结石模型大鼠肾乳头和肾皮质内布满草酸钙晶体 ,肾钙含量、2 4h尿草酸和尿钙分泌量分别为 13 8.3 9mg/g、82 .89μmol和97.3 5 μmol;对照组肾钙含量、2 4h尿草酸和尿钙分泌量分别为 1.5 4mg/g、2 4.2 2 μmol和3 .14 μmol,组间差异均有统计学意义(P <0 .0 1)。UPTF1mRNA在所有大鼠肾组织和肝组织中都有表达 ,但在正常大鼠和肾结石大鼠肾组织中的相对表达量分别为 1.73± 0 .2 5、1.86± 0 .19,两组差异无统计学意义意义 (P >0 .0 5 )。结论　尿液中的UPTF1来源于大鼠肾组织的生物合成 ,可能是草酸钙结石形成的生理性抑制因子 ,从而可以借助实验动物模型为研究UPTF1在尿结石形成中的作用提供了依据。     

单味中药金钱草、石韦、车前子对肾结石模型大鼠的预防作用

目的：观察单味中药金钱草、石韦、车前子对肾结石的预防作用。方法：90只大鼠随机分为5组,采用1.25%乙二醇和1%氯化铵制备大鼠肾结石模型,用上述3种单味中药的免煎剂型分别给各实验组大鼠肾结石模型灌胃,对照组用枸橼酸钾,4周后观察尿和肾组织中草酸钙结晶的形成情况。结果：单味中药金钱草组、石韦组、车前子组大鼠肾内草酸钙结晶明显少于模型组,而与西药枸橼酸钾组相当。金钱草组、石韦组、车前子组大鼠尿中草酸钙结晶的排泄均明显多于模型组。结论：单味中药金钱草、石韦、车前子对预防大鼠肾结石的形成有确切效果,并与西药枸橼酸钾相当,其作用可能主要是通过增加尿中草酸钙结晶的排泄而达到的。     

Effect of etidronate disodium (EHDP) on calcium oxalate renal stones induced by synthetic 1 alpha(OH) vitamin D3 and ethylene glycol in rats

Combination of 1 alpha(OH) D3(vit D) and ethylene glycol induced renal or ureteral stones or both consisting of calcium oxalate in male Wistar rats. This study investigates the effect of EHDP on calcium oxalate stone using the rat model. EHDP reduced the frequency of renal stone and calcium content in the kidney, and reduced the size of the stones in the renal pelvis and ureter. EHDP biochemically ameliorated renal injury induced by vit D and ethylene glycol. EHDP suppressed urinary excretion of calcium even though serum calcium slightly increased. EHDP had a phosphaturic action. EHDP elevated urinary excretion of magnesium. However, the severity of hypermagnesuria decreased in the rat which was not given EHDP concomitantly. Although EHDP slightly elevated urinary excretion of oxalate in the control rat, it did not affect the high level of urinary oxalate in the vit D/ethylene glycol rat. EHDP did not produce any histological change in the kidney or femoral bone. These data indicate that EHDP can suppress renal stone formation in the vit D/ethylene glycol rat. It is speculated that firstly, EHDP may physicochemically inhibit stone formation in the process of nidus, aggregation and crystal growth of calcium oxalate, under the supersaturated condition of calcium oxalate in the urine, and secondly, EHDP may endocrinologically inhibit production of 1,25 (OH)2 vit D in the kidney or inhibit 1, 25 (OH)2 vit D-mediated intestinal calcium absorption. It is suggested that in order to prevent stone recurrence, EHDP may be clinically applied not only to calcium phosphate stones but also to calcium oxalate stones and hypercalciuria mediated by an active form of vitamin D.     

肾结石大鼠肾组织bikunin mRNA的表达

目的 :研究bikunin在实验性肾草酸钙结石大鼠肾组织的表达及意义。方法 :采用乙二醇和氯化铵诱导大鼠肾草酸钙结石模型形成 ,检测各组大鼠肾功能、肾组织Ca2 + 含量和草酸钙晶体沉积、尿生化指标 ,并用逆转录聚合酶链反应 (RT PCR)检测bikuninmRNA在肾组织的表达情况。结果 :模型组大鼠的血清Cr、BUN、肾Ca2 + 含量、2 4h尿Ca2 + 、草酸 (Ox)分泌量和肾组织bikuninmRNA的表达均明显高于正常组 (P <0 .0 5 )。结论 :高草酸尿和草酸钙结晶的沉积能促使大鼠肾脏通过合成更多的bikunin来抑制大鼠肾组织草酸钙晶体的形成。     

N-乙酰半胱氨酸抑制大鼠肾草酸钙结石形成机制的研究

目的：探讨N一乙酰半胱氨酸（NAc）对大鼠肾草酸钙结石形成的影响及机制,为临床预防尿路结石提供理论依据。方法：将30只健康清洁成年雄性Wistar大鼠先在相同环境下适应性喂养1周,然后随机分为3组：A组（空白对照组）、B组（单纯诱石组）、C组（诱石＋NAC干预组）。A组饮去离子水,B组饮1％乙二醇的去离子水,C组饮1％乙二醇去离子水,并给予NAC100mg／（kg·d）灌胃。第4周处死大鼠,取出双肾,左肾纵向剖开,用10％甲醛固定,HE染色石蜡切片,在100、400倍光镜下观察肾组织草酸钙结晶沉积情况,并进行分级及评分。右肾皮质制成10％的匀浆,检测丙二醛（MDA）及超氧化物歧化酶（SOD）。全部实验数据通过SPSS17．0统计软件分析处理。结果：①肾脏结晶沉积情况分级及评分结果：与B组相比,C组的肾脏结晶沉积评分明显降低（P〈0．05）。②MDA检测结果：与B组相比,C组的MDA含量降低,差异有统计学意义（P〈0．05）。③SOD检测结果：与B组相比,C组的SOD含量增高,差异有统计学意义（P〈0．05）。④A、B、C三组肾组织结晶等级评分与s0D含量的相关系数为-0．499（P〈0．01）,结晶评分与MDA含量的相关系数为0．592（P〈0．01）。结论：NAc可以通过其抗氧化作用抑制大鼠肾草酸钙结石的形成。     

Inhibitory effect of fluoride on renal stone formation in rats.

The effect of fluorine (F) on stone formation induced by ethylene glycol (EG) was studied in rats. For different groups, the drinking water was supplemented with EG, sodium fluoride (NaF), EG+NaF, or nothing as control. An isotope-tracing method was used to evaluate experimental stone formation in the kidneys by introducing 45Ca intraperitoneally into rats and then measuring the radioactivity of the kidney. At the end of the 4-week experiment, rats of the EG+NaF group showed a significantly lower incidence of gross urinary stones and lower 45Ca activity in their kidneys compared to the EG group of rats. Both the EG group and EG+NaF group had markedly increased urinary oxalate excretion, with the latter significantly lower than the former (p less than 0.05). Urinary oxalate excretion was relatively lower in the NaF group than in the control group. This study indicates that NaF can inhibit renal stone formation induced by EG by decreasing oxalate synthesis and urinary oxalate excretion, and suggests a possible clinical therapeutic value of NaF in the prevention of oxalate kidney stones.     

The influence of sex hormones on renal osteopontin expression and urinary constituents in experimental urolithiasis

PURPOSE: To our knowledge the influence of sex hormones on urinary stone formation remains undetermined. We investigated the effect of castration on urinary lithogenic factors and renal osteopontin expression in rats previously treated with ethylene glycol. MATERIALS AND METHODS: Sprague-Dawley rats were divided normal males, castrated males, males with 2 weeks of 0.75% ethylene glycol treatment, castrated males with 2 weeks of 0.75% ethylene glycol treatment, normal females, castrated females, females with 2 weeks of 0.75% ethylene glycol treatment and castrated females with 2 weeks of 0.75% ethylene glycol treatment. We analyzed 24-hour urine samples for urinary constituents, such as calcium, oxalate, citrate, uric acid, phosphate, magnesium, sodium, potassium and creatinine. The kidneys were examined for osteopontin expression by Northern blot analysis and for crystal deposition by histological examination. RESULTS: In intact male rats calcium and citrate excretion decreased and oxalate excretion increased significantly after ethylene glycol treatment. Castrated male rats with ethylene glycol had greater calcium and less oxalate excretion than male intact rats with ethylene glycol. In intact female rats uric acid excretion decreased and only calcium excretion increased significantly after ethylene glycol treatment. Castrated female rats with ethylene glycol excreted significantly more oxalate and less calcium than intact female rats with ethylene glycol. Renal osteopontin expression was the same in male intact and castrated rats, and in female intact and castrated rats. In males with ethylene glycol expression was stronger in castrated than in intact rats. In females with ethylene glycol expression was weaker in castrated than in intact rats. No crystal deposits were found in the kidneys in any group. CONCLUSIONS: Testosterone appears to promote stone formation by suppressing osteopontin expression in the kidneys and increasing urinary oxalate excretion. Estrogen appears to inhibit stone formation by increasing osteopontin expression in the kidneys and decreasing urinary oxalate excretion.     

Effects of urinary pH and acid-base balance on the formation of calcium oxalate stone

The effects of urinary pH and acid-base balance on the calcium oxalate stone formation was investigated by two experiments. 24-hr urine samples were collected from 15 recurrent CaOx stone formers, 9 single stone formers and 6 age-matched controls. Inhibitory effect of 1% urine in various pH (4.0-9.0) were calculated by a seed crystal method. In the seed crystal system, there were no significant differences in the inhibitory activity of aggregation (Ia) and in the inhibitory activity of size (Is) for each pH of metastable solution between the stone former group and the control group. However, the value of Ia and Is showed a tendency of rise in proportion to a rise in pH. Rats model for calcium oxalate urolithiasis were fed with three different diets (1% NH4Cl, 5% NaHCO3 and 8% NaHCO3 diet) for three weeks. On the fourth week, 24-hr urine samples were collected. In the animal experiment, calcium oxalate stone formations were predominantly recognized in the kidney of the 1% NH4Cl diet group. The biochemical data showed an increase of urinary calcium and oxalate, and a decrease of urinary citrate. These results suggest that low urinary pH and metabolic acidosis are promoters of the calcium oxalate stone formation.     

Effect of acidosis on urine supersaturation and stone formation in genetic hypercalciuric stone-forming rats

BACKGROUND: We have successively inbred over 45 generations a strain of rats to maximize urine calcium excretion. The rats now consistently excrete 8 to 10 times as much calcium as controls and uniformly form poorly crystalline calcium phosphate kidney stones. In humans with calcium nephrolithiasis, consumption of a diet high in acid precursors is often cited as a risk factor for the development of calcium-based kidney stones; however, the effect of this diet on urinary supersaturation with respect to the common solid phases found in kidney stones has not been determined. METHODS: To determine the effect of the addition of an acid precursor on urine ion excretion, supersaturation, and stone formation, we fed these genetic hypercalciuric stone-forming (GHS) rats 13 g/day of a 1.2% calcium diet with 0.0, 0.5, 1.0, or 1.5% NH4Cl in the drinking water for 14 weeks (N = 8 for each). Urine was collected and analyzed every two weeks. RESULTS: As expected, the addition of dietary NH4Cl led to a progressive fall in urine pH and urine citrate, while urine ammonium increased. Urine calcium and phosphorus increased, while urine oxalate fell. Increasing dietary NH4Cl led to a fall in supersaturation with respect to CaHPO4 (brushite) and CaOx and a rise in supersaturation with respect to uric acid. In spite of differences in supersaturation, most rats in each group formed stones that contained calcium phosphate and not calcium oxalate. CONCLUSIONS: Thus, while the provision of additional dietary acids alters urinary ion excretion and lowers supersaturation with respect to CaHPO4 and CaOx, it does not change the character or rate of stone formation in the GHS rats.