| RongShi Granule, which is made up of field-balm, climbing fern spore, corydalis tuber, notoginseng, zedoary turmeric, earthworm and chicken's gizzard-skin, etc, was composed as the theory of Tradition Chinese medicine and experience in treatment of renal stone, and had good effects on prevention and treatment of renal stone in clinical practice that have already been proved by Tong-De Hospital of Zhejiang Province. The precise mechanism of renal stone formation still remains to be clarified because the causation of renal stone formation is complex, and there are also no specific for cure renal stone, so the operation and extrinsic lithotrity are still the primary treatment for renal stone at present, although they will cause certain injuries to the kidney and can not prevent relapse of the disease. With determining osteopontin content and urinary crystal components, the aim of our study was to evaluate the curative effect with method of modern pharmacology and to restate the mechanism of RongShi Granule on prevention and treatment of renal stone, which will provide scientific foundation for wide application of RongShi Granule.Experimental model rats of urinary stone were made with using oxalamide, ethylene glycol and ammonium chloride.1. The max administration dosage of RongShi Granule.We still assessed the security of Granule although the toxicity of Granule is so weak that its LD50 cannot be determined. The max administration dosage of RongShi Granule to mice is 3.84g/d, which is 192 times than that of adults in clinical administration (The clinical dosage for adults is 60 g/d.)2. Effects of RongShi Granule on the urinary stone formation in rats caused by oxalamide To observe the effects of the Granule on stones formation, rats of the stone group were givenoxalamide while the RongShi Granule groups were administrated RongShi Granule in addition to receiving oxalamide, the measured variables including the amount of animals with stone, stone rate, the contents of oxalic acid, calcium in kidney tissue and the percentage of crystal area. Theresults showed that all of the 5.8 g/kg> 11.6 g/kg and 17.4 g/kg dosage of RongShi Granule groups (the dose of low-dose group is nearly the clinical dosage for adults) could markedly reduce the oxalic acid and calcium concentrations and crystal area in kidney tissue (P<0.05) with an obvious dose-dependent manner, which indicated RongShi Granule could reduce the damage of the glomerular epithelium that caused by calcium oxalate crystal, so decrease the incidence of OPN adhering to disrepair epithelium and the location of crystal nucleation. The high dosage group could obviously decrease the amount of animals with stones in kidney, ureter and bladder (P<0.05).3. Extrinsic lithontriptic experiment of RongShi GranuleWe observed the dissolution function of Granule on calcium oxalate calculus in vitro. The result showed that the 5 % and 10 % RongShi Granule have a definite function of dissolving the calcium oxaiate calculus and the rate dissolution of calculus soaked with Granule after 10 days was 10.42 % and 18.64 %, respectively, and have significantly difference compared with those without Granule (PO.05).4. The influence of RongShi Granule to the expression of kidney osteopontin in the model rats with stone.Rats of urinary stone, those were molded with ethylene glycol and ammonium chloride, were continuously given RongShi Granule in 21 days (low-dose group, middle-dose group and high-dose group) and respectively obtained blood, urine and kidney samples every 7 days. Blood samples were utilized to be determined the concentration of urea nitrogen (BUN), creatinine (Cr), calcium and phosphorus, on the other hand, the concentration of uric calcium and oxalic acid were respectively measured by EDTA and photoelectric colorimetric method. The expression of kidney osteopontin and its mRNA were analyzed with immunohistochemical technology and RT-PCR.The expression of kidney osteopontin and its mRNA in model group were obviously higher than that of in solvent control group (.PO.01), and were the highest at 21 days, and 1.4 times (0.2813/0.2018) and 4 times (1.3975/0.3410) as that of in control, respectively. The expression of kidney osteopontin and its mRNA in all of the RongShi Granule groups were lower than that of in model group (?<0.05), with an obvious dose-dependent manner. The degree of the kidney calcium oxalate crystal of the rats in all the RongShi Granule groups was much lower than that of model group, and the uric calcium and oxalic acid of all of the RongShi Granule groups were much lower than that of model group (P<0.0\).5. Experimental research of RongShi Granule on anti-inflammatory, analgesic and diuretic effects.The ear-swelling mice model caused by xylene was used to investigate the anti-inflammatory effect of the Granule. The result that the degree of swelling was obviously lower in all of the RongShi Granule groups than in stone group (P<0.05), indicated RongShi Granule have markedly anti-inflammatory effect.To observe the analgesic effect of the Granule, the mice were injected with acetic acid inabdominal cavity and observed the twist reaction. The result indicated that all group of the RongShi Granule could markedly reduce the twist reaction (PO.05).To observe the diuretic effect of RongShi Granule with metabolic cage method, and the result showed the high and middle dosage groups could markedly increase the quantity of rats' emiction within 6h (P<0.05).In conclusion:l.The RongShi Granule could markedly inhibit the incidence of rats' total stone caused by oxalamide, reduce the amount of animals with stone, obviously decrease the oxalic acid, calcium content and crystal area ratio in the wet kidney tissue, ameliorate the damage of the glomerular epithelium that caused by calcium oxalate crystal, so decrease the incidence of OPN adhering to disrepair epithelium and the location of crystal nucleation.2.RongShi Granule could markedly alleviate the degree of ear-swelling caused by xylene, inhibit mice twist reaction caused by acetic acid and increase the quantity of rats' emiction within 6h, this proved the RongShi Granule have the anti-inflammatory, analgesic and diuretic function, could promote the stone to movement downwards and drainage, and had good anti-inflammation and analgesia to the inflammation and pain which may be caused by stone.3.The 5 % and 10 % RongShi Granule have some extent dissolution function to the calcium oxalate crystal. The uric calcium and oxalic acid of the high, middle and low dosage RongShi Granule groups are much less than that of model group, which indicated that the RongShi Granule could inhibit the stone development.4.The RongShi Granule could inhibit the expression of osteopontin and its mRNA in kidney stone rats, which prospected that the function mechanisms of Granule may relate with inhibiting the expression of non-phosphoric acid osteopontin, reducing the potential nucleation spot, and decreasing the calcium oxalate crystal formation of kidney in kidney stone rats.
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