| Objective:Urinary calculi are common diseases in the department of urinary surgery,which are characterized by complex etiology,unclear mechanism of stone formation,high incidence and easy recurrence,which has been troubling clinicians deeply.In this study,by evaluating the stone formation effects and advantages and disadvantages of the five main methods for establishing renal calcium oxalate stone model in rats,a solid foundation was laid for the subsequent experiments.A randomized controlled trial was designed to evaluate the effect of Fangshi Decoction with dififerent doses on the prevention and treatment of renal calcium oxalate stone in rats and preliminarily explore its action mechanism.Methods:Study 1:SD rats were divided into 6 groups(12 rats/group)according to the random number table,including normal saline control group,ethylene glycol+ammonium chloride group,ethylene glycol+ammonium chloride+calcium gluconate group,ethylene glycol+amnonium oxalate group,ethylene glycol+ ammonium oxalate+calcium gluconate group and diacetamide feed group.After the completion of modeling,the concentration of oxalic acid and calcium ion in urine and the content of serum creatinine were detected in the 24h urine of rats from each group.10%formalin solution was used to fix kidney tissue.Paraffin sections with HE staining were used to observe the damage degree of tissue structure and the formation of calcium oxalate crystal in renal tissue under microscope.Study 2:SD rats were divided into 6 groups(12 rats/group)according to the random number table:normal diet group,normal saline control group,Paishi granule control group,Fangshi Decoction with low dose group,Fangshi Decoction with middle dose group and Fangshi Decoction with high dose group.The renal calcium oxalate stone model of rats was duplicated by ethylene glycol+ammonium chloride method.After 28 days,a number of biochemical indexes in serum and 24 h urine were detected.10%formalin solution was used to fix the kidney tissue.Paraffin sections witth HE staining were used to observe the damage degree of tissue structure and the formation of calcium oxalate crystal in renal tissue under microscope.Results:Study 1:there was no significant difference in the overall rate of renal stoneformation for rats among the five modeling group(P>0.05);there was significant difference in the overall crystallinity with grade Ⅲ and grade IV for rats among the five modeling group(P<0.05);there was significant difference in the total excretion of oxalic acid and calcium ion in 24h urine and serum creatinine content for rats among the five modeling group(P<0.05);there was no significant difference in the total excretion of magnesium ion in 24h urine and calcium and magnesium ion in serum for rats among the five modeling group(P>0.05).Study 2:there was significant difference in the overall rate of renal stone formation between the six groups(P<0.05).Compared with the normal saline control group,there was significant difference in the Paishi granule control group,Fangshi Decoction with low,medium and high dose groups(P<0.001).Compared with the Paishi granule control group,there was significant difference in the Fangshi Decoction with middle dose group(P<0.05);there was no significant difference between Fangshi Decoction with low dose group and Fangshi Decoction with high dose group(P>0.05);there was significant difference in the total calcium oxalate crystal area ratio between six groups(P<0.05),compared with the normal saline control group,there was significant difference in Paishi granule control group,Fangshi Decoction with low,medium and high dose groups(P<0.05),compared with Paishi granule control group,there was significant difference in Fangshi Decoction with medium and high dose groups(P<0.05),there was no significant difference in the Fangshi Decoction with low-dose group(P>0.05);there was significant difference in the concentratiort of oxalic acid,calcium ion,citric acid and magnesium ion in 24h urine and the content of serum creatinine between the six groups(P<0.05).Compared with the normal saline control group,the Fangshi Decoction with low,middle and high-dose groups had significant difference in the concentration of oxalic acid,magnesium ion and citric acid in 24h urine and the content of serum creatinine(P<0.05);there was no significant difference in pH value in 24h urine and the content of serum calcium ion and magnesium ion for rats among the six groups(P>0.05).Conclusions:1.The five modeling methods can successfully duplicate the renal calcium oxalate stone model of rats,and the stone formation rate is similar.2.All the five modeling methods can promote the formation of calcium oxalate crystal by increasing the excretion of oxalic acid and calcium ion in the urine of rats and the damage to the kidney.Among them,ethylene glycol+ammonium chloride method has a satisfactory effect on the formation of calcium oxalate crystal,and the minimal damage to the kidney of rats.3.The five modeling methods were not related to the content of magnesium ion in urine and calcium ion and magnesium ion in blood.4.Fangshi Decoction with low,medium and high dose and Paishi granules can effectively prevent and treat the calcium oxalate stone in the kidney of rats,and reduce the degree of stone formation in the kidney of rats.Among them,the effect of Fangshi decoction with medium dose is the most obvious,followed by Fangshi decoction with low dose and Paishi granules,and the effect of Fangshi decoction with high dose is the worst,which may be caused by the aggravation of kidney damage caused by the high concentration of Fangshi decoction,but the therapeutic effect is reduced.5.Fangshi Decoction with three doses can increase the concentration of citric acid and magnesium ion in the urine of rats,reduce the concentration of oxalic acid and calcium ion in the urine and the content of serum creatinine,but have little relationship with the pH value in urine,urinary calcium ion,blood calcium ion and blood magnesium ion in rats.Considering that Fangshi decoction may exert therapeutic effect by increasing the number of oxalobacter flora in the intestine of rats or the decomposition of oxalate by oxalobacter flora in the intestine of rats,so as to reduce the absorption of oxalic acid into the blood through the intestinal tract,reduce the content of oxalic acid in the urine and finally achieve the purpose of inhibiting the formation of stones,its specific mechanism remains to be further studied. |
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