| Background:Glechoma longituba(Nakai)Kupr,China has rich resources,especially in North China,East China,Northeast China.There are records as early as two thousand years ago with Glechoma longituba(Nakai)Kupr has been used for the treatment of urinary stone disease,Chen Yuxian:Glechoma longituba(Nakai)Kupr can be used alone,this grass taste Xin,bitter,Liver and kidney,bladder by the power of good heat detoxification,dampness Tonglin,for hot shower,stone shower,damp heat jaundice embolism.Nanjing Tong Ren Tang development of the "row of stone particles”has served more than 40 years of domestic patients,20 million stones for the lifting of the pain of patients,the main medicine is Glechoma longituba(Nakai)Kupr,but the current domestic legislation on the prevention of calcium oxalate stone experimental study is not much in this study,the mechanism of calcium oxalate formation was discussed in vitro and in vivo and in vitro from the perspective of calcium oxalate formation.Objective:To observe the effect of Glechoma longituba(Nakai)Kupr on preventing the formation of calcium oxalate stone in vivo and in vitro,and to explore the mechanism of it to prevent the formation of calcium oxalate stone.Methods:(1)In the first part of the experiment,the nucleation inhibition rate was used as the kinetic index of calcium phosphate and calcium oxalate nucleus in vitro.Scanning electron microscopy was used to observe the effect of Glechoma longituba(Nakai)Kupr on calcium oxalate crystal growth;Mutual exclusion situation between crystals reflect indirectly by detecting the Zeta potential,to observe the effect of Glechoma longituba(Nakai)Kupr on calcium oxalate crystal aggregation ability;oxidative damage of renal tubular epithelial cell membrane by laser confocal microscopy and detection of hyaluronic acid,to observe the effect of Glechoma longituba(Nakai)Kupr on calcium oxalate crystals and renal tubular epithelial cell adhesion degree.(2)In the second part of the in vivo experiment,we constructed the rats' calcium oxalate animal model by using the lacquer agent,and used different concentrations of the extract,the potassium citrate and the Paishi-granule to influence the model construction.Serum calcium,phosphorus,potassium,creatinine and urea nitrogen and 24 h urine,calcium,phosphorus,oxalic acid and pH value.Renal histopathological damage and renal calcium deposition were observed by HE staining and von Kusa calcium staining.By detecting the contents of superoxide dismutase(SOD),lipid peroxide malondialdehyde(MDA)and catalase(CAT)in rat kidney,the effects of Glechoma longituba(Nakai)Kupr on oxidative stress in kidney were observed.The expression of osteopontin(OPN)mRNA and protein in the kidney tissues of rats were observed.The preventive effect of Glechoma longituba(Nakai)Kupr on calcium oxalate stones was observed.Results:(1)In the first part of the experiment,the maximum induction time of Glechoma longituba(Nakai)Kupr group was significantly lower than that of the blank control group,and the rate of nucleus formation was significantly higher than that of the blank control group,the difference was statistically significant(P<0.01).The maximum induction time of 1.5g/ml was significantly lower than that of potassium citrate group,and the rate of nucleus formation was significantly higher than that of potassium citrate group,the difference was statistically significant(P<0.05).Scanning electron microscope,irregular Glechoma longituba(Nakai)Kupr group of calcium oxalate crystal size,olive shape,round,square shape and even flake shaped crystals increase;different concentrations of Glechoma longituba(Nakai)Kupr group Zeta potential compared with the control group,the increase of 1.0g/ml and 1.5g/ml were significantly negative values,the difference was statistically significant(P<0.01).Compared with potassium citrate group 0.5g/ml and 1.0g/ml Glechoma longituba(Nakai)Kupr group was significantly negative values to reduce,the difference was statistically significant(P<0.01);The number of adherent cells of calcium oxalate crystal was obviously reduced in the Glechoma longituba(Nakai)Kupr group,and the hyaluronic acid was found to be significantly lower than that in the oxidized injury group,the difference was statistically significant(P<0.05).(2)The second part in vivo,Glechoma longituba(Nakai)Kupr and Paishi granule group and blank group,the urine volume increased,the difference was statistically significant(P<0.05);urinary oxalate excretion increased significantly compared with the control group,the difference was statistically significant(P<0.05),Glechoma longituba(Nakai)Kupr group compared with model urinary oxalate content significantly decreased,the difference was statistically significant(P<0.05),Glechoma longituba(Nakai)Kupr and Paishi granule group and potassium citrate group urinary oxalate was significantly increased,the difference was statistically significant(P<0.05);increased Glechoma longituba(Nakai)Kupr group compared with model of urinary citrate content,the difference was statistically significant(P<0.05);urinary calcium,phosphorus excretion and urinary pH were no significant differences between the groups.Glechoma longituba(Nakai)Kupr group,Paishi-granule group and model group of serum phosphorus decreased,the difference was statistically significant(P<0.05);0.5g/ml,1.5g/ml Glechoma longituba(Nakai)Kupr group,Paishi-granule group and potassium citrate group rat serum creatinine increased significantly,the difference was statistically significant(P<0.05).Compared with the control group,the renal HE staining showed that the edema of the proximal tubule cells was reduced,the lumen of tubules increased,the amount of crystal was small,and the inflammatory cells were less.Von Kossa calcium staining Glechoma longituba(Nakai)Kupr group:tubular lumen,crystal less,nipple tube black calcium salt deposition significantly reduced.Glechoma longituba(Nakai)Kupr group and Paishi-granule group compared with model group the activity of SOD increased,the difference was statistically significant(P<0.05),1.0g/ml,1.5g/ml of Glechoma longituba(Nakai)Kupr group compared with model group MDA content was significantly decreased,the difference was statistically significant(P<0.05),0.5g/ml,1.0g/ml and 1.5g/ml increased in Glechoma longituba(Nakai)Kupr group and the model group the content of CAT,the difference was statistically significant(P<0.05).The model of OPN mRNA was the highest,1.Og/ml,1.5g/ml of Glechoma longituba(Nakai)Kupr group and Paishi-granule group and calcium oxalate stone model group decreased expression of mRNA OPN(P<0.05),the expression of OPN protein decreased,the expression of mRNA is consistent with the group OPN situation.Conclusion:(1)In vitro,Glechoma longituba(Nakai)Kupr promoted the formation of calcium phosphate and calcium oxalate nuclei,inhibited the growth of calcium oxalate crystals,inhibited and inhibited the adhesion between calcium oxalate crystals and renal tubular epithelial cells,prevention of calcium oxalate formation.(2)Glechoma longituba(Nakai)Kupr reduced the intensity of oxidative stress in the kidneys,inhibited the expression of OPN,effectively controlled the formation of calcium oxalate stones in rats,and its mechanism may be induced by inhibition of oxalic acid and calcium oxalate of renaloxidative stress injury. |
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