排石冲剂对草酸钙肾结石大鼠p47phox和TGF-β1表达的影响

Effect of Paishi Granule on Expression of p47phox and TGF-β1 in Rats with Calcium Oxalate Kidney Stones

目的观察排石冲剂对草酸钙肾结石大鼠p47phox、转化生长因子(TGF)-β1表达的影响,探讨排石冲剂治疗肾结石的作用机制。方法Wistar大鼠60只,随机分为6组:正常对照组、模型对照组、枸橼酸钾组和排石冲剂小、中、大剂量组(1.2,2.4,3.6 g·mL^-1),每组各10只。分别检测各组大鼠尿草酸、尿钙的含量;用Vonkossa钙结晶染色观察肾组织中草酸钙结晶,并采用形态分级评分法比较各组草酸钙结晶的量化评分;通过荧光定量-聚合酶链反应(RT-PCR)法检测大鼠肾组织中p47phox、TGF-β1、白细胞介素(IL)-6 mRNA的表达水平。结果与正常对照组比较,其余各组大鼠尿草酸及尿钙含量均显著升高(P<0.01);与模型对照组比较,排石冲剂小剂量组的尿草酸及尿钙含量无明显差异(P>0.05),排石冲剂中剂量组尿草酸及尿钙含量显著降低(P<0.05);排石冲剂大剂量组、枸橼酸钾组的尿草酸及尿钙显著降低(P<0.01),但两组之间差异无统计学意义。与正常对照组比较,模型对照组、排石冲剂小剂量组草酸钙结晶评分显著升高(P<0.01);与模型对照组比较,排石冲剂中剂量组评分显著降低(P<0.05),排石冲剂大剂量组、枸橼酸钾组评分显著降低(P<0.01)。与正常对照组比较,其余各组p47phox、TGF-β1、IL-6 mRNA表达水平均显著增强;与模型对照组比较,排石冲剂小、中剂量组无明显差异,排石冲剂大剂量组、枸橼酸钾组显著降低(P<0.05),但两组之间无明显差异。结论排石冲剂能降低大鼠尿草酸、尿钙的排泄,能抑制大鼠肾组织的钙质沉积,降低肾组织中p47phox、TGF-β1、IL-6 mRNA的水平。排石冲剂能抑制结石的形成,其作用机制可能与抑制草酸钙结石对肾小管上皮细胞的氧化炎症损伤有关。

Objective To study the effects of Paishi granule on the expression of p47phox and TGF-β1 in rats with calcium oxalate kidney stones,and the mechanism of Paishi granule in reducing the formation of kidney stones.Methods Sixty Wistar rats were randomly divided into 6 groups:normal control group,model control group,potassium citrate treatment group and 3 groups of different doses of Paishi granule(1.2,2.4 and 3.6 g·mL^-1).The contents of oxalic acid and calcium in urine of each groups were measured.Calcium oxalate crystals in renal tissue were observed by using Vonkossa calcium crystal staining.Expression levels of p47phox,TGF-β1,IL-6 mRNA in kidney tissue were detected by RT-PCR.Results Compared with the normal control group,the contents of oxalic acid and calcium in urine of the other 5 groups were significantly increased(P<0.01).Compared with the model control group,there was no significant difference in the content of uric oxalic acid and urinary calcium in the low-dose group of the Paishi granule(P>0.05),and there was a significant decrease in uric acid and urinary calcium in the medium-dose group(P<0.05).In the high dose group of Paishi granule and potassium citrate group,there were significant decreases both in uric oxalic acid and calcium(P<0.01).Calcium oxalate crystals were not found in the kidney tissue of control rats.Large quantities of black calcium crystals were deposited in the model control group,and scattered calcium crystals were found in the kidney tissues of low and medium dose Paishi granule groups.The renal structure of high-dose group and potassium citrate group was close to normal,and no obvious crystallization deposition was found.Compared with nromal control group,model control group and low-dose group had a very significant increase in crystal score(P<0.01).Compared with model control group,the crystal score of medium-dose group,high dose group and potassium citrate treatment group were very significantly reduced(P<0.05 in medium-dose group,P<0.01 in high-dose and potassium citrate group).The RT-PCR test showed that the levels of p47phox,TGF-β1,and IL-6 mRNA in other groups were significantly higher than those of control groups,however,there was no significant difference between low-dose group,medium-dose group,and model control group.The mRNA levels of high-dose group and potassium citrate group were significantly lower than model control group(P<0.05).There was no significant difference between the high-dose group and potassium citrate group.Conclusion Paishi granule can reduce the excretion of uric acid and calcium in rats,inhibit calcium deposition in rat kidney tissue,and reduce the levels of p47phox,TGF-β1,IL-6 mRNA in kidney tissue.Paishi granule can inhibit the formation of stones,and its mechanism may be related to the oxidative inflammation of renal tubules epithelial cells by inhibiting calcium oxalate stones.