酶化石复合菌粉抑制大鼠肾结石形成及对肾功能的保护作用

Study on the effect of meihuashi complex bacteria product on renal stone formation and renal function in rats

目的探讨酶化石复合菌粉抑制大鼠肾脏草酸钙结石形成及对肾功能的保护作用。方法选用24只SPF级、10周龄雄性大鼠,随机分为对照组(正常饮水和饲料,n=6)、模型组(1%乙二醇饮水,n=6)、枸橼酸氢钾钠治疗组(1%乙二醇饮水+枸橼酸氢钾钠0.9 g/kg灌胃,n=6)和酶化石复合菌粉治疗组(1%乙二醇饮水酶化石复合菌粉0.4 g/kg灌胃,n=6)。全自动生化分析仪检测第0、28天血清中的尿素、肌酐和Ca^(2+)浓度,以及尿液中的草酸、Ca^(2+)浓度;采用HE染色观察肾脏组织病理情况,免疫组化检测肾组织中NF-κB p65、STAT3、TNF-α、IL-1β表达;采用ELISA检测大鼠血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)指标。结果HE染色结果证实模型组肾小管管腔内充满草酸钙结晶;与模型组比较,酶化石复合菌粉治疗组和枸橼酸氢钾钠组肾小管草酸钙结晶大量减少。第0天尿液Ca^(2+)和草酸及血清中的尿素、肌酐、Ca^(2+)在各组之间无显著性差异。对照组相比,模型组第28天尿液Ca^(2+)、草酸及血清中的尿素、肌酐显著升高(P<0.05);与模型组比较,酶化石复合菌粉治疗组、枸橼酸氢钾钠组的尿液Ca^(2+)、草酸及血清中的尿素、肌酐均下降(P<0.05)。与对照组比较,模型组血清CAT、GSH-Px和SOD指标水平下降,而MDA指标水平升高(P<0.05);与模型组比较,酶化石复合菌粉治疗组和枸橼酸氢钾钠组中CAT、GSH-Px和SOD指标水平升高,而MDA指标下降(P<0.05)。与对照组比较,模型组肾脏NF-κB p65、STAT3、TNF-α、IL-1β在组织中的表达上升(P<0.05);与模型组比较,酶化石复合菌粉治疗组和枸橼酸氢钾钠组中NF-κB p65、STAT3、TNF-α、IL-1β表达降低(P<0.05)。结论酶化石复合菌粉能够降低尿草酸、Ca^(2+)含量,增强抗氧化应激,具有抑制肾结石形成及对肾功能的保护作用。

Objective To investigate the effect of Meihuashi complex bacteria product(MCBP)on the inhibition of calcium oxalate stone formation and the protection of renal function in rats.Methods Twenty four SPF grade male rats(10 weeks old)were randomly divided into the control group(normal drinking water and feed,n=6),model group(1%ethylene glycol,n=6),potassium sodium hydrogen citrate(PSHC)treatment group(1%ethylene glycol+0.9 g/kg sodium potassium hydrogen citrate,n=6)and MCBP treatment group(1%ethylene glycol+0.4 g/kg enzyme fossil compound bacteria powder,n=6).On days 0 and 28,the concentration of urea,creatinine and Ca^(2+)in serum as well as the concentration of oxalic acid and Ca^(2+)in urine were measured.HE and immunohistochemical staining were used to detect factors(NF-κB P65,STAT3,TNF-αand IL-1β)in the renal tissues.Other indicators were also measured,including malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)in serum.Results The results of HE staining demonstrates that in the model group,lumina of the renal tubules was filled with calcium oxalate crystals,while the condition was greatly improved in the MCBP and PSHC treatment group.There was no significant difference in the concentration of urea,creatinine and Ca^(2+)in serum as well as the concentration of oxalic acid and Ca^(2+)in urine among all groups on D 0.However,on D 28,such indicators of the model group was greatly elevated in comparison with those of the control group(P<0.05).Compared with the model group,the concentration of oxalic acid and Ca^(2+)in urine and the concentration of urea and creatinine in serum in the MCBP and PSHC treatment group was all decreased with statistical difference(P<0.05).Compared with the control group,CAT,GSH-Px and SOD levels in the model group were all decreased,while the MDA level was increased(P<0.05).Compared with the model group,CAT,GSH-Px and SOD levels were increased in the MCBP and PSHC treatment group(P<0.05),but the MDA levels was decreased.Compared with the control group,the expression of NF-κB p65,STAT3,TNF-αand IL-1βin renal tissues was increased in the model group(P<0.05).Compared with the model group,the expression of NF-κB p65,STAT3,TNF-αand IL-1βwas decreased in the MCBP and PSHC treatment group(P<0.05).Conclusion Meihuashi complex bacteria product can reduce oxalic acid and Ca^(2+)content in urine and enhance antioxidant activity.It has inhibitory effects on kidney stone formation and protective effects on renal function.