黄芪-丹参药对干预梗阻大鼠肾小管上皮细胞转分化的研究

Effect of Herbal Pair of Astragali Radix and Salviae Miltiorrhizae Radix Et Rhizoma on Expression of α-Smooth Muscle Actin and E-cadherin in Ureteral Urinary Obstructive Rat Kidney

目的:观察黄芪-丹参药对及其组分对单侧输尿管梗阻(UUO)大鼠肾组织α-平滑肌肌动蛋白(α-SMA)、上皮型钙黏蛋白(E-cadherin)表达的影响,分析其对肾小管上皮细胞的干预作用。方法:66只SD大鼠随机分为7组,正常组、模型组、Fosinopril组、丹参总酚酸组、黄芪总皂苷组、黄芪-丹参颗粒剂配伍组、黄芪-丹参组分配伍组。造模后第2天各组按剂量给予相应药物ig治疗14 d。福辛普利组按6.6 mg.kg-1给药,丹参总酚酸组按40 mg.kg-1给药,黄芪总皂苷组按37.5 mg.kg-1给药,黄芪丹参组分配伍组中黄芪总皂苷、丹参总酚酸分别按37.5 mg.kg-1和40 mg.kg-1给药,黄芪丹参颗粒剂组黄芪、丹参颗粒均按10 g.kg-1给药,第15天腹主动脉采血检测肾功能(肌酐、尿素氮)检测及代谢笼收集尿液测定视黄醇结合蛋白(RBP)水平。免疫组化染色,光学显微镜下观察肾组织α-SMA,E-cadherin表达变化。结果:肾功能结果分析显示,UUO组大鼠术后血清Cr达(52.34±7.63)μmol.L-1,显著高于正常组(43.29±4.13)μmol.L-1(P<0.05),但除福辛普利组外,各给药组大鼠Cr水平与模型组相比差异无统计学意义。各组大鼠血清BUN水平差异无统计学意义;各造模组大鼠尿RBP水平较正常组显著升高(P<0.05),其中,福辛普利组、组分配伍组大鼠尿RBP分别为(0.41±0.05),(0.41±0.04)μmol.L-1,与模型组(0.49±0.09)μmol.L-1相比有显著性差异(P<0.05)。免疫组化结果均以模型组病变最严重;病变最轻的为福辛普利组及组分配伍组。结论:黄芪丹参药对及其有效成分对UUO大鼠肾功能、肾小管功能有一定的保护作用,其机制可能与其通过减少模型鼠肾组织α-SMA表达,增加E-cadherin表达,干预肾小管上皮细胞转分化而防治肾间质纤维化有关。

Objective：To observe effect of effective constituents of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma on the expression of α-SMA and E-cadherin in UUO rat kidney.Method：Sixty-six SD rats were randomly divided into 7 groups,normal group,model group,fosinopril group,salvianolic acids group,astragalus saponins group,granules combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma group,constituents combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma group.Each group was treated with different does of drugs after UUO surgery for 14 days.The doesage of fosinopril group was 6.6 mg·kg-1,salvianolic acids group was 40 mg·kg-1,astragalus saponins group was 37.5 mg·kg-1,constituents combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma group was given salvianolic acids 40 mg·kg-1 and astragalus saponins group was 37.5 mg·kg-1,granules combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma group was given Astragali Radix granules and Salviae Miltiorrhizae Radix et Rhizoma granules 10 g·kg-1.Blood was withdrawn from abdominal aortic to determine the renal function（creatinine,urea nitrogen）and metabolic cages were used to collect urine for retinol binding protein（RBP）determination,the obstructed kidney was taken out and fixed by 10% paraform and made into paraffin section by dehydration and embedding which were used for immunohistochemistry experiment.Result： The level of serum Cr in UUO rats reached（52.34±7.63） μmol·L-1,was higher than that of（43.29±4.13） μmol·L-1 in the normal group,however,except fosinopril,there was no significant difference between the treatment group and the model group No significant differences were found in the serum BUN.The standard of urinary RBP expression in every UUO group was significantly higher than normal group（P0.05）,compared with the model group that its figure was（0.49±0.09）μmol·L-1,the fosinopril group and constituents group RBP levels were（0.41±0.05）,（0.41±0.04）μmol·L-1,of which had obvious significance（P0.05）.Immunohistochemical results showed that the most severely pathological changes was in the model group,the lightest pathological changes were found in fosinopril group and the group with constituents combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma.Conclusion：Renal function has been damaged in UUO rats,Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma effective constituents and their combination could ameliorate renal function and reduce the expression of tubular protein RBP in urine.Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma and their active constituents intervention could alleviate renal fibrosis at a certain degree.