复肾功方对慢性肾衰竭大鼠Klotho蛋白及mRNA表达的影响

目的探讨复肾功方(Fushengong decoction,FSGD)对慢性肾衰竭(Chronic renal failure,CRF)大鼠Klotho蛋白及mRNA表达的影响。方法将55只雄性SD大鼠随机分为正常组、模型组及复肾功方低、中、高剂量组,每组11只。采用腺嘌呤含量为0.5%的饲料建立CRF模型,连续喂养21 d。模型复制成功后,所有大鼠改用常规饲料喂养;复肾功方低、中、高剂量组按照体质量分别以含生药4、8、16 g·kg^-1的复肾功方水煎剂灌胃给药,正常组及模型组给予同体积生理盐水,给药体积20 mL·kg^-1·d^-1,每日1次,连续灌胃给药30 d。实验结束后,取各组大鼠血清检测血肌酐(SCr)、尿素氮(BUN)的含量;HE染色法检测大鼠肾脏组织的病理形态学变化;采用Western Blot法和免疫组化法检测肾脏组织Klotho蛋白的表达;Real-time PCR法检测肾脏组织Klotho mRNA的表达。结果与正常组比较,模型组大鼠血清SCr、BUN的含量均显著升高(P<0.01),肾脏组织病理损害明显,肾脏Klotho蛋白及mRNA的表达显著降低(P<0.01);与模型组比较,复肾功方各剂量组大鼠的肾脏组织病理损害明显减轻,血清SCr、BUN含量显著降低(P<0.05,P<0.01),大鼠肾脏组织Klotho蛋白及mRNA的表达明显升高(P<0.05)。结论复肾功方能够降低CRF大鼠血清SCr、BUN的含量,减轻肾损伤,可能与上调肾脏组织的Klotho蛋白及mRNA的表达有关。

Effects of Fushengong Decoction on Expression of Klotho Protein and mRNA in Kidney of Rats with Chronic Renal Failure

Objective To explore the effects of Fushengong decoction(FSGD)on the expression of Klotho protein and mRNA in chronic renal failure(CRF)rats. Methods A total of 55 male Sprague-Dawley rats were randomly divided into normal group,model group,and low,medium and high dose groups of Fushengong decoction,with11 rats in each group. Rats were fed with fodder containing 0.5% adenine continuously for 21 days to establish CRF models. After successful modeling,all rats were switched to conventional feed. The low,medium and high dose groups rats were given intragastric administration of Fushengong decoction once a day for 30 days at dosages of 4,8,and 16 g·kg^-1(crude drug/body mass),respectively. The normal group and the model group rats were given the same volume of normal saline(NS)at 20 mL·kg^-1·d^-1. After the experiment,the serum of rats were collected to detect the contents of SCr and BUN;pathomorphological change of renal tissues of the rats were observed by hematoxylin-eosin staining. Protein expression of Klotho was detected by Western Blot and immunohistochemistry.The expression of Klotho mRNA in the kidney was measured by Real-time PCR. Results Compared with the normal group,the contents of serum SCr and BUN of the model group rats significantly increased(P < 0.01)and the renal tissues were obviously damaged;the expression of renal Klotho protein and Klotho mRNA significantly decreased(P < 0.01). Compared with the model group rats, the pathological damage of renal tissues of all Fushengong decoction group rats alleviated distinctly and the contents of serum SCr and BUN reduced significantly(P < 0.05,P < 0.01);the expression of renal Klotho protein and Klotho mRNA increased obviously(P < 0.05). Conclusion Fushengong decoction can reduce the contents of serum SCr and BUN and alleviate renal damage in CRF rats,which may be related to up-regulating the expression of Klotho protein and Klotho mRNA.