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重庆医科大学 中医药学院,附属第一医院,重庆 400016
张洋,在读硕士,从事肾病中医基础与临床研究,E-mail:942118290@qq.com
* 黄学宽,教授,博士生导师,从事内分泌代谢疾病及肾病中医理论与临床研究,E-mail:xkhuang2002@cqmu.edu.cn
收稿日期:2019-12-26,
网络出版日期:2020-10-16,
纸质出版日期:2020-12-20
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张洋,黄学宽,沈清等.复肾功方对慢性肾衰竭大鼠Wnt/β-catenin信号途径的影响[J].中国实验方剂学杂志,2020,26(24):89-95.
ZHANG Yang,HUANG Xue-kuan,SHEN Qing,et al.Effect of Fushengong Prescription on Wnt/β-catenin Signaling Pathway in Kidney of Rats with Chronic Renal Failure[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):89-95.
张洋,黄学宽,沈清等.复肾功方对慢性肾衰竭大鼠Wnt/β-catenin信号途径的影响[J].中国实验方剂学杂志,2020,26(24):89-95. DOI: 10.13422/j.cnki.syfjx.20202436.
ZHANG Yang,HUANG Xue-kuan,SHEN Qing,et al.Effect of Fushengong Prescription on Wnt/β-catenin Signaling Pathway in Kidney of Rats with Chronic Renal Failure[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):89-95. DOI: 10.13422/j.cnki.syfjx.20202436.
目的
2
观察复肾功方对慢性肾衰竭(CRF)大鼠分泌型糖蛋白(Wnt)/
β-
连环蛋白(
β
-catenin)信号途径的影响,探讨其减少细胞外基质(ECM)的沉积,抗肾小管间质纤维化(TIF)及延缓CRF进展的作用机制。
方法
2
将55只雄性SD大鼠随机将分为正常组,模型组,复肾功方低、中、高剂量组,每组11只。正常组常规饲养,其余4组大鼠则用腺嘌呤含量为0.5%的饲料建立CRF模型,连续喂养21 d。造模成功后所有大鼠改用常规饲料,正常组和模型组大鼠给予20 mL·kg
-1
生理盐水灌胃;复肾功方低、中、高剂量组大鼠则按体质量分别灌胃4,8,16 g·kg
-1
复肾功方水煎剂,1次/d,连续灌胃30 d。实验结束后,马松(Masson)染色观察大鼠肾组织病理形态学变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测肾组织Wnt4,
β
-catenin mRNA的表达,蛋白免疫印迹法(Western blot)检测肾组织Wnt4,
β
-catenin,基质金属蛋白酶-7(MMP-7)蛋白的表达,免疫组化检测肾组织Wnt4,
β
-catenin蛋白的表达。
结果
2
与正常组比较,模型组大鼠肾小管间质纤维化明显增多,肾组织Wnt4,
β
-catenin mRNA及Wnt4,
β
-catenin,MMP-7蛋白的表达均显著升高(
P
<
0.01);与模型组比较,复肾功方各组大鼠肾组织Wnt4,
β
-catenin mRNA及Wnt4,
β
-catenin,MMP-7蛋白表达均明显降低(
P
<
0.05)。
结论
2
复肾功方减少ECM的沉积、抗肾小管间质纤维化、延缓CRF进展可能与抑制Wnt/
β
-catenin信号途径的激活有关。
Objective
2
To observe the effects of Fushengong prescription on secretory glycoprotein (Wnt)/
β
-serial protein (
β
-catenin) signaling pathway in kidney of rats with chronic renal failure (CRF),and to further explore its mechanism of releasing the aggregation of extracellular matrix(ECM),inhibiting renal tubule interstitial fibrosis (TIF) and prolonging the progression of CRF.
Method
2
A total of 55 SD male rats were randomly divided into the normal group,the model group,and the low, medium and high dose groups of Fushengong prescription,with 11 rats in each group.The normal group was routinely reared and the other 4 groups of rats were used to establish CRF model with 0.5% adenine fodder, fed them continuously for 21 d. After successful modeling,all model rats were switched to conventional feed. Normal saline (NS) was given the normal group and the model group by 20 mL·kg
-1
·d
-1
, the low, middle and high dose groups rats of Fushengong prescription were given intragastric administration Fushengong prescription according to the body weight of 4, 8, 16 g·kg
-1
,once a day,continuous gavage for 30 d. After the experiment,the pathomorphism change of renal tissues of rats was measured by Masson staining, the expression of Wnt4 and
β
-catenin mRNA in the kidney tissues were observed by the method of Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), the expression of Wnt4,
β
-catenin and matrix metalloproteinase-7(MMP-7) protein of renal tissues were detected by the methods of Western blot. The expression of Wnt4,
β
-catenin protein of renal tissues were detected by the methods of immunohistochemistry (IHC).
Result
2
Compared with normal group,renal tubule interstitial fibrosis of renal tissues increased distinctly and the expression of Wnt4,
β
-catenin and MMP-7 protein increased significantly in the model group. Wnt4 and
β
-catenin mRNA also increased significantly in model group(
P
<
0.01). Compared with model group, the expression of Wnt4,
β
-catenin and MMP-7 protein in the Fushengong prescription groups decreased obviously (
P
<
0.05). The expression of Wnt4 and
β
-catenin mRNA in Fushengong prescription groups also decreased obviously.
Conclusion
2
The mechanism of Fushengong prescription can release the aggregation of ECM,inhibit TIF and delay the progression of CRF,which may be related with the activation of Wnt/
β
-catenin signal pathway.
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