基于“水火相济”理论探讨慢性心力衰竭对肾功能和肾脏纤维的影响及益气温阳活血利水法的干预研究

Influence of Chronic Heart Failure on Kidney Function and Fibrosis andIntervention of Benefiting Qi-Warming Yang-Activating Blood Circulation-Promoting Urination Method Based on Theory of“Harmonization of Water and Fire”

目的观察慢性心力衰竭对肾功能和肾脏纤维化的影响及益气温阳活血利水法的干预作用,探讨"水火相济"理论内涵。方法随机将50只C57BL/6J小鼠分为假手术组、模型组、中药低、中、高剂量组,通过主动脉缩窄术建立慢性心力衰竭模型小鼠,造模后中药低、中、高剂量组给予益气温阳活血利水中药对应剂量灌胃,假手术组和模型组给予等量生理盐水灌胃,连续给药8周后,小动物超声检测小鼠心脏射血分数、收缩分数变化。于末次给药2 h后取材,全自动生化分析仪检测血清肌酐、尿素氮水平,微量酶标法检测血清GSH含量,冰冻切片检测肾脏组织ROS水平,免疫组化法检测肾脏CollagenⅠ、CollagenⅢ、p53、COX2、FTH1、GPX4蛋白表达情况,RT-PCR法检测肾脏组织P53、SLC7A11、GPX4、PTGS2、NOX1、FTH1 mRNA表达水平。结果与假手术组比较,模型组小鼠心脏EF、FS、GSH水平显著降低,SCr、BUN、ROS水平显著升高,肾脏CollagenⅠ、CollagenⅢ、JNK、p53、PTGS2、NOX1蛋白或mRNA水平显著升高,GPX4、FTH1、SLC7A11蛋白或mRNA水平显著降低;与模型组比较,经治疗后各项指标水平明显改善,差异具有统计学意义。结论益气温阳活血利水法在改善CHF小鼠心功能的同时能够改善肾脏损伤,减少纤维化,其机制可能与抑制JNK/p53信号通路表达,进一步调控SLC7A11/GPX4/GSH和PTGS2/NOX1信号通路进而降低ROS水平减少细胞铁死亡。

Objective To observe the effects of chronic heart failure on the kidney and the intervention of benefiting Qi-warming Yang-activating blood circulation-promoting urination methodandexplore the theoretical"harmonization of water and fire".Methods Fifty C57 BL/6 J mice were randomly divided into sham operation group,model group,low,medium and high dose groups of traditional Chinese medicine.The chronic heart failure model was set up by aortic coarctation.After the model establishment,the low,medium and high doses of traditional Chinese medicine were given to the corresponding doses of benefiting Qi-warming Yang-activating blood circulation-promoting urination prescription.Thesham operation group and model group were given the same amount of saline.After 8 weeks of continuous administration,the changes of cardiac ejection fraction and contraction fraction of the mice were detected.After 2 hours of the last administration,the material was taken out,and the serum creatinine and urea nitrogen levels were detected by an automatic biochemical analyzer.The serum GSH content was determined by microenzyme labeling method.The frozen sections were used for detecting ROS levels.Immunohistochemistry was used to detect the expressiosn of Collagen I,Collagen III,p53,COX2,FTH1 and GPX4 proteins.The mRNA expression levels of P53,SLC7 A11,GPX4,PTGS2,NOX1 and FTH1 were detected by RT-PCR.Results Compared with the sham operation group,the levels of EF,FS and GSH in the model group were significantly decreased.The levels of SCr,BUN and ROS were significantly increased,and the levels of Collagen I,Collagen III,JNK,p53,PTGS2,NOX1 protein or mRNA were significantly increased.The levels of GPX4,FTH1,SLC7 A11 protein or mRNA were significantly decreased.Compared with the model group,the levels of various indicators were significantly improved after treatment,and the difference was statistically significant.Conclusion Benefiting Qi-warming Yang-activating blood circulation-promoting urination method can improve kidney function and reduce fibrosis while improving heart function in chronic heart failure mice.The mechanism may be related to inhibiting the expression of JNK/p53 signaling pathway and further regulating SLC7 A11/GPX4/GSH and PTGS2/NOX1 signaling pathways so as to decrease ROS levels reduces cellular ferroptosis.