温阳振衰颗粒对心肾综合征模型大鼠血浆外泌体及心、肾组织p38MAPK表达的影响

Effects of Wenyang Zhenshuai Granules (温阳振衰颗粒) on the Expression of p38MAPK in the Plasma Exosomes and the Tissues of Heart and Kidney of Cardio-Renal Syndrome Model Rats

目的探讨温阳振衰颗粒对心肾综合征大鼠心功能、肾功能的影响及可能作用机制。方法 40只SD大鼠随机分为空白组、模型组、温阳振衰组、依那普利组,每组10只。除空白组外其余各组大鼠采用阿霉素3mg/kg腹腔注射,每周1次,共6周建立心肾综合征大鼠模型。造模3周后空白组和模型组大鼠给予生理盐水每次1ml/100g灌胃,温阳振衰组每次给予0.72g/kg温阳振衰颗粒灌胃,依那普利组每次给予0.9mg/kg依那普利片灌胃。每天给药2次,连续4周。HE染色观察肾组织病理改变,TUNEL法检测心肌及肾组织细胞凋亡阳性率,检测心功能、肾功能及血清N端前脑钠素(NT-pro BNP)、肾损伤分子1(KIM-1)含量,检测心脏和肾脏p38丝裂原活化蛋白激酶(p38MAPK)和磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)蛋白及mRNA表达,检测血浆外泌体定量,心脏、肾脏组织及血浆外泌体miR-155表达。结果与空白组比较,模型组大鼠心功能、肾功能损伤,心肌及肾组织细胞凋亡阳性率增加,血清NT-pro BNP、KIM-1含量升高,心脏和肾脏p-p38MAPK蛋白、p38MAPK mRNA表达均升高,心脏、肾脏和血浆外泌体miR-155均降低(P<0.05或P<0.01);与模型组比较,温阳振衰组和依那普利组上述指标均不同程度改善(P<0.01)。结论温阳振衰颗粒可改善心肾综合征模型大鼠心功能、肾功能,其作用可能与增加外泌体miR-155抑制p-p38MAPK表达,从而抗细胞凋亡有关。

Objective To explore the effects and possible mechanism of Wenyang Zhenshuai Granules(温阳振衰颗粒) on cardiac and renal function. Methods A total of 40 SD rats were randomly divided into blank group, model group, Wenyang Zhenshuai group, and enalapril group, with 10 rats in each group. Rat except for the blank group were injected intraperitoneally with 3 mg/kg doxorubicin, once a week for 6 weeks to establish rat model of cardio-renal syndrome. After 3 weeks of modeling, rats in the blank and model groups were given 1 ml/100 g of saline by gavage each time, the Wenyang Zhenshuai group was given Wenyang Zhenshuai Granules by gavage 0.72 g/kg every time, and the enalapril group was given enalapril tablets intragastrically 0.9 mg/kg each time for gavage. All the groups were administrated twice a day for 4 weeks. HE staining was used to observe the pathological changes of renal tissue. The positive rate of apoptosis in myocardium and renal tissue was detected by TUNEL method. Heart function, renal function, serum N-terminal pro-brain natriuretic peptide(NT-pro BNP) and renal injury molecule 1(KIM-1) content were detected, heart and kidney p38 mitogen-activated protein kinase(p38 MAPK), phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK) protein and mRNA expression were detected. Quantitation of plasma exosomes, expression of plasma exosome miR-155 in the heart and kidney were detected. Results Compared with the blank group, damaged heart function and kidney function, increased positive rates of apoptosis in myocardium and kidney tissue, increased serum NT-pro BNP and KIM-1 levels, increased p-p38 MAPK protein and p38 MAPK mRNA in the heart and kidney was increased, and decreased exosomes miR-155 in the heart, kidney and plasma were found(P<0.05 or P<0.01). Compared with the model group, the above indicators were improved to different degree in the Wenyang Zhenhuai group and the enalapril group(P<0.01). Conclusion Wenyang Zhenshuai Granules can improve the cardiac and renal function of rats with cardio-renal syndrome. The action may be related to increasing exosome miR-155 to inhibit the expression of p-p38 MAPK, thus exerting anti-apoptotic effects.