Effect of Wenyang Zhenshuai Granules on autophagy and apoptosis of myocardial cells in septic rats via regulating miR-132-3p/UCP2 expression / 中国中药杂志

This study aimed to investigate the effect of Wenyang Zhenshuai Granules(WYZSG) on autophagy and apoptosis of myocardial cells in rats with sepsis via regulating the expression of microRNA-132-3p(miR-132-3p)/uncoupling protein 2(UCP2). Sixty SD rats were randomly divided into modeling group(n=50) and sham operation group(n=10). The sepsis rat model was constructed by cecal ligation and perforation in the modeling group. The successfully modeled rats were randomly divided into WYZSG low-, medium-and high-dose groups, model group and positive control group. Rats in the sham operation group underwent opening and cecum division but without perforation and ligation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of rat myocardial tissue. Myocardial cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL) assay. Real-time quantitative polymerase chain reaction(RT-qPCR) was performed to detect the expression of miR-132-3p and the mRNA expressions of UCP2, microtubule-associated protein light chain 3(LC3-Ⅱ/LC3-Ⅰ), Beclin-1 and caspase-3 in rat myocardial tissue. The protein expressions of UCP2, LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 in myocardial tissue were detected by Western blot. Dual luciferase reporter assay was used to verify the regulatory relationship between miR-132-3p and UCP2. The myocardial fibers of sepsis model rats were disordered, and there were obvious inflammatory cell infiltration as well as myocardial cell edema and necrosis. With the increase of the WYZSG dose, the histopathological changes of myocardium were improved to varying degrees. Compared with the conditions in the sham operation group, the survival rate and left ventricular ejection fraction(LVEF) of rats in the model group, positive control group and WYZSG low-, medium-and high-dose groups were decreased, and the myocardial injury score and apoptosis rate were increased. Compared with the model group, the positive control group and WYZSG low-, medium-and high-dose groups had elevated survival rate and LVEF, and lowered myocardial injury score and apoptosis rate. The expression of miR-132-3p and the mRNA and protein expressions of UCP2 in myocardial tissue in the model group, positive control group and WYZSG low-, medium-and high-dose groups were lower, while the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 were higher than those in the sham operation group. Compared with model group, the positive control group and the WYZSG low-, medium-and high-dose groups had an up-regulation in the expression of miR-132-3p and the mRNA and protein expressions of UCP2, while a down-regulation in the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3. WYZSG inhibited excessive autophagy and apoptosis of myocardial cells in septic rats and improved myocardial injury, possibly by regulating the expression of miR-132-3p/UCP2.

Content Determination of Ginsenoside Rg1, Ginsenoside Re and Ginsenoside Rb1 in Wenyang Zhenshuai Granules by HPLC / 中国中医药信息杂志

s: Objective To establish an HPLC method to determine the contents of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 in Wenyang Zhenshuai Granules.Methods The separation was preformed on Agilent TC-C18 column (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitril-0.05% aqueous solution of phosphoric acid with gradient elution (0–35 min, 19% acetonitrile; 35–58 min, 19%–29% acetonitrile; 58–70 min, 28% acetonitrile; 70–100 min, 29%–40% acetonitrile). The flow rate was 1.0 mL/min; the wavelength of detector was 203 nm; the temperature of column was 35℃.Results The calibration curves showed good linearity in the range of 0.22–2.2 μg (ginsenoside Rg1,r=0.9998), 0.22–2.2 μg (ginsenoside Re,r=0.9998) and 0.26–2.6 μg (ginsenoside Rb1,r=0.9991), respectively. The average recoveries of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 were 98.04%, 96.58% and 96.75%, respectively.Conclution The method is accurate, hypersensitized and reproducible, which can be applied to the quality control of Wenyang Zhenshuai Granules.

Effect of Wenyang Zhenshuai Granules on the Expression of Myocardial MEF2 in Model Rats with Chron-ic Heart Failure / 中国药房

OBJECTIVE:To study the effect of Wenyang zhenshuai granules on myocardial MEF2 in model rats with chronic heart failure(CHF). METHODS:10 rats were randomly selected in 60 rats as normal control group(normal saline),the remain-ing 50 rats were randomly divided into model control group (normal saline),positive control group [enalapril,1.8 mg/(kg·d)], Wenyang zhenshuai granules low-dose,medium-dose,high-dose groups [0.72,1.44,2.88 g/(kg·d)] after induced to CHF models by adriamycin,intragastrically administrated twice a day,for 4 weeks. After administration,heart function indexes [left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS),left ventricular end-diastolic diameter (LVSD) and left ventricular end-diastolic diameter(LVDD)] of rats in each group were detected,protein expressions of myocardial MEF2 and phos-phorylated MEF2 (p-MEF2) and mRNA expression of MEF2 were determined. RESULTS:Compared with normal control group, LVEF,LVFS of rats in model control group and each administration group were significantly reduced (P<0.05),LVSD,LVDD were significantly increased (P<0.05);protein expression of myocardial p-MEF2 was significantly down-regulated (P<0.05), and there were no statistical significances in protein or mRNA expressions of MEF2 (P>0.05). Compared with model control group,heart function indexes of rats in each administration group were significantly improved (P<0.05);protein expression of myocardial p-MEF2 was significantly up-regulated (P<0.05),and there were no statistical significances in protein or mRNA ex-pressions of MEF2(P>0.05). CONCLUSIONS:Wenyang zhenshuai granule can antagonize the down-regulation effect of adriamy-cin-induced MEF2 protein phosphorylation,which may be one of the mechanisms of its treatment for CHF.