Danlou Tablet Improves Chronic Intermittent Hypoxia-Induced Dyslipidemia and Arteriosclerosis by HIF-1α-Angptl4 mRNA Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To detect whether Danlou Tablet (DLT) regulates the hypoxia-induced factor (HIF)-1α-angiopoietin-like 4 (Angptl4) mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia (CIH)-induced dyslipidemia and arteriosclerosis.@*METHODS@#The mature adipocytes were obtained from 3T3-L1 cell culturation and allocated into 8 groups including control groups (Groups 1 and 5, 0.1 mL of cell culture grade water); DLT groups (Groups 2 and 6, 0.1 mL of 1,000 µg/mL DLT submicron powder solution); dimethyloxalylglycine (DMOG) groups (Groups 3 and 7, DMOG and 0.1 mL of cell culture grade water); DMOG plus DLT groups (Groups 4 and 8, DMOG and 0.1 mL of 1,000 µg/mL DLT submicron powder solution). Groups 1-4 used mature adipocytes and groups 5-8 used HIF-1 α-siRNA lentivirus-transfected mature adipocytes. After 24-h treatment, real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1 α and Angptl4. In animal experiments, the CIH model in ApoE-/- mice was established. Sixteen mice were complete randomly divided into 4 groups including sham group, CIH model group [intermittent hypoxia and normal saline (2 mL/time) gavage once a day]; Angptl4 Ab group [intermittent hypoxia and Angptl4 antibody (30 mg/kg) intraperitoneally injected every week]; DLT group [intermittent hypoxia and DLT (250 mg/kg) once a day], 4 mice in each group. After 4-week treatment, enzyme linked immunosorbent assay was used to detect the expression levels of serum total cholesterol (TC) and triglyceride (TG). Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.@*RESULTS@#Angptl4 expression was dependent on HIF-1 α, with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1 α -transfected cells. DLT inhibited HIF-1 α and Angptl4 mRNA expression (P<0.05 or P<0.01) and reduced HIF-1 α and Angptl4 protein expressions with DMOG in mature adipocytes (all P<0.01), as the effect on HIF-1 α protein also existed in the presence of siHIF-1 α (P<0.01). ApoE-/- mice treated with CIH had increased TG and TC levels (all P<0.01) and atherosclerotic plaque. Angptl4 antibody and DLT both reduce TG and TC levels (all P<0.01), as well as reducing atherosclerotic plaque areas, narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.@*CONCLUSION@#DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1 α-Angptl4 mRNA signaling pathway.

Based on "intestinal flora-inflammation" pathway to investigate effect and mechanism of Danlou Tablet on atherosclerosis of ApoE-/- mice / 中草药

Objective: To investigate the efficacy and mechanism of Danlou Tablet against atherosclerosis model of ApoE-/- mice fed with high fat diet. Methods: C57BL/6J mice were used as controls and ApoE-/- mice were randomly divided into two groups after 24 weeks of high fat feeding, including the model group received saline and the treatment group received Danlou Tablet. Animals were executed after 8 weeks of treatment and serum was collected to measure blood lipids; Plaque formation in the aorta was observed by red O and HE staining; 16 S rRNA sequencing was used to analyze changes in intestinal flora, and GC-MS test for detection fecal SCFAs content, ELISA for the determination of serum LPS, and real time PCR for detection of mRNA expression. Results: Compared with the control group, the blood lipid levels were increased; intestinal flora was imbalance with increased harmful bacteria and reduced beneficial bacteria; The level of serum LPS and inflammation around the aorta were increased in the model group. Compared with the model group, the contents of TG, TC, LDL-C and the plaque area of Danlou Tablet group were decreased (P < 0.01); Danlou Tablet group can regulate intestinal flora, thus effectively reducing serum LPS and inflammatory factors TNF-α, ICAM-1 and IL-1β levels around the aorta (P < 0.01). Conclusion: Danlou Tablet exerts an anti-atherosclerosis action with favorable efficacy through restructing the intestinal flora stucture, inhibiting endotoxin releasing and constraining the inflammatory response induced by dysbacteriosis.

Protective Effects of Danlou Tablet () against Murine Myocardial Ischemia and Reperfusion Injury In Vivo / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the in vivo effect of Danlou Tablet (, DLT) on myocardial ischemia and reperfusion (I/R) injury.</p><p><b>METHODS</b>DLT effects were evaluated in mouse heart preparation using 30-min coronary occlusion followed by 24-h reperfusion and compared among sham group (n=6), I/R group (n=8), IPC group (ischemia preconditioning, n=6) and DLT group (I/R with DLT pretreatment for 3 days, 750 mg•kg•day, n=8). The effects of DLT were characterized in infarction size (IS) compared with risk region (RR) and left ventricle using the Evans blue/triphenyltetrazolium chloride double dye staining method in vivo. Furthermore, the dose-dependent effect of DLT on I/R injury was evaluated by double staining method. Five different concentrations of DLT (0.625, 1.25, 2.5, 5 and 10 g•kg•day) were chosen in this study, and dose-response curve of DLT was obtained on these data.</p><p><b>RESULTS</b>The ratio of IS to left ventricle was significantly smaller in the DLT and IPC groups than the I/R group (P<0.05 or P<0.01), the ratio of IS to RR was also reduced in the DLT and IPC groups (P<0.01), while there were no differences in RR among the four groups (P>0.05). Experiments showed incidence of arrhythmias was reduced in the DLT group (P<0.01). Furthermore, DLT produced a dose-dependent inhibitory effect with a half maximal inhibitory concentration of 1.225 g•kg•day.</p><p><b>CONCLUSIONS</b>Our research concluded that DLT was effective in reducing I/R injury in mice, and provided experimental supports for the clinical use of DLT.</p>

Effect of Danlou Tablet () on peri-procedural myocardial injury among patients undergoing percutaneous coronary intervention for non-ST elevation acute coronary syndrome: A study protocol of a multicenter, randomized, controlled trial / 中国结合医学杂志

<p><b>BACKGROUND</b>It has been shown that administration of statins reduced the risk of peri-procedural myocardial damage. However, it remains unclear whether Chinese medicine Danlou Tablet (), similar to statins, may protect patients undergoing percutaneous coronary intervention (PCI) from peri-procedural myocardial damage.</p><p><b>OBJECTIVE</b>To demonstrate the hypothesis whether treatment with Danlou Tablet would improve clinical outcome in patients undergoing selective PCI with non-ST elevation acute coronary syndrome (NSTE-ACS) in China.</p><p><b>METHODS</b>Approximately 220 patients with unstable angina or non-ST-segment elevation myocardial infarction undergoing PCI will be enrolled and randomized to Danlou Tablet treatment (4.5 g/day for 2 days before intervention, with a further 4.5 g/day for 90 days thereafter) or placebo. All patients will not receive Danlou Tablet before procedure. The primary end point is to evaluate the incidence of cardiac death, myocardial infarction or unplanned re-hospitalization and revascularization after 30 days in patients undergoing selective PCI treated with Danlou Tablet compared with placebo. Secondary endpoints include the incidence of peri-procedural myocardial injury, 3-month clinical outcomes, the quality of life and Chinese medicine syndromes assessment.</p><p><b>CONCLUSION</b>This study protocol will provide important evidence of Danlou Tablet treatment on the peri-procedural myocardial injury in patients with NSTE-ACS undergoing selective PCI, which may support a strategy of routine Danlou Tablet therapy to improve the clinical outcomes.</p>

Establishment of a rat model of atherosclerosis with syndrome of intermingled phlegm and blood stasis and intervention experiment with a Chinese medicine prescription Danlou tablet / 中国比较医学杂志

ObjectiveToestablisharatmodelofatherosclerosiscombinedwithsyndromeofintermingledphlegm and blood stasis , to observe the inflammatory reactions and the treatment effect of prescription ( Danlou tablet ) on the rat model.Methods Thirty-two healthy male SD rats were randomly and equally divided into four groups , namely, normal control group, model control group, atorvastatin group (ATV group), and Danlou group (DLP group).The normal control group was given basic forage , while other three groups were given high fat forage plus intraperitoneal injection of vitamin D 3 and balloon injury of the left common carotid artery to build rat atherosclerosis model combined with syndrome of intermingled phlegm and blood stasis , and then received intragastric administration of saline , atorvastatin suspension and Danlou tablets suspension for 4 weeks, respectively.After intervention, both serum lipid and hs-CRP, IL-6, TNF-α, and LP-PLA2 levels were determined by ELISA , pathological alterations in the thoracic aorta was analyzed using HE staining , the expressions of IL-6, TNF-αand LP-PLA2 mRNA in the thoracic aorta tissue were assessed by real-time fluorescent quantitativePCRtechnology.Results ①Comparedwiththenormalcontrolgroup,thereweresignificantincreasesin serum TC, LDL-C, hs-CRP, IL-6, TNF-α, and LP-PLA2 levels (P0.05).There were also reduced aortic intimal hyperplasia , macrophage infiltration and plaque area compared with those of the model group .Conclusions Rat model of atherosclerosis combined with syndrome of intermingled phlegm and blood stasis can be established by high fat diet feeding combined with the intraperitoneal injection of vitamin D 3 and balloon injury of carotid artery .The prescription ( Danlou tablet ) can inhibit the inflammatory reaction and ameliorate atherosclerotic changes in the rat models .