Shexiang Tongxin Dropping Pill Promotes Angiogenesis through VEGF/eNOS Signaling Pathway on Diabetic Coronary Microcirculation Dysfunction

Objective:To study the effect of Shexiang Tongxin Dropping Pill(STDP)on angiogenesis in diabetic cardiomyopathy mice with coronary microcirculation dysfunction(CMD).Methods:According to a random number table,6 of 36 SPF male C57BL/6 mice were randomly selected as the control group,and the remaining 30 mice were injected with streptozotocin intraperitoneally to replicate the type 1 diabetes model.Mice successfully copied the diabetes model were randomly divided into the model group,STDP low-dose group[15 mg/(kg·d)],medium-dose group[30 mg/(kg·d)],high-dose group[60 mg/(kg·d)],and nicorandil group[15 mg/(kg·d)],6 in each group.The drug was given by continuous gavage for 12 weeks.The cardiac function of mice in each group was detected at the end of the experiment,and coronary flow reserve(CFR)was detected by chest Doppler technique.Pathological changes of myocardium were observed by hematoxylin-eosin staining,collagen fiber deposition was detected by masson staining,the number of myocardial capillaries was detected by platelet endothelial cell adhesion molecule-1 staining,and the degree of myocardial hypertrophy was detected by wheat germ agglutinin staining.The expression of the vascular endothlial growth factor(VEGF)/endothelial nitric oxide synthase(eNOS)signaling pathway-related proteins in myocardial tissue was detected by Western blot.Results:Compared with the model group,medium-and high-dose STDP significantly increased the left ventricular ejection fraction and left ventricular fraction shortening(P<0.01),obviously repaired the disordered cardiac muscle structure,reduced myocardial fibrosis,reduced myocardial cell area,increased capillary density,and increased CFR level(all P<0.01).Western blot showed that high-dose STDP could significantly increase the expression of VEGF and promote the phosphorylation of vascular endothelial growth factor receptor 2,phosphoinositide 3-kinase,protein kinase B,and eNOS(P<0.05 or P<0.01).Conclusion:STDP has a definite therapeutic effect on diabetic CMD,and its mechanism may be related to promoting angiogenesis through the VEGF/eNOS signaling pathway.

Shexiang Tongxin Dropping Pill Allieviates Heart Failure via Extracellula Matrix-Receptor Interaction Pathways Based on RNA-Seq Transcriptomics and Experimental Studies

Objective:To investigate the protective mechanisms of Chinese medicine Shexiang Tongxin Dropping Pills(STDP) on heart failure(HF).Methods:Isoproterenol(ISO)-induced HF rat model and angiotensin Ⅱ(Ang Ⅱ)-induced neonatal rat cardiac fibroblast(CFs) model were used in the present study.HF rats were treated with and without STDP(3 g/kg).RNA-seq was performed to identify differentially expressed genes(DEGs).Cardiac function was evaluated by echocardiography.Hematoxylin and eosin and Masson’s stainings were taken to assess cardiac fibrosis.The levels of collagen Ⅰ(Col Ⅰ) and collagen Ⅲ(Col Ⅲ) were detected by immunohistochemical staining.CCK8 kit and transwell assay were implemented to test the CFs’ proliferative and migratory activity,respectively.The protein expressions of α-smooth muscle actin(α-SMA),matrix metalloproteinase-2(MMP-2),MMP-9,Col I,and Col Ⅲ were detected by Western blotting.Results:The results of RNA-seq analysis showed that STDP exerted its pharmacological effects on HF via multiple signaling pathways,such as the extracellular matrix(ECM)-receptor interaction,cell cycle,and B cell receptor interaction.Results from in vivo experiments demonstrated that STDP treatment reversed declines in cardiac function,inhibiting myocardial fibrosis,and reversing increases in Col Ⅰ and Col Ⅱ expression levels in the hearts of HF rats.Moreover,STDP(6,9 mg/mL) inhibited the proliferation and migration of CFs exposed to Ang Ⅱin vitro(P<0.05).The activation of collagen synthesis and myofibroblast generation were markedly suppressed by STDP,also the synthesis of MMP-2 and MMR-9,as well as ECM components Col Ⅰ,Col Ⅲ,and α-SMA were decreased in Ang Ⅱ-induced neonatal rats’ CFs.Conclusions:STDP had anti-fibrotic effects in HF,which might be caused by the modulation of ECM-receptor interaction pathways.Through the management of cardiac fibrosis,STOP may be a compelling candidate for improving prognosis of HF.

Effect of Shexiang Tongxin Dropping Pills(麝香通心滴丸) on the Immediate Blood Flow of Patients with Coronary Slow Flow

Objective: To observe the immediate effect and safety of Shexiang Tongxin dropping pills(麝香通心滴丸, STDP) on patients with coronary slow flow(CSF), and furthermore, to explore new evidence for the use of Chinese medicine in treating ischemic chest pain. Methods: Coronary angiography(CAG) with corrected thrombolysis in myocardial infarction(TIMI) frame count(CTFC) was applied(collected at 30 frames/s). The treatment group included 22 CSF patients, while the control group included 22 individuals with normal coronary?ow. CSF patients were given 4 STDP through sublingual administration, and CAG was performed 5 min after the medication. The immediate blood ?ow frame count, blood pressure, and heart rate of patients before and after the use of STDP were compared. The liver and kidney functions of patients were examined before and after treatments. Results: There was a signi?cant difference in CTFC between groups(P<0.05). The average CTFC values of the vessels with slow blood ?ow in CSF patients were, respectively, 49.98±10.01 and 40.42±11.33 before and after the treatment with STDP, a 19.13% improvement. The CTFC values(frame/s) measured before and after treatment at the left anterior descending coronary artery, left circumflex artery, and right coronary artery were, respectively, 48.00±13.32 and 41.80±15.38, 59.00±4.69 and 50.00±9.04, and 51.90±8.40 and40.09±10.46, giving 12.92%, 15.25%, and 22.76% improvements, respectively. The CTFC values of vessels with slow ?ow before treatment were signi?cantly decreased after treatment(P<0.05). There were no apparent changes in the heart rate, blood pressure, or liver or kidney function of CSF patients after treatment with STDP(all P>0.05). Conclusions: The immediate effect of STDP in treating CSF patients was apparent. This medication could signi?cantly improve coronary ?ow without affecting blood pressure or heart rate. Our ?ndings support the potential of Chinese medicine to treat ischemic chest pain.

Shexiang Tongxin Dropping Pill(麝香通心滴丸)Reduces Coronary Microembolization in Rats via Regulation of Mitochondrial Permeability Transition Pore Opening and AKT-GSK3βPhosphorylation

Objective To investigate the protective effects of Shexiang Tongxin Dropping Pill(麝香通心滴丸,STDP)following sodium laurate-induced coronary microembolization(CME)in rats.Methods Forty rats were divided into 4 groups:the control(sham)group,CME group,low-dose STDP pretreatment group(20 mg·kg^(−1)·d^(−1)),and high-dose STDP pretreatment group(40 mg·kg^(−1)·d^(−1)).The rats were intragastric administrated with STDP 2 weeks before operation.Moreover,the histopathological alterations were observed using optical microscopy and transmission electron microscopy.Antioxidant biomarkers were analyzed by enzyme-linked immunosorbent assay.Mitochondrial functions including the mitochondrial permeability transition pore(mPTP)mtDNA copy number were determined and proteins of AKT/GSK3βwere analyzed by Western blot.Results The rats in the CME group showed a significant increase in the fibrinogen-like protein 2 expression level and mitochondrial dysfunction and a decrease in the expression level of antioxidant biomarkers(superoxide dismutase and catalase,P<0.01 for all).In contrast,the rats in the low-and high-dose STDP pretreatment groups showed a significant decrease in coronary microthrombi(P<0.05);moreover,STDP restored the antioxidant-related protein activities and mitochondrial function,inhibited mPTP opening,decreased AKT-Ser473 phosphorylation,and increased GSK3β-Ser9 phosphorylation(P<0.05 or P<0.01).Conclusion STDP may be useful for treatment of CME,possibly via regulation of mPTP opening and AKT/GSK3βphosphorylation.

Shexiang Tongxin dropping pill(麝香通心滴丸)protects against sodium laurate-induced coronary microcirculatory dysfunction in rats

OBJECTIVE:To investigate the protective effects of Shexiang Tongxin dropping pill(麝香通心滴丸,STDP)in a rat model of coronary microcirculatory dysfunction(CMD).METHODS:Sprague-Dawley rats were allocated randomly into four groups:sham,CMD model,STDP,and nicorandil.After 4 weeks of treatment,CMD was induced by injection of sodium laurate(0.2 m L,2 g/L)into the left ventricle while obstructing the ascending aorta.Rats in the sham group underwent an identical surgical procedure but were administered physiological(0.9%)saline(0.2 m L).Twenty-four hours after surgery,blood samples were collected for biochemical analyses and enzyme-linked immunosorbent assays.Heart tissues were removed for histopathology staining;apoptosis and inflammatory cytokines were examined by Western blotting.RESULTS:The STDP group had a lower level of creatine kinase-myocardial band,lactate dehydrogenase,and cardiac troponin-I than that in the CMD model group.Infiltration of inflammatory cells,myocardial ischaemia,and microthrombosis were relieved in the STDP group compared with CMD model group.Levels of endothelin-1,nuclear factor-kappa B,tumour necrosis factor-α,interleukin-6,interleukin-1β,malondialdehyde,B-cell lymphoma(Bcl)-2-associated X protein,and caspase-3 were lower,and levels of nitric oxide,Bcl-2,and superoxide dismutase were higher,in the STDP group in comparison with the CMD model group.CONCLUSION:STDP pretreatment improved the CMD induced by sodium laurate via anti-inflammatory,anti-apoptosis,and anti-oxidant mechanisms.

Antiplatelet and myocardial protective effect of Shexiang Tongxin Dropping Pill in patients undergoing percutaneous coronary intervention:A randomized controlled trial

Background:High on-clopidogrel platelet reactivity could be partially explained by loss-of-function alleles of CYP2 C19,the enzyme that converts clopidogrel into its active form.Shexiang Tongxin Dropping Pill(STDP)is a traditional Chinese medicine to treat angina pectoris.STDP has been shown to improve blood flow in patients with slow coronary flow and attenuate atherosclerosis in apolipoprotein E-deficient mice.However,whether STDP can affect platelet function remains unknown.Objective:The purpose of this study is to examine the potential effects of STDP on platelet function in patients undergoing percutaneous coronary intervention(PCI)for unstable angina.The interaction between the effects of STDP with polymorphisms of CYP2 C19 was also investigated.Design,participants and intervention:This was a single-center,randomized controlled trial in patients undergoing elective PCI for unstable angina.Eligible subjects were randomized to receive STDP(210 mg per day)plus dual antiplatelet therapy(DAPT)with clopidogrel and aspirin or DAPT alone.Main outcome measures:The primary outcome was platelet function,reflected by adenosine diphosphate(ADP)-induced platelet aggregation and platelet microparticles(PMPs).The secondary outcomes were major adverse cardiovascular events(MACEs)including recurrent ischemia or myocardial infarction,repeat PCI and cardiac death;blood biomarkers for myocardial injury including creatine kinase-MB isoenzyme(CK-MB)and high-sensitive troponin I(hs Tn I);and biomarkers for inflammation including intercellular cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),monocyte chemoattractant protein-1(MCP-1)and galectin-3.Results:A total of 118 subjects(mean age:[66.8±8.9]years;male:59.8%)were included into analysis:58 in the control group and 60 in the STDP group.CYP2 C19 genotype distribution was comparable between the 2 groups.In comparison to the control group,the STDP group had significantly lower CKMB(P<0.05)but similar hs Tn I(P>0.05)at 24 h after PCI,lower ICAM-1,VCAM-1,MCP-1 and galectin-3 at 3 months(all P<0.05)but not at 7 days after PCI(P>0.05).At 3 months,the STDP group had lower PMP number([42.9±37.3]vs.[67.8±53.1]counts/μL in the control group,P=0.05).Subgroup analysis showed that STDP increased percentage inhibition of ADP-induced platelet aggregation only in slow metabolizers(66.0%±20.8%in STDP group vs.36.0%±28.1%in the control group,P<0.05),but not in intermediate or fast metabolizers.The rate of MACEs during the 3-month follow-up did not differ between the two groups.Conclusion:STDP produced antiplatelet,anti-inflammatory and cardioprotective effects.Subgroup analysis indicated that STDP inhibited residual platelet reactivity in slow metabolizers only.