Combination of Panax notoginseng saponins and aspirin potentiates platelet inhibition with alleviated gastric injury via modulating arachidonic acid metabolism.

High platelet reactivity and gastric mucosal injury after aspirin (ASA) treatment are associated with poor compliance and an increased risk of cardiovascular events. Panax notoginseng saponins (PNS) have been widely used for the treatment of coronary heart disease (CHD) in addition to antiplatelet drugs in China; however, the joint effect and possible mechanism of PNS in addition to ASA on platelet activation and gastric injury remain unclear. This study was designed to investigate the combinational effects of PNS with ASA, and to explore the underlying mechanism via arachidonic acid (AA) metabolism pathway using lipidomic analysis. In a randomized, assessor-blinded trial, 42 patients with stable coronary heart disease (SCHD) and chronic gastritis were randomly assigned to receive ASA (n = 21) or PNS + ASA (n = 21) for 2 months. Compared with ASA alone, PNS + ASA further inhibited CD62p expression, GPIIb-IIIa activation and platelet aggregation and led to increased platelet inhibition rate. PNS + ASA suppressed the activity of platelet cyclooxygenase (COX)-1, and decreased the production of TXB2, PGD2, PGE2, 11-HETE, the downstream oxylipids of AA/COX-1 pathway in platelets, compared with ASA alone. The severity of dyspepsia assessment (SODA) results showed that patients in PNS + ASA group exhibited relieved dyspeptic symptoms as compared with those in ASA group, which might be associated with enhanced secretion of gastrin and motilin. In vivo study of myocardial infarction rats demonstrated that PNS attenuated ASA-induced gastric mucosal injury, which was related to markedly boosted gastric level of 6,15-diketo-13,14-dihydro-prostaglandin (PG)F1α, 13,14-dihydro-15-keto-PGE2 and PGE2 from AA/PG pathway in response to PNS + ASA compared with ASA alone. In summary, our study demonstrated that the combination of PNS and ASA potentiated the antiplatelet effect of ASA via AA/COX-1/TXB2 pathway in platelets, and mitigated ASA-related gastric injury via AA/PG pathway in gastric mucosa.

Panax quinquefolius saponins combined with dual antiplatelet drug therapy alleviate gastric mucosal injury and thrombogenesis through the COX/PG pathway in a rat model of acute myocardial infarction.

OBJECTIVES: Previous studies have found that Panax quinquefolius saponins (PQS) combined with dual antiplatelet therapy (DAPT) of aspirin and clopidogrel enhances antithrombotic effects while reducing gastric mucosal injury induced by DAPT. We investigated the effects of the combined drug therapy (PQS+DAPT) through the COX/PG pathways. METHODS: Acute myocardial infarction (AMI) was induced in Wistar rats by ligation of the left anterior descending (LAD) coronary artery, and the animals were randomly divided into Model, DAPT, and PQS+DAPT groups. Rats in the sham group did not undergo artery ligation. They were intragastrically treated for 14 days. Myocardial infarct size; myocardial pathology; platelet aggregation rate, CD62p activation, concentrations of thromboxane B2 (TXB2), 6-keto-PGF1α, tissue plasminogen activator (t-PA), and plasminogen activator inhibitor (PAI), the TXB2/6-keto-PGF1α ratio were measured. The ultrastructure of the gastric mucosa was observed by scanning electron microscopy. The expression of PGE2 and 6-keto-PGF1α in gastric mucosa was measured by radioimmunoassay, and levels of COX-1, COX-2, and VEGF in gastric mucosa were assessed using immunohistochemistry. RESULTS: The addition of Panax quinquefolius saponins (PQS+DAPT) to standard DAPT therapy significantly decreased the myocardial infarct area, degree of myocardial lesions, TXB2 and PAI levels, and the TXB2/6-keto-PGF1α ratio, while increasing 6-keto-PGF1α and t-PA levels and reducing the degree of gastric mucosal injury. Expression of PGE2, 6-keto-PGF1α, COX-2, and VEGF in the gastric mucosa was upregulated in the PQS+DAPT group compared with the standard DAPT group. CONCLUSION: PQS increases the degree of DAPT inhibition of myocardial necrosis and antiplatelet effects in AMI rats, as well as reducing damage to the gastric mucosa caused by DAPT. The mechanism may be related to inhibition of TXB2 and PAI activity and elevation of 6-keto-PGF1α and t-PA levels in blood, and may be associated with upregulated expression of COX-2, PGE2, PGI2, and VEGF in gastric tissue.

rs5911 and rs3842788 Genetic Polymorphism, Blood Stasis Syndrome, and Plasma TXB2 and hs-CRP Levels Are Associated with Aspirin Resistance in Chinese Chronic Stable Angina Patients.

The identification of single nucleotide polymorphisms (SNPs) related to aspirin resistance (AR) is of great significance for the explanation why some individuals demonstrate an incomplete response to aspirin and for optimizing the antiplatelet therapy strategy. The study was designed to investigate the possible associated genetic markers and clinical factors of AR for Chinese patients with chronic stable angina after PCI and to analyze the association between TXA2, PGI2, hs-CRP level, AR, and gene polymorphisms. Totally 207 chronic stable angina patients who received 100 mg maintenance dose daily of aspirin for more than 7 days were enrolled. The inhibition of platelets was assessed using light transmittance aggregometry. TXB2, 6-keto-PGF1α, and hs-CRP were measured by radioimmunoassay. Genotyping was performed using Taqman probe technique (rs5787 and rs5911) and gene sequencing technology (rs3842788). By using binary logistic regression analysis, the impact of clinical and genetic determinants on AR was evaluated. The prevalence of AR and aspirin semiresistance (ASR) was 3.86% and 20.76%, respectively, in Chinese chronic stable angina patients. rs5911 A/C and C/C versus A/A genotype (OR = 5.546, 95% CI = 1.812-11.404), rs3842788 A/G versus G/G genotype (OR = 8.358, 95% CI = 2.470-28.286), and blood stasis syndrome (BSS, OR = 10.220, 95% CI = 4.242-24.621) were associated with AR, but rs5787 variants were all homozygous of G/G genotype. Plasma TXB2 and hs-CRP increased significantly in AR and ASR group, while 6-keto-PGF1α showed no difference, and TXB2 level was significantly higher in carriers of the rs3842788 A/G genotype. According to our results, rs5911 and rs3842788 are proved to be specific genetic markers of AR in Chinese chronic stable angina patients for the first time, and BSS was also proved to be a remarkable determinant for AR. The AR and ASR patients were with increased plasma TXB2 and hs-CRP levels, and the TXB2 level was influenced by the variation of rs3842788 genotype.

[Dynamic Visible Microcirculation and Activating Blood Removing Stasis Research].

The technology of dynamic visualization microcirculation is to observe changes of mi- crocirculation by using visualization technology, combined with fluorescence labeling, high-speed video, electronic microscopy and other methods, converting rest images into dynamic visual graphics. In the past ten years, domestic researchers used this technology, and carried out researches on the heart, piamater/mesenteric microcirculation, dynamically observed effects of activating blood removing stasis (ABRS) research monomer, single herb and compound on arteriole/venula diameter, and the velocity of red blood cell, white blood cell adhesion, platelet aggregation, microvascular permeability, superoxide production, and plasma albumin leakage, etc. Microcirculation detection technologies were further opti- mized. Exerting direct advantages of dynamic visualization microcirculation, systematically evaluating ac- tion mechanism of Chinese herbs (multi-sites and multi-targets) , and interaction mechonism between Chinese herbs and Western drugs are important directions for future researches on microcirculation.

Panax quinquefolium saponin combined with dual antiplatelet drugs inhibits platelet adhesion to injured HUVECs via PI3K/AKT and COX pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Panax quinquefolium saponin (PQS) is the active component extracted from traditional Chinese medicine Panax quinquefolius L. and has been widely used as a supplement to dual antiplatelet drugs (DA) for treatment of coronary artery disease (CAD) for two decades; however, the efficacy of PQS combined with DA against platelet adhesion to endothelial cells (ECs), an essential step in thrombosis, remains unclear. AIM OF THE STUDY: To compare PQS combined with DA and DA alone in inhibiting platelet adhesion to injured human umbilical vein endothelial cells (HUVECs) and to explore the possible mechanisms focusing on PI3K/AKT, COX-2/6-keto-PGF1α, and COX-1/TXB2 pathways. METHODS: HUVECs injured by oxidized low-density lipoprotein (ox-LDL) were randomly allocated into control, model, DA, PQS+DA (P+DA), LY294002 (a PI3K inhibitor)+DA (L+DA), and LY294002+PQS+DA (LP+DA) groups. HUVEC apoptosis, platelet adhesion to injured HUVECs, and platelet CD62p expression were assayed by fluorescence activated cell sorting (FACS). The concentrations of 6-keto-PGF1α and TXB2 in the supernatant were measured by radioimmunoassay. Protein expression of phosphorylated-PI3K, PI3K, phosphorylated-AKT, AKT, COX-1, and COX-2 in both platelets and HUVECs was evaluated by western blot. RESULTS: Compared to DA alone, PQS combined with DA reduced platelet adhesion to HUVECs and HUVEC apoptosis more potently, increased the concentration of supernatant 6-keto-PGF1α and up-regulated phospho-AKT protein in HUVECs. LY294002 mitigated the effects of PQS on HUVEC apoptosis and platelet adhesion. CONCLUSIONS: These findings show that PQS as a powerful supplement to DA, attenuated HUVEC apoptosis and improved the DA-mediated reduction of platelet adhesion to injured HUVECs and the underlying mechanisms may be associated with PI3K/AKT and COX pathways in HUVECs and platelets. PQS might provide a new complementary approach to improve the prognosis of thrombotic diseases in future.

Pretreatment with a combination of ligustrazine and berberine improves cardiac function in rats with coronary microembolization.

AIM: We have shown that a combination of ligustrazine and berberine produces more effective inhibition on platelet activation and inflammatory reactions in rat acute myocardial infarction compared with either agent alone. In this study we evaluated the beneficial effects of a combination of ligustrazine and berberine in a rat model of coronary microembolization (CME). METHODS: SD rats were treated with ligustrazine, berberine, ligustrazine+berberine, or clopidogrel for 2 weeks. When the treatment completed, CME was induced by injection of sodium laurate into the left ventricular, while obstructing the ascending aorta. All rats were intubated for hemodynamic measurements. Blood samples were collected for biochemical analyses, flow cytometry, and ELISAs. Heart tissues were isolated for histopathology and subsequent protein analyses. RESULTS: Pretreatment with the combination of ligustrazine (27 mg·kg(-1)·d(-1)) and berberine (90 mg·kg(-1)·d(-1)) significantly improved cardiac function, and decreased myocardial necrosis, inflammatory cell infiltration, microthrombosis and serum CK-MB levels in CME rats. In addition, this combination significantly decreased plasma ET-1 levels and von Willebrand factor, inhibited ADP-induced platelet activation, and reduced TNFα, IL-1ß, ICAM-1 and RANTES levels in serum and heart tissues. The protective effects of this combination were more prominent than those of ligustrazine or berberine alone, but comparable to those of a positive control clopidogrel (6.75 mg·kg(-1)·d(-1)). CONCLUSION: The combination of ligustrazine and berberine significantly improved cardiac function in rat CME model via a mechanism involving antiplatelet and anti-inflammatory effects.

[Chinese Herbal Compounds for Supplementing Qi and Activating Blood Circulation Combined with Dual Antiplatelet Drugs Alleviated Human Umbilical Vein Endothelial Cell Injury and Platelet Adhe- sion via Up-regulation of P13K/Akt Pathway].

OBJECTIVE: To observe the effect and underlying mechanism of Chinese herbal com- pound (CHC) for supplementing qi and activating blood circulation (SQABC) combined with dual antiplatelet drugs (DA) on oxidized low density lipoprotein (ox-LDL) induced human umbilical vein endothelial cell (HUVEC) injury and platelet adhesion evoked by injured endothelial cells (ECs) based on P13K/Akt signaling pathway. METHODS: HUVECs were randomly divided into 5 groups, i.e., the blank control group, the model group (80 mg/L ox-LDL) , the DA group (15 µg/mL aspirin +10 µg/ mL clopidogrel +80 mg/L ox-LDL) , the Panax Quinquefolium saponins ( PQS, 160 µLg/mL) + Panax Notoginseng saponins (PNS, 160 µg/mL) +DA group, the LY294002 (30 µg/mL) + PQS + PNS + DA group. HUVEC apoptosis rate and platelet adhesion to HUVECs were detected by flow cytometry. Concentration of lactate dehydrogenase ( LDH) in HUVEC supernatant was detected by biochemical assay. Concentration of intercellular adhesion molecular ([CAM) was detected by radioimmunoassay. Protein expressions of p-P13K and p-Akt in HU- VECs were detected by Western blot. RESULTS: Compared with the blank control group, the apoptosis rate of HUVECs, mean fluorescence indicator ( MFI) , concentrations of both LDH and ICAM increased (P <0. 05) , and p-Akt protein expression decreased (P <0. 05) in the model group. Compared with the model group, the apoptosis rate of HUVECs and LDH concentration increased (P <0. 05), concentrations of MFI and ICAM obviously decreased (P <0. 05) in the DA group. The apoptosis rate, MFI, concentrations of both LDH and ICAM all decreased in the PQS + PNS + DA group (P <0. 05). p-Akt protein. expres- sion in HUVECs obviously increased in the PQS + PNS + DA group (P <0. 05). Compared with the DA group, HUVEC apoptosis rate, MFI, concentrations of both LDH and ICAM in supernatant obviously decreased, p-Akt expression in HUVECs increased in the PQS + PNS + DA group (all P <0. 05). p-Akt protein expression in HUVECs was inhibited after adding specific P13K inhibitor LY294002. Protection men- tioned above all disappeared in the PQS + PNS + DA group (P <0. 05). CONCLUSION: CHC for SQABC combined with DA could alleviate ox-LDL induced apoptosis of endothelial cells and reduce injured ECs e- voked platelet adhesion via up-regulation of P13K/Akt pathway in ECs.

The Effect of Xuefuzhuyu Oral Liquid on Aspirin Resistance and Its Association with rs5911, rs5787, and rs3842788 Gene Polymorphisms.

Aspirin should be continued indefinitely in patients after interventional therapy, but 10% to 40% of patients experience recurrent vascular events despite adequate aspirin therapy, a condition known as aspirin resistance (AR). Xuefuzhuyu oral liquid, derived from the classic recipe Xuefuzhuyu decoction, has been well documented to inhibit platelet aggregation and to improve hemorheology. The aims of this study were to investigate the effects of Xuefuzhuyu oral liquid on AR in patients with chronic stable angina after percutaneous coronary intervention (PCI) and the possible genetic markers related to the drug response. 43 patients diagnosed as having aspirin resistance or semi-resistance were randomly divided into control and treatment groups after screening 207 stable CHD patients. Platelet aggregation rate was determined using turbidimetry. Three single nucleotide polymorphisms in COX-1 (rs5787, rs3842788) and GP IIb (rs5911) were genotyped in whole blood samples using ABI PRISM 7900 HT Fast Real-Time instrument and ABI PRISM 3730 DNA Sequencer. The results showed that Xuefuzhuyu oral liquid could effectively improve blood stasis syndrome and AR by inhibiting ADP-induced platelet aggregation and that patients with the rs5911 genetic variant exhibited better drug response upon treatment with Xuefuzhuyu oral liquid, which suggests Xuefuzhuyu oral liquid as a new possible drug for the prevention of AR.