Network pharmacology-based approach to research the effect and mechanism of Si-Miao-Yong-An decoction against thromboangiitis obliterans.

BACKGROUND: Si-Miao-Yong-An decoction (SMYAD) is a conventional therapeutic formula for treat thromboangiitis obliterans (TAO), consisting of four Chinese herbs: Lonicerae japonicae Thunb. (Jinyinhua), Scrophularia ningpoensis Hemsl. (Xuanshen), Angelica sinensis (Oliv.) Diels (Danggui) and Glycyrrhiza uralensis Fisch. (Gancao). However, the mechanism of SMYAD in TAO treatment remains unclear. METHODS: Components, as well as potential targets of SMYAD in TAO therapy, were downloaded from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Subsequently, with the Database for Annotation, Visualization, and Integrated Discovery (DAVID) server, the gene ontology (GO) biological processes and the Kyoto encyclopedia of genes and genomes (KEGG) signalling pathways of the targets enrichment were performed. Next, based on STRING online database, the protein interaction network of vital targets was built and analysed. Molecular docking and calculation of the binding affinity were performed using AutoDock. The PyMOL software was employed to observe docking outcomes of active compounds and protein targets. Based on the predicted outcomes of network pharmacology, in vivo and in vitro tests were performed for validation. In vivo experiment, the TAO rats model was established using sodium laurate injection into the femoral artery. The symptoms as well as pathological changes of the femoral artery were observed. Besides, the predicted targets were verified by the RT-qPCR, in vitro experiment. The cell viability in LPS-induced human umbilical vein endothelial cells (HUVECs) was detected using CCK-8 kit, and the predicted targets were also verified by the RT-qPCR. RESULTS: In the network pharmacology analysis, we obtained 105 chemical components in SMYAD and 24 therapeutic targets. We found that the mechanism SMYAD in TAO therapy was primarily associated with inflammation and angiogenesis by constructing multiple networks. Quercetin, vestitol and beta-sitosterol were important compounds, and interleukin-6 (IL6), MMP9, and VEGFA were key targets. According to molecular docking, active compounds (quercetin, vestitol and beta-sitosterol) and targets (IL6, MMP9 and VEGFA) showed good binding interactions. In in vivo experiment, SMYAD ameliorated the physical signs and pathological changes, inhibited the expression of IL6 and MMP9, and enhanced the expression of VEGFA. In an in vitro experiment, SMYAD increased the cell viability of LPS-induced HUVECs and the expression of VEGFA, and reduced the expression of IL6 and MMP9. CONCLUSIONS: This study showed that SMYAD improves TAO symptoms and inhibits the development of TAO. The mechanism could be associated with anti-inflammatory and therapeutic angiogenesis.

Protective Effects and Potential Mechanism of Tongxinluo on Mice with Thromboangiitis Obliterans Induced by Sodium Laurate.

OBJECTIVE: To investigate the effects of Tongxinluo (TXL) on thromboangiitis obliterans (TAO) and the underlying mechanisms. METHODS: Ninety male C57/BL6J mice were randomly divided into 6 groups according to a random number table: the sham group, TAO model group, Compound Danshen Tablet (CDT) group, and the high-, medium-, and low-dose TXL groups. All mice except the sham group were injected with sodium laurate (0.1 mL, 5 mg/mL) in the femoral artery to establish TAO mouse model. After modeling, mice in the sham and TAO model groups were intragastrically administered 0.5% (w/v) sodium carboxymethylcellulose, mice in the CDT group were intragastrically administered 0.52 g/kg CDT, and mice in the TXL-H, TXL-M, and TXL-L groups were intragastrically administered 1.5, 0.75, and 0.38 g/kg TXL, respectively. After 4 weeks of gavage, the recovery of blood flow in the lower limbs of mice was detected by Laser Doppler Imaging. The pathological changes and thrombosis of the femoral artery were observed by morphological examination. The expressions of tumor necrosis factor α (TNF-α) and inducible nitric oxide synthase (iNOS) in the femoral artery wall were detected by HE staining. Levels of thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α), endothelin-1 (ET-1), interleukin (IL)-1ß and IL-6 were measured using enzyme-linked immunosorbent assay (ELISA). Levels of activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (FIB) were detected by a fully automated biochemical analyzer. RESULTS: TXL promoted the restoration of blood flow in the lower limbs, reduced the area of thrombosis in the femoral artery, and alleviated the pathological changes in the femoral artery wall. Moreover, the levels of TXB2, ET-1, IL-6, IL-1ß, TNF-α and iNOS were significantly lower in the TXL groups compared with the model group (P<0.05 or P<0.01), while the level of 6-keto-PGF1α was significantly higher (P<0.01). In addition, APTT, PT, and TT were significantly prolonged in TXL groups compared with the model group (P<0.05 or P<0.01), and FIB levels were significantly decreased compared with the model group (P<0.01). CONCLUSIONS: TXL had a protective effect on TAO mice, and the mechanism may involve inhibition of thrombosis and inflammatory responses. TXL may be a potential drug for the treatment of TAO.

Integrated pharmacokinetics and pharmacometabolomics to reveal the synergistic mechanism of a multicomponent Chinese patent medicine, Mailuo Shutong pills against thromboangiitis obliterans.

BACKGROUND: Mailuo Shutong Pills (MLST) have displayed pharmacological activity against thromboangiitis obliterans (TAO). However, the active ingredients and therapeutic mechanism of MLST against TAO remained to be further clarified. PURPOSE: The aim of this study was to explore the active components of MLST and their synergistic mechanism against TAO by integrating pharmacokinetics (PK) and pharmacometabolomics (PM). METHODS: TAO model rats were established by sodium laurate solution. Firstly, the efficacy of MLST was evaluated by gangrene score, blood flow velocity, and hematoxylin-eosin (H&E) staining. Secondly, PK research was conducted on bioavailable components to characterize their dynamic behaviors under TAO. Thirdly, multiple plasma and urine metabolic biomarkers for sodium laurate-induced TAO rats were found by untargeted metabolomics, and then variations in TAO-altered metabolites following MLST treatment were analyzed utilizing multivariate and bioinformatic analysis. Additionally, metabolic pathway analysis was performed using MetaboAnalyst. Finally, the dynamic link between absorbed MLST-compounds and TAO-associated endogenous metabolites was established by correlation analysis. RESULTS: MLST significantly alleviated gangrene symptoms by improving the infiltration of inflammatory cells and blood supply in TAO rats. Significant differences in metabolic profiles were found in 17 differential metabolites in plasma and 24 in urine between Sham and TAO rats. The 10 bioavailable MLST-compounds, such as chlorogenic acid and paeoniflorin, showed positive or negative correlations with various TAO-altered metabolites related to glutamate metabolism, histidine metabolism, arachidonic acid metabolism and so on. CONCLUSION: This study originally investigated the dynamic interaction between MLST and the biosystem, providing unique insight for disclosing the active components of MLST and their synergistic mechanisms against TAO, which also shed light on new therapeutic targets for TAO and treatment.

Identifying quality markers of Mailuoshutong pill against thromboangiitis obliterans based on chinmedomics strategy.

BACKGROUND: Mailuoshutong pill (MLSTP) is a traditional Chinese medicine (TCM) for the treatment of Thromboangiitis obliterans (TAO, Buerger's disease) which is a segmental non-atherosclerotic inflammatory occlusive disorder. However, the mechanism and quality standards of MLSTP have not been sufficiently studied. PURPOSE: This work aims to investigate the potential mechanisms and quality markers (Q-markers) of MLSTP treating TAO based on the chinmedomics strategy. METHODS: The therapeutical effect of MLSTP on TAO rats was evaluated by changes in body weight and clinical score, regional blood flow velocity and perfused blood vessel distribution, hematoxylin-eosin (H&E) staining, serum metabolic profile. Moreover, both endogenous metabolites and exogenous components were simultaneously detected in serum based on ultra-high performance liquid chromatography coupled with a Q Exactive hybrid quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS), and multivariate analysis was applied to identify the biomarkers, as well as the dynamic changes of metabolites were observed to explore the mechanism of action of MLSTP. In addition, the pharmacodynamic material basis were identified by correlation analysis between biomarkers and absorbed constituents. Finally, the Q-markers of MLSTP were determined according to the screening principles of Q-marker and validated the measurability. RESULTS: MLSTP treatment alleviated disease severity of TAO, reduced inflammatory infiltration, and ameliorated vascular function. 26 potential biomarkers associated with glutamate metabolism, linoleic acid metabolism, arachidonic acid metabolism and so on were identified. Besides, 27 prototypical components were identified in serum, 16 of which were highly correlated with efficacy and could serve as the pharmacodynamic material basis of MLSTP against TAO. In addition, 7 compounds, namely, sweroside, chlorogenic acid, calycosin-7-glucoside, formononetin, paeoniflorin, liquiritigenin and 3-butylidenephthalide, were considered as potential Q-markers of MLSTP. Ultimately, the measurability of the seven Q-markers was validated by rapid identifcation and quantifcation. CONCLUSION: This study successfully clarified the therapeutic effect and Q-markers of MLSTP by chinmedomics strategy, which is of great significance for the establishment of quality standards. Furthermore, it provides a certain reference for the screening of Q-markers in TCM prescriptions.

Herbal therapy treatment in thromboangiitis obliterans: a retrospective clinical study.

BACKGROUND: Critical limb ischemia in patients with thromboangiitis obliterans (TAO, Buerger's disease) is associated with refractory rest pain, gangrene, and increased rates of amputation. Tuoju lotion was prepared by the Pharmacy Department of Dongfang Hospital. The focus of the study is to elicit the efficacy of the addition of Herbal therapy treatment to conventional treatment in TAO patients with severe extremity pain and to assess any statistically significant benefits in patient's pain control at rest. We fund that the addition of herbal therapy treatment can augment conventional treatments in TAO patients by improving or eliminating intermittent claudication symptoms, prolonging claudication distance, and reducing total blood viscosity. At the same time, Tuoju lotion can improve microcirculation status in the short term. The purpose of this study was to investigate the effect of topical Herbal therapy treatment on patient outcomes in patients with TAO. METHODS: Seventy patients with TAO treated between January 2009 and July 2019 were included in a retrospective analysis of a single university hospital vascular center. Forty patients received topical herbal treatment in addition to conventional therapy and were compared to a control group who received standard treatment alone (n=30). RESULTS: Patients in both, the experimental and control group, were matched according to age and gender. There was no significant difference in course of disease and past medical history between the two groups. The mean ankle brachial index (ABI), toe pressure, and blood viscosity were also similar in both groups. Rest pain score (baseline VAS 4.76±2.87, post-treatment 3.32±1.29) and walking distance (baseline 169.7±23.6 m, post-treatment 284.5±32.3 m) significantly improved in the herbal treatment group. ABI values improved and total blood viscosity decreased in both groups with no significant difference between the herbal and conservative treatment arms. However, the arterial blood pressure ratio in the lower extremity stage showed no difference between the superficial femoral artery and the popliteal artery. CONCLUSIONS: The addition of Herbal therapy treatment to conventional treatment in TAO patients with severe extremity pain was associated with a reduction of rest pain and intermittent claudication.

Protective effects and potential mechanism of salvianolic acid B on sodium laurate-induced thromboangiitis obliterans in rats.

BACKGROUND: The root of Salvia miltiorrhiza f. alba (RSMA) (Lamiaceae) is used for the treatment of patients with thromboangiitis obliterans (TAO) in traditional Chinese medicine. Previously, a mixture of phenolic acids extracted from RSMA has shown significant protective effects on TAO rats. PURPOSE: This study investigates the inhibitory effects of salvianolic acid B on TAO induced by sodium laurate injection in rats to explore the effective constituents of RSMA in TAO treatment. METHODS: TAO rats were developed using injected sodium laurate. After treatment with ligustrazine hydrochloride (15 mg/kg) and various doses of salvianolic acid B (10, 20, 40 mg/kg) by tail intravenous injection, levels of thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α) and endothelin-1 (ET-1) in plasma were determined using enzyme-linked immunosorbent assay. The right femoral arteries were studied by hematoxylin and eosin staining and immunohistochemical analysis to determine pathological changes and overexpression of tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS) in the femoral artery walls of TAO rats. RESULTS: Salvianolic acid B significantly decreased the expressions of TXB2 and ET-1 and increased the expression of 6-keto-PGF1α in plasma, and significantly inhibited the overexpression of TNF-α and iNOS in the femoral artery walls of TAO rats at medium and high doses. CONCLUSION: Salvianolic acid B has a protective effect on TAO rats. The mechanism may involve inhibition of thrombosis and TAO-associated inflammatory responses, which may explain the success of RSMA treatment of TAO in humans in traditional Chinese medical practice. Hence, it may be a potential drug for TAO treatment in conventional medicine.

Conservative treatment of patients with thromboangiitis obliterans or cannabis-associated arteritis presenting with critical lower limb ischaemia.

BACKGROUND: Proper management of patients with thromboangiitis obliterans (TAO) or cannabis-associated arteritis (CAA), presenting with critical lower limb ischaemia (CLI) remains controversial, and data are limited. PATIENTS AND METHODS: Patients with TAO or CAA presenting with CLI between 2011 and 2016 were retrospectively evaluated. Patients requiring primary intervention were excluded. Conservative treatment included: (a) weight-adjusted bemiparin plus six hours/day intravenous iloprost for 28 days, (b) aspirin (100 mg/day) plus cilostazol (100 mg twice/day) after discharge, and (c) strict recommendations/monitoring for smoking cessation. Main outcomes included symptom recession, ankle-brachial index (ABI) improvement, and healing of lesions at the time of discharge as well as amputation, revascularization, and abstinence rate during follow-up. RESULTS: Overall, 23 patients (TAO: 15; CAA: 8) were included within six years, none of the patients reported any other factor than smoking. All patients presented with rest pain and 12 patients with ulcer or necrotic lesions. Mean ABI measurement at presentation was 0.46 ± 0.2, after 28 days of treatment, all patients showed improvement regarding clinical picture and ABI measurement (0.54 ± 0.1; p < 0.05). During follow-up, only three patients underwent bypass surgery and two patients underwent major amputation, although the smoking abstinence rate was very low (13 %). CONCLUSIONS: Intravenous iloprost plus bemiparin for 28 days together with per os aspirin plus cilostazol seem to produce promising results in patients with TAO/CAA, treated for CLI, even with a low smoking abstinence rate. However, larger series are needed to further evaluate inter-group differences and potential prognostic factors.

Integrated treatment for lower-limb stage II thromboangiitis obliterans by interventional therapy and oral administration of Chinese medicine: a randomized controlled clinical trial.

METHODS: Ninety lower-limb stage II or worse TAO patients were randomly divided into three groups: group A (30 cases) treated by intervention and oral administration of Chinese medicine; group B (30 cases) treated by intervention alone; and group C (30 cases) treated only with oral administration of Chinese medicine. Therapeutic effects were observed, including the cure rate; the recurrence rate after one month, three months, six months, nine months, and one year; the ankle brachial indexes; the incidence of complications; and the level of C-reactive protein and erythrocyte sedimentation rate. RESULTS: Group A had significantly better clinically curative effects, related indexes, and outcomes during the long-term follow-up survey, than that of groups B and C. CONCLUSION: Integrated treatment is more effective for treating lower-limb stage II or worse TAO. OBJECTIVE: To observe if integrated treatment is better than other therapies for lower-limb stage II thromboangiitis obliterans (TAO).

Effect of Ocimum basilicum L. on cyclo-oxygenase isoforms and prostaglandins involved in thrombosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Ocimum basilicum L. (OBL) is a plant used in traditional Uyghur medicine for the treatment and prevention of cardiovascular disease. In previous studies we had found an antihypertensive and antithrombotic effect suggestive of an effect on prostaglandins, which we attempt to document here. MATERIALS AND METHODS: 6-keto-PGF1α, the metabolite of prostacyclin, and PGE2 were measured in the supernatant of human umbilical vein endothelial cells (HUVEC) and basal or LPS-stimulated mouse coeliac macrophage cultures exposed to OBL ethanol (OBL-E) extracts and petroleum ether, chloroform, ethylacetate and butanol (PE, C, EA, B) fractions. In addition, 6-keto-PGF1α and thromboxane B2 (TXB2) were measured in a rat model of thromboangiitis obliterans exposed or not to OBL. RESULTS: Short-term exposure to OBL-E dose-dependently increased 6-keto-PGF1α from HUVEC, and long-term (24h) exposure decreased it. OBL-C and OBL-B increased 6-keto-PGF1α, whereas the other fractions tended to decrease it after 24h exposure. The extract and all fractions decreased basal and stimulated PGE2 production, but only OBL-EA and OBL-B reduced PGE2 in stimulated cultures to concentrations below the unstimulated values (P<0.05). In vivo OBL increased 6-keto-PGF1α and decreased TXB2. CONCLUSION: OBL and its extracts increased 6-keto-PGF1α and reduced PGE2 and TXB2 production in a dose and time-related manner. This could indicate simultaneous inhibition of COX-2 and stimulation of endothelial COX-1. The butanol fraction seemed most promising in this respect.

[Effect of Mailuoning injectable powder on experimental vascular occlusion angeitides in rats].

OBJECTIVE: To investigate the effect of Mailuoning injectable powder on experimental vascular occlusion angeitides in rats. METHODS: Rats were injected laurostearic acid into arteria cruralis to induce the model of experimental vascular occlusion angeitides, then we observed the changes of objective sign of rats, and analysed throm ranking through pathological section under electro-microscope. RESULTS: Mailuoning injectable powder could decrease the quantity of throm in blood vessel, and improve hemorrheoiogy. CONCLUSION: The results show that Mailuoning injectable powder has obvious therapeutical effect on experimental vascular occlusion angeitides in rats, and its mechanism may be related to the anti-throm in blood vessel and improving hemorrheology.

[Buerger's disease starting in the upper extremity. A favorable response to nifedipine treatment combined with stopping tobacco use]. / Enfermedad de Buerger de inicio en extremidad superior. Respuesta favorable al tratamiento con nifedipina asociado a la supresión del tabaco.

A case of a 38-year-old smoker male who had vasoconstriction and instep claudication of the right hand, is presented. After a year of evolution, he experienced the same alteration on the left foot. He was diagnosed as suffering from thromboangiitis obliterans, by means of angiography. After oral nifedipine treatment, combined with cessation of smoking, all symptoms and trophics regressed.

[Formation of extrinsic blood thrombosis in 102 cases of thromboangiitis obliterans].

The formation of extrinsic blood thrombosis in 102 cases of thromboangiitis obliterans and 54 healthy persons were observed with chandler method and their clinical manifestations. All the 102 patients were male, of which 13 cases were in ischemic stage, 45 cases in dystrophic stage, and 44 cases in necrotic stage. According to the differentiation of TCM, 8 belong to Yin-cold type, 50 belong to downward flowing of damp-heat type, 42 belong to blood stasis type, and 2 others. Combining method of TCM-WM was adopted to give 45 cases systemic treatment, that is, taking the promotion of blood flow to remove its stasis as the dominant factor, accompanied by intravenous drip and operation. Results showed that the length, wet weight and dry weight of extrinsic blood thrombosis were higher apparently in the cases of thromboangiitis obliterans than those in healthy persons (P less than 0.001), and there were significant differences between the values of extrinsic blood thrombosis in different stages and different degrees. After systemic treatment with TCM-WM, the values of extrinsic blood thrombosis decreased remarkably, and their symptoms and signs improved or disappeared. The clinical total effective rate was 88.89%, and the decreasing rate of the values of extrinsic blood thrombosis was 98.9%. This suggested that the high blood coagulation and extrinsic thrombosis should draw special attention and may act as an approach reflecting the severity of the disease and evaluating the effects of treatment.

[Study of thromboangiitis obliterans treated with "vascular no. 3" using Doppler ultrasound].

A clinical study was carried out in 31 patients (39 limbs) with thromboangiitis obliterans (TAO, Buerger's Disease) by Chinese herbs "Vascular No.3". Doppler ultrasound flow velocity tracing at the femoral, popliteal, posterior tibial and dorsalis pedis levels and blood pressure at the ankle level were measured. The results showed that TAO patients treated with "Vascular No.3" had a total effective rate of 92.3% and a distinct improvement of 76.9%. The results of Doppler ultrasound proved that "Vascular No.3" significantly decreased arterial lumen stenosis level, increased blood velocity and local flow, promoted developing arterial collaterals and improved vaso-functional conditions.