Pharmacodynamic and targeted amino acid metabolomics researches on the improvement of diabetic retinopathy with Fufang Xueshuantong component compatibility.

The Fufang Xueshuantong capsule (FXT) has significant preventive and therapeutic effects on diabetic retinopathy(DR), but the compatibility of its active components remains to be thoroughly explored. In this study, a zebrafish diabetic retinopathy model was established using high-mixed sugars, and the optimal ratios of notoginseng total saponins, total salvianolic acid, astragaloside, and harpagide were selected through orthogonal experiments. Furthermore, we used UPLC-QqQ/MS to detect the changes in amino acid content of DR zebrafish tissues after administration of FXT and its compatible formula to analyze the effects of FXT and its compatible formula on amino acid metabolites. The results showed that the final compatibility ratios of the components were 8: 5: 1: 6.6 by comprehensive evaluation of the indicators. FXT and its compatibility formula had beneficial effects on retinal vasodilatation, lipid accumulation in the liver, total glucose, and VEGF levels in DR zebrafish, and all of them could call back some amino acid levels in DR zebrafish. In this research, we determined the compatible formulation of the active ingredients in the FXT and investigated their efficacy in DR zebrafish for further clinical applications.

Umbilical Cord Mesenchymal Stem Cells Combined with Fufang Xueshuantong Capsule Attenuate Oxidative Stress and Vascular Lesions in Diabetic Rats by Activating Nrf-2/HO-1 Signaling Pathway.

BACKGROUND: Macrovascular lesions are the main cause of death and disability in diabetes mellitus, and excessive accumulation of cholesterol and lipids can lead to long-term and repeated damage of vascular endothelial cells. Umbilical cord mesenchymal stem cells (UCMSCs) can attenuate vascular endothelial damage in type 1 diabetic mice, while Fufang Xueshuantong capsule (FXC) has a protective effect on endothelial function; however, whether FXC in combination with UCMSCs can improve T2DM macrovascular lesions as well as its mechanism of action are not clear. Therefore, the aim of this study was to reveal the role of FXC + UCMSCs in T2DM vasculopathy and their potential mechanism in the treatment of T2DM. METHODS: The control and T2DM groups were intragastrically administered with equal amounts of saline, the UCMSCs group was injected with UCMSCs (1×106, resuspended cells with 0.5 mL PBS) in the tail vein, the FXC group was intragastrically administered with 0.58 g/kg FXC, and the UCMSCs + FXC group was injected with UCMSCs (1×106) in the tail vein, followed by FXC (0.58 g/kg), for 8 weeks. RESULTS: We found that FXC+UCMSCs effectively reduced lipid levels (TG, TC, and LDL-C) and ameliorated aortic lesions in T2DM rats. Meanwhile, Nrf2 and HO-1 expression were upregulated. We demonstrated that inhibition of Nrf-2 expression blocked the inhibitory effect of FXC+UCMSCs-CM on apoptosis and oxidative stress injury. CONCLUSION: Our data suggest that FXC+UCMSCs may attenuate oxidative stress injury and macroangiopathy in T2DM by activating the Nrf-2/HO-1 pathway.

A review on the chemical profiles, quality control, pharmacokinetic and pharmacological properties of Fufang Xueshuantong Capsule.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Xueshuantong Capsule (FXC) is a traditional Chinese medicine (TCM) formula composed of four herbs including Panax notoginseng, Astragalus membranaceus, Salvia miltiorrhiza, and Scrophularia ningpoensis. Long-term and extensive clinical applications have confirmed that FXC could exert significant effects on fundus, cardiovascular and cerebrovascular occlusive diseases. AIM OF THE REVIEW: To systematically analyze and summarize the existing researches involving quality and efficacy re-evaluation of FXC, point out the typical problems, and further propose some opinions to contribute to future study. MATERIALS AND METHODS: Literatures concerning FXC were collected from online scientific databases including China National Knowledge Infrastructure, WanFang Data, PubMed, Science Direct, Scopus, Web of Science, Springer Link up to June 2020. All eligible studies are analyzed and summarized in this review. RESULTS: This review outlines the chemical profiles, quality control, pharmacokinetic and pharmacological properties of FXC based on reported results. Some problems are pointed out for FXC: the quality control needs further improvement, the pharmacokinetic properties have not been comprehensively investigated, and in-depth and systematic mechanism researches are scarce. Hereon we propose several directions for future study: (a) establishment of feasible HPLC or LC-MS based quantitative methods for simultaneous determination of multiple components to monitor the overall quality; (b) pharmacokinetic studies concerning humans, drug-drug interactions, and correlation with pharmacodynamics; (c) pharmacological mechanism researches integrating multi-omics technologies (gut microbiome, metabolomics, etc.). CONCLUSIONS: This review highlights the researches on quality and efficacy re-evaluation of FXC, and points out some typical problems. Further in-depth studies should focus on the promotion of quality control, pharmacokinetic properties, and pharmacological mechanism.

Distribution of chemical compounds of Fufang Xueshuantong capsule in eyes and kidney of beagle dogs / 中国中药杂志

Diabetic retinopathy(DR) and diabetic nephropathy(DN) are the most common complications of diabetes, and the main causes of death and disability of diabetes. Clinical reports showed that Fufang Xueshuantong capsule(FXT) had effective curative effect on DR and DN, but there was no report on the distribution of chemical compounds of FXT in beagle dog eyes and kidneys. In this study, FXT was given by gavage administration in Beagle dogs for 3 days, and then their eyeballs and kidneys were taken. The chemical compounds in beagle dog eyes and kidneys were detected by HPLC LTQ-Orbitrap technology. Furthermore, by comparing with the data from retrieving literature and references, the chemical compounds were identified by the accurate mass, retention time (tR), and MS/MS. Fourteen and 19 types of notoginsenosides were respectively identified in eyeballs and kidneys in this study, and these results could lay foundation for clarifying the effective ingredients of FXT in treatment of DR and DN.

[Distribution of chemical compounds of Fufang Xueshuantong capsule in eyes and kidney of beagle dogs].

Diabetic retinopathy(DR) and diabetic nephropathy(DN) are the most common complications of diabetes, and the main causes of death and disability of diabetes. Clinical reports showed that Fufang Xueshuantong capsule(FXT) had effective curative effect on DR and DN, but there was no report on the distribution of chemical compounds of FXT in beagle dog eyes and kidneys. In this study, FXT was given by gavage administration in Beagle dogs for 3 days, and then their eyeballs and kidneys were taken. The chemical compounds in beagle dog eyes and kidneys were detected by HPLC LTQ-Orbitrap technology. Furthermore, by comparing with the data from retrieving literature and references, the chemical compounds were identified by the accurate mass, retention time (tR), and MS/MS. Fourteen and 19 types of notoginsenosides were respectively identified in eyeballs and kidneys in this study, and these results could lay foundation for clarifying the effective ingredients of FXT in treatment of DR and DN.

A combination of the main constituents of Fufang Xueshuantong Capsules shows protective effects against streptozotocin-induced retinal lesions in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Xueshuantong Capsule, an herbal formula licensed for clinical use in China, which is composed of Panax notoginseng (Burkill) F.H. Chen, Salvia miltiorrhiza Bunge, Astragalus membranaceus (Fisch.) Bunge, and Scrophularia ningpoensis Hemsl, has proven effective for the treatment of diabetic retinopathy. However, its bioactive constituents are still ambiguous. In this study, the therapeutic effects of a combination of the main constituents of Fufang Xueshuantong Capsule (cFXT) were evaluated in streptozotocin (STZ)-induced retinal lesions to identify the bioactive constituents. METHODS: Sprague-Dawley rats, except for those in the control group (vehicle+vehicle), were administered a single injection of 60mg/kg STZ. One-week later, STZ-treated rats were randomly divided into three groups-one STZ group (STZ+vehicle) and two cFXT treatment groups (STZ+cFXT). The rats in the latter two groups received cFXT 44.8mg/kg or cFXT 22.4mg/kg by intragastric gavage once per day, for 24 consecutive weeks. The rats in the control and STZ groups received the vehicle in the same way. Body weights and fasting blood glucose levels were recorded every four weeks. After treatment, hemorheological tests were performed to record the erythrocyte aggregation indexes, blood viscosity, and plasma viscosity. The trypsin digestion method was used to observe pericyte and acellular capillary counts in the retina. Ultraviolet spectrophotometry was utilized to measure the activity of aldose reductase (AR) by measuring the nicotinamide adenine dinucleotide phosphate (NADPH) consumption at 340nm. An immunohistochemical assay was used to observe the expressions of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in the retina. The expression levels of intercellular adhesion molecule-1 (ICAM-1), endothelin-1 (RT-1),and occludin in the retina were tested by the western blot assay. RESULTS: cFXT is composed of 991.44mg/g saponins of Panax notoginseng, 1.62mg/g harpagoside, 0.70mg/g cryptotanshinone, 0.74mg/g tanshinone I, and 5.50mg/g astragaloside A. Although it showed no effects on the increased body weight and blood glucose levels induced by STZ in rats. However, it showed a tendency to attenuate the increase in erythrocyte aggregation, plasma viscosity, and acellular vessel and pericyte loss, paralleled with a reversal of the hyper-activation of AR, the hyper-expression of VEGF, ICAM-1, and ET-1, and the hypo-expression of PEDF and occludin in the retinas of STZ-treated rats. CONCLUSION: The saponins of Panax notoginseng, harpagoside, cryptotanshinone, tanshinone I, and astragaloside A are the main bioactive constituents of Fufang Xueshuantong Capsule and contribute to the attenuation of STZ-induced retinal lesions in rats. These constituents can be used as the base to optimize a new drug for the treatment of diabetic retinopathy, and can be selected for quality control of Fufang Xueshuantong Capsules.

Study on effect of Fufang Xueshuantong capsule on serpine1,α-serpine1 protein and KLF2 in patients with lower extremity venous thrombosis / 中国生化药物杂志

Objective To explore the effect of serpine1, α-serpine1 protein and KLF2 in patients with lower extremity venous thrombosis by Fufang Xueshuantong capsule.Methods 44 patients were diagnosed with lower extremity venous thrombosis were collected, and randomly divided into experimental group and control group, 22 cases in each group, giving the conventional antiplatelet therapy, on the basis of this, the experimental group was given Fufang Xueshuantong capsule 1.5 g, three times a day orally, and the control group was given context Shutong granules 20 g, three times a day orally.2 weeks for a course of treatment, three courses of treatment.Before and after treatment in the detection of two groups of patients with serum serpine1, α-serpine1 KLF2 and protein contents, blood rheology, coagulation function, and carries on the analysis.ResuIts Compared with the control group, experimental group patients has better curative effect, the performance of: serum serpine1and α-serpine1 protein content were significantly decreased(P<0.05).KLF2 expression was significantly higher (P<0.05).Blood viscosity, erythrocyte aggregation index, erythrocyte sedimentation rate decreased significantly (P<0.05).Fib, D-D content significantly decreased (P<0.05).ConcIusions Fufang Xueshuantong capsule can reduce serum serpine1, α-serpine1 protein content and promote KLF2 expression, decrease the blood viscosity, erythrocyte sedimentation rate, improve patients with hypercoagulable state in patients with lower extremity venous thrombosis, so as to achieve good clinical efficacy.

Network pharmacology analyses of the antithrombotic pharmacological mechanism of Fufang Xueshuantong Capsule with experimental support using disseminated intravascular coagulation rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Xueshuantong (FXST) Capsule is developed on a traditional Chinese medicine remedy, with a four-herb formula of Panax notoginseng, Radix astragali, Salvia miltiorrhizae and Radix scrophulariaceae. It has been used for treatment of the clinic cardiovascular disease for many years. MATERIALS AND METHODS: Due to its complexity of compositions and polypharmacological effects, it often complicates understanding of the mechanisms of action. In the present work, we have constructed an integrated model of system pharmacology to investigate the polypharmacological mechanisms of FXST formulation for treatment of thrombosis disease. RESULTS: The predicted results showed that 22 ingredients in FXST were closely associated with 41 protein targets related to blood coagulation, fibrinolysis and platelet aggregation. Through analysis of the compound-protein target association, significant cross-targets between each herb indicated the multiple active chemical ingredients might interact with the same target simultaneously and thus explained the synergistic mechanisms of the principle of Traditional Chinese medicines (TCMs) as ''Jun (emperor) - Chen (minister) - Zuo (adjuvant) - Shi (courier)''. To validate the polypharmacological effects predicted by our network pharmacology (NetPharm) analysis, we have carried out experimental investigation the effects of FXST on the disorders of the blood coagulation system in a lipopolysaccharide-induced disseminated intravascular coagulation (DIC) rat model. The results showed that FXST could significantly ameliorate the activation of coagulation system, which is congruent with the cross-target prediction by NetPharm approach. CONCLUSIONS: The combined investigations provide more insight into better understanding of the pharmacological mechanisms of FXST, and may also offer an alternative avenue to further explore the chemical and pharmacological basis of TCMs.

Simultaneous Determination of Eight Effective Compounds of Fufang Xueshuantong ;Capsule in Rat Plasma by LC-MS/MS / 中国中医药信息杂志

Objective To establish a sensitive and specific LC-MS/MS method for measurement of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rd, tanshinone Ⅰ, astragaloside Ⅳ and harpagosidein of Fufang Xueshuantong Capsule in rat plasma. Methods The HPLC separation was performed on Thermo Hypersil GOLD column (2.1 mm× 100mm, 5 μm) at 30 ℃, injecting 10 μL and using acetonitrile-water (0.1% formic acid) as the mobile phrase (B was acetonitrile, A was 0.1%formic acid;0-10 min, 25%-55%B;10-20 min, 55%-70%B) with the flow rate of 0.2 mL/min. Detection was performed on a tandem quadrapole mass spectrometer using positive electrospray ionization, SRM scan mode. Results The eight compounds showed good linearity in wide ranges (notoginsenoside R1 1.00-800 ng/mL, ginsenoside Rg1 0.950-760 ng/mL, ginsenoside Re 1.44-1440 ng/mL, ginsenoside Rb1 1.33-1330 ng/mL, ginsenoside Rd 9.90-990 ng/mL, harpagosidein 1.01-1010 ng/mL, astragaloside Ⅳ 1.16-928 ng/mL, tanshinone Ⅰ 10.0-800 ng/mL). In addition, the accuracy and recovery were around 85%-115%and 50%-70%. The RSD of intra and inter day precision were lower than 15%. Conclusion The method is specific, rapid and sensitive. Therefore, it can be applied to pharmacokinetic study of eight effective compounds in Fufang Xueshuantong Capsule.

The clinical research on early diabetic nephropathy treated with Fufang-Xueshuantong capsule / 国际中医中药杂志

Objective To observe the clinical effect of early diabetic nephropathy treated with Fufang-Xueshuantong capsule. Methods 90 hospitalized patients from September 2011 to October 2012 with early diabetic nephropathy were recruited in Shangqiu First people's hospital endocrinology of Henan Province. All patients were divided into two groups randomly, with 45 patients in each group. The control group was treated with diabetes education, blood glucose control, blood pressure control, and blood lipid regulation, while the treatment group was treated with Fufang-Xueshantong capsule on the basis of the control group. The change of blood glucose, HbA1c, urine microalbumin, and Ualb/Cr were measured before and 12 weeks after the treatment. Results ① Compared the urine microalbumin, Ualb/Cr of the two groups before and after treatment:the urine microalbumin, Ualb/Cr of the two groups after the treatment [Treatment group was(24.5±9.1)mg/L,(2.4± 1.6)mg/mmol, Control group was(85.9±8.6)mg/L,(5.6±2.1)mg/mmol] were significantly reduced than before the treatment (P0.05). Conclusion Fufang-Xueshuantong capsule had good effect in treating early diabetic nephropathy, it can also delay the progress of renal damage.

Application of quantitative analysis of multi-components by single-marker in detection of eight components of Fufang Xueshuantong capsule / 国际药学研究杂志

Objective: To set up a new method - quantitative analysis of multi-components by single-marker(QAMS) to determine eight components - notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and Rd, harpagoside, cryptotanshinone and tanshinone I in Fufang Xueshuantong(FX) capsule simultaneously, and validate its accuracy and feasibility for application in preparation. Methods: FX capsule was used as the research object, notoginsenoside R1 was selected as marker under 203 nm to evaluate the quality of ginsenoside Rg1, Re and Rb1, Rd, and cryptotanshinone as marker under 270 nm to evaluate the quality of harpagoside and tanshinone I. The relative correction factors (RCF) of components were calculated. The method was validated by comparison of the quantitative results between external standard method and QAMS method. Results: No significant differences were found in the quantitative analysis of components by external standard method and QAMS method. Conclusion: It is a convenient and accurate method to determine multi-components when authentic standard substances are not available. It can be used to control the quality of FX capsule.

Chinese medicinal formula Fufang Xueshuantong capsule could inhibit the activity of angiotensin converting enzyme.

Fufang Xueshuantong (FXST) capsule, a Chinese medicinal formula composed of four herbals - Panax notoginseng, Radix Astragali, Radix Salvia Miltiorrhizae and Radix Scrophulariaceae, has been used to treat cardiovascular diseases for many years, but the pharmacological mechanisms underlying its effects has not been clarified. This study investigates if a connection between FXST and angiotensin converting enzyme (ACE) might be an explanation for its pharmacological effects. ACE inhibition assay was performed on FXST capsule, 50% ethanol extracts from the four herbals and three selected saponins most abundant in P. notoginseng (Ginsenoside Rg1, Ginsenoside Rb1 and Notoginsenoside R1) using a biochemical test. Reversed-phase high-performance liquid chromatography of liberated hippuric acid from the ACE assay was conducted to determine the inhibitory effect. As a result, FXST and extracts from P. notoginseng showed a significant and dose-dependent inhibition on ACE activity with the IC50 values of 115 µg/ml and 179 µg/ml, respectively. But extracts from the other three herbals and the three selected saponins had no significant effect on ACE inhibition. Compared to other reported plant extracts, FXST could be considered as an effective ACE inhibitor. The inhibition of ACE activity supports the traditional use of FXST on blood circulation and the inhibitory property of FXST is mainly caused by P. notoginseng.