Study on mechanisms and molecular verification of Buyang Huanwu decoction in treating ischemic stroke from the perspective of cuproptosis

Background:Ischemic stroke(IS)is a global health issue and the current treatment options for IS are inadequate.Buyang Huanwu decoction(BHD)has demonstrated effectiveness in treating IS.However,the mechanisms by which BHD treats IS remain unclear,and no studies have been conducted to analyze these mechanisms from the perspective of Cuproptosis.In order to investigate the potential of BHD to intervene in IS through Cuproptosis,this study employed a systematic pharmacological approach and molecular docking verification.Methods:To investigate the mechanism of BHD in treating IS through Cuproptosis,relevant information on the structure,targets,and major biological functions and pathways of compounds related to BHD was collected from databases such as PubChem,PharmMapper,UniProt,and GeneCards.The results were then visualized using Cytoscape3.6.1,Ledock,and pymol software.Results:BHD is composed of 7 Chinese medicines,which contain 82 compounds,including 10 core compounds.These compounds are associated with 241 genes,of which 97 are common to BHD,IS,and Cuproptosis.The 97 common genes,including 10 core genes,are involved in biological processes such as proteolysis,regulation of apoptosis and cholesterol storage,as well as cellular components and molecular functions.The common genes among BHD,IS,and Cuproptosis,including 10 core genes,participate mainly in Kyoto Encyclopedia of Genes and Genomes pathways such as pathways in cancer,PI3K-Akt signaling pathway,and Estrogen signaling pathway.According to molecular docking results,linolenic acid showed good docking scores with 9 out of 10 core genes,except for SRC.13-hydroxy-9,11-octadecadienoic acid also demonstrated good docking scores with EGFR,MAPK14,and F2.Similarly,senkyunone also had good docking scores with EGFR,MAPK14,and F2,all of which had docking energy greater than−5 kcal.Conclusion:In this study,the potential of BHD for treating IS through Cuproptosis and its underlying mechanisms were explored and partially validated through molecular docking.However,due to the limitations of the systems pharmacology research method,further validation through cell experiments,animal experiments,and clinical trials may be necessary to confirm these findings.

The effects of Buyang Huanwu decoction combined with Western medicine in diabetic foot treatment:a systematic review and meta-analysis

Objective:Systematically evaluate the effect of Buyang Huanwu decoction(BHD)combined with Western medicine(WM)in treating diabetic foot(DF).Methods:Randomized controlled trials that examined the effects of BHD on DF were collected from various databases,including CBM,PubMed,EMBASE,Web of Science,CNKI,Wanfang,and Weipu.Two researchers evaluated the quality of the literature and extracted relevant data,which was then analyzed using RevMan 5.3 for meta-analysis.The analysis was carried out in accordance with the 2020 PRISMA checklist.Results:The study analyzed 19 articles involving 1284 patients.According to the meta-analysis,BHD combined with WM improved clinical efficacy and reduced traditional Chinese medicine syndrome scores,glycosylated hemoglobin,and fibrinogen levels in DF patients.Although the BHD combined with WM showed a trend towards reducing wound area,the difference was not significant.Additionally,the incidence of adverse reactions did not show a significant decrease with BHD combined with WM.Conclusion:The findings of the study indicated that the use of BHD combined with WM yielded better therapeutic outcomes than WM alone,supporting the recommendation of using BHD with WM for DF treatment.This approach is suggested for further clinical application and promotion.

Neuroprotective effects of Buyang Huanwu decoction on cerebral ischemia-induced neuronal damage

Among the various treatment methods for stroke, increasing attention has been paid to tradi- tional Chinese medicines. Buyang Huanwu decoction is a commonly used traditional Chinese medicine for the treatment of stroke. This paper summarizes the active components of the Chinese herb, which is composed of Huangqi （Radix Astragali seu Hedysari）, Danggui （Radix Angelica sinensis）, Chishao （Radix Paeoniae Rubra）, Chuanxiong （Rhizoma Ligustici Chuanx- iong）, Honghua （Flos Carthami）, Taoren （Semen Persicae） and Dilong （Pheretima）, and identifies the therapeutic targets and underlying mechanisms that contribute to the neuroprotective prop- erties of Buyang Huanwu decoction.

Buyang Huanwu Decoction regulates neural stem cell behavior in ischemic brain

The traditional Chinese medicine Buyang Huanwu Decoction has been shown to improve the neu- rological function of patients with stroke. However, the precise mechanisms underlying its effect remain poorly understood. In this study, we established a rat model of cerebral ischemia by middle cerebral artery occlusion and intragastrically administered 5 g/kg Buyang Huanwu Decoction, once per day, for 1, 7, 14 and 28 days after cerebral ischemia. Immunohistochemical staining revealed a number of cells positive for the neural stem cell marker nestin in the cerebral cortex, the subven- tricular zone and the ipsilateral hippocampal dentate gyrus in rat models of cerebral ischemia. Buyang Huanwu Decoction significantly increased the number of cells positive for 5-bromodeoxyuridine （BrdU）, a cell proliferation-related marker, microtubule-associated protein-2, a marker of neuronal differentiation, and growth-associated protein 43, a marker of synaptic plasticity in the ischemic rat cerebral regions. The number of positive cells peaked at 14 and 28 days after intragastric administration of Buyang Huanwu Decoction. These findings suggest that Buyang Huanwu Decoction can promote the proliferation and differentiation of neural stem cells and en- hance synaptic plasticity in ischemic rat brain tissue.

Buyang Huanwu Decoction fraction protects against cerebral ischemia/reperfusion injury by attenuating the inflammatory response and cellular apoptosis

Buyang Huanwu Decoction fraction extracted from Buyang Huanwu Decoction contains saponins of Astragalus, total paeony glycoside and safflower flavones. The aim of this study was to demonstrate the neuroprotective effect and mechanism of Buyang Huanwu Decoction fraction on ischemic injury both in vivo and in vitro. In vivo experiments showed that 50-200 mg/kg Buyang Huanwu Decoction fraction reduced infarct volume and pathological injury in ischemia/reperfusion rats, markedly inhibited expression of nuclear factor-KB and tumor necrosis factor-a and promoted nestin protein expression in brain tissue. Buyang Huanwu Decoction fraction （200 mg/kg） exhibited significant effects, which were similar to those of 100 mg/kg Ginkgo biloba extract. In vitro experimental results demonstrated that 10-100 mg/L Buyang Huanwu Decoction fraction significantly improved cell viability, decreased the release of lactate dehydrogenase and malondialdehyde levels, and inhibited the rate of apoptosis in HT22 cells following oxygen-glucose deprivation. Buyang Huanwu Decoction fraction （100 mg/L） exhibited significant effects, which were similar to those of 100 mg/L Ginkgo biloba extract. These findings suggest that Buyang Huanwu Decoction fraction may represent a novel, protective strategy against cerebral ischemia/reperfusion injury in rats and oxygen-glucose deprivation-induced damage in HT22 cells in vitro by attenuating the inflammatory response and cellular apoptosis.

Microvessel changes in the gerbil hippocampus after cerebral ischemia and reperfusion by Buyang Huanwu decoction pretreatment

Buyang Huanwu decoction （BYHWD） is a classic recipe for the prevention and treatment of ischemic cerebrovascular disease. Gerbils were pretreated with BYHWD, and then subjected to cerebral ischemia and reperfusion. Microvascular changes were determined with laser Doppler monitoring, tannic acid-ferric chloride mordant, and electron microscopy. Results showed that BYHWD pretreatment could enhance the function of hippocampal microvessels, prevent injury, and increase microvasular density and microvasular area density. Thus, these results suggest that BYHWD pretreatment could prevent microvascular occlusion, enhance the capacity of microvascular reperfusion, increase cerebral blood flow, and inhibit neuronal damage, and may be an effective therapy against brain ischemic injury.

Buyang Huanwu decoction increases vascular endothelial growth factor expression and promotes angiogenesis in a rat model of local cerebral ischemia

Angiogenesis in the infarct periphery can improve blood flow. Vascular endothelial growth factor （VEGF） has been considered a potential therapeutic target for stroke. Buyang Huanwu decoction （BYHWD） is a classic traditional formula in traditional Chinese medicine and is used to treat stroke; in addition, the promotion effects on VEGF protein expression have been confirmed. However, little is known about how BYHWD regulates angiogenesis, or about the effects of BYHWD on VEGF mRNA expression. For this reason, the present study measured microvessel density in rats with cerebral ischemia using immunohistochemistry. In addition, VEGF expression was measured by re-verse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay to determine the effects of BYHWD on angiogenesis and VEGF expression in rats with cerebral ischemia. Results demonstrated that microvessel density, as well as VEGF mRNA and protein expression, increased after 7 and 14 days of BYHWD treatment, which suggests that BYHWD promoted angiogenesis following cerebral ischemia and upregulated VEGF mRNA and protein expression in ischemic cerebral regions.

The active principle region of Buyang Huanwu decoction induced differentiation of bone marrow-derived mesenchymal stem cells into neural-like cells Superior effects over original formula of Buyang Huanwu decoction

The present study induced in vitro-cultured passage 4 bone marrow-derived mesenchymal stem cells to differentiate into neural-like cells with a mixture of alkaloid, polysaccharide, aglycone, glycoside, essential oils, and effective components of Buyang Huanwu decoction （active principle region of decoction for invigorating yang for recuperation）. After 28 days, nestin and neuron-specific enolase were expressed in the cytoplasm. Reverse transcription-PCR and western blot analyses showed that nestin and neuron-specific enolase mRNA and protein expression was greater in the active principle region group compared with the original formula group. Results demonstrated that the active principle region of Buyang Huanwu decoction induced greater differentiation of rat bone marrow-derived mesenchymal stem cells into neural-like cells in vitro than the original Buyang Huanwu decoction formula.

Effects of Buyang Huanwu decoction on cell proliferation and differentiation in the hippocampal dentate gyrus of aged rats following cerebral ischemia/reperfusion

BACKGROUND： The mobilization of endogenous stem cells is an effective way to promote repair following ischemic brain damage. Buyang Huanwu decoction （BHD） can effectively improve cerebral blood flow and protect against cerebral ischemia/reperfusion damage. OBJECTIVE： To study the effects of BHD on cell proliferation and differentiation in the hippocampal dentate gyrus of rats following cerebral infarction, to investigate the protective effects of BHD against cerebral infarction, and to analyze the dose-effect relationship. DESIGN, TIME AND SETTING： This randomized, controlled, animal study was performed at the Laboratory of Department of Physiology, Henan College of Traditional Chinese Medicine, China from June 2007 to February 2008. MATERIALS： A total of 36 male, Sprague Dawley rats, aged 20-21 months, were equally and randomly assigned to the following groups： sham operation, model control, and nimodipine, as well as high-dose, moderate-dose, and low-dose BHD. BHD was composed of milkvetch root, Chinese angelica, red peony root, earthworm, peach seed, safflower, and Szechwan Iovage rhizome, which were provided by the Outpatient Department, Henan College of Traditional Chinese Medicine, China. METHODS： The Chinese medicinal ingredients described above were decocted. The external carotid artery was ligated in rats from the sham operation group. Rat models of focal cerebral infarction were established by middle cerebral artery occlusion in the model control and nimodipine groups, as well as the high-dose, moderate-dose, and low-dose BHD groups. The drugs were administered by gavage 5 days, as well as 2 hours, prior to model induction. Rats in the nimodipine group were daily administered a 6 mg/kg nimodipine suspension by gavage. Rats in the high-dose, moderate-dose, and low-dose BHD groups were administered daily 26, 13, and 6.5 g/kg BHD, respectively. Rats in the sham operation and model control groups were treated with an equal volume of saline. MAIN OUTCOME MEASURES： The effects of BHD on neurological dysfunction score, brain water content, cell proliferation and differentiation in the hippocampal dentate gyrus, and pathological changes in the ischemic brain hemisphere were measured in cerebral infarction rats. RESULTS： Compared with the sham operation group, the neurological dysfunction score, brain water content, number of BrdU-positive cells, BrdU/NeuN-positive cells, and BrdU/GFAP-positive cells in the hippocampal dentate gyrus significantly increased in the model control group （P 〈 0.01 ）. Compared with the model control group, neurological dysfunction score and brain water content were significantly decreased （P 〈 0.01 or 0.05）, as were the number of BrdU-positive and BrdU/NeuN-positive cells （P 〈 0.01 or 0.05）. The number of BrdU/GFAP-positive cells was significantly reduced （P 〈 0.05） in the nimodipine group, high-dose, moderate-dose, and low-dose BHD groups. Compared with the nimodipine group, the neurological dysfunction score was significantly reduced in the moderate-dose BHD group （P 〈 0.05）. However, the number of BrdU-positive cells was significantly increased in the rat hippocampal dentate gyrus in the high-dose and moderate-dose BHD groups （P 〈 0.01 or 0.05）. The following was determined by microscopy： slightly disarranged neural cells, mild vascular dilatation, inflammatory cell infiltration, and light tissue edema were observed in the nimodipine group; inflammatory celt infiltration was reduced in the low-dose BHD group; cerebral edema and inflammatory cell infiltration were significantly reduced in the high-dose and in the moderate-dose BHD group. Electron microscopy revealed lipofuscin, slightly swollen mitochondria, and normal rough endoplasmic reticulum in the high-dose and moderate-dose BHD groups. Improvement was best in the moderate-dose BHD group. CONCLUSION： Cerebral ischemia activated proliferation of neural stem cells in the rat hippocampal dentate gyrus. The actions of BHD against cerebral ischemia/reperfusion damage correlated with proliferation and differentiation of neural stem cells in the hippocampal dentate gyrus. A moderate-dose of BHD resulted in the most effective outcome.

Buyang Huanwu decoction enhances cell membrane fluidity in rats with cerebral ischemia/reperfusion

After bilateral carotid artery occlusion for 30 minutes and reperfusion for 2 hours, distinct pathological changes presented in the cerebral cortex and cerebellum of rats. Compared with normal rats, nerve cell membrane fluidity significantly decreased in ischemia/reperfusion rats as detected by spin-labeling electron spin resonance, consistent with order parameter S and rotational correlation time TC measurements. Brain nerve cells from rats with ischemia/reperfusion injury were cultured with 1-100 mg/mL Buyang Huanwu decoction. Results showed that Buyang Huanwu decoction gradually increased membrane fluidity dose-dependently to normal levels, and eliminated hydroxide （OH＇） and superoxide （O2＇） free radicals dose-dependenUy. These findings suggest that Buyang Huanwu decoction can protect against cell membrane fluidity changes in rats with ischemia/ reperfusion injury by scavenging free radicals.

Mitogen activated protein kinase signaling pathways participate in the active principle region of Buyang Huanwu decoction-induced differentiation of bone marrow mesenchymal stem cells

Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell differentiation into neurons. Mitogen-activated protein kinase signaling was identified as one of the key pathways underlying this differentiation process. The present study shows phosphorylated extracellular signal-regulated protein kinase and phosphorylated p38 protein expression was increased after differentiation. Cellular signaling pathway blocking agents, PD98059 and SB203580, inhibited extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways respectively, mRNA and protein expression of the neuronal marker, neuron specific enolase, and neural stem cell marker, nestin, were decreased in bone marrow mesenchymal stem cells after treatment with the active principle region of Buyang Huanwu decoction. Experimental findings indicate that, extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways participate in bone marrow mesenchymal stem cell differentiation into neuron-like cells, induced by the active principle region of Buyang Huanwu decoction.

Buyang Huanwu decoction up-regulates Notch1 gene expression in injured spinal cord

Expression of genes in the Notch signaling pathway is altered in the injured spinal cord, which indicates that Notch participates in repair after spinal cord injury. Buyang Huanwu decoction, a traditional Chinese herbal preparation, can promote the growth of nerve cells and nerve fibers; however, it is unclear whether Buyang Huanwu decoction affects the Notch signaling pathway in injured spinal cord. In this study, a rat model was established by injuring the T10 spinal cord. At 2 days after injury, rats were intragastrically administered 2 m L of 0.8 g/m L Buyang Huanwu decoction daily until sacrifice. Real-time reverse transcription polymerase chain reaction analysis demonstrated that at 7, 14 and 28 days after injury, the expression of Notch1 was increased in the Buyang Huanwu decoction group compared with controls. These findings confirm that Buyang Huanwu decoction can promote the expression of Notch1 in rats with incomplete spinal cord injury, and may indicate a mechanism to promote the repair of spinal cord injury.

Plasma metabolomics study of Buyang Huanwu Decoction improving learning andmemory in D-gal model mice based on GC-MS

Objective:To study the protective effect of Buyang Huanwu Decoction on the learning and memory ability of D-gal induced aging mice using GC-MS metabolomics method.Methods:Twenty-four three-month-old kunming animals were selected as experimental samples and randomly divided into control group,model group and Buyang Huanwu Decoction according to their body weight.The memory level of experimental animals was detected by novel body recognition test,and the neuron structure of experimental animals was detected by HE staining.The plasma of each group of experimental animals was quantitatively analyzed by gas chromatography-mass spectrometry,and the important metabolites and main metabolic pathways in the process of pathological changes were traced by using plasma metabolism.Results:In HE staining,compared with blank group,hippocampal neurons in model group were disordered and morphologically abnormal.Compared with the model group,the hippocampal neurons in Buyang Huanwu Decoction group arranged regularly and had normal morphology.Compared with blank group,the index of new object recognition in model group was significantly decreased(P<0.05);Compared with model group,the new object recognition index of Buyang Huanwu Decoction group was significantly increased(P<0.05).Five different metabolites of AD were identified by GC-MS,which were L-pyroglutamate,lysine,pyrophosphoric acid,creatinine andα-lactose.Conclusion:Buyang Huanwu Decoction has a significant effect on D-gal model mice,and can effectively improve their learning and memory level.The mechanism may be related to glutathione metabolism and aminophyl biosynthesis.

Effect of Buyang Huanwu Decoction on the proliferation,migration and adhesion of endothelial progenitor cells in rats with acute myocardial infarction

Objective:To investigate the intervention effect of Buyang Huanwu Decoction on endothelial progenitor cell function(EPCs),and to explore the therapeutic mechanism of Buyang Huanwu Decoction.Methods:This research take 54 rats were divided into blank group,the control group,model group,Chinese medicine,western medicine group and combine traditional Chinese and western medicine group,tonifying Yang also five decoction and atorvastatin calcium for acute myocardial infarction(AMI)rats group intervention,by using the method of density gradient centrifugation separation training each rat endothelial progenitor cells,using tetramethyl azo salt trace enzyme reaction colorimetry,used in the determination of adhesion ability and improved Boyden chamber,the method of analysis and comparison between groups of endothelial progenitor cells proliferation,adhesion and migration.Results:In 7 days,14 days and 4 weeks,acute myocardial infarction model of rat peripheral blood EPCs count,proliferation,migration and adhesion ability were compared with control group were significantly decline,tonifying Yang also five decoction group,western medicine control group and combine traditional Chinese and western medicine group of peripheral blood EPCs count,proliferation,migration and adhesion ability compared with model group were significantly increased(P<0.05),and combine traditional Chinese and western medicine group is the traditional Chinese medicine and western medicine group on the improvement of the function of EPCs effect more significantly(P<0.05).Conclusion:Statins combined with Buyang Huanwu Decoction can better improve the endothelial function of AMI rats than traditional Chinese medicine or western medicine.Further clinical studies on statins may better improve the endothelial function of AMI patients,and provide a new research entry point for improving the long-term prognosis of AMI patients.

Effects of Buyang Huanwu decoction and Astragalus mongholicus on platelet activating factor receptor activity in rabbits in vitro

BACKGROUND: The pharmacological action of traditional Chinese medicine compound is the comprehensive effect of the various ingredients, and the interactions of various ingredients are closely correlated with the final effect. In order to reveal the compatibility mechanism of BHD's prescription in treating and preventing ischemic cerebrovascular disease, we needed explore the effect and relation of ingredients in the prescription. OBJECTIVE: To observe the effect of Buyang Huanwu decoction (BHD) and Astragalus mongholicus on the activity of platelet activating factor receptor (PAFR) in the platelet of rabbits in vitro, and investigate the mechanism of Astragalus mongholicus. DESIGN: A decomposed recipes study. SETTING: Guangzhou University of Traditional Chinese Medicine. MATERIALS: Five New Zealand rabbits, weighing 2-3 kg, both sexes, were used. BHD was composed of Sheng Huang Qi 120 g, Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g, Hong Hua 3 g. The prescription for activating blood circulation consisted of Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g and Hong Hua 3 g. The prescription for invigorating qi consisted of 120 g Sheng Huang Qi. The prepared herbal pieces were purchased from the traditional Chinese medicine Dispensary of Foshan Second People's Hospital, and appraised by Professor Xu from Science of Chinese Materia Medica College, Guangzhou University of Traditional Chinese Medicine. 3H-PAF was supplied by Amersham Co., Ltd. (specific activity: 6. 475 TBq/mmol; batch number: 200402); PAF standard by Biomol Co., Ltd. (batch number: P1318V). METHODS: The experiments were carried out in the Laboratory of Nuclear Medicine, Guangzhou University of Traditional Chinese Medicine from September to December 2004. ① Injections of BHD, prescriptions for activating blood circulation and invigorating qi were prepared by the decoction and alcohol sedimentation technique. Rabbit common carotid artery blood (40 mL) was drawn via intubation to prepare platelet suspension of (0.8-1.0)×1010 L-1. ② Determination of 3H-PAF and washed PAFR binding: The general combination tube (T) contained washed platelet-rich plasma (WPRP) 380 μL + 3H-PAF (0.35 nmol/L)10 μL+distilled water 5 μL; The nonspecific binding tube (P) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+cold PAF (1 μmol/L) 5 μL; The sample tube (Y) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+experimental medicine (injection of BHD, prescriptions for activating blood circulation or invigorating qi) 5 μL. The test was conducted for three times for each sample in the same way as mentioned above. The samples were shaken on the oscillator for 30 s, then bathed at 25 ℃ for 40 minutes, and the reaction was terminated with cold Tris buffer containing 0.1% BSA, multichannel cell detachment separator was used for vacuum suction to filter the separated free 3H-PAF, and the filter paper was washed with cold Tris buffer for four times, then dried in the baking oven (80 ℃) for 1 hour, and placed in xylol liquid scintillator, and the radioactivity was determined automatically by the liquid scintillation detector. The mean of the three parallel tubes was calculated. The specific binging inhibition rate was calculated: SBIR=[(T-Y)/(T-P)]×100%]. ③ Univariate analysis of variance was conducted. And for comparison of each paired groups, the q test was adopted. MAIN OUTCOME MEASURES: Effect of BHD whole prescription, prescriptions for activating blood circulation and invigorating qi on the specific binding inhibition rate of 3H-PAF and PAFR. RESULTS: BHD, prescriptions for activating blood circulation and invigorating qi were all able to inhibit the specific binding of 3H-PAF to PAFR, the specific blinding inhibition rates were (45.90±7.50)%, (97.90±1.84)% and (26.75±2.48)%, respectively, and there were significant differences between every two groups (P < 0.01). CONCLUSION: Single Astragalus mongholicus (120 g) can inhibit the specific blinding of PAFR in the platelet of the rabbit with 3H-PAF, but the combination of Astragalus mongholicus with the drugs for activating blood circulation in BHD can significantly decrease the inhibiting action of the latter on PAFR activity of the platelet, reflecting the combined mechanism of 'removing blood stasis without injuring the vital qi' in BHD.

Acupuncture combined with Buyang Huanwu decoction in the treatment of sequelae of cerebrovascular accident:a systematic review and meta-analysis

Background:Cerebrovascular accident has high morbidity,high disability,and high mortality,possibly causing several serious sequelae and bringing a heavy burden to the patient,the family,and the society.Buyang Huanwu decoction,a classic ancient prescription of traditional Chinese medicine,and acupuncture are often used in combination to treat the sequelae of cerebrovascular accident.In the present study,we systematically evaluated the clinical efficacy and safety of acupuncture combined with Buyang Huanwu decoction in treating the sequelae of cerebrovascular accident.Methods:Relevant randomized controlled trials of acupuncture combined with Buyang Huanwu decoction in treating the sequelae of cerebrovascular accident were comprehensively retrieved from electronic databases from inception to May 8,2020,including PubMed,Embase,The Cochrane Library,China National Knowledge Infrastructure,WanFang Data,and China Science and Technology Journal Database.The main outcomes of clinical efficacy,degree of nerve function impairment,daily self-care capacity,Fugl-Meyer locomotor function score,life quality score,and occurrence of adverse reactions were subjected to meta-analysis.Results:The 28 randomized controlled trials included 2,341 patients with the sequelae of cerebrovascular accident.Systematic review and meta-analysis indicated that compared with acupuncture alone,its combined treatment with Buyang Huanwu decoction significantly increased clinical efficacy(RR=1.19,95%CI(1.14,1.23),P<0.001),improved nerve function impairment(WMD=−4.89,95%CI(−8.25,−1.53),P=0.004),promoted daily self-care capacity(WMD=15.87,95%CI(11.22,20.51),P<0.001),strengthened Fugl-Meyer locomotor function(WMD=20.89,95%CI(13.79,27.98),P<0.001),enhanced physical functioning(WMD=16.99,95%CI(6.01,27.96),P<0.001)and mental health(WMD=16.91,95%CI(9.14,24.69),P<0.001)in terms of life quality score(SF-36 scale)of patients with sequelae of cerebrovascular accident,with few adverse reactions.Conclusion:Compared with acupuncture alone,its combination with Buyang Huanwu decoction had better comprehensive effects on sequelae of cerebrovascular accident without obvious adverse reactions.

Meta-analysis of Buyang Huanwu decoction in animal model of cerebral ischemia

Objective:To evaluate the clinical efficacy of Buyang Huanwu decoction in treating animal model of cerebral ischemia.Methods:We searched PubMed,EMbase,Cochrane,CNKI,VIP,WanFang(1983 to 2021)for randomized control trials about Buyang Huanwu decoction treating animal model of cerebral ischemia.Results:According to the inclusion and exclusion criteria and assessment the quality of included trials(RCTs),the extraction of effective data,7 randomized clinical trials and their efficiency were evaluated,statistical analysis was conducted by using RevMan 5.3 software.The outcome measures assessed were neurological score and/or infarction volume.Meta-analysis showed that the neurological behavior scores of Buyang Huanwu decoction on animal model of cerebral ischemia was lower than the control group(SMD=2.06,95%CI(1.75,2.37),P<0.00001);The cerebral infarct volume of Buyang Huanwu decoction on animal model of cerebral ischemia were smaller than the control group(SMD=4.08,95%CI(3.57,4.58),P<0.00001).Conclusion:It was effective by using Buyang Huanwu decoction to reduce neurological behavior scores and the volume of animal model of cerebral ischemia.

Effects of Buyang Huanwu Decoction on Intestinal Barrier, Intestinal Flora, and Trimethylamine Oxide in Rats with Heart Failure

Objective: To explore the mechanisms of Buyang Huanwu Decoction(BYHWD) modulating the gut microbiome and trimethylamine oxide(TAMO) to exert cardioprotective effects. Methods: Ligation of the left anterior descending coronary artery was performed in rats to induce heart failure(HF). Except for the shamoperation group(n=10), 36 operation-induced models were randomized into 3 groups using a random number table(n=12 in each group): the model group, the BYHWD group(15.02 g/kg BYHWD), and the positive group(4.99 g/kg metoprolol succinate). After 4-week treatment(once daily by gavage), echocardiography was applied to evaluate the cardiac function and the Tei index(the ratio of ventricular isovolumic contraction time(IVCT)and isovolumic diastolic time(IVRT) to ejection time(ET)) was calculated;hematoxylin-eosin(HE) staining was observed to characterize the pathology of the myocardium and small intestinal villi. D-lactic acid was detected by an enzyme-linked immunosorbent assay(ELISA). Expressions of occludin, claudin-1, and zonula occludens(ZO-1) were detected by Western blot. 16S ribosomal ribonucleic acid(16S rRNA) sequencing was used to explore the changes in the intestinal flora. TMAO was detected via liquid chromatography-tandem mass spectrometry(LC-MS/MS). Results: In the echocardiography, the Tei index was considerably lower in the positive and BYHWD groups compared with the model group(P<0.05). Besides, BYHWD improved the pathology of myocardium and small intestine of HF rats and lowered the D-lactic acid content in the serum, when compared with the model group(P<0.05). BYHWD also improved the expression of occludin and claudin-1(P<0.05);in the gut microbiota analysis, BYHWD slowed down modifications in the structure distribution of gut microbiota and regulated the diversity of intestinal flora in HF rats. The content of TMAO in the serum was significantly lowered by BYWHT compared with the model group(P<0.05). Conclusion: BYHWD may delay progression of HF by enhancing the intestinal barrier structure, and regulating intestinal flora and TAMO.

Efficacy and safety of Buyang Huanwu decoction(补阳还五汤)for diabetic peripheral neuropathy:a systematic review and Meta-analysis

OBJECTIVE:To evaluate the efficacy and safety of Buyang Huanwu decoction(BYHWD)in treating diabetic peripheral neuropathy(DPN).METHODS:Eight electronic databases,including China National Knowledge Infrastructure Database,Wanfang Database,China Science and Technology Journal Database,Chinese Biomedical Literature Database,Cochrane Library,Embase,Web of Science,and PubM ed,were searched for randomized controlled trials(RCTs)of BYHWD to treat DPN.We identified all RCTs related to BYHWD and those on the treatment of DPN with the combination of mecobalamin.RevMan software was used for the statistical analysis.RESULTS:Twenty-one RCTs with a total of 1945 patients were included.The methodological quality of the literature included was low.Meta-analysis showed that the efficacy of the treatment group was significantly better than that of the control group in the treatment of DPN with BYHWD[risk ratio(RR)=0.33,95%CI(0.27,0.40),Z=11.25,P<0.00001].The median nerve of median motor nerve conduction velocity(MNCV)[mean difference(MD)=4.16,95%CI(1.35,6.98)]and median sensory NCV(SNCV)[(MD=3.28,95%CI(2.35,4.22)]were improved in the treatment group.The MNCV in the common peroneal nerve[(MD=1.63,95%CI(0.39,2.87)]and SNCV[(MD=4.56,95%CI(3.16,5.97)]were significantly higher than those in the control group(P<0.01).Plasma viscosity[(MD=-0.15,95%CI(-0.20,-0.09),Z=5.17,P<0.01)],whole blood high shear[(MD=-0.83,95%CI(-1.56,-0.11),Z=2.26,P=0.02)]and whole blood low shear[(MD=-1.61,95%CI(-2.28,-0.94),Z=4.68,P<0.01)]decreased significantly after treatment.There was no significant difference in fasting blood glucose[(MD=-0.42,95%CI(-0.89,0.05),Z=1.76,P=0.08)]between the treatment and control groups;postprandial blood glucose[(MD=-0.62,95%CI(-1.19,0.05),Z=2.12,P=0.03)]decreased significantly.No significant difference was found in the blood lipid levels between the treatment and control groups,including triglycerides[(MD=-0.21,95%CI(-0.52,0.10),Z=1.34,P=0.18)]and cholesterol[(MD=-0.13,95%CI(-0.27,0.00),Z=1.92,P=0.06)].Of the 21 RCTs,only five reported adverse reactions,and four studies reported the length of follow-up.No serious adverse events were reported.None of the studies reported the quality of life and economic conditions.CONCLUSIONS:Our study suggests that BYHWD has a significant therapeutic effect on DPN.High-quality,largescale RCTs are needed to provide more reliable evidence.

Buyang Huanwu Decoction Ameliorates Damage of Erectile Tissue and Function Following Bilateral Cavernous Nerve Injury

Objective To verify the effect of Buyang Huanwu Decoction(BHD)in ameliorating erectile dysfunction(ED)after radical prostatectomy(RP).Methods The composition of BHD was verified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS/MS)analysis.Bilateral cavernous nerve crush injury(BCNI)in rats was used to mimic the neurovascular injury occurring after RP.By the envelope method,forty rats were randomly divided into 4 groups as follows:sham(cavernous nerves exposed only),model(BCNI),low-dosage BHD[LBHD,12.8 g/(kg·d)],and high-dosage BHD[HBHD,51.2 g/(kg·d)]groups,10 rats in each group,feeding for 3 weeks respectively.Erectile function was evaluated by measuring intracavernosal pressure(ICP).Changes in the histopathology of corpus cavernosum(CC)were examined by hematoxylin-eosin staining.Meanwhile,the fibrosis of CC was measured by Masson’s trichrome staining and Western blot was used to detect the expressions of collagen I,transforming growth factor beta 1(TGF-β1)andα-smooth muscle actin(α-SMA).Apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)and Western blot for determining the expressions of B-cell lymphoma 2(Bcl-2)and Bcl-2-associated X(Bax).The oxidative stress in the CC were assessed by the superoxide dismutase(SOD),malondialdehyde(MDA)and reactive oxygen species(ROS)levels.The proteins expression of c-Jun N-terminal kinase(JNK)and c-Jun were detected by Western blot.In addition,the expression ofα-SMA and p-c-Jun in the CC was observed by double immunofluorescence staining.Results The UPLC-QTOF-MS/MS analysis showed that BHD contained calycosin-7-O-β-D-glucoside,ononin,calycosin and formononetin.Compared with the model group,LBHD and HBHD treatment improved the ICP and the circumference,area,and weight of CC(P<0.05 or P<0.01).Furthermore,LBHD and HBHD treatments increased CC smooth muscle content and decreased apoptosis index(P<0.05 or P<0.01).LBHD and HBHD also elevated SOD and expression level ofα-SMA and Bcl-2,and reduced MDA and ROS levels,as well as expression of TGF-β1,collagen I,Bax,p-c-JNK,p-JNK in the CC compared with the model group(P<0.05 or P<0.01).The double immunofluorescence staining showed that the fluorescence degree of p-c-Jun in both LBHD and HBHD treatment groups was significantly reduced,whereas theα-SMA expression increased(P<0.05 or P<0.01).Conclusions BHD can improve ED of rats with BCNI,which is related to inhibiting fibrosis,apoptosis,and oxidative stress of CC.The ROS/JNK/c-Jun signaling pathway may play an important role in the process.