Effects of Tongluo Jiedu prescription on immune function and oxidative stress in patients with oral cancer

BACKGROUND Oral cancer,which is caused by mucous membrane variation,represents a prevalent malignant tumor in the oral and maxillofacial region,posing a significant threat to patients’lives and safety.While surgical intervention stands as a cornerstone treatment for oral cancer patients,it carries the risk of incomplete treatment or high rates of postoperative recurrence.Hence,a multifaceted approach incorporating diverse treatment modalities is essential to enhance patient prognosis.AIM To analyze the application effect of Tongluo Jiedu prescription as adjuvant therapy and its influence on patient prognosis in patients with oral cancer.METHODS Eighty oral cancer patients in our hospital were selected and divided into the observation group and control group by a random number table.The control group was treated with continuous arterial infusion chemotherapy of cisplatin and 5-fluorouracil.The observation group was additionally given Tongluo Jiadu prescription.The inflammatory stress level,peripheral blood T-cell subsets,and immune function of the two groups were subsequently observed.SPSS 21.0 was used for data analysis.RESULTS The observation group demonstrated lower levels of interleukin-6 and C-reactive protein,and a higher level of tumor necrosis factor in comparison to the control group.After treatment,the immune function in the observation group was significantly better than in the control group.CONCLUSION Tongluo Jiedu prescription can improve the immune function and oxidative stress level of patients with oral cancer and accelerate the recovery process.

Advancing oral cancer care:Insights from Tongluo Jiedu prescription

This editorial highlights the study which investigated the potential of traditional Chinese medicine(TCM)as an adjunctive therapy in oral cancer management.The study included 80 patients with oral cancer to evaluate the effects of the Tongluo Jiedu prescription on inflammatory stress markers,peripheral blood T-cell subsets,and overall immune function.The results indicated that Tongluo Jiedu substantially enhances immune function and reduces oxidative stress,thereby aiding in patient recovery and potentially minimizing treatment-related complications.This editorial discusses the broader implications of these findings for oral cancer care and emphasizes the importance of integrating TCM principles into modern oncology practices.

Yiqi Tongluo capsule has protective effects against non-alcoholic fatty liver disease via regulating PI3K/AKT signaling

Background:Oxidative stress is one of the key elements in the progression of non-alcoholic fatty liver disease(NAFLD),and Yiqi Tongluo capsule(YTC)have a variety of physiological activities which include antioxidant.The purpose of this investigation was to discover the potential mechanisms of YTC ameliorates NAFLD.Methods:In this investigation,a high-fat diet(HFD)was adopted to establish a NAFLD mouse model.Liver samples were stained for oil red O and hematoxylin and eosin staining.The levels of total cholesterol(TC),triglyceride(TG),malondialdehyde(MDA),and superoxide dismutase(SOD)in the tissues were also detected.Network pharmacology was analyzed to filter out the key ingredients and targets of effect of YTC for the therapy of NAFLD.Subsequently,free fatty acids(FFA)was applied to induce Aml12 cells for in vitro experiments,and the cell samples were stained with oil red O and assayed for TC,TG,MDA,and SOD contents.At last,the Western blot technique was used to illuminate the pathway by which YTC plays a protective role against NAFLD.Results:Histopathological results demonstrated that YTC ameliorated tissue damage in the HFD-induced mouse model.At the same time,it also reduced the contents of TC,TG,MDA and increased the expression of SOD in the liver tissue of NAFLD mouse model.All of these findings demonstrate that YTC can play a role in the treatment of NAFLD by ameliorating oxidative stress.Network analysis of YTC ameliorates NAFLD mainly by modulating the PI3K-Akt signaling pathway.Follow-up in vitro experiments revealed that FFA caused lipid accumulation in Aml12 cells,which was dramatically reduced by YTC.Meanwhile,YTC could remarkably reduce the FFA-induced elevation of TC,TG,and MDA contents,and reverse the FFA-induced reduction of SOD contents.Western blot was verified for the PI3K-Akt signaling pathway.It was found that FFA could remarkably decrease the expression of p-PI3K,p-Akt,and p-GSK-3βproteins,which could be significantly increased after YTC treatment.Conclusions:The combination of network analysis prediction and experimental verification was used to identify the therapeutic effect of YTC on NAFLD.The protective effect was achieved by YTC through upregulation of PI3K-Akt-GSK-3βpathway.

Effect of Zhutan Tongluo Tang on fibrinolytic activity following intracerebral hemorrhage in rats

The neuroprotective effect of Zhutan Tongluo Tang is associated with the activities of the fibrinolytic system. Thus the present study was designed to investigate therapeutic effects of Zhutan Tongluo Tang on intracerebral hemorrhage in rats, induced by injecting collagenase into one side of the caudate nucleus. Fibrinolytic indices including tissue plasminogen activator, plasminogen activator inhibitor-1 and D-Dimer were determined. The results obtained demonstrated that Zhutan Tongluo Tang treatment could alleviate the neural and behavioral impairments of intracerebral hemorrhaging rats. Increased frequencies of placing their forelimb correctly and decreased frequencies of turning left were observed. Enzyme linked immunosorbent assay showed that Zhutan Tongluo Tang could significantly elevate the concentration of plasma tissue plasminogen activator and D-Dimer, while depress plasminogen activator inhibitor-1. Immunohistochemistry demonstrated increased levels of catalase and glutathione in whole brain rat tissue following intracerebral hemorrhage. In addition, elevated expression of Bcl-2 in various hippocampal regions was seen. These findings describe the ability of Zhutan Tongluo Tang to activate the fibrinolytic system, and suggests that antioxidant and apoptosis inhibitory mechanisms may be playing a crucial role in protecting against collagenase-induced intracerebral hemorrhage.

The Chinese herbal formula Tongluo Jiunao promotes expression of brain-derived neurotrophic factor/tropomyosin-related kinase B pathways in a rat model of ischemic brain injury

The neurotrophin-Trk receptor pathway is an intrinsic pathway to relieve damage to the central nervous system. The present study observed the effects of Tongluo Jiunao （TLJN）, which comprises Panax Notoginseng and Gardenia Jasminoides, on expression of brain-derived neurotrophic factor （BDNF） and tropomyosin-related kinase B （TrkB） in a rat model of focal cerebral ischemic injury. Xue Sai Tong （XST）, comprising Panax Notoginseng, served as the positive control. Mechanisms of neuroprotection were analyzed following TLJN injection. Following establishment of the middle cerebral artery occlusion models, TLJN and XST were intraperitoneally injected, and 2, 3 5-triphenyltetrazolium chloride staining results revealed that TLJN injection reduced infarct volume, suggesting that TLJN exerted a neuroprotective effect. Enzyme-linked immunosorbent assay results showed that TLJN elevated BDNF and growth associated protein-43 expression in ischemic brain tissues, as well as serum BDNF levels. Reverse-transcription polymerase chain reaction and western blot results showed that TLJN injection did not affect TrkB expression in the ischemic brain tissues of rats. These results suggested that TLJN injection reduced damage to ischemic brain tissues and increased BDNF expression. In addition, TLJN injection resulted in better promoting effects on neurotrophic factor expression compared with XST.

Clinical efficacy of angina pectoris after pci in patients with coronary heart disease complicated with type 2 diabetes by Yiqi Yangyin and Huatan Tongluo recipe

Objective:To evaluate the clinical efficacy of Yiqi Yangyin and Huatan Tongluo Recipe in improving the symptoms of angina pectoris and cardiac function in patients with coronary heart disease complicated with type 2 diabetes after PCI.Methods Sixty-two patients with coronary heart disease complicated with type 2 diabetes after PCI were randomly divided into treatment group and control group,with 31 patients in each group.Both groups were treated with western medicine.The treatment group added Yiqi Yangyin and Huayu Tongluo Recipe granules.The changes of metabolic equivalent(METs),angina symptoms,TCM symptoms,BNP,hypersensitive C-reactive protein(hs-CRP),Interleukin-6(IL-6),Tumor necrosis factor-(TNF-α)and left ventricular ejection fraction(LVEF)were analyzed after 1 course of treatment and Corrected TIMI Frame Count(CTFC)after six month.Results The improvement of METS and LVEF in the treatment group was better than that in the control group(P<0.05);the improvement of BNP,hs-CRP,IL-6,TNF-αand CTFC in the treatment group was better than that in the control group;the curative effect of angina pectoris in the treatment group was better than that in the control group;the effective rate of TCM syndrome improvement in the treatment group was better than that in the control group.Conclusion Yiqi Yangyin and Huatan Tongluo Recipe can effectively alleviate angina pectoris symptoms in patients with coronary heart disease complicated with type 2 diabetes after PCI,suggesting cardiac function,inhibiting inflammatory response,and improving microcirculation disturbance.

Guben Tongluo Formula Protects LPS-induced Damage in Lamina Propria B Lymphocytes Through TLR4/MyD88/NF-κB Pathway

Objective The main pathological feature of immunoglobulin A nephropathy(IgAN),an autoimmune kidney disease,is the deposition of IgA immune complexes,accompanied by mesangial cell proliferation and elevated urine protein.The Guben Tongluo formula(GTF)is a traditional Chinese medicine prescription,which has predominant protective effects on IgAN.However,the therapeutic mechanism of the GTF in IgAN remains elusive.The present study aimed to determine the effects of GTF in treating IgAN via regulating the TLR4/MyD88/NF-κB pathway.Methods In the present study,lamina propria B lymphocytes were treated with different concentrations of lipopolysaccharide(LPS)(0,1,5,10 and 20 ng/mL).Flow cytometry was used to define positive CD86+CD19+cells.CCK-8 assay was used to examine cell proliferation.RNAi was used to induce TLR4 silencing.qRT-PCR and Western blotting were used to determine gene expression.Results It was found that the LPS dose-dependently increased the content of IgA and galactose-deficient IgA1(Gd-IgA),the levels of TLR4,Cosmc,MyD88 and phosphorylated(p)-NF-κB,and the ratio of CD86+CD19+and IgA-producing B cells.However,the TLR4 knockdown reversed the role of LPS.This suggests that TLR4 mediates the effects of LPS on lamina propria B lymphocytes.Furthermore,the GTF could dose-dependently counteract the effects of LPS and TLR4 overexpression on lamina propria B lymphocytes through the TLR4/MyD88/NF-κB pathway.Conclusion Collectively,these results demonstrate that the GTF can regulate the TLR4/MyD88/NF-κB pathway to treat IgAN model lamina propria B lymphocytes stimulated by LPS.

Multicenter Randomized Double-Blind Controlled Clinical Study of Huoxue Tongluo Recipe(活血通络方)External Washing in the Treatment of Chemotherapy-Induced Peripheral Neuropathy

Objective: To objectively evaluate the efficacy of Huoxue Tongluo Recipe(活血通络方) in the treatment of chemotherapy-induced peripheral neuropathy(CIPN).Methods: A multicenter, randomized, double-blind, block, and parallel-controlled study was conducted in which 184 subjects who met the inclusion criteria were randomized into a trial group(92 cases), a control group(46 cases), a placebo group(46 cases) at a 2:1:1 ratio, respectively.Huoxue Tongluo Recipe(活血通络方) granules, Huangqi Guizhi Wuwu Decoction(黄芪桂枝五物汤) instant granules, placebo instant granules were given for hand and foot soaking, with water temperature 38 ℃-45 ℃, soaking for 20 minutes and the intervention time was 14 days.Results:(1) The total effective test drug was 41.30%, the control group was 32.61%, and the placebo group was 32.61%.There was no significant difference in the efficiency of the 3 groups by CMH chi-square test(P>0.05);(2) In the statistics of previous chemotherapy regimens including oxaliplatin or paclitaxel as stratification factors, the overall efficiency of the trial group was significantly different from the other 2 groups.Conclusion: Huoxue Tongluo Recipe(活血通络方) external washing can improve the peripheral nerve injury caused by chemotherapy to some extent, especially in patients with previous chemotherapy regimen including oxaliplatin and paclitaxel.

Effector mechanism of Yunnan medicine “Yiqi Tongluo soup” in alleviating diabetic peripheral neuropathy by promoting autophagy

Objective:This study aimed to evaluate the effector mechanism of Yiqi Tongluo soup in alleviating diabetic peripheral neuropathy.Methods:Thirty male SD rats were divided into the control group,mode1 group,and Yiqi Tongluo group.Each group consists of 10 rats.Except for the normal group,a high-sugar and high-fat diet combined with streptozocin injection were administered to the groups to constnuct the type 2 diabetes mellitus model.Then,6.48 g/kg of Yiqi Tongluo soup was administered to the Yiqi Tongluo group every day by gavage.An equal volume of physiological saline was administered to the control and model groups every day by gavage.During the experiment,changes in blood glucose and nerve conduction velocity were monitored in all groups.After the experiment,hematoxylin and eosin and luxol fast blue staining were carried out,and morphological changes in the sciatic nerve and myelin sheath in all groups were observed.Western blot was used to observe changes in expression levels of autophagy related proteins,Beclin-1,LC3,and P62.Results:Blood glucose significantly increased,and nerve conduction velocity significantly decreased in the model group.Yiqi Tongluo soup significantly decreased blood glucose level while simultaneously increasing nerve conduction velocity.Hematoxylin and eosin and luxol fast blue myelin sheath staining results showed that Yiqi Tongluo soup could improve sciatic nerve disorder,irregular myelin sheath morphology,and demyelination in type 2 diabetes mellitus rats.Western blot results showed that Beclin-1 and LC3I/LC3I expression levels in the model group significantly decreased,while P62 level significantly increased.Compared with the model group,Beclin-1 and LC3II/LC3I expression levels significantly increased and P62 levels significantly decreased in the Yiqi Tongluo group.Conclusion:Yiqi Tongluo soup can inhibit diabetic peripheral neuropathy by increasing autophagy.

Clinical Efficacy of Ningxin Tongluo Decoction(宁心通络汤)Combined with Dapagliflozin on Patients of Type 2 Diabetes Mellitus Complicated with Ventricular Premature Beats

Objective:To explore the clinical efficacy of Ningxin Tongluo Decoction(宁心通络汤)combined with Dapagliflozin in the treatment of type 2 diabetes mellitus complicated with ventricular premature beats.Methods:Ninety-two patients with type 2 diabetes mellitus complicated with ventricular premature beats in our hospital were randomly divided into control group(n=46)and observation group(n=46).The control group was treated with routine Western medicine,and the observation group was treated with Ningxin Tongluo Decoction on the basis of the control group.The clinical efficacy of the two groups before and after treatment,TCM syndrome score,blood glucose levels[fasting blood glucose(GLU),glycosylated hemoglobin(Hb A1c)],glucose metabolism levels[fasting insulin(FINS),insulin resistance index(HOMA-IR)],the number of ventricular premature beats detected by 24-hour ambulatory electrocardiogram,heart rate turbulence(HRT)[initial oscillation(TO),oscillation slope(TS)],and heart rate variability(HRV)standard deviation of two normal R-R intervals(SDNN),standard deviation of average normal R-R intervals every 5 minutes(SDANN)and root mean square of adjacent normal R-R intervals(RMSSD)were observed.Results:After treatment,the total effective rate of the observation group was 89.13%,significantly higher than that of the control group(73.91%),and the difference was statistically significant(P<0.05).Both groups could reduce TCM syndrome score,blood glucose level,glucose metabolism level,reduce the number of ventricular premature beats,improve HRT and HRV levels,and the observation group was superior to the control group,and the difference was statistically significant(P<0.05).Conclusion:Ningxin Tongluo Decoction combined with Dapagliflozin is effective in treating type 2 diabetes mellitus patients with ventricular premature beats.It can reduce blood glucose level,improve HRV and HRT,improve autonomic nerve function and exert antiarrhythmic effect.

Clinical Study on Application of Xinrun Tongluo Method Based on the Theory of Collateral Diseases in Treating Androgenic Baldness of Blood Heat Wind Dryness Syndrome

[Objectives]To explore the intervention effect of the representative formula of Xinrun Tongluo method,Liangxue Xiaofeng Powder,on the incidence of androgenic alopecia in the syndrome of blood heat and wind dryness.[Methods]A total of 72 patients with androgenic alopecia in Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine from October,2022 to June,2023 were randomly divided into a control group(36 cases,treated with Western medicine)and a treatment group(36 cases,treated with Chinese herbal formula+Western medicine).The short-term and long-term efficacy of the two groups of patients was compared.[Results]The hair microscopic signs and short-term and long-term efficacy of the treatment group were significantly better than those of the control group before and after treatment,with a statistically significant difference(P<0.05).[Conclusions]The representative formula of Xinrun Tongluo method is Liangxue Xiaofeng Powder,which has better clinical efficacy as an auxiliary Western medicine in the treatment of androgenic alopecia patients with blood heat and wind dryness syndrome,and is worthy of further promotion and application in clinical practice.

Study on Preparation Process and Quality Standard of Tianlu Tongluo Granules

[Objectives]This study was conducted to optimize the preparation process of Tianlu Tongluo Granules and establish its quality standard.[Methods]Tianlu Tongluo Granules was prepared by an orthogonal test.TLC was used to identify Radix Astragali preparata,Radix Paeoniae Alba,Scutellariae Radix and Acori Tatarinowii Rhizoma in the granules,and the content of paeoniflorin in the finished product was determined by HPLC.[Results]The extraction process of Tianlu Tongluo Granules was as follows:adding 6 times of water,and decocting twice,1 h/time.In the forming process,the optimal ratio of thick paste to excipients as 10∶8.6,and the optimal ratio of excipients dextrin:starch:colloidal silicon dioxide was 6.5∶0.5∶1.6.The results showed that TLC spots were obvious,the separation was significant,and the specificity was strong.The paeoniflorin content had a good linear relationship in the concentration range of 5.9048-590.48 mg/L(r=0.9999).The average sample recovery was 101.10%,and the RSD was 1.18%.[Conclusions]The preparation process is stable and reasonable,and the established quality control method is specific and reproducible,and can be used for the quality control of Tianlu Tongluo Granules.

Effect of Tongluo Shenggu Prescription on ARCO Stage Ⅱ non-traumatic femoral head necrosis: A prospective study

Objective:To observe the clinical results and therapeutic characteristics of Tongluo Shenggu Decoction in the treatment of ARCOⅡnon-traumatic femoral head necrosis.Methods:120 patients with ARCOⅡnon-traumatic femoral head necrosis from January 2016 to January 2017 were studied by prospective matched and controlled design.60 cases in the treatment group were treated with Tongluo Shenggu Formula.The control group(60 cases)received high energy extracorporeal shock wave therapy.The imaging stability rate,Harris score and SF-36 score were used as clinical efficacy indicators,followed up for 24 months,and the follow-up results were statistically analyzed.Results:Among the 120 patients,5 fell off and the rest were followed up.The imaging stability rate is heavy.The imaging stability rate of the patients in the treatment group is 78.95%,while that in the control group is 72.41%.The two groups have the same stability rate(P>0.05).In Harris score,the Harris score of the patients in the treatment group was 92.678.45 points,significantly higher than that of the control group(80.3924.65 points),the difference was statistically significant(p<0.05),and the pain degree,joint function and joint deformity score of the treatment group were better than those of the control group(p<0.05).In the SF-36 score,the scores of physiological function,physiological function,physical pain,overall health,social function and emotional function in the treatment group were significantly higher than those in the control group,the difference was statistically significant(p<0.05).the incidence of adverse events in the treatment period of the two groups was lower and the patients could relieve themselves in the later stage.Conclusion:Tongluo Shenggu prescription has definite curative effect in treating ARCOⅡnon-traumatic femoral head necrosis,especially in improving hip joint function,relieving hip joint pain and improving quality of life.To observe the clinical results and therapeutic characteristics of Tongluo Shenggu Decoction in the treatment of ARCOⅡnon-traumatic femoral head necrosis.Methods:120 patients with ARCOⅡnon-traumatic femoral head necrosis from January 2016 to January 2017 were studied by prospective matched and controlled design.60 cases in the treatment group were treated with Tongluo Shenggu Formula.The control group(60 cases)received high energy extracorporeal shock wave therapy.The imaging stability rate,Harris score and SF-36 score were used as clinical efficacy indicators,followed up for 24 months,and the follow-up results were statistically analyzed.Results:Among the 120 patients,5 fell off and the rest were followed up.The imaging stability rate is heavy.The imaging stability rate of the patients in the treatment group is 78.95%,while that in the control group is 72.41%.The two groups have the same stability rate(P>0.05).In Harris score,the Harris score of the patients in the treatment group was 92.678.45 points,significantly higher than that of the control group(80.3924.65 points),the difference was statistically significant(p<0.05),and the pain degree,joint function and joint deformity score of the treatment group were better than those of the control group(p<0.05).In the SF-36 score,the scores of physiological function,physiological function,physical pain,overall health,social function and emotional function in the treatment group were significantly higher than those in the control group,the difference was statistically significant(p<0.05).the incidence of adverse events in the treatment period of the two groups was lower and the patients could relieve themselves in the later stage.Conclusion:Tongluo Shenggu prescription has definite curative effect in treating ARCOⅡnon-traumatic femoral head necrosis,especially in improving hip joint function,relieving hip joint pain and improving quality of life.

Mechanism of Bushen Tongluo granule on osteoarthritis based on network pharmacology and molecular docking technology

Objective:In this study,we used network pharmacology and molecular docking technology to analyze the mechanism of Bushen Tongluo granule in the treatment of osteoarthritis.Methods:The main active components and corresponding targets of Bushen Tongluo granule were screened from thetraditional Chinese medicine systems pharmacology database.The targets related to osteoarthritis were collected from the Online Mendelian Inheritance in Man,Therapeutic Target database,GeneCards,Pharmacogenomics Knowledgebases and Drugbank databases.Cytoscape3.9.0 software was used to construct the action network diagram of“Bushen Tongluo Granule-Active Component-Target”.Builded a protein-protein interaction network from the STRING database.The Bioconductor platform and R language 4.0.3 were used for Gene Ontologyfunction andKyoto Encyclopedia of Genes and Genomespathway enrichment analysis.Then,selected the pathway most associated with osteoarthritis for specific analysis.Finally,the core genes were screened and verified by molecular docking using AutoDockTools software.Results:71 principal components of Bushen Tongluo granule and 183 potential therapeu-tic targets for osteoarthritis were obtained.Twenty-eight key targets of Bushen Tongluo granulein the treatment of osteoarthritis are enriched in 158 pathways.Among them,the tumor necrosis factor signaling pathway,interleukin-17 signaling pathway,Toll-like receptor signaling pathway,T helper cell 17 cell differentiation and hypoxia-inducible factor-1signaling pathway involved in key targets are closely related to osteoarthritis.The relevant vital targets were involved in the regulation of DNA transcription factor activity,the response to chemical stress,the response to reactive oxygen species,the response to oxidative stress,the proliferation of muscle cells,the proliferation of epithelial cells and the biological processes such as responses to lipopolysaccharides,responses to molecules of bacterial origin.Molecular docking showed that protein kinase B 1-β-sitosterol,the tumor necrosis factor-naringenin,interleukin-6-luteolin,mitogen-activated protein kinase-quercetin,vascular endothelial growth factor A-quercetin and prostaglandin-endoperoxide synthase 2-luteolin have strong docking activities.Conclusions:Bushen Tongluo granule can reduce the inflammatory response and inhibit articular cartilage angiogenesis in the treatment of osteoarthritis,which may be achieved by regulating the inflammatory signaling pathway and hypoxia-inducible factor-1 signaling pathway.

Tongluo Jiedu as an adjuvant therapy for oral cancer

Oral cancer is one of the malignant neoplasms that present major global health challenge.It is the sixth most prevalent type of cancer in the world,with a high incidence and mortality rate.This letter is a review of the study by Yin et al which was published in the World Journal of Clinical Cases(2024).The study evaluated the effect of Tongluo Jiedu as an adjuvant treatment for oral cancer.Over the years,there has been a continuous search for effective and less invasive treatments for oral cancer.This article emphasizes and discusses various therapeutic options currently available,and it highlights that early intervention and multidisciplinary management are crucial for improving outcomes.Traditional Chinese medicine,particularly Tongluo Jiedu,presents potential complementary approach to co-nventional oral cancer therapies.Future research on Tongluo Jiedu should be focused on validation of its efficacy and safety through large,well-designed clinical trials,as well as better understanding of the molecular mechanisms in-volved,and optimization of therapeutic combinations.Additionally,continuous education of health professionals is key to the effective and safe integration of this traditional medicine into clinical practice.Continuous research is essential for optimization of therapeutic strategies and for addressing the challenges presented by this neoplasm.

温肾通络止痛方对小鼠H型骨微血管内皮细胞/骨髓间充质干细胞共培养体系的影响

背景:骨骼依赖血管与骨细胞之间的密切连接来维持完整性,骨骼处在生理低氧环境中,因此在低氧环境下研究血管生成与骨生成具有重要意义。目的:探究在低氧环境下温肾通络止痛方对H型骨微血管内皮细胞/骨髓间充质干细胞共培养体系的影响及相关机制。方法:运用酶消化法及流式分选技术,分选小鼠H型血管特异型骨微血管内皮细胞;应用骨髓贴壁法分离获取小鼠骨髓间充质干细胞;采用Transwell小室以及缺氧培养工作站建立两种细胞低氧共培养体系,使用50,100μg/m L质量浓度温肾通络止痛方冻干粉进行干预;通过划痕迁移实验与管形成实验评估共培养体系内H型骨微血管内皮细胞的成血管功能;通过碱性磷酸酶染色与茜素红染色评估共培养体系内骨髓间充质干细胞的成骨分化能力;使用Western Blotting与q-PCR检测共培养体系内H型骨微血管内皮细胞中PDGF/PI3K/AKT信号轴相关分子的蛋白表达及mRNA表达。结果与结论:(1)与常氧组相比,低氧组H型骨微血管内皮细胞划痕迁移率及新生血管长度均显著升高(P<0.05),骨髓间充质干细胞成骨分化能力增强(P<0.05);PDGF/PI3K/AKT信号轴相关分子的蛋白、mRNA表达升高(P<0.05);(2)与低氧组相比,经不同质量浓度温肾通络止痛方干预后H型骨微血管内皮细胞划痕迁移率及新生血管长度进一步提高(P<0.05),骨髓间充质干细胞成骨分化能力更强(P<0.05),PDGF/PI3K/AKT信号轴相关分子的蛋白、mRNA表达也进一步升高(P<0.05)。结果表明:低氧条件下H型血管生成及骨生成可能与PDGF/PI3K/AKT信号轴有关,而温肾通络止痛方可能通过激活PDGF/PI3K/AKT信号轴促进“H型血管生成-骨生成”作用从而防治骨质疏松症。

Tongluo Digui decoction(通络地龟汤)treats renal injury in diabetic rats by promoting autophagy of podocytes

OBJECTIVE:To investigate the effects of Tongluo Digui decoction on renal injury and streptozotocin-induced podocyte autophagy in diabetic rats.METHODS:Male Sprague-Dawley rats were randomly divided into six groups:normal,model,Tongluo Digui decoction(high,medium,and low dose)and valsartan.Streptozotocin was injected intraperitoneally to replicate the diabetic animal model.After 8 weeks,proteinuria was evaluated to establish the diabetic nephropathy model.Treatments were administered daily via the intragastric route.At 16 weeks after gavage,we determined 24 h urine protein concentration,and blood glucose,serum creatinine,and urea nitrogen concentrations.Then,rats were sacrificed,and kidneys were harvested and stained with periodic acid-Schiff to evaluate the pathological changes in glomeruli,including glomerular podocytes by transmission electron microscopy.Western blot analysis was used to determine the expression of nephrin,podocin,p62,beclin-1,LC3Ⅱ/Ⅰ,and p-m TOR/m TOR protein in kidney tissues.RESULTS:Compared with the model group,Tongluo Digui decoction was associated with decreases in 24 h urine protein concentration,and blood glucose,hemoglobin A1 c,serum creatinine,urea nitrogen concentrations,total serum protein and albumin.Concurrently,mesangial mesenteric broadening and fusion of foot processes were reduced,the glomerular basement membrane was not significantly thickened,and the number of podocytes and the number of autophagosomes in the podocytes was increased.Further,expression of nephrin,podocin,LC3Ⅱ,and beclin-1 protein in kidney tissue was up-regulated,while expression of p62 protein was down-regulated and m TOR phosphorylation was inhibited.CONCLUSION:Tongluo Digui decoction may inhibit the progression of diabetic nephropathy by inhibiting m TOR phosphorylation,thereby increasing autophagy to protect podocytes and reducing proteinuria.

Effects of Huatan Tongluo decoction on vascular endothelial growth factor receptor 2 expression in synovial tissues of rats with collagen-induced arthritis

OBJECTIVE: To determine the therapeutic effect and potential mechanism of Huatan Tongluo decoction on rats with collagen-induced arthritis.METHODS: Forty specific pathogen-free Wistar rats were selected, and 10 were randomly selected as the control(group 1). The remaining rats were injected intradermally with emulsified type Ⅱ bovine collagen at the tail base and back, followed by a booster 7 d post first immunization. After establishing collagen-induced arthritis(CIA), rats were randomly divided into three groups(n = 10). The rats were treated orally for 30 d as follows: group 1, saline; group 2, model(saline); group 3, tripterygium polyglycoside(TP; 7.81 mg/kg, positive control);group 4, Huatan Tongluo decoction(HTTL; 7.5 g/kg). Body weight, ankle swelling and arthritis index were measured over the course of the study. The rats were sacrificed 30 d after treatment. Morphological changes in the synovium were observed by hematoxylin and eosin staining. Pannus formation and synovial thickness in the left ankle were observed by color Doppler ultrasoundVascular endothelial growth factor(VEGF) and VEGFR2 protein levels were measured by immunohistochemistry.VEGF/VEGFR2 mRNA levels were measured by real-time quantitative polymerase chain reaction.RESULTS: Compared with the model group, a significantly lower arthritis index was observed in the positive control group(P < 0.05) and HTTL group(P < 0.01), after treatment. Both positive control and HTTL reduced intra-articular pannus formation and synovial thickening. Furthermore, VEGF mRNA,and VEGFR2 protein and mRNA levels were significantly downregulated(P < 0.05) in the treatment groups.CONCLUSION: Inhibition of the expression of VEGF and VEGFR2 in synovial tissues and the formation of pannus and synovial hyperplasia may be part of the mechanism of HTTL for relieving the symptoms of rheumatoid arthritis in CIA rats.

Efficacy of Getong Tongluo Capsule(葛酮通络胶嚢)for Convalescent-Phase of Ischemic Stroke and Primary Hypertension:A Multicenter,Randomized,Double-Blind,Controlled Trial

Objective:To evaluate whether the efficacy of Getong Tongluo Capsule(葛酮通络胶囊,GTC,consisted of total flavone of Radix Puerariae)on improving patients'quality of life and lowering blood pressure are superior to the extract of Ginkgo biloba(EGB)for patients with convalescent-phase ischemic stroke and primary hypertension.Methods:This randomized,positive-drug-and placebo-controlled,double-blind trial was conducted from September 2015 to October 2017.Totally 477 eligible patients from 18 hospitals in China were randomly assigned in a 2:1:1 ratio to the following interventions,twice a day for 12 weeks:(1)GTC 250 mg plus EGB-matching placebo 40 mg(237 cases,GTC group),(2)EGB 40 mg plus GTC-matching placebo 250 mg(120 cases,EGB group)or(3)GTC-matching placebo 250 mg plus EGB-matching placebo 40 mg(120 cases,placebo group).Moreover,all patients were orally administered aspirin enteric-coated tablets 100 mg,once a day fo r 12 weeks.The primary outcome was the Barthel Index(Bl).The secondary outcomes included the control rate of blood pressure and National Institutes of Health Stroke Scale(NIHSS)scores.The incidence and severity of adverse events(AEs)were calculated and assessed.Results:The Bl relative independence rates,the clinical recovery rates of NIHSS,and the total effective rates of NIHSS in the GTC and EGB groups were significantly higher than the placebo group at 12 weeks after treatment(P<0.05),and no statistical significance was found between the GTC and EGB groups(P>0.05).The control rate of blood pressure in the GTC group was significantly higher than the EGB and placebo groups at 12,18 and 24 weeks after treatment(P<0.01).There were no statistically significant differences in the incidences of AEs,adverse drug reactions,or serious AEs among the 3 groups(P>0.05).Conclusion:GTC exhibited significant efficacy in improving patients'quality of life as well as neurological function and controlling hypertension.

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

Objective:The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule(LTC)on ischemic stroke rats.Methods:Twenty-six rats were randomly divided into four groups,including sham group,sham+LTC group,MCAO group,and MCAO+LTC group.Ischemic stroke rats were simulated by middle cerebral artery occlusion(MCAO),and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days.LTC therapy was validated in terms of neurobehavioral abnormality evaluation,cerebral infarct area,and histological assessments.The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis,aiming to finding the molecules in respond to the surgery or LTC treatment.Results:Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats.The interstitial edema,atrophy,and pyknosis of glial and neuronal cells occurred in the infarcted area,core area,and marginal area of cerebral cortex were improved after LTC treatment.A total of 13 potential biomarkers were observed,and Youden index of 11 biomarkers such as LysoPC,SM,and PE were more than 0.7,which were involved in neuroprotective process.The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism,together with nicotinate and nicotinamide metabolism.Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.Conclusion:The present study could provide evidence that metabolomics,as systematic approach,revealed its capacity to evaluate the holistic efficacy of TCM,and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.