Chinese Herbal Formulation Research Articles

Rhein attenuates inflammation through inhibition of NF-κB and NALP3 inflammasome in vivo and in vitro.

Rhein is an important component in traditional Chinese herbal medicine formulations for gastrointestinal disorders, including inflammatory bowel diseases such as ulcerative colitis. In this study, we investigated the beneficial effects of rhein in inflammation models in the transgenic zebrafish line TG (corolla eGFP), in which both macrophages and neutrophils express eGFP and RAW264.7 macrophages. We found that the tail-cutting-induced migration of immune cells was significantly reduced in transgenic zebrafish treated with rhein. In addition, the production of proinflammatory cytokines, including IL-6, IL-1β, and tumor necrosis factor-α, were significantly reduced in lipopolysaccharide (LPS)-induced RAW264.7 macrophages treated with rhein. Parallel to the inhibition of proinflammatory cytokines, rhein significantly reduced phosphorylation levels of NF-κB p65 and inducible nitric oxide synthase, as well as COX-2 protein expression levels. Furthermore, rhein significantly reduced NALP3 and cleaved IL-1β expression in LPS + ATP-induced RAW264.7 macrophages. Thus, the present study demonstrates that rhein may exhibit its anti-inflammatory action via inhibition of NF-κB and NALP3 inflammasome pathways.

Abstract P4-13-04: Efficacy of dipsacus asperoides (DA) in a model for triple negative breast cancer

Abstract Background: The triple negative breast cancer (TNBC) is characterized by the absence of estrogen receptor-α (ER-α), Progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER-2). This molecular subtype responds only to conventional cytotoxic chemotherapy and to small molecule based targeted therapy. These therapeutic options are associated with long-term systemic toxicity and acquired tumor resistance leading to compromised efficacy. These limitations emphasize identification of efficacious non-toxic agents for secondary prevention/therapy of TNBC. Dipsacus asperoides (DA) is a Chinese herb of a nutritional nature. The root of this herb represents an ingredient that is commonly included in the traditional Chinese herbal formulations for health management purposes. The present study examines the growth inhibitory effects of DA and identifies possible mechanistic leads for their efficacy in a preclinical cell culture model for TNBC. Experimental Model, Herbal Extract and Biomarkers: The human mammary carcinoma derived ER-α -, PR - and HER-2 - MDA-MB-231 cell linerepresents the model for TNBC. Non-fractionated aqueous extract from DA represents the test agent. Anchorage dependent growth, anchorage independent (AI) colony formation, cell cycle progression and relevant pathway specific mechanistic assays represent the quantitative biomarkers for efficacy. Results: Treatment of MDA-MB-231 cells with DA extract induces a dose dependent cytostatic growth arrest (IC50: 0.0015%, IC90: 0.0030%), and strongly inhibits AI colony formation. Within the cytostatic concentration range, DA treatment inhibits cell cycle progression via progressive G2/M arrest, suggesting abrogation of the G2/M checkpoint affecting DNA damage repair pathway. The induction of cellular apoptosis by DA is indicated by a dose dependent up-regulation of pro-apoptotic Caspase 3/7 activity. Conclusion: These data identify potential mechanistic leads for the efficacy of DA in the present model. The present study validates a mechanism based approach to prioritize efficacious non-toxic herbal extracts for secondary prevention/therapy of the TNBC molecular subtype of clinical breast cancer. Citation Format: Telang NT, Nair HB, Wong GYC. Efficacy of dipsacus asperoides (DA) in a model for triple negative breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P4-13-04.

Pharmacokinetic interaction of aconitine, liquiritin and 6-gingerol in a traditional Chinese herbal formula, Sini Decoction

1. This study aimed to investigate the pharmacokinetic interaction of the three ingredients in a traditional Chinese herbal formulation, Sini Decoction, and provide evidence for its compatibility mechanism.2. First, the effect of liquiritin and 6-gingerol on the pharmacokinetic parameters of aconitine was investigated in rats by using a sensitive and reliable LC–MS/MS method. Then the Caco-2 cell monolayer model and Rhodamine-123 uptake assay were used to investigate the effect of liquiritin and 6-gingerol on the absorption of aconitine and the activity of P-gp.3. The Cmax of aconitine increased significantly (p < 0.05) from 10.34 ± 1.99 to 17.68 ± 2.65 ng/mL with the pretreatment of liquiritin (20 mg/kg), and to 17.43 ± 0.96 ng/mL with 6-gingerol (20 mg/kg). When aconitine was co-administered with liquiritin and 6-gingerol, the Cmax and AUC(0–t) of aconitine increased approximately twofold, and while t1/2 only increased 1.2-fold. The Caco-2 cell monolayer model and Rhodamine-123 uptake assay indicated that both liquiritin and 6-gingerol could increase the absorption of aconitine by inhibiting the activity of P-gp.4. These results indicated that both liquiritin and 6-gingerol could promote the absorption of aconitine and increase its drug concentration in blood by inhibiting the activity of P-gp, and it could also provide evidence for compatibility mechanism of the traditional Chinese herbal formula, Sini Decoction.

Stephania Tetrandra and Ginseng-Containing Chinese Herbal Formulation NSENL Reverses Cisplatin Resistance in Lung Cancer Xenografts.

Chinese Herbal Formulation, supplement energy and nourish lung (SENL), effectively enhances chemotherapeutic efficacy in lung cancer treatment and reverses multi-drug resistance (MDR) in lung cancer cells in vitro. The present study is designed to assess the effect of a New SENL (NSENL, modification of SENL) formulation on resistance to chemotherapy of cisplatin (DDP)-resistant human lung cancer cell line (A549/DDP) xenografts in nude mice. We assessed six constituents in NSENL by high performance liquid chromatography (HPLC). BALB/c nude mice harboring A549/DDP cell xenografts were established to assess the antitumor effect of NSENL and its impact on the expression of MDR related genes. The six constituents in NSENL, including ginsenoside Rg1, ginsenoside Rb1, ginsenoside Rg3, astragaloside IV, ophiopogonin D and tetrandrine were quantitated simultaneously by HPLC. The combination of NSENL with DDP significantly inhibited tumor growth at a rate of up to 66.8% ([Formula: see text]). In addition, NSENL as monotherapy or combined with DDP downregulated multidrug resistance-associated protein 1 (MRP1), basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor 1 (FGFR1) at both the mRNA and protein levels ([Formula: see text]), reduced glutathione S-transferase π (GST-π) protein expression and tumor microvascular density as well as decreased phosphorylation of protein kinase B (Akt) and mammalian target of rapamycin (mTOR) ([Formula: see text]). These findings demonstrated that NSENL can reverse MDR in A549/DDP cells in vivo, an effect possibly associated with downregulation of MDR-associated genes as well as inhibition of bFGF/FGFR and phosphatidylinositol 3-kinase (PI3K)/Akt/mTOR signaling pathways.

Pharmacokinetic evaluation of Shenfu Injection in beagle dogs after intravenous drip administration

Shenfu Injection (SFI) is a well-defined Chinese herbal formulation that is obtained from red ginseng and processed aconite root. The main active constituents in SFI are ginsenosides and aconitum alkaloids. In this work, ginsenosides (ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Rc) and aconitum alkaloids (benzoylmesaconine and fuziline) were used as the index components to explore the pharmacokinetic behavior of SFI. A selective and sensitive HPLC–MS/MS method was developed for the quantification of ginsenosides and aconitum alkaloids in dog plasma and was used to characterize the pharmacokinetics of the five index components after intravenous drip of three different dosages of SFI in beagle dogs. The pharmacokinetic properties of the index components were linear over the dose range of 2–8mL/kg.

Angiogenesis in Ischemic Stroke and Angiogenic Effects of Chinese Herbal Medicine.

Stroke is one of the major causes of death and adult disability worldwide. The underlying pathophysiology of stroke is highly complicated, consisting of impairments of multiple signalling pathways, and numerous pathological processes such as acidosis, glutamate excitotoxicity, calcium overload, cerebral inflammation and reactive oxygen species (ROS) generation. The current treatment for ischemic stroke is limited to thromolytics such as recombinant tissue plasminogen activator (tPA). tPA has a very narrow therapeutic window, making it suitable to only a minority of stroke patients. Hence, there is great urgency to develop new therapies that can protect brain tissue from ischemic damage. Recent studies have shown that new vessel formation after stroke not only replenishes blood flow to the ischemic area of the brain, but also promotes neurogenesis and improves neurological functions in both animal models and patients. Therefore, drugs that can promote angiogenesis after ischemic stroke can provide therapeutic benefits in stroke management. In this regard, Chinese herbal medicine (CHM) has a long history in treating stroke and the associated diseases. A number of studies have demonstrated the pro-angiogenic effects of various Chinese herbs and herbal formulations in both in vitro and in vivo settings. In this article, we present a comprehensive review of the current knowledge on angiogenesis in the context of ischemic stroke and discuss the potential use of CHM in stroke management through modulation of angiogenesis.

Synergistic anti-liver fibrosis actions of total astragalus saponins and glycyrrhizic acid via TGF-β1/Smads signaling pathway modulation

Ethnopharmacological relevanceHuangqi decoction (HQD) is a well-known traditional Chinese herbal formulation, It is an effective treatment for consumptive disease and chronic liver diseases. It consists of Radix Astragali (Astragalus membranceus(Fisch.) Bge. Root, Huangqi) and Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch., root and rhizome, Gancao). Total astragalus saponins (AST) is a main component of Radix Astragali and glycyrrhizic acid(GA) is a main component of Radix Glycyrrhizae. Our primary results showed that the combination of AST and GA had an obvious synergistic effect in reducing liver collagen deposition and decreasing serum alanine aminotransferase (ALT) activity in dimethylnitrosamine (DMN)-induced liver fibrosis. Aim of the studyThrough in vivo and in vitro experiments, we aimed at investigating the key anti-fibrosis signal pathway TGF-β1/Smads to further explore the synergistic mechanism of AST and GA. Material and methodsTwo hepatic fibrosis animal models, bile duct ligation-induced (BDL) and DMN-induced, were utilized. Rats were treated orally with AST, GA or AST/GA, with the effects evaluated via liver histopathology, hydroxyproline (Hyp) levels, and α-SMA expression. In the hepatic stellate cell line JS-1, cells were treated with AST/GA for 24h, followed by a cell viability assessment using Cell Counting Kit-8(CCK-8) and Real-time PCR and Western blot analysis of α-SMA, ColⅠ and TGF-β1/Smads signaling pathway related components. ResultsThe AST/GA combination attenuated liver tissue inflammation, collagen deposition, Hyp levels, and α-SMA expression in both BDL-and DMN-stimulated hepatic fibrosis rats. In vitro results showed that the AST/GA combination significantly inhibited JS-1 cell viability, significantly suppressed α-SMA, ColⅠ, TGF-β1, Smad2 and Smad3 mRNA and protein expression, as well reduced p-Smad2/3. Compared with AST or GA treatment alone, the AST/GA combination significantly reduced Smad3 mRNA expression levels and TGF-β1, Smad3, and p-Smad2/3 protein levels. ConclusionsAST and GA synergistically alleviated both BDL-and DMN-induced hepatic fibrosis via TGF-β1/Smads signaling pathway inhibition in hepatic stellate cells.

Screening of effective ingredients of Yinzhihuang injection against cholestasis and their mechanism

Chinese herbal medicinal formulation Yinzhihuang injection is widely used in clinic for jaundice and chronic liver diseases in eastern Asian countries. However, the pharmacologically active components and the underlying mechanism are unclear. In this study, 30 male ICR mice were randomly assigned into 6 groups： normal control, model control, chlorogenic acid group, geniposide group, baicalin group and wogonoside group. The liver function, liver pathological changes and bile acid metabolism-related gene expression in mice were assayed. The serum levels of ALT, AST, ALP, TBA in chlorogenic acid group (15.89±2.53), (18.32±2.56), (26.38±9.87) U•L⁻¹, (40.63±7.67) μmol•L⁻¹, respectively and geniposide group (20.54±2.36), (24.28±5.19), (35.09±5.03) U•L⁻¹, (42.86±7.11) μmol•L⁻¹, respectively were lower than those in the model control group (59.52±10.94), (128.37±17.97),(169.52±9.62) U•L⁻¹, (132.50±33.00) μmol•L⁻¹, respectively. Hematoxylin-eosin staining showed necrosis, infiltration of inflammation cell in chlorogenic acid group and geniposide group were milder than those in the model control group. Q-PCR analysis revealed the expression of bile acid metabolism-related genes was normalized after treatment with chlorogenic acid or geniposide. Chlorogenic acid and geniposide improved the liver injury and cholestasis effectively, and reversed the mRNA expression of bile acid metabolism-related genes induced by ANIT. So, chlorogenic acid and geniposide were protective for cholestasis, suggesting their pharmacodynamic effect in Yinzhihuang injection.

Abstract P3-09-04: Growth inhibitory effects of Cornus officinalis on a model for triple negative breast cancer

Abstract Background: The triple negative breast cancer (TNBC) molecular subtype is characterized by the absence of estrogen receptor-α (ER-α), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER-2) expressions. TNBC is refractory to endocrine and to HER-2 selective therapy, while options for the treatment of choice involving high dose Anthracyclin/Taxol based conventional chemotherapy and/or small molecule inhibitor based targeted therapy are associated with systemic toxicity and acquired tumor resistance, leading to compromised efficacy. These limitations emphasize a need to identify efficacious non-toxic agents for secondary prevention/therapy of TNBC. Cornus officinalis (CO) is a Chinese herb of a nutritional nature in the form of a fruit. It is a major ingredient herb in some well-known Chinese herbal formulations for health management purposes. Growth inhibitory efficacy of CO in a model for ER positive Luminal A molecular subtype of clinical breast cancer is associated with abrogation of estradiol promoted growth via generation of anti-proliferative estradiol metabolites [Telang et al: Mol. Med. Rep. 5: 20-28, 2012].The present study examines the growth inhibitory effects of CO, and identifies possible molecular targets for its efficacy in a pre-clinical cell culture model for TNBC. Experimental model, Herbal Extract and Biomarkers: The human mammary carcinoma-derived ER-α negative, PR negative and HER-2 negative MDA-MB-231 cell line represents the model for TNBC. Non-fractionated aqueous extract from CO represents the test agent. Anchorage dependent growth, anchorage independent (AI) colony formation, cell cycle progression, cellular apoptosis and relevant pathway specific mechanistic assays represent the biomarkers for efficacy. Results: Treatment of MDA-MB-231 cells with CO extract induces substantial dose dependent cytostatic growth arrest (IC50:0.1%; IC90:0.5%), and strongly inhibits AI colony formation. Within the cytostatic range CO inhibits G1 to S phase transition leading to G1 arrest that is accompanied by decreased expressions of Cyclin D1 and p-RB. Cellular apoptosis induced by CO is associated with modulation of the BAX-BCL-2 pathway, and up-regulation of the pro-apoptotic Caspase 3/7 activity. Conclusions: These data provide a mechanistic evidence for efficacy of CO as a naturally occurring nutritional substance in a cell culture model for TNBC. The present study validates a mechanism based approach to prioritize efficacious non-toxic herbal extracts for secondary prevention/therapy of the TNBC molecular subtype of clinical breast cancer. Citation Format: Telang NT, Nair HB, Wong GYC. Growth inhibitory effects of Cornus officinalis on a model for triple negative breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-09-04.

Synergism of Chinese Herbal Medicine: Illustrated by Danshen Compound.

The primary therapeutic effects of Chinese herbal medicine (CHM) are based on the properties of each herb and the strategic combination of herbs in formulae. The herbal formulae are constructed according to Chinese medicine theory: the “Traditional Principles for Constructing Chinese Herbal Medicinal Formulae” and the “Principles of Combining Medicinal Substances.” These principles of formulation detail how and why multiple medicinal herbs with different properties are combined together into a single formula. However, the concept of herbal synergism in CHM still remains a mystery due to lack of scientific data and modern assessment methods. The Compound Danshen Formula (CDF) is a validated formula that has been used to treat a variety of diseases for hundreds of years in China and other countries. The CDF will be employed to illustrate the theory and principle of Chinese herbal medicine formulation. The aim of this review is to describe how Chinese herbal medicinal formulae are constructed according to Chinese medicine theory and to illustrate with scientific evidence how Chinese herbs work synergistically within a formula, thereby supporting Chinese medicine theory and practice.

From "Kidneys Govern Bones" to Chronic Kidney Disease, Diabetes Mellitus, and Metabolic Bone Disorder: A Crosstalk between Traditional Chinese Medicine and Modern Science.

Although traditional Chinese medicine (TCM) and Western medicine have evolved on distinct philosophical foundations and reasoning methods, an increasing body of scientific data has begun to reveal commonalities. Emerging scientific evidence has confirmed the validity and identified the molecular mechanisms of many ancient TCM theories. One example is the concept of “Kidneys Govern Bones.” Here we discuss the molecular mechanisms supporting this theory and its potential significance in treating complications of chronic kidney disease (CKD) and diabetes mellitus. Two signaling pathways essential for calcium-phosphate metabolism can mediate the effect of kidneys in bone homeostasis, one requiring renal production of bioactive vitamin D and the other involving an endocrine axis based on kidney-expressed Klotho and bone-secreted fibroblast growth factor 23. Disruption of either pathway can lead to calcium-phosphate imbalance and vascular calcification, accelerating metabolic bone disorder. Chinese herbal medicine is an adjunct therapy widely used for treating CKD and diabetes. Our results demonstrate the therapeutic effects and underlying mechanisms of a Chinese herbal formulation, Shen-An extracts, in diabetic nephropathy and renal osteodystrophy. We believe that the smart combination of Eastern and Western concepts holds great promise for inspiring new ideas and therapies for preventing and treating complications of CKD and diabetes.

Effect of the herbal formulation Jianpijiedu on the TCRVβCDR3 repertoire in rats with hepatocellular carcinoma and subjected to food restriction combined with laxative.

The aim of this study was to investigate the effects of the Chinese herbal formulation Jianpijiedu (JPJD) in a rat model of orthotopic hepatocellular carcinoma (OHC). The tumor-bearing rats underwent food restriction combined with laxative (FRL) treatment in order to model the nutritional and digestive symptoms of patients with hepatocellular carcinoma. In addition, the study aimed to elucidate the effect of JPJD on the T cell receptor Vβ-chain complementarity-determining region 3 (TCRVβCDR3) repertoire and the underlying mechanism. The FRL rat model was established by alternate-day food restriction and the oral administration of Glauber's salt (sodium sulfate), based on which the OHC model was then established. Subsequently, the FRL-OHC induced animals received JPJD or thymopentin-5 (TP5) for 17 days. Differences in the TCRVβCDR3 repertoire in the rat thymus, liver and hepatocellular carcinoma tissues were analyzed by polymerase chain reaction. Compared with the FRL-OHC model animals without any treatment, those treated with JPJD exhibited significantly inhibited hepatocellular carcinoma growth (P<0.05), reduced weight loss (P<0.01) and stable visceral indices (P<0.05). Furthermore, the JPJD treatment appeared to improve Simpsons diversity index (Ds) values and the quasi-Gaussian distribution rate of the TCRVβCDR3 repertoire in the thymus, liver and hepatocellular carcinoma tissues. However, no anti-hepatoma effects were evident in the rats treated with TP5. In addition, TP5 increased the Ds values and the quasi-Gaussian distribution rate of the TCRVβCDR3 repertoire in hepatocellular carcinoma tissues compared with those in the JPJD-treated group. The anti-hepatoma effects of JPJD in FRL-OHC-induced animals may be due to the promotion of the Ds values of the TCRVβCDR3 repertoire.

INTERVENTION EFFECT AND DOSE-DEPENDENT RESPONSE OF TANREQING INJECTION ON AIRWAY MUCUS HYPERSECRETION IN LIPOPOLYSACCHARIDE-INDUCED RATS

Background: Tanreqing injection, a Chinese herbal formulation comprising Radix Scutellariae, Fructus Forsythiae, Flos Lonicerae, Antelope horn, and Bear bile powder, has been used to treat bronchitis and pneumonia for many years in China. However, its anti-mucus-hypersecretion mechanism has yet not been fully interpreted. We aim to assess the effect and dose-response relationships of Tanreqing injection on lipopolysacchaide (LPS) induced airway mucus hypersecretion in rats.&#x0D; Material and methods: Forty-eight male rats were randomly divided into four groups (12 per group). A rat model of airway mucus hypersecretion was generated with LPS. Tanreqing injection was given by intratracheal instillation, and bronchoalveolar lavage fluid (BALF) from the right lung was collected. BALF total protein was determined by bicinchoninic acid disodium assay (BCA). Muc5ac was measured using enzyme linked immunosorbent assay (ELISA) and immunohistochemistry. The expression of muc5ac mRNA was detected by real-time polyermase chain reaction (RT-PCR). The middle lobe of the right lung was stained with alcian blue-periodic acid sthiff (AB-PAS) and positive staining relative shading area examined.&#x0D; Results: LPS caused airway mucus hypersecretion, The LPS-induced airway mucus hypersecretion increased beginning at 24 hr, and peaked at 96 hr. Tanreqing injection could inhibit airway mucus hypersecretion, and suppress the expression of muc5ac mRNA.&#x0D; Conclusion: Tanreqing injection inhibits airway mucus hypersecretion in a certain dose-dependent trend.

The efficiency of adjusted-da-chai-ling-tang in radiation-induced brain edema in patients with brain tumors

Background: Brain edema induced by radiotherapy is a common complication in patients with brain tumors, for which medical treatment is the treatment of choice. Adjusted-Da-Chai-Ling-Tang, a Chinese herbal formulation, has been confirmed to be protective against the radiation-induced edema. In this study, we investigated the efficiency of adjusted-Da-Chai-Ling-Tang in radiation-induced brain edema in patients with brain tumors. Materials and Methods: A total of 46 patients with brain tumors treated with radiotherapy alone or combined with surgery were enrolled. These patients were divided into two groups: The experimental group with adjusted-Da-Chai-Ling-Tang and the control group with conventional medical treatment. Clinical data including symptoms and serologic results were collected pretreatment and on the 4^th, 7^th and 10^th day posttreatment. Magnetic resonance imaging of the brain was performed to investigate changes in brain edema. Results: Clinical symptoms including headache, dizziness, nausea/vomiting and fatigue significantly improved in the experimental group (P＜0.05). No difference in serological results was observed. Brain edema was significantly reduced in the experimental group in magnetic resonance imaging (P＜0.05). Conclusion: Adjusted-Da-Chai-Ling-Tang is effective in the treatment of radiation-induced brain edema in patients with brain tumors. No obvious side effects were observed.

Huangqin-Tang Ameliorates TNBS-Induced Colitis by Regulating Effector and Regulatory CD4(+) T Cells.

Huangqin-Tang decoction (HQT) is a classic traditional Chinese herbal formulation that is widely used to ameliorate the symptoms of gastrointestinal disorders, including inflammatory bowel disease (IBD). This study was designed to investigate the therapeutic potential and immunological regulatory activity of HQT in experimental colitis in rats. Using an animal model of colitis by intrarectally administering 2,4,6-trinitrobenzenesulfonic acid (TNBS), we found that administration of HQT significantly inhibited the severity of TNBS-induced colitis in a dose-dependent manner. In addition, treatment with HQT produced better results than that with mesalazine, as shown by improvedweight loss bleeding and diarrhoea scores, colon length, and intestinal inflammation. As for potential immunological regulation of HQT action, the percentages of Th1 and Th17 cells were reduced, but those Th2 and Treg cells were enhanced in LPMCs after HQT treatment. Additionally, HQT lowered the levels of Th1/Th17-associated cytokines but increased production of Th2/Treg-associated cytokines in the colon and MLNs. Furthermore, we observed a remarkable suppression of the Th1/Th17-associated transcription factors T-bet and ROR-γt. However, expression levels of the Th2/Treg-associated transcription factors GATA-3 and Foxp3 were enhanced during treatment with HQT. Our results suggest that HQT has the therapeutic potential to ameliorate TNBS-induced colitis symptoms. This protective effect is possibly mediated by its effects on CD4+ T cells subsets.

Effects of er-miao-san extracts on TNF-alpha-induced MMP-1 expression in human dermal fibroblasts.

BackgroundVarious health benefits have been attributed to Er-Miao-San (EMS), a traditional Chinese herbal formulation that contains equal amounts of cortex phellodendri (Phellodendron amurense Ruprecht) and rhizoma atractylodis (Atractylodes lancea D.C). However, its effect on the anti-inflammatory activity in human dermal fibroblasts (HDFs) and the mechanism underlying this effect are unknown.ResultsThis study investigated the effects of EMS on TNF-α-induced MMP-1 expression in HDFs. Our data show that EMS inhibited TNF-α-induced MMP-1 expression in a concentration-dependent manner. Interestingly, EMS maintained IκB content without inhibiting the phosphorylation of MAPKs, which are well-established upstream kinases of NF-κB. Moreover, EMS reduced the level of nuclear p65 protein in HDFs. Luciferase assay revealed that EMS inhibits the transcriptional activity of NF-κB by stabilizing IκB. Our results show that EMS exerts its anti-inflammatory effect by inhibiting NF-κB-regulated genes such as IL-1β and IL-8. Moreover, EMS effectively inhibited TNF-α-induced expression of MMP-1 via the NF-κB pathway.ConclusionsTaken together, our data suggest that EMS could potentially be used as an anti-inflammatory and anti-aging treatment.

Hepatoprotective effects of a Chinese herbal formulation, Yingchen decoction, on olaquindox-induced hepatopancreas injury in Jian carp (Cyprinus carpio var. Jian).

In order to identify effective hepatoprotective herbs for clinical application in fish farming, 200 mg/kg olaquindox (OLA) was added to a basal diet (group 1, control) to form OLA diet (group 2), then 1.35, 2.7 and 5.4 % (w/w) of a Chinese herbal formulation, Yingchen decoction (YCD), were added to the OLA diet to form three additional diets for groups 3, 4 and 5, respectively. A total of 375 juvenile Jian carp (Cyprinus carpio var. Jian) (52.12 ± 2.95 g/tail) were divided into five groups (triplicates per group) and fed the five diets mentioned above, respectively, for 6 weeks. At the termination of feeding experiment, serum biochemical indexes, viability of hepatocytes and the hepatopancreas microstructure for each group were detected and observed. The results showed that serum ALT and AST in group 2 were significantly higher than the control (P < 0.05). Plasma membranes hepatocyte nuclei in group 2 were found to be mostly indistinct, compared to group 1, and gradually recovered with the increasing supplementation of YCD in group 3, 4 and 5. The viability of isolated hepatocytes in group 2 was the lowest and gradually recovered with the increasing supplementation of YCD in group 3, 4 and 5. The results suggest that YCD protected the Jian carp hepatopancreas against injury from OLA, and that 5.4 % YCD would be the optimum dosage in a Jian carp diet.

Validated high-performance thin-layer chromatographic method for the determination of dibenzyl cyclooctadiene lignans fromSchisandra grandiflora

A simple, accurate, and rapid high-performance thin-layer chromatographic (HPTLC) method has been established and validated for the simultaneous quantification of the four biologically active dibenzyl cyclooctadiene lignans, i.e., schisandrin (1), gomisin B (2), deoxyschisandrin (3), and gomisin N (4) from the hexane extract of Schisandra grandiflora (Wall.) Hook. f. & Thoms. Separation was performed on silica gel 60 F254 plates using a saturated mixture of toluene-ethyl acetate-methanol (6:1:1 v/v) as the mobile phase. Quantitation was performed using densitometric absorption-reflection mode at 225 nm. The method was validated for its selectivity, linearity, precision, and accuracy. HPTLC was hyphenated with a mass spectrometer as an additional tool for the confirmation of the markers using an interface. The developed method is simple and convenient which can be applied for the regular quality analysis of the raw plant material used in Chinese traditional herbal formulations.

Er-Miao-San, a traditional herbal formula containing Rhizoma Atractylodis and Cortex Phellodendri inhibits inflammatory mediators in LPS-stimulated RAW264.7 macrophages through inhibition of NF-κB pathway and MAPKs activation

Ethnopharmacological relevanceEr-Miao-San (EMS) is a traditional Chinese herbal formulation that contains combinations of Rhizoma Atractylodis (RA) and Cortex Phellodendri (CP). It exhibits analgesic and anti-inflammatory activities and have been used for the treatment of various “Bi Zheng” for thousand years in China. The aims of the present study were to investigate the anti-inflammatory activities of EMS and elucidate the underlying mechanisms with regard to its molecular basis of action for the best combination. Materials and methodsThe anti-inflammatory effects of EMS were studied by using lipopolysaccharide (LPS)-stimulated activation of nitric oxide (NO) and pro-inflammatory cytokine production in mouse RAW264.7 macrophages. Expression of inducible NO synthase (iNOS), mitogen-activated protein kinases (MAPKs) phosphorylation, p65 phosphorylation, inhibitor-κBα (IκBα) degradation, and NF-κB DNA-binding activity were further investigated. ResultsThe present study demonstrated that EMS could suppress the production of NO in LPS-stimulated RAW264.7 macrophages. However, CP and RA did not have significant inhibitory effect on them. EMS also inhibited the production of tumor necrosis factor-alpha, interleukin-1 beta and macrophage chemotactic protein-1. Further investigations showed EMS could suppress iNOs expression and p38 phosphorylation. EMS significantly decreased the content of IκBα, reduced the level of phosphorylated p65 and suppressed the NF-κB DNA-binding activity. All these results suggested the inhibitory effects of EMS on the production of inflammatory mediators through the inhibition of the NF-κB pathway. ConclusionsOur results indicated that EMS inhibited inflammatory events and iNOS expression in LPS-stimulated RAW264.7 cells through the inactivation of the MAPK and NF-κB pathway. This study gives scientific evidence validating the use of EMS in treatment of patients with “Bi Zheng” in clinical practice in traditional Chinese medicine.

Development of a microdialysis system to monitor lamivudine in blood and liver for the pharmacokinetic application in herbal drug interaction and the gene expression in rats

The aim of study is to develop a novel multiple microdialysis technique coupled to a validated chromatographic system for the measurement of protein-unbound form lamivudine and investigation of its herb–drug interaction in rat blood and liver. Furthermore, gene expression changes of drug metabolizing enzymes in rat were evaluated by microarray analysis after being treated with a traditional Chinese herbal formulation, Long-Dan-Xie-Gan-Tang (LDXGT). The analyte was separated by a reverse-phase C18 column using the mobile phase comprising methanol and 10mM KH2PO4 (15:85, v/v, adjusted to pH 6.0 with NaOH) with the flow rate of 0.8mL/min, and the UV wavelength was set at 270nm. The processes of method validation followed Food and Drug Administration (FDA) guidelines. The pharmacokinetic data demonstrated that the area under the concentration–time curve (AUC) of the lamivudine alone and the LDXGT pretreated group were 532±37.6 and 550±44.2minμg/mL in rat blood after lamivudine administration (10mg/kg, i.v.) and 682±196 and 642±153minμg/mL in rat liver, respectively. The herb–drug pharmacokinetic interaction showed that with either lamivudine alone or in combination with pretreated with LDXGT, the pharmacokinetic parameters were not significantly changed except the apparent volume of distribution (Vd) at a high dose of lamivudine (30mg/kg). In addition, microarray analysis showed that among 70 altered genes (selection criteria: |Fold change|≧2 and p<0.05), only 11 genes were involved in drug metabolism and indicated that a relatively small portion of drug metabolizing genes in liver were altered at the genome level after the therapeutic dose of LDXGT treatment. In conclusion, these studies provide constructive information to interpret the herb–drug interactions between lamivudine and a popular Chinese herbal formulation.