Runhui Liu

Acylated Iridoids with Cytotoxicity from Valeriana jatamansi

Thirteen new acylated iridoids, jatamanvaltrates A-M (1-13), together with nine known valepotriates (14-22), were isolated from the whole plants of Valeriana jatamansi (syn. Valeriana wallichii). The structures of these new compounds were assigned by detailed interpretation of spectroscopic data. Jatamanvaltrates D (4) and H (9) are the first examples of naturally occurring valepotriates containing an o-hydroxybenzoyloxy moiety at C-10. All isolated compounds were tested for their cytotoxicity against lung adenocarcinoma (A549), metastatic prostate cancer (PC-3M), colon cancer (HCT-8), and hepatoma (Bel7402) cell lines.

Huang-Lian-Jie-Du-Tang exerts anti-inflammatory effects in rats through inhibition of nitric oxide production and eicosanoid biosynthesis via the lipoxygenase pathway

Objectives Huang‐Lian‐Jie‐Du‐Tang (HLJDT) is a traditional Chinese medicine with a long history of anti‐inflammatory use, but its pharmacological effects have not been thoroughly investigated. This study aimed to evaluate the anti‐inflammatory activity of HLJDT in vivo and in vitro.

Metabolomic analysis of biochemical changes in the plasma and urine of collagen-induced arthritis in rats after treatment with Huang-Lian-Jie-Du-Tang

ETHNOPHARMACOLOGICAL RELEVANCE Huang-Lian-Jie-Du-Tang (HLJDT oren-gedoku-to in Japanese), a classical traditional Chinese medicine (TCM) formula, is well known for the treatment of inflammatory-related diseases such as gastritis, dermatitis, and ulcerative colitis. Our previous studies have indicated that HLJDT has therapeutic potential in rheumatoid arthritis treatment. To investigate the therapeutic mechanism of a traditional Chinese medicine formula Huang-Lian-Jie-Du-Tang (HLJDT oren-gedoku-to in Japanese) and its constituents combination for collagen-induced arthritis in rats using a metabolomics approach. MATERIALS AND METHODS Rats were divided into 9 groups, and drugs were administered from on the day after the onset of arthritis (day 12) until day 31 of the experiment once daily continuously. Urine and plasma were analyzed by reversed-phase liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). Partial least-squares discriminate analysis (PLS-DA) models were built to evaluate the therapeutic effects of HLJDT and its constituents combination. 15 identified CIA biomarkers were investigated to explain its therapeutic mechanism. RESULTS Administration of HLJDT and its constituents combination in CIA rats not only significantly reduced arthritic scores and serum levels of IL-1β but also improved histopathologic changes in joint architecture. Urinary and plasma metabolic profiling revealed that perturbation of energy metabolism, lipid metabolism, oxidative injury and some amino acids metabolism occurred in collagen-induced arthritis (CIA). Our results also indicated that the disturbed urinary levels of succinic acid, citric acid, creatine, uridine, pantothenic acid, carnitine, phenylacetylglycine, allantoin and plasma levels of phenylpyruvic acid in model rats were gradually restored to normal after administration of HLJDT. The treatment of constituents combination of HLJDT group was able to restore to normal the disturbed urinary levels of citric acid, creatine, pantothenic acid, carnitine, pantothenic acid, phenylacetylglycine and plasma levels of uric acid, L-histidine, and l-phenylalanine in model rats. CONCLUSIONS Our study indicates that HLJDT and its constituents combination treatment can ameliorate CIA through partially regulating the perturbed energy metabolism. Our work demonstrated that metabonomics-based approach is a promising new tool to evaluate the therapeutic effects and mechanism of complex TCM prescriptions.

Plasma pharmacochemistry based approach to screening potential bioactive components in Huang-Lian-Jie-Du-Tang using high performance liquid chromatography coupled with mass spectrometric detection

ETHNOPHARMACOLOGICAL RELEVANCE Huang-Lian-Jie-Du-Tang (HLJDT), a classic prescription of traditional Chinese medicine (TCM), has been used in clinical over 1700 years for the treatment of gastrointestinal disorders, cardiovascular diseases and Alzheimer disease. But the active components of HLJDT were ambiguous, which seriously restricted its clinical application. MATERIALS AND METHODS The methodology of plasma pharmacochemistry was applied to screen the bioactive components in HLJDT. A reliable LC/MS system was established for detecting the prototype compounds and metabolites in dosed plasma after oral administration of HLJDT. By comparative analysis of the chemical profiles of HLJDT extracts, blank plasma and dosed plasma, potential bioactive compounds in HLJDT may be discovered. RESULTS By comparing the retention time, MS and MS/MS spectra with those of reference standard and literature data, 30 components including 22 prototype compounds and 8 metabolites from HLJDT were discovered as potential bioactive components in rat plasma. CONCLUSIONS A reliable and effective method was established to screen the potential bioactive components in the formula of HLJDT, which provided useful information for the further study of action mechanism of HLJDT.

The inhibitory activities of the components of Huang-Lian-Jie-Du-Tang (HLJDT) on eicosanoid generation via lipoxygenase pathway.

AIM OF THE STUDY Huang-Lian-Jie-Du-Tang (HLJDT) is a traditional Chinese medicine with anti-inflammatory use. In the present study, the effects of its component herbs and pure components were observed on eicosanoid generation to find out the contributory components and their precise targets on arachidonic acid (AA) cascade. MATERIALS AND METHODS By monitoring leukotriene B(4) (LTB(4)), 5-hydroxyeicosatetraenoic acid (5-HETE), and 12-hydroxy-5,8,10-heptadecatrienoic acid (12-HHT), we compared the effects of HLJDT, HLJDT free of one or two component herbs, and water extract of four single component herbs of HLJDT (Rhizoma coptidis, Radix scutellariae, Cortex phellodendri and Fructus gardeniae) on eicosanoid generation in rat elicited peritoneal macrophages. In addition, thirteen pure compounds from HLJDT (baicalin, baicalein, wogonoside, wogonin, berberine, magnoflorine, phellodendrine, coptisine, palmatine, jateorrhizine, crocin, chlorogenic acid, and geniposide) were tested in the macrophages. Furthermore, the efficacies of these thirteen compounds were evaluated on cell-free purified enzymes: leukotriene A(4) hydrolase (LTA(4)H), 5-, 15-lipoxygenase (5-, 15-LO), and cyclo-oxygenase-1/2 (COX-1/2). Moreover, the possible synergetic effect on LO pathway derived LTB(4) generation between the active components was also tested in rat peritoneal macrophages. RESULTS Our experiments showed that Rhizoma coptidis and Radix scutellariae were responsible for the suppressive effect of HLJDT on eicosanoid generation. Some of the pure components including baicalein, baicalin, wogonoside, wogonin, coptisine, and magnoflorine inhibited eicosanoid generation in rat macrophages via LO pathway of AA cascade. Further experiments on cell-free purified enzymes confirmed that Radix scutellariae derived baicalein and baicalin showed significant inhibition on 5-LO and 15-LO, while Rhizoma coptidis derived coptisine showed medium inhibition on LTA(4)H. On the other hand, no significant inhibition of thirteen components on COX-1/2 was observed. Moreover, the slight synergetic inhibition on LTB(4) between baicalein and coptisine was proved in the rat peritoneal macrophages. CONCLUSIONS Baicalein and coptisine, the active components of HLJDT, for the first time are found to interfere with arachidonic acid cascade via inhibition on different points of LO pathway. This finding makes the mechanism of HLJDT clearer and achieves its safer therapeutic application.

Berberine induces dendritic cell apoptosis and has therapeutic potential for rheumatoid arthritis

OBJECTIVE To investigate the effects of berberine on dendritic cell (DC) apoptosis and its potential as a therapeutic agent in rheumatoid arthritis (RA). METHODS Bone marrow (BM)-derived myeloid DCs (MDCs) and plasmacytoid DCs (PDCs) were generated by culturing BM cells with granulocyte-macrophage colony-stimulating factor/interleukin-4 or flt3L, respectively. Splenic DCs, T cells, and B cells were purified using a magnetic-activated cell sorting system. In vitro apoptosis was assessed by annexin V/propidium iodide or Hoechst 33258 staining. The in vivo effects of berberine were examined in mice with collagen-induced arthritis (CIA). Immune responses against type II collagen (CII) were determined by assaying serum antibody levels, lymphocyte proliferation, and cytokine production. The proportions of DCs and apoptosis of different immune cell subsets in spleens and lymph nodes were analyzed by flow cytometry and immunohistochemistry after subset-specific surface marker labeling and TUNEL staining. RESULTS Exposure of MDCs to berberine during BM cell differentiation reduced cell recovery by inducing apoptosis. Sensitivity to berberine-induced apoptosis was acquired starting on day 3 of DC differentiation, and mature DCs were more sensitive to berberine than immature DCs. Murine peritoneal macrophages, RAW 264.7 cells, and Jurkat cells were insensitive to berberine-induced apoptosis. Splenic DCs were more sensitive to berberine than T and B cells. Susceptibility of PDCs to berberine-induced apoptosis was similar to that of MDCs. In mice with CIA, berberine treatment ameliorated arthritis, suppressed CII-specific immune responses, and selectively increased the incidence of apoptosis in DCs within spleens and lymph nodes. CONCLUSION These findings show that berberine selectively induces apoptosis in DCs. Berberine may thus represent a novel therapeutic agent for RA.

Biomarkers in the early period of acute myocardial infarction in rat serum and protective effects of Shexiang Baoxin Pill using a metabolomic method.

ETHNOPHARMACOLOGICAL RELEVANCE To identify the biomarkers in early period of acute myocardial infarction (AMI) in rat serum and reveal the effective mechanism of a Traditional Chinese Medicine (TCM) named Shexiang Baoxin Pill (SBP). MATERIAL AND METHOD A metabolomic approach using reversed-phase liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was developed. RESULTS Fourteen biomarkers in the early period of acute myocardial infarction (AMI) in rat serum were identified. These biomarkers include 5-methylcytosine, cystathionine ketimine, 2-oxoadipic acid, thymidine, epinephrine, homocystine, uric acid, 12(S)-hydroperoxyeicosatetraenoic acid (12s-HPETE), 11-dehydrocorticosterone, 12(S)-hydroxyeicosatetraenoic acid (12s-HETE), deoxycorticosterone, corticosterone, aldosterone and cortisol. Through pathway analysis of these biomarkers, inflammation, hypertrophy and oxidative injury were considered the most relevant pathological changes in early period of AMI. CONCLUSION Identification of AMI biomarkers not only supplied a systematic view of the progression of AMI in the early period but also provided the theoretical basis for the prevention or treatment of AMI. The results demonstrated that SBP pretreatment could offer protective effects for AMI through regulating the pathway of steroid hormone biosynthesis.

Alkaloids from Corydalis saxicola and Their Anti‐Hepatitis B Virus Activity

Eight protoberberine‐type alkaloids and two indole alkaloids were isolated from the MeOH extracts of the herb Corydalis saxicola Bunting (Papaveraceae). Their structures were identified as dehydrocavidine (1), dehydroapocavidine (2), dehydroisoapocavidine (3), berberine (4), dehydroisocorypalmine (5), coptisine (6), tetradehydroscoulerine (7), berbinium (8), 1‐formyl‐5‐methoxy‐6‐methylindoline (9), and 1‐formyl‐2‐hydroxy‐5‐methoxy‐6‐methylindoline (10). Compounds 3, 9, and 10 are new alkaloids. All compounds were tested for anti‐HBV activity against the 2.2.15 cell line in vitro. Dehydrocavidine (1), dehydroapocavidine (2), and dehydroisoapocavidine (3) exhibited inhibitory activity against HBsAg and HBeAg, but no cytotoxicity against the 2.2.15 cell line.

Chemical constituents of Crinum asiaticum L. var. sinicum Baker and their cytotoxic activities.

A phytochemical investigation of the bulbs of Crinum asiaticum L. var. sinicum Baker resulted in the isolation of two new alkaloids, asiaticumines A and B (1 and 2, resp.), together with 21 known compounds, including nine alkaloids, four amides, five phenolic compounds, and three flavonoids. All 23 compounds were isolated for the first time from Crinum asiaticum L. var. sinicum Baker. Their structures were elucidated by spectroscopic methods. In addition, ten alkaloids, 1–10, were evaluated for their cytotoxic activities against human tumor cell lines A549, LOVO, HL‐60, and 6T‐CEM. Compounds 3, 4, and 7–10 selectively showed remarkable inhibition against one or more of the tested cell lines.

Structural Characterization and Identification of Major Constituents in Jitai Tablets by High-Performance Liquid Chromatography/Diode-Array Detection Coupled with Electrospray Ionization Tandem Mass Spectrometry

In the present study a universally applicable HPLC-DAD/ESI-MS/MS method was developed for carrying out the comprehensive characterization of Jitai tablets (JTT). Based on the ESI-MSn fragmentation patterns of the reference standards, a total of 101 components were identified or tentatively characterized by comparing their retention times, UV and MS spectra with those of reference standards or through the matching of empirical information with those of published components in the in-house library. The characteristic fragmentation pattern of alkaloids, phenolic acids, tanshinones, flavonoid glycosides, cyanogenic glycosides, ginsenosides, 2-(2-phenylethyl) chromones, phthalides and gingerol-related compounds were tentatively elucidated using structurally-relevant product ions. It was observed that neutral losses of C9H10O3 and C9H8O2 were the characteristic product ions of scopola alkaloids. Neutral fragment mandelonitrile was the characteristic ion of cyanogenic glycosides. To our knowledge, tropylium ion and C4H2O unit were the characteristic ions of 2-(2-phenylethyl) chromone, which resulted from the Retro-Diels-Alder (RDA) cleavage of the C ring. The results indicated that the developed analysis method could be employed as a rapid, effective technique for structural characterization of chemical constituents in TCM. This work is expected to provide comprehensive information for the quality evaluation and pharmacokinetic studies of JTT.