Integrated network pharmacology and pharmacological investigations to explore the potential mechanism of Ding-Chuan-Tang against chronic obstructive pulmonary disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Ding-Chuan-Tang (Abbreviated as DCT) is frequently prescribed for treatment of respiratory diseases, including chronic obstructive pulmonary disease (COPD), which is characterized by coughing, wheezing, and chest tightness in traditional Chinese medicine (TCM). However, the potential mechanism of DCT has not been investigated. AIM OF STUDY: The aim of the study is to explore the efficiency of DCT in the treatment of COPD in vivo and in vitro, and to illustrate the possible mechanism against COPD. METHODS: COPD model was induced by exposure of mice to cigarette smoke (CS) for 16 weeks. Enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, Western blot, etc., were used to explore the efficiency and mechanisms of DCT. Network pharmacology analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, etc., was performed to explore the potential targets in the treatment of DCT on COPD. RESULTS: DCT significantly alleviated pulmonary pathological changes in mouse COPD model, and inhibited inflammatory response induced by CS and LPS in vivo and in vitro. Network pharmacology analysis suggested that DCT alleviated COPD via inhibiting inflammation by regulating PI3K-AKT pathway. In cell-based models, DCT suppressed the phosphorylation of PI3K and AKT, which further regulated its downstream targets Nrf2 and NF-κB, and inhibited inflammatory response. CONCLUSIONS: DCT effectively attenuated COPD in the mouse model induced by CS. The therapeutic mechanism of DCT against COPD was closely associated with the regulation of PI3K-AKT pathway and its downstream transcription factors, Nrf2 and NF-κB.

Methotrexate Chemotherapy Causes Growth Impairments, Vitamin D Deficiency, Bone Loss, and Altered Intestinal Metabolism-Effects of Calcitriol Supplementation.

Vitamin D deficiency or insufficiency is prevalent in childhood cancer patients and survivors after chemotherapy; further studies are needed to investigate the underlying aetiology and effectiveness of vitamin D supplementation in preventing chemotherapy-induced bone loss. This study used a rat model of treatment with antimetabolite methotrexate to investigate whether methotrexate chemotherapy causes vitamin D deficiency and if vitamin D supplementation attenuates the resultant bone loss. Methotrexate treatment (five daily injections) decreased serum vitamin D levels (from 52 to <30 ng/mL), reduced body and bone lengthening and tibial trabecular bone volume, and altered intestinal vitamin D metabolism, which was associated with intestinal mucosal damage known to cause malabsorption of nutrients, including dietary vitamin D and calcium. During the early stage after chemotherapy, mRNA expression increased for vitamin D activation enzyme CYP27B1 and for calcium-binding protein TRPV6 in the intestine. During the intestinal healing stage, expression of vitamin D catabolism enzyme CYP24 increased, and that of TRPV6 was normalised. Furthermore, subcutaneous calcitriol supplementation diminished methotrexate-induced bone loss due to its effect suppressing methotrexate-induced increased bone resorption. Thus, in young rats, methotrexate chemotherapy causes vitamin D deficiency, growth impairments, bone loss, and altered intestinal vitamin D metabolism, which are associated with intestinal damage, and vitamin D supplementation inhibits methotrexate-induced bone loss.

Identification of novel Nrf2 activators from Cinnamomum chartophyllum H.W. Li and their potential application of preventing oxidative insults in human lung epithelial cells.

Human lung tissue, directly exposed to the environmental oxidants and toxicants, is apt to be harmed to bring about acute or chronic oxidative insults. The nuclear factor erythroid 2-related factor 2 (Nrf2) represents a central cellular defense mechanism, and is a target for developing agents against oxidative insult-induced human lung diseases. Our previous study found that the EtOH extract of Cinnamomum chartophyllum protected human bronchial epithelial cells against oxidative insults via Nrf2 activation. In this study, a systemic phytochemical investigation of the aerial parts of C. chartophyllum led to the isolation of thirty chemical constituents, which were further evaluated for their Nrf2 inducing potential using NAD(P)H: quinone reductase (QR) assay. Among these purified constituents, a sesquiterpenoid bearing α, ß-unsaturated ketone group, 3S-(+)-9-oxonerolidol (NLD), and a diphenyl sharing phenolic groups, 3, 3', 4, 4'-tetrahydroxydiphenyl (THD) significantly activated Nrf2 and its downstream genes, NAD(P)H quinone oxidoreductase 1 (NQO-1), and γ-glutamyl cysteine synthetase (γ-GCS), and enhanced the nuclear translocation and stabilization of Nrf2 in human lung epithelial cells. Importantly, NLD and THD had no toxicities under the Nrf2 inducing doses. THD also demonstrated a potential of interrupting Nrf2-Keap1 protein-protein interaction (PPI). Furthermore, NLD and THD protected human lung epithelial cells against sodium arsenite [As(III)]-induced cytotoxicity. Taken together, we conclude that NLD and THD are two novel Nrf2 activators with potential application of preventing acute and chronic oxidative insults in human lung tissue.

Screening of traditional Chinese medicines with therapeutic potential on chronic obstructive pulmonary disease through inhibiting oxidative stress and inflammatory response.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a major public health problem and gives arise to severe chronic morbidity and mortality in the world. Inflammatory response and oxidative stress play dominant roles in the pathological mechanism of COPD, and have been regarded to be two important targets for the COPD therapy. Traditional Chinese medicines (TCMs) possess satisfying curative effects on COPD under guidance of the TCM theory in China, and merit in-depth investigations as a resource of lead compounds. METHODS: One hundred ninety-six of TCMs were collected, and extracted to establish a TCM extract library, and then further evaluated for their potency on inhibitions of oxidative stress and inflammatory response using NADP(H):quinone oxidoreductase (QR) assay and nitric oxide (NO) production assay, respectively. RESULTS: Our investigation observed that 38 of the tested TCM extracts induced QR activity in hepa 1c1c7 murine hepatoma cells, and 55 of them inhibited NO production in RAW 264.7 murine macrophages at the tested concentrations. Noteworthily, 20 of TCM extracts simultaneously inhibited oxidative stress and inflammatory responses. CONCLUSION: The observed bioactive TCMs, particularly these 20 TCMs with dual inhibitory effects, might be useful for the treatment of COPD. More importantly, the results of the present research afford us an opportunity to discover new lead molecules as COPD therapeutic agents from these active TCMs.

Steamed root of Rehmannia glutinosa Libosch (Plantaginaceae) alleviates methotrexate-induced intestinal mucositis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Intestinal mucositis induced by chemotherapy is a severe clinical problem in cancer patients that currently lack effective interventions. In traditional Chinese medicine, chemotherapeutic toxicity is diagnosed as Qi and Yin deficiency, and steamed rehmannia root (SRR) is frequently prescribed to these patients. Whether SRR can prevent the adverse effects remains to be confirmed experimentally. The present study used a rat model to investigate potential efficacy and action mechanisms of SRR in attenuating the adverse effects caused by chemotherapy. MATERIALS AND METHODS: Intraperitoneal injection of a single dose of anti-metabolite methotrexate (MTX, 25mg/kg) was given to adult Wistar rats, which also received oral gavage of water or SRR (1.08g/kg twice daily 3 days before and 4 days after MTX treatment), or calcium folinate (CF, a clinically used MTX antidote as a comparison, at 1mg/kg twice daily 36h after MTX treatment), or SRR and CF in combination. Animals were sacrificed 4 days after MTX treatment. Complete blood cell counting was carried out. Jejunum was analyzed histologically for mucosal damage, immunohistochemically for proliferating cell nuclear antigen (PCNA), and biochemically for thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH), as well as for tumor necrosis factor alpha (TNF-α). RESULTS: MTX treatment led to weight loss, leucopenia, polycythemia, increase in large thrombocyte ratio, intestinal villus atrophy, crypt loss and reduction in PCNA positive crypt cells, increases in mucosal TBARS and TNF-α and decrease in GSH. All these alterations were inhibited by SRR administration except leucopenia, and the effects of CF or CF plus SRR supplementation were found to be inferior to those of SRR. CONCLUSIONS: SRR can alleviate MTX-induced gut mucositis, which may be achieved by inhibiting MTX-induced oxidative stress and inflammatory response. These findings support the application of SRR in chemotherapy but not the combined application of SRR and CF.

[Discussion on the quality marker of rehmanniae radix praeparata in the Chinese Pharmacopoeia].

OBJECTIVE: In the Chinese Pharmacopoeia (2010 edition), the minimum limit of verbascoside (acteoside) in Rehmanniae Radix Praeparata (RRP) was set at 0.20%, the rationality of the standard was evaluated in this paper. METHODS: 10 samples of Rehmanniae Radix (RR) and RRP were collected or prepared. According to the traditional method, different samples of RRP were prepared from the same batch of RR, and the content of verbascoside was determined by HPLC. RESULTS: The average content of verbascoside in RR and RRP was 0.0226% and 0.0097%, respectively, and their difference was up to the level of P < 0.01. The content of verbascoside was found decreased with processing time increasing. Long-time storage also results in substantial loss of verbascoside. CONCLUSION: Processing results in verbascoside decreasing significantly. It is unreasonable that RR and RRP have the same standard in the content of verbascoside. It is suggested that the maximum limit of verboscoside should be set in the new version of Chinese pharmacopoeia.

Antidepressant effect of Shudihuang on mice exposed to unpredictable chronic mild stress.

AIM OF THE STUDY: Depression is a severe mood disorder. It was treated with Shudihuang, the steamed roots of Rehmannia glutinota Libosch. (SRG), in traditional Chinese medicine. The present paper was designed to verify its antidepressant effect. MATERIALS AND METHODS: A mouse model of depression was established though unpredictable chronic mild stress (UCMS). Low and high doses of SRG were administered orally. Fur state, body and organ weight, and gastric ulcers were examined. Locomotion was assayed in open field test. Liver antioxidant indexes were measured spectrophotometrically. RESULTS: Fur state, body and organ weight were found to be insensitive to UCMS. The locomotion reduced by UCMS was restored by low dose of SRG (2.5 g/kg BW) but not by high dose (5 g/kg BW). UCMS resulted in aggravated gastric ulceration, elevated liver malondialdehyde, together with reduced total antioxidant capability, glutathione content, and superoxide dismutase and catalase activities. The alterations were improved by SRG in a dose-dependent manner. The differences in the activity of glutathione peroxidase were statistically nonsignificant among groups. Clomipramine the positive drug was similar to SRG especially in antioxidation. CONCLUSION: SRG is of therapeutic value for depression-like disorders, and antioxidation may be one of the mechanisms underlying its antidepressant action.

[Advances in research on pharmacodynamics and chemical conversion of catalpol].

Catalpol is a active component in Rehmannia Root, and its pharmacological effects are extensive. In this paper its effects and chemical changes were summarized. This will offer the reference for further research work catalpol.

[Extraction and salting-out purification of alpha-galactosidase and beta-glucosidase from fresh roots of Rehmannia glutinosa].

OBJECTIVE: To extract and preliminarily purify alpha-galactosidase and beta-glucosidase from the fresh roots of Rehmannia glutinosa. METHODS: With the enzyme activity as a criterion, the best procedure of extraction was selected though orthogonal design method, and the desired saturation of ammonium sulfate in two-step salting-out was settled by gradient sedimentation of root extract according to enzyme activity and protein content. RESULTS: Temperature and solvent volume affect the extraction of alpha-galactosidase significantly, while solvent type to beta-glucosidase. Therefore the procedure for extracting two enzymes was decided as mixing comminuted fresh root with 3 times phosphate buffer, and placing the mixture in refrigeratory at 4 degrees C for 4 hours, and then obtaining the enzyme liquid by centrifuging at 4 degrees C. 30% and 60% saturation was defined as the lower and upper point for two-step ammonium sulfate salting-out of the two enzymes. CONCLUSION: alpha-Galactosidase and beta-glucosidase exist in the fresh roots, and can be preliminarily purified through two-step salting-out.

[Advances in study on chemical constituents from plants of genus Pfaffia and their bioactivities].

The progress in the studies on chemical constituents and pharmacological activity of the genus Pfaffia is summarized in recent 20 years. These plants contain various chemical constituents and have broad bioactivities such as sthenic, anti-tumor, analgesic and anti-inflammatory and should be further investigated.

[Preservation of germplasm of Rehmannia glutinosa in vitro].

OBJECTIVE: To establish a method for the germplasm preservation of R. glutinosa. METHOD: Plantlets of different cultivars obtained by tip culture were inoculated into test tubes with MS medium supplemented with 0.2 mg.L-1 BA and 0.02 mg.L-1 NAA, and preserved at 4-6 degrees C in dark. At the same time, different media (A. distilled water + 10 g.L-1 agar; B. 1/2 MS + 5 g.L-1 agar; C. MS + 10 g.L-1 agar; D. 1/2 MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar; E. MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar) were set to conserve plantlets of "85-5" on the same condition. RESULT: 4 months later, the death rate of "Xinggeda" was 73%, "Tucheng" 60%, "85-5" 33%, and "Beijing 1" 9%. All of the plantlets of "85-5" in different media kept alive. 10 months later, most of the preserved plantlets browned and wilted except those on medium A. CONCLUSION: The low temperature endurance of R. glutinosa is cultivar-dependent. Medium A can preserve "85-5" for more than 10 months in vitro.

[Photosynthetic pigment contents of different germplasm of Rehmannia glutinosa and the relationship between pigments and leaf color].

OBJECTIVE: To evaluate the germplasm of Rehmannia glutinosa on the basis of photosynthetic pigment contents (PPC). METHOD: 20 cultivars were planted on the same condition. On Oct. 23 and Sept. 25, 3 leaves per cultivar were collected on different plants, and 80 mg mesophyll was collected among upper lateral veins and was ground in 96% alcohol, and the supernatant was subjected to measure on a spectrophotometer (Angilent 8453). RESULT: The PPCs among cultivars were significantly different at a P < or = 0.01 level. The results of the measurements were similar. Chlolophyll a was the most abundant pigment, but varied to a great extent among different cultivars. 20 cultivars were divided into 9 homogeneous groups according to the contents of chlorophyll a by Duncan's multiple range test at P < or = 0.05. In addition, the content of chlorophyll a was closely related to leaf color. The cultivars with higher chlolophyll a had deep green leaves, and those with lower had yellow green or pale green leaves. CONCLUSION: PPC was an inherent character and an important index for the germplasm evaluation of R. glutinosa.