Effect of allyl isothiocyanate on oxidative stress in COPD via the AhR / CYP1A1 and Nrf2 / NQO1 pathways and the underlying mechanism.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is currently the third leading cause of death globally. Oxidative stress affects various molecular mechanisms and is the main driving factor of COPD. Ally isothiocyanate (AITC) is an effective component of Semen Sinapis Albae, which has favorable effects for the treatment of COPD, but its mechanism has not been fully elucidated. PURPOSE: This study aimed to elucidate the antioxidant effect of AITC on COPD and its molecular mechanism, and preliminarily determine the role of AhR in the progression of COPD. STUDY DESIGN: The COPD rat model was established by smoking combined with intratracheal instillation of lipopolysaccharide. Different doses of AITC, positive control drug acetylcysteine, AhR inhibitor alpha-naphthoflavone, and agonist beta-naphthoflavone were administered by gavage. Human bronchial epithelial cells induced by cigarette smoke extract (CSE) were used in an in vitro model to explore the molecular mechanisms of AITC. METHODS: The effects of AITC on lung function and oxidative stress in rats were evaluated in vivo using the respiratory function test, white blood cell count, enzyme-linked immunosorbent assay, and histological staining. The changes in protein expression in the lung tissue were detected by immunohistochemistry and Western blotting. RT-PCR, western blotting, and immunofluorescence were used to explore the molecular mechanisms of AITC. Enzyme-linked immunosorbent assay, reactive oxygen species probing, and flow cytometry were used to determine the antioxidant effect of AITC. RESULTS: AITC can improve the lung function of rats with COPD, restore lung tissue structure, improve oxidative stress, reduce inflammation, and inhibit lung cell apoptosis. AITC reversed the upregulation of AhR and CYP1A1 and the down-regulation of Nrf2 and NQO1 in the lung tissues of rats with COPD. CSE stimulation can increase the expressions of AhR and CYP1A1 and decrease the expressions of Nrf2 and NQO1 in 16HBE cells, leading to severe oxidative stress and inflammatory response and, ultimately, apoptosis. AITC inhibited AhR and CYP1A1 expressions, induced Nrf2 and NQO1 expressions, promoted Nrf2 nuclear translocation, and improved CSE-induced toxicological effects. CONCLUSION: AITC may improve lung oxidative stress by inhibiting the AhR / CYP1A1 and activating the Nrf2 / NQO1 pathways, thereby delaying the pathological progression of COPD.

[Comparative study on serum transitional components of Huatan Jiangqi Capsules in rats under pathological and physiological status].

The differences of transitional components and metabolic processes of Huatan Jiangqi Capsules(HTJQ) in rats under normal physiological and pathological conditions of COPD were analyzed by UPLC-Q-TOF-MS. The rat COPD model was established by passive smoking and intratracheal instillation of lipopolysaccharide. After the normal and COPD model rats were douched with HTJQ, the blood was collected from hepatic portal vein and the drug-containing serum samples were prepared by methanol precipitation of protein. Then, 10 batches of drug-containing serum samples of HTJQ were prepared and analyzed by UPLC serum fingerprint to evaluate the quality and stability of drug-containing serum samples. UPLC-Q-TOF-MS was used to collect the mass spectrometric information of the transitional components. Twenty-eight transitional components of HTJQ in normal rats and 25 transitional components of HTJQ in COPD model rats were identified by UPLC-Q-TOF-MS. Under pathological and physiological conditions, there were not only the same transitional components in rat serum, but also corresponding differences. Further studies showed that there were also differences in the metabolic process of transitional components between the two conditions. In normal rats, most of the metabolic types of transitional components were phase I reactions. In COPD model rats, phase Ⅰ reactions decreased and phase Ⅱ reactions increased correspondingly. With UPLC-Q-TOF-MS technology, the differences of transitional components and the metabolism process of HTJQ in rats under normal physiological and pathological conditions were analyzed. The results showed that types of transitional components and the activity of some metabolic enzymes would be changed in COPD pathological state, which would affect the metabolic process of bioactive components in vivo. It laid a foundation for further elucidating the metabolic process and pharmacodynamic substance basis of HTJQ.

The application of biotechnology in medicinal plants breeding research in China.

Breeding is not only an important area of medicinal plants research but also the foundation for the superior varieties acquirement of medicinal plants. The rise of modern biotechnology provides good opportunities and new means for medicinal plants breeding research in China. Biotechnology shows its technical advantages and new development prospects in breeding of new medicinal plants varieties with high and stable yield, good quality, as well as stress-resistance. In this paper, we describe recent advances, problems, and development prospects about the application of modern biotechnology in medicinal plants breeding research in China.

[Influence of huatanjiangqi prescription on multidrug resistance-associated protein 1 in human bronchial epithelial cells].

OBJECTIVE: To investigate the effect of Huatanjiangqi prescription (Sinapis Semen, Perillae Fructus, Cynanchi Stauntonii Rhizoma et Radix, Inulae Herba, Angelicae Sinensis Radix, Honey-fried Ephedrae Herba) on multidrug resistance-associated protein 1 (MRP1) in human bronchial epithelial cells. METHODS: The human bronchial epithelial cells line 16HBE140- was used to analyze the in vitro effect of Huatanjiangqi prescription on MRP1 transport. 5-CFDA was used as a model MRP1 substrate and was measured with flow cytometry. The mRNA expression of MRP1 was detected by real-time PCR. RESULTS: Huatanjiangqi prescription could promote the proliferation of human bronchial epithelial cells 16HBE140- in a certain range of concentration; Compared with the control group (5-CFDA), low, medium and high concentration (100, 1 000, 2 000 microg/mL) of Huatanjiangqi prescription on MRP1 function were increased by 22.59%, 47.14% and 68.36%, respectively; Huatanjiangqi prescription could concentration-dependently induce the expression of MRP1 mRNA, medium and high concentration could induce a significant difference. CONCLUSION: Huatanjiangqi prescription can improve MRP1 efflux function and mRNA levels in a concetration-dependent manner.

Germplasm preservation in vitro of Polygonum multiflorum Thunb.

BACKGROUND: The root of Polygonum multiflorum Thunb. is a common traditional Chinese medicine. In recent years, the wild resources of P. multiflorum have been seriously broken, and the cultivated varieties have been degrading. The germplasm resources of P. multiflorum need protection and preservation. So far, no in vitro germplasm preservation of P. multiflorum has been reported. OBJECTIVE: To explore a method for the in vitro germplasm preservation of P. multiflorum. MATERIALS AND METHODS: A large number of buds from seed explants were induced by tissue culture. The single buds were used as experimental materials to study the effects of plant growth regulator, temperature, and osmotic pressure on the preservation time, growth recovery, and genetic stability. RESULTS: When the buds were inoculated onto Murashige and Skoog (MS) basal media containing 4% w/v sucrose, 2% w/v mannitol, and 1% w/v sorbitol, supplemented with paclobutrazol (PP333) 1.0 mg/l, abscisic acid (ABA) 5.0 mg/l, and daminozide (B9) 30.0 mg/l in an illuminated chamber under a 16 h photoperiod of 1500 lx light intensity at 15°C for 10 months, the survival rate was over 70% with good growth recovery and genetic stability. CONCLUSION: The results of this study can be used for medium-term in vitro germplasm preservation of P. multiflorum, and meeting actual needs of research and production.

[Study on inducing embryogenic callus of Dioscorea zingiberensis by orthogonal design].

OBJECTIVE: To select the suitable medium to induce embryogenic callus of Dioscorea zingiberensis. METHODS: Plantlet of Dioscorea zingiberesis in vitro was obtained by using apical meristem as explant. The different parts of the plantlets were cultured to select the best explant used for inducing callus and embryoids. Growing rate and diosgenin content were calculated in orthogonal test to optimize combination of phytohormones for inducing embryogenic callus. RESULTS: The leaves were suitable explants to induce callus and embryoid. The inducing rate of callus and embryoids reached 92.5% and 42.5%, respectively. The optimal medium for inducing embryogenic callus was MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L + 2,4-D 1.0 mg/L. CONCLUSION: The results of this study can be used for effective induction of embryogenic callus of Dioscorea zingiberensis, and lay the foundation for the subsequent research of artificial seeds.

[Effects of huatan jiangqi capsule on the levels of multi-drug resistance-associated protein 1 in the bronchial epithelial cells of model rats with chronic obstructive pulmonary disease].

OBJECTIVE: To observe the effects of huatan jiangqi capsule (HJC) on the expression levels and functions of multi-drug resistance-associated protein 1 (MRP1) in the bronchial epithelial cells of chronic obstructive pulmonary disease (COPD) model rats, and to explore the mechanism of HJC for treating COPD. METHODS: Twenty-four male wistar rats were randomly divided into the normal control group, the model group, and HJC group. Except the normal control group, the COPD rat model was established in the rest groups using quantitative stimulation with tobacco, SO2, and caroid aerosol rebreathing method. The indices of the post-treatment lung functions, the cell counts of bronchoalveolar lavage fluid (BALF), the pathological features of the lung tissue were observed. The concentration of LTC, in lung tissues was also examined by ELISA. The expression of MRP1 of the pulmonary tracheal epithelium was detected using immunohistochemical assay. RESULTS: (1) The pulmonary compliance, the forced expiratory volume at 0. 3 second (FEV 0.3%)/the forced vital capacity (FVC), the peak expiratory flow, the maximum mid expiratory flow decreased more significantly in the model group than in the normal control group (P < 0.05). The aforesaid pulmonary function indices obviously increased in the HJC group when compared with the model group (P < 0.05). (2) The air inflammation was aggravated with obvious emphysema in the model group. The inflammation and emphysema occurred in the HJC group in a milder degree. (3) Compared with the normal control group, the levels of LTC4 significantly increased in the lung tissue of the model group and HJC group (P < 0.01). Compared with the model group, the levels of LTC4 significantly decreased in the lung tissues of the HJC group (P < 0.05). (4) Compared with the normal control group, the protein expression of the bronchial epithelial MRP1 significantly decreased in the model group (P < 0.01). Compared with the model group, the protein expression of the bronchial epithelial MRP1 were significantly enhanced in the HJC group (P < 0.05). CONCLUSION: HJC could effectively alleviate the lung inflammation, postpone the occurrence and development of COPD possibly through effecting the functions and expressions of MRP1 in COPD rats.

[Quality evaluation of prepared slices of Paeonia lactiflon--determination of paeoniflorin by HPLC].

OBJECTIVE: To establish the quality criteria of the prepared slices of Paeonia lactiflon. METHOD: RP-HPLC was used for the determination of paeoniflorin in 10 lots of samples by ultrasound-assisted extraction. RESULT: The samples were extracted with 50% methanol. Seperation of the solution was performed on an ODS column with a mobile phase of acetonitrile-water (18:82), detected at 230 nm. CONCLUSION: The method is simple, repeatable, accurate and applicable.