Tuberostemonine A

Morphological and chemical variation of Stemona tuberosa from southern China - Evidence for heterogeneity of this medicinal plant species.[Pubmed:28580601]

Plant Biol (Stuttg). 2017 Sep;19(5):835-842.

The occurrence of bioactive alkaloids and tocopherols was studied in 15 different provenances of Stemona tuberosa Lour. collected in southern China, to examine chemical variation of individuals that show notable differences in flower characteristics. Morphological variations stimulated examination of chemical characteristics of these individuals. Methanolic root extracts of 15 individuals of S. tuberosa were comparatively assessed with HPLC-UV-DAD/ELSD. Five of seven compounds were co-chromatographically identified. Two compounds were isolated and their structure elucidated using NMR and MS. Amounts of alkaloids and tocopherols were determined using HPLC-UV-DAD/ELSD with the external standard method. Five alkaloids, tuberostemonine (1), Tuberostemonine A (2), neotuberostemonine (3), tuberostemonine N (4), stemoninine (5) and two 3,4-dehydrotocopherol derivatives were identified. Within S. tuberosa alkaloid accumulation tends either towards tuberostemonine (1) or stemoninine (5). All individuals show a notable co-occurrence of compounds 1 or 5 and 3,4-dehydro-delta-tocopherol (6). These results coincide with differences in flower morphology of S. tuberosa. Stemona tuberosa, as defined in the Flora of China, shows a remarkable variation in flower morphology and additionally in the accumulation of alkaloids. The obtained data show the need for future species delimitation to either species or subspecies level.

Tuberostemonine reverses multidrug resistance in chronic myelogenous leukemia cells K562/ADR.[Pubmed:28529625]

J Cancer. 2017 Apr 9;8(6):1103-1112.

Objective: To investigate the reversal effect of tuberostemonine on MDR in myelogenous leukemia cells K562/ADR. Methods: Human myelogenous leukemia cells K562 and their adriamycin-resistance cells K562/ADR were used. The growth curve of cells treated by Tuberostemonine And the Non-toxic concentration of tuberostemonine were determined by MTT, Cell apoptosis was determined by MTT and flow cytometry. The expression of MDR1, Survivin and Livin was detected by RT-PCR. The activity of P-gp was detected by flow cytometry. Western blot was used to detect the expression of NF-kappaB and Survivin. Results: The effect of tuberostemonine on K562/ADR showed a dose-dependence, and 350mug/mL and 500mug/mL of tuberostemonine could inhibit the expression of MDR1 (P<0.05). While no function difference of P-gp was detected. With the increased concentration of tuberostemonine, the inhibitory effect were enhanced to the expression of NF-kappaB. Tuberostemonine combined with adriamycin could time-dependently inhibit the cell proliferation (P<0.05) and obviously promoted the cell apoptosis (P<0.05). Also the tuberostemonine could inhibit the expression of Survivin. Conclusion: There are no direct relations between Tuberostemonine And P-gp, but tuberostemonine could reverse the multidrug resistance of K562/ADR via down-regulating the expression of Nf-kappaB and inhibiting th1e expression of Survivin.

Metabolic profiles of neotuberostemonine and tuberostemonine in rats by high performance liquid chromatography/quadrupole time-of-flight mass spectrometry.[Pubmed:28448890]

J Pharm Biomed Anal. 2017 Jul 15;141:210-221.

Neotuberostemonine (NS) and tuberostemonine (TS), a pair of stereoisomers, are the active components contained in Stemona tuberosa, an antitussive herbal medicine in China. Two isomers have different pharmacological efficacies, which will be related with their in vivo disposition. However, the metabolic fates of NS and TS remain unknown. A method of high performance liquid chromatography/quadrupole time-of-flight mass spectrometry coupled with mass detect filter technique was established to investigate the metabolites in rat plasma, bile, urine, and feces after oral administration of the equal doses of NS and TS. The results showed that NS produced 48 phase I metabolites, including NS, 3 hydrolyzed, 14 hydroxylated, 20 monohydrolyzed+hydroxylated and 10 dihydrolyzed+hydroxylated metabolites. The number of detected NS metabolites was 11, 39, 22 and 30 in plasma, bile, urine and feces. TS yielded 23 phase I metabolites, including TS, 3 hydrolyzed, 7 hydroxylated, 9 monohydrolyzed+hydroxylated and 3 dihydrolyzed+hydroxylated metabolites. Besides, TS yielded 9 phase II metabolites, including 1 glucuronic acid and 2 glutathione conjugates, and the later further degraded and modified into cysteine-glycine, cysteine and N-acetylcysteine conjugates. The number of detected TS metabolites was 9, 24, 24 and 15 in plasma, bile, urine and feces. Different metabolic patterns may be one of the main reasons leading to different pharmacological effects of NS and TS.

The therapeutic effects of tuberostemonine against cigarette smoke-induced acute lung inflammation in mice.[Pubmed:26849941]

Eur J Pharmacol. 2016 Mar 5;774:80-6.

Chronic obstructive pulmonary disease (COPD) is mainly caused by cigarette smoking and is characterized by the destruction of lung parenchyma, structural alterations of the small airways, and systemic inflammation. Tuberostemonine (TS) is an alkaloid-type phytochemical from Stemona tuberosa. In the present study, we evaluated the anti-inflammatory effect of TS in a cigarette smoke (CS)-induced mouse model of acute lung inflammation. The mice were whole-body exposed to CS or fresh air for 7 days. TS was administered by an intraperitoneal (i.p.) injection 1h before exposure to CS. To test the effects of TS, the numbers of total cells, neutrophils, macrophages and lymphocytes in the bronchoalveolar lavage (BAL) fluid were counted. Furthermore, we measured the levels of several chemokines, such as GCP-2, MIP-3alpha, MCP-1 and KC, in the lung tissue. The cellular profiles and histopathological analysis demonstrated that the infiltration of peribronchial and perivascular inflammatory cells significantly decreased in the TS-treated groups compared with the CS-exposure group. The TS treatment significantly ameliorated the airway epithelial thickness induced by CS exposure and caused a significant decrement in the production of chemokines in the lung. These results suggest that TS has anti-inflammatory effects against CS-induced acute lung inflammation.

[Comparative study of four alkaloids contents and antitussive activities of Stemona tuberosa from different habitats of Guangxi Province].[Pubmed:26027114]

Zhong Yao Cai. 2014 Nov;37(11):1956-60.

OBJECTIVE: To compare the contents of four alkaloids and antitussive activities of Stemona tuberosa from different habitats of Guangxi Province. METHODS: The HPLC separation was performed on a Merck Purospher STAR RP18 (250 mm x 4. 6 mm, 5 microm) column by gradient elution using 0. 05% ammonia-acetonitrile as the mobile phase. The flow rate was 1. 0 mL/min, the dectection wave-length was set at 210 nm,and the column temperature was 40 degrees C. The antitussive potency of total alkaloids of Stemonae Radix from different habitats was evaluated on guinea pigs with citric acid aerosol to induce cough. RESULTS: The range of recoveries of this mehtod was 98. 24% ~ 101. 21%, with all the constituents showing good linearity(the correlation coefficents above 0. 999). The major chemotype of Stemonae Radix in Guangxi was stemoninine, following by Tuberostemonine And croomine, and finally neotuberostemonine. The antitussive activitiy of Stemona tuberosa was in a concentration-dependent manner. CONCLUSION: Stemonae Radix from Dongxing, Fangcheng can reduce cough times and prolong cough incubation period, and thus Dongxing, Fangcheng is the best habitat in Guangxi in the present experiments.

Suppression of IL-2 production and proliferation of CD4(+) T cells by tuberostemonine O.[Pubmed:25491339]

Chem Biodivers. 2014 Dec;11(12):1954-62.

Tuberostemonine stereoisomers are natural alkaloids found in Stemona tuberosa, that are known to have anti-inflammatory and anti-infective properties. Tuberostemonine Alkaloids inhibit inflammation by suppressing the expression of inflammatory mediators such as cyclooxygenase and nitric oxide synthase. However, the direct immunomodulatory properties of Tuberostemonine Alkaloids in T cells have not been elucidated so far. In this study, the activities in T cells of tuberostemonine N (TbN) and a novel alkaloid, tuberostemonine O (TbO), isolated from S. tuberosa, were investigated. Although TbN did not have a significant effect on cytokine production in splenic T cells, TbO selectively suppressed interleukin (IL)-2 production. Moreover, TbO, but not TbN, significantly inhibited IL-2 production by primary CD4(+) T cells and delayed the T-cell proliferation in a dose-dependent manner. Addition of excess recombinant IL-2 restored the decreased cell-division rates in TbO-treated CD4(+) T cells to control levels. Collectively, these findings suggest that the immunomodulatory effects of TbO occurred by the suppression of IL-2 expression and IL-2-induced T-cell proliferation, suggesting a potential beneficial role of Tuberostemonine Alkaloids for the control of chronic inflammatory and autoimmune diseases caused by hyperactivated T cells.

TLC-image analysis of non-chromophoric tuberostemonine alkaloid derivatives in Stemona species.[Pubmed:24079167]

Nat Prod Commun. 2013 Aug;8(8):1065-8.

A simple, selective, precise, and accurate thin-layer chromatographic (TLC) image analytical method was developed and validated for simultaneous quantification of the major components in the root extracts of Stemona tuberosa (tuberostemonine, tuberostemonine N and neotuberostemonine)), and S. phyllantha (Tuberostemonine And Tuberostemonine A). The analysis was performed by TLC on silica gel 60 F254 aluminum plates using a mixture of dichloromethane: ethyl acetate: methanol: ammonium hydroxide (50:45:4:1) as mobile phase. Post-derivatization was employed by dipping the TLC plate into Dragendorff's reagent to visualize the spots. Image analysis of the scanned TLC plate was performed to detect the contents of tuberostemonine derivatives. The polynomial regression data for the calibration plots showed good linear relationships within the concentration range of 2-7 microg/spot. The method gave satisfactory precision, accuracy, selectivity and could simultaneously quantify tuberostemonine, Tuberostemonine A, tuberostemonine N and neotuberostemonine. Dried powdered roots of S. tuberosa grown in Thailand contained 1.31 +/- 0.28, 1.63 +/- 0.18 and 1.24 +/- 0.27% tuberostemonine, tuberostemonine N, and neotuberostemonine (dry weight), respectively, while S. phyllantha roots contained 1.39 +/- 0.14% Tuberostemonine And 0.39 +/- 0.08% Tuberostemonine A (dry weight). The proposed method was simple, inexpensive, and more accessible to apply for many local authorities and small laboratories.