Jaridon 6, a new diterpene from Rabdosia rubescens (Hemsl.) Hara, can display anti-gastric cancer resistance by inhibiting SIRT1 and inducing autophagy.

Tumor resistance is the main cause of treatment failure and is associated with many tumor factors. Jaridon 6, a new diterpene extracted from Rabdosia rubescens (Hemsl.) Hara, which has been previously extracted by our research team, has been tested having more obvious advantages in resistant tumor cells. However, its mechanism is unclear. In this study, we studied the effect and the specific mechanism of Jaridon 6 in resistant gastric cancer cells. Cytotoxicity test, colony test, western blotting, and nude test verified the anti-drug resistance ability of Jaridon 6 in the MGC803/PTX and MGC803/5-Fu cells. Jaridon 6 has shown obvious inhibitory effects in the sirtuin 1 (SIRT1) enzyme test. Transmission electron microscopy and immunofluorescence tests further proved the autophagic action of Jaridon 6. Jaridon 6 could inhibit the proliferation of the resistant gastric cancer cell in vivo and in vitro. Jaridon 6 inhibited SIRT1 enzyme and induced autophagy by inhibiting the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway. Thus, it may be considered for treating gastric cancer resistance by individual or combined administration, as an SIRT1 inhibitor and autophagy inducer.

Wenyang Huazhuo Tongluo formula inhibits fibrosis via suppressing Wnt/ß-catenin signaling pathway in a Bleomycin-induced systemic sclerosis mouse model.

BACKGROUND: Systemic sclerosis (SSc) is an autoimmune disease characterized by fibrosis of the skin and internal organs. So far, no Western medicine treatment can completely inhibit or reverse the progress of SSc, while at the same time, our previous series of studies have shown that the treatment of SSc by the Wenyang Huazhuo Tongluo formula (WYHZTL), a Chinese herbal decoction, shows a delightful prospect. The aim of this study is to further investigate the mechanism of anti-fibrosis of WYHZTL formula in SSc mouse model. METHODS: The Bleomycin-induced SSc mouse model was treated with saline (BLM), high-dosage of WYHZTL formula (WYHZTL-H), medium-dosage of WYHZTL formula (WYHZTL-M), low-dosage of WYHZTL formula (WYHZTL-L) and XAV-939, a small molecule inhibitor of Wnt/ß-catenin signaling pathway, by the intragastric administration and intraperitoneal injection, respectively. The mRNA and protein levels of Wnt/ß-catenin signaling pathway associated genes, fibrosis markers and histopathology were detected by reverse transcription-quantitative polymerase chain reaction, Western blotting and hematoxylin/eosin-staining. The levels of Wnt1, CTGF and DKK1 protein in serum were detected by enzyme-linked immunosorbent assay. RESULTS: Compared with BLM group, the WYHZTL formula and XAV-939 could significantly inhibit the thickness of the skin tissue of the SSc mouse model. The mRNA expression levels of GSK3ß and DKK1 in the WYHZTL formula and XAV-939-treated group were significantly higher than those in the BLM group, while Wnt1, ß-catenin, TCF4, cyclin D1, survivin, VEGF, CTGF, FN1, collagen I/III were decreased. Compared with BLM group, the protein expression levels of GSK3ß and DKK1 in the WYHZTL formula and XAV-939-treated group were upregulated, while Wnt1, ß-catenin, cyclin D1, survivin, CTGF, FN1, collagen I/III were downregulated. WYHZTL formula and XAV-939 could inhibit expression of Wnt1 and CTGF, but promoted DKK1 in serum. Furthermore, WYHZTL-H seemed more effective than WYHZTL-M and/or XAV-939 on regulating Wnt1, ß-catenin, TCF4, GSK3ß, DKK1, cyclin D1, survivin, VEGF, CTGF, FN1 and collagen I/III. CONCLUSION: This present study demonstrates that WYHZTL formula has anti-fibrosis effect in Bleomycin-induced SSc mouse model in a dosage-dependent manner, and the molecular mechanism may be related to the inhibition of Wnt/ß-catenin signaling pathway.

Wenyang Huazhuo Tongluo formula, a Chinese herbal decoction, improves skin fibrosis by promoting apoptosis and inhibiting proliferation through down-regulation of survivin and cyclin D1 in systemic sclerosis.

BACKGROUND: Fibrosis is a major contributor to systemic sclerosis (SSc)-related morbidity, and rapid, progressive skin involvement predicts later mortality. Western medicine therapies for SSc cannot produce satisfactory effects currently, while Traditional Chinese Medicine (TCM), such as the Wenyang Huazhuo Tongluo (WYHZTL) formula, a Chinese herbal decoction, has shown amazing anti-fibrosis efficacy on SSc in clinical applications. This study is aiming to investigate the anti-fibrotic mechanism of WYHZTL formula for the treatment of SSc. METHODS: Fibroblasts from primary culture of skin lesions of SSc patients were exposed to rat medicated sera containing WYHZTL or XAV939, a small-molecule inhibitor of both tankyrase 1/2 and Wnt/ß-catenin pathway. Cell counting kit-8 assay and Annexin V FITC/PI apoptosis kit were used to analyze cell proliferation and apoptosis in fibroblasts, respectively. Reverse transcription-polymerase chain reaction (RT-PCR) and western blotting were used to detect the mRNA and protein levels of cyclin D1 and survivin. RESULTS: After 28, 48 and 72 h of incubation, the proliferative ability of the fibroblasts cells was obviously reduced by the sera containing WYHZTL compared with that in the control group; the percentage of apoptotic cell population in the sera containing WYHZTL treated fibroblasts cells was significantly higher than that in those treated with the control sera, and was about similar to that in those treated with XAV939. The sera containing WYHZTL could down-regulate both mRNA and protein levels of cyclin D1 and survivin, compared with the control group. CONCLUSIONS: The present study demonstrates the antiproliferative and pro-apoptotic actions of WYHZTL formula against fibroblasts and the effect may be related to the down-regulation of mRNA and protein levels of cyclin D1 and survivin in SSc.

Two new sesquiterpene lactones from the fruits of Illicium jiadifengpi.

Two new seco-prezizaane-type sesquiterpenes, 3,4-dehydroneomajucin (1) and 1,2,3,4-tetradehydroneomajucin (2), were isolated from the fruits of Illicium jiadifengpi. The structure of these compounds was determined using 1D and 2D NMR and ESI-MS. The isolates were evaluated for their anti-hepatitis B virus activities on the Hep G2.2.15 cell line. The inhibitory rates of compounds 1 and 2 on the HBeAg and HBsAg expression were 30.08 ± 3.09% and 11.43 ± 1.92% at a concentrations of 68.00 µM and 7.88 ± 1.21% and 16.96 ± 4.24% at a concentration of 68.50 µM, respectively.

High performance liquid chromatography time of flight electrospray ionization mass spectrometry for quantification of sesquiterpenes in Chrysanthemi indici Flos active extract.

BACKGROUND: Chrysanthemi indici Flos, a traditional herbal medicine is used to clearing heat-toxicity, removing the liver fire, and improving eyesight. In our preliminary work, an active extract of CTC in C. An indici Flos with anti-hepatitis B virus and liver protective activity was found by HepG2.2.1.5 test and experiment of protein synthesis in mice's injured liver. In this work, we aimed to study the active faction CTC further by qualitative and quantitative analysis method. MATERIALS AND METHODS: High performance liquid chromatography time of flight electrospray ionization mass spectrometry (HPLC TOF ESI-MS) analysis method of the CTC was established. Cumambrin A and angeloylcumambrin B in CTC were analyzed by high performance liquid chromatography-ultraviolet-evaporative light scattering detector (HPLC-UV-ELSD) analysis methods. A binary gradient elution program was conducted for chromatographic separation with acetonitrile (A) and ultrapure water (B) as follows: 0-10 min, 42-46% A; 10-20 min, 46-55% A; 20-25 min, 55-60% A; and 25-35 min, 60-75% A. The column temperature and UV wavelength were set at 30°C and 205 nm. RESULT: Ten constituents including (3R, 5R, 6S, 7S, 10R)-7-(2-hydroxy-2-propyl)-10-methyl-4-methyleneperhy, dronaphthal-ene-3, 5, 6-triol acetone solvate; (+)-edusmance-4, (14)-ene-11, 13-diol; linarin; luteolin; apigenin; tricin; 5, 3',4'- trimethyl-6,7-dimethoxy flavones; cumambrin A; acacetin; and angeloylcumambrin B in CTC were identified by HPLC TOF ESI-MS. The contents of sesquiterpenes in CTC were decreased by storing years. CONCLUSIONS: The results showed that both UV and ELSD methods were feasible, accurate, and the determination results were in good consistency. The contents of two sesquiterpenes decreased with storing years. Two sesquiterpenes could be used as quality control for C. indici flos CTC.

A new sesquiterpene lactone from the fruits of Illicium henryi.

AIM: To study the chemical constituents of the fruits of Illicium henryi. METHOD: Chromatographic separations on silica gel, Sephadex LH-20 gel and MCI gel were used to isolate the compounds. The structures were elucidated based on extensive spectroscopic data analyses. RESULTS: Seven compounds were obtained and their structures were identified as 10-benzoyl-cycloparvifloralone (1), cycloparvifloralone (2), 2α-hydroxycycloparviforalone (3), henrylactone B (4), merrillianone (5), henrylactone C (6) and 7, 14-ortholactone- 3-hydroxyfloridanolide (7). CONCLUSION: Compound 1 is a new sesquiterpene lactone. The tested compounds showed weak anti-HBV activities on HBV surface antigen (HBsAg) secretion and HBV e antigen (HBeAg) secretion using Hep G2.2.15 cell line.

Two new monoterpenes from the fruits of Illicium lanceolatum.

Two new monoterpenes, p-mentha-1(7),8-dien-2-O-ß-D-glucoside and trans-2,4-dihydroxy-2,4-dimethyl-trans-1-acetic acid γ-lactone were isolated from the fruits of Illicium lanceolatum along with trans-2,4-dihydroxy-2,4-dimethyl-cis-1-acetic acid γ-lactone, (1R,2R,4R)-8-p-menthen-1,2-diol, trans-sobrerol, (1S,2S,4R)-p-menthane-1,2,8-triol and (1S, 2S, 4R, 8R)-p-menthane-1,2,9-triol. The structures of the isolates were confirmed by spectroscopic analysis and they showed no inhibitory effects on the in vitro growth of microbial organisms (Escherichia coli, Staphyloccocus aureus, Bacillus subtilis) at less than 1.0 mg/mL.

Diterpenoid alkaloids from Aconitum kirinense.

A new C(18)-diterpenoid alkaloid, kirinenine A (1), was isolated from the root of Aconitum kirinense, along with eight known diterpenoid alkaloids. The structures of all compounds were characterized on the basis of extensive NMR and MS analyses and by comparison with literature values, and the new one was further confirmed by X-ray crystallographic diffraction. All the compounds were isolated from the title plant for the first time.

Two new compounds from Urena lobata L.

Two new compounds 1 and 2 have been isolated from the aerial parts of Urena lobata L. The structures of the two new compounds were established as ceplignan-4-O-ß-d-glucoside (1) and 2,5-dihydroxy benzoic acid-7-(2,6-dimethyl-6-hydroxy-2,7-octadienoic acid) anhydride-5-O-ß-d-apiofuranosyl(1 â†’ 2)-ß-d-glucoside (urenoside A) (2), on the basis of chemical and spectral evidence, including 1D and 2D NMR spectroscopic data as well as mass spectrometry (HR-ESI-MS).

[The isolation and identification of a new lignanoside from Selaginella tamariscina (Beauv.) Spring].

AIM: To study the chemical constituents of Selaginella tamariscina (Beauv.) Spring. METHODS: The compounds were isolated and purified by macroporous adsorption resin, Sephadex LH-20 and silica gel column chromatography and identified on the basis of their physicochemical and spectral data. RESULTS: Four compounds were obtained from the n-BuOH fraction of 70% acetone extracts. Their structures were elucidated as (7S, 8R)-7, 8-dihydro-7-(4-hydroxy-3,5-dimethoxyphenyl)-8-hydroxymethyl-[1'-( 7'-hydroxyethyl)-5' methoxyl] benzofuran-4-O-beta-D-glucopyranoside (tamariscinoside C, I), D-mannitol (II), tyrosine (II), shikimic acid (IV). CONCLUSION: Compound I is a new compound, compounds II and III were obtained from the genius for the first time, compound IV was yielded from the plant for the first time.

[Study on the chemical constituents of Selaginella tamariscina (Beauv.) Spring].

AIM: To study the chemical constituents of Selaginella tamariscina (Beauv.) Spring. METHODS: Various chromatographic techniques were used to separate and purify the chemical constituents. Their physico-chemical properties and spectral data were used to elucidate the structures. RESULTS: Four compounds were isolated from the n-BuOH fraction of the water-extracts. Their structures were identified as 1-hydroxy-2-[2-hydroxy-3-methoxy-5-(1-hydroxyethyl)-phenyl]-3-(4-hydroxy-3,5-dimethoxy)-propane-1-O-beta-D-glucopyranoside (tamariscinoside B, I), adenosine (II), guanosine (III), arbutin (IV). CONCLUSION: Tamariscinoside B (I) is a new compound, while the others were isolated from Selaginella for the first time.

[Isolation and structure identification of the chemical constituents from pine needles of Pinus massoniana Lamb].

AIM: To study the chemical constituents of the water-extracts of the pine needles of Pinus massoniana Lamb.. METHODS: Pine needles were collected in Xixia country and extracted with boiling water for two times and the water-extracts were concentrated. The crude extract of pine needles was fractionated into four fractions by Et2O, EtOAc and n-BuOH. Various column chromatography with D-101 macroreticular resin, Toyopearl HW-40 and silica gel were employed for the isolation and purification of compounds from the n-BuOH fraction of pine needles. The structures of the compounds were identified by physiochemical properties and spectral analysis (UV, IR, MS, 1HNMR, 13CNMR, DEPT, HMQC, HMBC, etc.). RESULTS: Four compounds were isolated from the n-BuOH fraction of water-extracts, and their structures were identified as 3-methoxyl-9'-O-alpha-L-rhamnopyranosyl-4':7,5':8-diepoxyneoligan-4,9-diol (massonianoside E, I), 4,4',8,8',9-pentahydroxyl-3,3'-dimethoxyl-7,9'-monoepoxylignan (II), umbelliferon (III), 4-(4'-hydroxyl-3'-methoxylbenzyl)-2-butanone (IV). CONCLUSION: Compound I is a new compound, and compounds II, III and IV were isolated from this plant for the first time.

[Isolation and structural identification of chemical constituents from Selaginella tamariscina (Beauv.) Spring].

AIM: To study the chemical constituents of the water-extracts of Selaginella tamariscina (Beauv.) Spring. METHODS: Various chromatographic techniques were used to separate and purify the constituents. Their physico-chemical properties and spectral data were used to elucidate the structures. RESULTS: Nine compounds were isolated and identified as (2R,3S)-dihydro-2- (3',5'-dimethoxy-4'-hydroxyphenyl)-7-methoxy-5-acetyl-benzofuran (1), 3-hydroxy-phenpropionic acid-(2'-methoxy-4'-carboxy-phenol) ester (tamariscina ester A, 2), sygringaresinol (3), 1-(4'-hydroxyl-3'-methoxyphenyl)glycerol (4), ferulic acid (5), caffeic acid (6), vanillic acid (7), syringic acid (8), and umbelliferone (9). CONCLUSION: Compound 1 and 2 are new compounds, and the others were isolated from Selaginella for the first time.

[Two new phenylethanoid glycosides from Corallodiscus flabellata].

AIM: To study the chemical constituents from Corallodiscus flabellata. METHODS: Fresh plant of Corallodiscus flabellata was extracted twice with boiling water, filtered to remove insoluble materials, concentrated under reduced pressure at temperature 55 degrees C to a small volume. The concentrated liquor was subjected to solvent-solvent partitioning using ether, ethyl acetate, and n-butanol (saturated with water). The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content. Their fractions from resin were repeatedly chromatographed over Sephadex LH-20, Toyopearl HW-40, gel MCI, Gel CHP-20 and silica gel column. Structures of compounds obtained were identified on the basis of their spectral data, hydrolysis and chemical correlation. RESULTS: Two phenylethanoid glycosides (I, II) and three phenolic acids were obtained from the EtOAc fraction of water-extracts. Their structures were identified as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->2)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[(5-O- Vanilloyl)-beta-D-apiofuranosyl(1-->2)]-beta-D-glucopyranoside (II), vanillic acid (III), syringic acid (IV) and ferulic acid (V). CONCLUSION: I and II are new compounds. Compounds III, IV and V were isolated from this plant for the first time.

[Studies on the lignan chemical constituents from pine needles of Pinus massoniana Lamb].

AIM: To study the chemical constituents of the pine needles from Pinus massoniana Lamb.. METHODS: Various chromatographic techniques were used for the separation and purification. The physicochemical properties and spectral data were used to elucidate the structures. RESULTS: Three compounds were isolated from the n-BuOH fraction of water-extracts. The structures were identified as (7S,8R)-4,9'-dihydroxyl-3, 3'-dimethyoxyl-7, 8-dihydrobenzofunan-1'-propanolneolignan-9-O-alpha- L-rhamnopyranoside (massonianoside D, 7), 4,4',9,9'-tetrahydroxyl-3,3'-dimethyoxyl-tetrahydrocyclolignan ((+)-isolariciresinol, 8) and 4,4',9'-trihydroxyl-3,3'-dimethyoxyl-tetrahydrocyclolignan-9-O-beta-D- xylopyranoside (isolariciresinol-9-O-xyloside, 9) on the basis of spectral data (ORD, UV, IR, MS, 1HNMR, 13CNMR, 1H-1HCOSY, HMQC and HMBC etc.). CONCLUSION: Compound 7 is a new compound, while compounds 8 and 9 were isolated from this plant for the first time.

[Isolation and structural identification of phenylethanoid glycosides from Corallodiscus flabellata].

AIM: To study the chemical constituents from Corallodiscus flabellata. METHODS: The compounds were isolated and purified by macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data. RESULTS: Three phenylethanoid glycosides (I-III) were obtained from the n-BuOH fraction of water-extracts. Their structures were elucidated as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->3)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[4-O-trans-caffeoyl-beta-D-apiofuranosyl (1-->3)-beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (II) and 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl(1-->3)-beta- D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (III). CONCLUSION: Compounds I, II and III are new compounds.

[Isolation and structure identification of lignans from pine needles of pinus massoniana Lamb].

AIM: To study the chemical constituents from the water-extracts of pine needles of Pinus massoniana Lamb. METHODS: Chromatographic techniques were used to separate and purify compounds. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1H, 13C-1H NMR, DEPT, HMBC etc.) were used to elucidate the structures. RESULTS: Three lignans were isolated from the n-BuOH fraction of water-extracts. Their structures were identified as (7S,8R)-3',4,9'-tridihydroxy-4-methoxy- 9-O-shikimoyl-7,8-dihydrobenzofuran-1'-propylneolignan (massonianoid A, I), (7S,8R)-4,9-dihydroxy-3,3'-dimethoxy-7,8- dihydrobenzofuran-1'-propylneolignan (II) and 4,4',8-trihydroxy-4,4'-dimethoxy-9-lignanolide (III). CONCLUSION: Compound I is a new compound. While II and III were isolated from this plant for the first time.

[Isolation and structure identification of ligan glycosides from pine needles of Pinus massoniana lamb].

AIM: To study the chemical constituents of the pine needles of Pinus massoniana lamb.. METHODS: Various chromatographic techniques were used to separate and purify. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1 H COSY, HMQC, DEPT, HMBC and ORD ect.) were measured for structure elucication. RESULTS: Three compounds were isolated from the n-BuOH fraction of water-extracts. Their structures were identified as massonianoside A (4), massonianoside A: (7S, 8R)-3, 4, 9'-trihydroxyl-3-methyoxyl-7, 8-dihydrobenzofunan-1'-propanolneoligan-9-O-alpha-L-rhamnopyranoside, massonianoside C (5), (7S, 8R)-9,9'-dihydroxyl-3,3'-dimethyoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-alpha-L-rhamnopyranoside and cedrusin-4-O-beta-glucoside (6), (7S, 8R)-3',9,9'-trihydroxyl-3-methoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-beta-D-glucopyranoside. CONCLUSION: Compound 4 and 5 are new compounds.

[Studies on the chemical constituents from herba of Corallodiscus flabellata].

OBJECTIVE: To study the chemical constituents from Corallodiscus flabellata. METHOD: The compounds were isolated with macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data. RESULT: Six compounds were obtained and identified as vanillic acid, 3,4-dihydroxyphenylethyl-8-O-beta-D-glucopyranoside, syringic acid, caffeic acid, isoacteoside, ferulic acid. CONCLUSION: All the compounds were isolated from this plant for the first time.