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Objective:Rosin alcohol two glycosidase (pinores inoldiglucoside, PDG) effect on the improvement of the osteoporosis in mice and the possible mechanism of action.Methods:A mouse model of osteoporosis was established by removing bilateral ovaries. According to the random number table method, 50 mice were divided into model (MOD) group (equal volume of normal saline), positive control (PC) group (0.2 mg/kg estradiol), PDG-low dose (L) group (5 mg/kg) PDG), PDG-high dose (H) group (10 mg/kg PDG by gavage) and PDG-H+ML385 group (10 mg/kg PDG by gavage after intramuscular injection of 30 mg·kg-1 ML385), 10 mice in each group; Another 10 mice without any ovarian treatment were selected as sham group (intragastric administration of the same amount of normal saline). After 3 weeks of drug intervention, the serum alkaline phosphatase (ALP), tartrate resistant acid phosphatase (TRAP), interleukin-6 (IL-6) and tumor necrosis factor were detected by enzyme-linked immunosorbent assay (ELISA) -α (TNF-α). The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by dual energy X-ray. The mRNA and protein expression levels of nuclear factor related factor 2 (Nrf2), oxidase heme oxygenase 1 (HO-1), quinone oxidoreductase 1 (NQO1) were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively.Results:The serum ALP levels in Sham, MOD, PC, PDG-L, PDG-H and PDG-H + ML385 groups were (25.88±3.42), (47.42±5.32), (36.20±3.37), (38.95±3.24), (29.66±2.64), (39.57±2.06) U/dL, respectively; The serum trap levels were (2.18±0.40), (4.69±0.83), (3.58±0.38), (3.67± 0.48), (2.93±0.26), (3.81±0.49) U/L, respectively; BMD (153.04±12.96), (86.25±6.71), (126.53±8.99), (119.77±8.84), (139.18±15.94), (92.09±4.17) mg/cm 2; The expression levels of Nrf2 mRNA were 1.00±0.00, 0.35±0.04, 0.67±0.05, 0.54±0.03, 0.82±0.08 and 0.48±0.04, respectively; The expression levels of HO-1 mRNA were 1.00±0.00, 0.25±0.03, 0.56±0.03, 0.47± 0.03, 0.71±0.04 and 0.37±0.04, respectively; The expression levels of nqo1mrna were 1.00±0.00, 0.38± 0.02, 0.63±0.03, 0.58±0.04, 0.79±0.05 and 0.44±0.03, respectively; Nrf2 protein expression levels were 0.98±0.08, 0.26±0.04, 0.52 ±0.06, 0.46±0.03, 0.86±0.07, 0.45±0.05. HO-1 protein expression levels were 0.39±0.02, 0.09±0.01, 0.15 ±0.03, 0.17±0.03, 0.26±0.03, 0.12±0.02. NQO1 protein expression levels were 0.53±0.03, 0.21±0.02, 0.38±0.04, 0.29±0.02, 0.55±0.03, 0.24±0.04, respectively; The levels of serum IL-6 were (104.25±11.35), (515.38±74.48), (351.78± 65.12), (364.73±36.64), (246.18±17.52), (408.93±32.56) pg/ml, respectively; The levels of serum TNF-α were (33.79±3.55), (170.11±19.24), (76.09±8.99), (84.95±6.12), (66.98±3.73), (119.04±9.75) pg/mL, respectively; The serum SOD levels were (258.47±19.25), (72.15±8.12), (187.60±14.63), (152.61±12.36), (204.22±19.65), (138.01±13.62) U/mL, respectively; The serum MDA levels were (2.02±0.27), (4.75±0.73), (3.19±0.46), (3.7±0.49), (2.91±0.42), (4.10 ±0.25) μmol/L respectively. There were significant differences between MOD group and Sham group, or PC, PDG-L, PDG-H, PDG-H+ML385 group and MOD group,or PDG-H+ML385 group and PDG-H ( P<0.05) . Conclusion:PDG can reduce bone inflammation and oxidative stress by activating Nrf2 pathway and improve the state of osteoporosis.
ABSTRACT
From the fungus Trichoderma sp., we isolated seven novel 18-residue peptaibols, neoatroviridins E-K (1-7), and six new 14-residue peptaibols, harzianins NPDG J-O (8-13). Additionally, four previously characterized 18-residue peptaibols neoatroviridins A-D (14-17) were also identified. The structural configurations of the newly identified peptaibols (1-13) were determined by comprehensive nuclear magnetic resonance (NMR) and high-resolution electrospray ionization tandem mass spectrometry (HR-ESI-MS/MS) data. Their absolute configurations were further determined using Marfey's method. Notably, compounds 12 and 13 represent the first 14-residue peptaibols containing an acidic amino acid residue. In antimicrobial assessments, all 18-residue peptaibols (1-7, 14-17) exhibited moderate inhibitory activities against Staphylococcus aureus 209P, with minimum inhibitory concentration (MIC) values ranging from 8-32 μg·mL-1. Moreover, compound 9 exhibited moderate inhibitory effect on Candida albicans FIM709, with a MIC value of 16 μg·mL-1.
Subject(s)
Peptaibols/chemistry , Trichoderma/metabolism , Tandem Mass Spectrometry/methods , Anti-Infective Agents/pharmacology , Spectrometry, Mass, Electrospray Ionization/methodsABSTRACT
Background: Cathartic colon belongs to the category of 'constipation' in Traditional Chinese Medicine, and its pathogenesis is related to deficiency of kidney temperament and weak promotion ability, which has become a hot spot of global medical attention. Aims: To investigate the effect and possible mechanism of Jiawei Shenqi-wan (JSQW) on intestinal transmission function and pathological changes of interstitial cells of Cajal (ICCs) in rats with cathartic colon. Methods: Forty rats were randomly divided into blank group, model group, prucalopride group and JSQW group. Fecal moisture content, fecal particle number and intestinal transit rate were detected. The pathological changes of ICCs were observed under transmission electron microscope. RT-qPCR was used to detect the mRNA expressions of water channel proteins (AQP3 and AQP9) and SCF/c-kit pathway. Immunohistochemistry was used to measure the expressions of α-smooth muscle actin (α-SMA), inducible nitric oxide synthase (iNOS) and 5-hydroxytryptamine (5-HT)
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OBJECTIVE:To establish the method for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules. METHODS:HPLC switching walvelength method was adopted. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-0.45％ phosphoric acid solution-triethylamine(50:49:1,V/V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 265 nm(berberine hydrochloride)and 280 nm(baicalin). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride and baicalin were 60.3-312.8 ng(r=0.9997)and 81.5-368.9 ng(r=0.9999). The limits of quantitation were 0.6668,0.7740 ng,andthe limits of detection were 0.2226,0.2580 ng,respectively. RSDs of intermediate precision,stability and repeatability tests were all lower than 1.0％. The recoveries were 96.48％-99.30％(RSD=1.06％,n=6) and 95.20％-99.39％(RSD=1.66％,n=6), respectively. RSDs of durability test were all lower than 2.0％. CONCLUSIONS:The method is simple, precise, stable, reproducible,accurate and durable. It can be used for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules.