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Objective: To observe effect of Mori Folium-containing serum on glucose consumption and cell activity of fat cell line 3T3-L1 insulin resistance (IR) model, in order to screen out the optimal concentration of drug-containing serum, detect effect of Mori Folium on the content of inflammatory factors, and explore the possible mechanism. Method: 3T3-L1 preadipocytes in logarithmic growth phase were selected, and induced with 10 mg·L-1 insulin (Ins), 0.25 mmol·L-1 dexamethasone (DEX) and 0.5 mmol·L-1 3-isobutyl-methylxanthine(IBMX) for 48 h and then with 10 mg·L-1 Ins for 48 h. After the cells were differentiated into mature adipocytes, they were induced with 1 μmol·L-1 DEX for 96 h to establish IR model. Glucose content in the supernatant of cells was detected by glucose oxidase after serum containing Mori Folium cultured for 12,24,36, 72 h. Methyl-thiazdyl-tetrazolium(MTT) was used to detect the effect of serum containing Mori Folium on IR cells activity. The content of tumor necrosis factor-α(TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, the effects of inflammatory factors on the expressions of insulin signaling pathway proteins insulin receptor (InsR), insulin receptor substrate (IRS), p-IRS1 and glucose transporter 4 (GLUT4) were determined by Western blot. Result: Serum containing Mori Folium could significantly increase the glucose consumption rate and cell activity of IR cells (Pα (PPPConclusion: Mori Folium can significantly improve IR status of 3T3-L1 cells, and its mechanism may be related to inhibiting TNF-α and promoting the expressions of insulin signaling pathway proteins.
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Aim Toobservetheeffectofconcisepre-scriptions of Chinese medicine Huannao Yicong Decoc-tion(HYD)on regulatory pathway of secretase in APP/PS1 double transgenic cell line(HEK293),and to in-vestgateitsmechanism.Methods Theproliferationof AD cell model and the toxicity of each investigational drugs were ebserved by CCK-8;the changes in micro-scopic structure of each group were observed by(Trans-mission electron microscope,TEM);the activities of gamma-secretes was observed by Dual Luciferase Re-porter Gene Assay Kit ,and then the expression of pre-senilin 1(PS1),carboxyl terminus of Hsc70-interacting protein(CHIP),GTP binding protein (CDC42 ),ante-rior pharynx defective-1α(APH-1α),Hypoxia induc-ible factor-1α(HIF-1α) were detected by Western blot.Results 15%HYDserumincreasedthecellac-tivity compared to blank serum (P 0. 05 );compared to control group,HYD directly group inhibited the HIF-1αprotein ex-pression after 48h medication(P<0. 05);compared to 0h midicaiton,DAPT group inhibited the HIF-1αpro-tein expression at the point of 24 h (P<0. 05 ).Con-clusions HYDcantreatADthroughprotectingthe mitochondrial function,reducing the formation of lipo-fuscin,inhibiting the activity of γsecretase by down-regulating the activity of HIF-1α,decreasing the stabil-ity and activity of PS1 by promoting the expression of CDC42.This shows that HYD has good research and development prospect as an effective drug for preven-tion and treatment of AD.
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OBJECTIVE: To establish the fingerprint of the serum containing drug of Yinchenhao Decoction (YCHD), and analyze the drug-originated constituents in serum. METHODS: An HPLC method was set up on a Symmetry C18(4.6 mm × 250 mm, 5 μm) column with a Security Guard Cartridges C18 (4.0 mm × 3.0 mm) guard column by gradient elution of acetonitrile-0.1% phosphoric acid in water at the flow rate of 1.0 mL · min-1. The column temperature was maintained at 30℃. The detection was carried out at 238 and 440 nm. SD rats were taken as serum donors. Ten batches of serum containing drugs were prepared after ig administration of YCHD. The fingerprints of these serum samples were determined, the common modes were established, and the similarities between fingerprints were evaluated. Drug-originated constituents in blood were distinguished by comparing the fingerprint of serum containing YCHD with the one of control serum. The sources of drug-originated constituents were analyzed by comparing the fingerprint of serum containing YCHD with the ones of serum containing individual herbs of YCHD. Drug-originated constituents in blood were identified by comparing the retention time and UV spectra of peaks in fingerprint of serum containing YCHD with the ones of YCHD and reference substances, respectively. RESULTS: The similarities between the fingerprints of ten batches of serum containing YCHD and common models at 238 and 440 nm were greater than 0.904 and 0.903, respectively. Twenty common peaks of drug-originated constituents were identified in the fingerprints of serum containing drug of YCHD at 238 and 440 nm. Out of them, six were the original compounds, the other 14 were metabolites. Three constituents out of six original form ones were identified as geniposidic acid and genipo-side (iridoids originated from GF), and rhein (anthraquinone from RRR), and the other three were identified as an anthraquinone from RRR and two crocetin derivatives from FG. Out of 14 metabolites, one was identified as the metabolite of iridoid, four were from anthraquinone, and nine were from crocetin derivatives. CONCLUSION: The established method is accurate, reliable and can be used for the analysis of drug-originated constituents in serum containing drug of YCHD.
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A rapid and validated UPLC-MS method was developed for investigating the absorbed components of Paederia scandens (Lour.) Merrill (P. scandensy) in rat plasma. The bioactive constituents in plasma samples from rats administrated orally with P. scandens extract were analyzed by Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Four prototype compounds were identified in rat serum as potential bioactive components of P. scandens by comparing their retention times and mass spectrometry data or by mass spectrometry analysis and retrieving the reference literatures. Glucuronidation after deglycosylation was the major metabolic pathway for the iridoid glycosides in P. scandens. These results showed that the methods had high sensitivity and resolution and were suitable for identifying the bioactive constituents in plasma after oral administration of P. scandens. providing helpful chemical information for further pharmacological and mechanistic researched on the P. scandens.
Subject(s)
Animals , Male , Rats , Administration, Oral , Chromatography, Liquid , Methods , Drugs, Chinese Herbal , Metabolism , Iridoid Glycosides , Blood , Rats, Wistar , Rubiaceae , Chemistry , Sensitivity and Specificity , Spectrometry, Mass, Electrospray Ionization , MethodsABSTRACT
Objective: To investigate the effects of serum containing dioscin on sodium current (INa) and reveal the mechanisms of cardioprotection and antiarrhymias. Methods: Serum was obtained from the aortaventralis from Wistar rats after ig administration 4 d of dioscin 300 mg/kg, twice daily for 4 d. Single ventricular myocytes were isolated from adult rat hearts by enzymatic dissociation and the effects of dioscin on INa were observed by whole-cell patch clamp. Results: Serum containing dioscin shifted downward the I-V curve with increased peak current density. With the effects of 1, 10, and 100 μL serum containing dioscin, the peak current density was dose-dependently changed from (-52.10 ± 3.80) pA/pF to (-76.44 ± 4.09) pA/pF and (-81.96 ± 4.70) pA/pF, respectively (P < 0.01 vs control); Dioscin facilitated the activation process with the inactivation process unchanged. The half activation potential was changed from (-57.69 ± 1.86) mV to (-59.71 ± 2.57) mV, ( -66.56 ± 1.32) mV (P < 0.01 vs control), and (-68.52 ± 3.91) mV (P < 0.01 vs control), respectively; The recovery process of sodium channel was accelerated by 1, 10, and 100 μL serum containing dioscin with the recovery constant τ changed from (150.73 ± 21.49) ms to (143.19 ± 13.88) ms, (84.83 ± 18.03) ms (P < 0.01 vs control), and (80.63 ± 13.89) ms (P < 0.01 vs control). Conclusion: Serum containing dioscin could increase the sodium current by facilitating the activation process and accelerating the recovery process of sodium channel.
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Objective To observe the effect of serum containing Qiangjijianli oral liquid in-vitro proliferation of rat mesenchymal stem cells (MSCs). Methods Rat mesenchymal stem cells dissociated from the bone marrow by density gradient method were cultured. MSCs were identified by marking of bromodeoxyuridine (Brdu) and staining of CD44, CD45. Experimental group was cultivated with serum containing Qiangjijianli oral liquid and control group with blank serum. Optical absorption value of MSCs was stained by methyl thiazolyl tetrazolium (MTT). Results Serum containing Qiangjijianli oral liquid at different concentrations could promote the proliferation of MSCs, the difference being significant in comparison with the control group (P
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Objective To compare the content of main constituents of Compound Wurenchun Capsules dissolved in serum and plasma of rats. Methods Serum and plasma containing drug were prepared after ig the preparation to rats. Lichrosphere C_ 18 (250 mm?4.6 mm, 5 ?m) column and Phenomenex Description C_ 18 (4.0 mm?3.0 mm) protective column were used. The mobile phase consisted of methanol-water, eluted in gradient mode. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 254 nm. Results The linear ranges of schisandrin and schisandrin B were 0.051 2 .614 4 and 0.039 8—0.477 6 ?g. The average relative recoveries of schinsandrin and schisandrin B were 96.72%, 101.06%, 102.05%, and 99.03%, 100.18%, 100.28% in low, middle, and high concentrations, respectively. The average contents of schisandrin in serum and plasma were 11.063 2 and 12.883 7 ?g/mL, schisandrin B were 7.490 9 and 12.590 8 ?g/mL, respectively. Conclusion The main constitu-ents of Compound Wurenchun Capsules contained in plasma account for higher than the ones in serum.
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Objective To determine schisandrin,schisandrol B,deoxyschizandrin,and schisandrin B in serum containing drug of Compound Wurenchun Capsula.Methods An HPLC method was set up.Li-chrosphere C18 column(250 mm ?4.6 mm,5 ?m) and Phenomenex Description C18(4.0 mm?3.0 mm)protective column were used.Acetonitrile-water was used as gradient mobile phase.The flow rate was 1.0 mL/min.The column temperature was 30 ℃ and the detection wavelength was 210 nm.Results The linear ranges of schisandrin,schisandrol B,deoxyschizandrin,and schisandrin B were within 0.051 2-0.768 0 ?g(r=0.999 5),0.054 0-0.810 0 ?g(r=0.999 6),0.012 3-0.184 5 ?g(r=0.999 8),and 0.039 8-0.597 0 ?g(r=0.999 6),respectively.The average concentration of these four lignans in serum containing drug were 8.021 1,6.231 0,0.530 8,and 5.851 0 ?g/mL,respectively.Conclusion This method is easy,sensitive,specific,and accurate for the assaying of the four lignans in serum containing drug of Compound Wurenchun Capsula.
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AIM: To illuminate the therapeutic basis of Compound Wurenchun Capsules in combination with serum pharmacology,the lignans of this preparation migrating to blood of rats after oral administration having been accomplished. METHODS: By UPLC-MS/MS method, lignans migrating to blood were affirmed by comparing the extracted ion chromatography (EIC) of the serum containing drug with the ones of Compound Wurenchun Capsules, the control serum and control articles, correlated ion peak in mass-spectrogram were analyzed at the same time. RESULTS: Five lignans migrating to blood have been found, they are schisandrin, schisandrol B, schisantherin, deoxyschizandrin and schisandrin B. CONCLUSION: Five lignans above mentioned are likely the effective substances of Compound Wurenchun Capsules in human body. More study by means of combining with serum pharmacology will illuminate the therapeutic basis of this preparation.