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This study was aimed to develop a simultaneous determination of zinc and cadmium with carbon fiber electrode by linear sweep stripping voltammetry. Effects of analytical base solution, accumulation potential, ac-cumulation time and scanning velocity were also investigated on the determination. The results showed that in 1 mol·L-1 H2SO4 medium solution, zinc and cadmium had good electrochemical response on the carbon fiber electrode and appeared sensitive anodic stripping peaks at-0.97 V and-0.77 V, respectively. The anodic stripping peak cur-rents and concentrations of zinc and cadmium showed good linear relationships, with correlation coefficients R2 of 0.996 5 and 0.995 4, respectively. The detection limits of zinc and cadmium (S/N=3) were 1.5×10-9 mol·L-1 and 1.0×10-10 mol·L-1, respectively. The average recoveries of zinc and cadmium were 98.9%and 98.1%with the rela-tive standard deviations (RSD) of 2.72%and 2.45%(n=6), respectively. It was concluded that the method of simul-taneous determination of zinc and cadmium with carbon fiber electrode was simple, sensitive and accurate.
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Hypoxia-ischemia leads to serious neuronal damage in some brain regions and is a strong risk factor for stroke. The aim of this study was to investigate the neuroprotective effect of tanshinone I (TsI) derived from Danshen (Radix Salvia miltiorrhiza root extract) against neuronal damage using a mouse model of cerebral hypoxia-ischemia. Brain infarction and neuronal damage were examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin histochemistry, and Fluoro-Jade B histofluorescence. Pre-treatment with TsI (10 mg/kg) was associated with a significant reduction in infarct volume 1 day after hypoxia-ischemia was induced. In addition, TsI protected against hypoxia-ischemia-induced neuronal death in the ipsilateral region. Our present findings suggest that TsI has strong potential for neuroprotection against hypoxic-ischemic damage. These results may be used in research into new anti-stroke medications.
Subject(s)
Animals , Mice , Brain , Brain Infarction , Abietanes , Drugs, Chinese Herbal , Eosine Yellowish-(YS) , Fluoresceins , Hematoxylin , Hypoxia-Ischemia, Brain , Neurons , Neuroprotective Agents , Risk Factors , Salvia miltiorrhiza , Stroke , Tetrazolium SaltsABSTRACT
Objective To investigate the influences of Radix salvia miltiorrhiza (RSM) preconditioning on the expressions of phosphorylated eukaryotic initiation factor 2α (p-eIF-2ot) and caspase12 in hepatic ischemia reperfusion in rats.Methods Eighty Wistar rats were randomly divided into the control group (5 rats),sham operation group (25 rats),ischemia reperfusion (IR) group (25 rats) and RSM pretreated group (25 rats).Rats in the sham operation group,IR group and RSM pretreated group were subdivided into 5 groups at different time intervals (0,3,12,24 and 72 hours).Mter midline laparotomy,all structures in the hepatic portal were clamped for 45 minutes followed by different periods of reperfusion.Rats in the control group did not receive any treatment; rats in the sham operation group only received anatomy of the hepatic portal without clamping; rats in the RSM pretreated group received RSM by intravenous injection 30 minutes before ischemia at a dose of 6 ml/kg.Rats in the sham operation group and the IR group received a dose of normal saline as RSM pretreated group.The protein expressions of p-eIF-2α and caspase12 in the hepatic tissues of each group were detected by the Western blot,and the pathological changes of hepatic tissues in each group were detected by hematoxylin and eosin staining.All data were analyzed using the analysis of variance,LSD test or Games-Howell method.Results The relative expression of p-eIF-2α in the control group was 0.296 ± 0.038,and the relative expressions of p-eIF-2α in the sham operation group at different time points were 0.304 ± 0.048,0.298 ± 0.038,0.272 ± 0.042,0.266 ± 0.076 and 0.296 ± 0.043,with no significant difference between the 2 groups (P ＞ 0.05).The relative expression of p-eIF-2α in the IR group at 0 hour was 0.310 ± 0.034,which had no significant difference compared with the control group and the sham operation group (F =0.15,P ＞0.05).The relative expressions of p-eIF-2α in the IR group at 3,12,24 hours were 0.386 ± 0.021,0.710 ± 0.034,0.474 ± 0.017,which had no significant difference compared with the control group and the sham operation group (F =11.90,211.52,25.15,P ＜ 0.05).The relative expression of p-eIF-2α in the IR group at 72 hours was 0.336 ± 0.043,which was back to the level of the control group and the sham operation group (F =1.57,P ＞ 0.05).The relative expressions of p-eIF-2α in the RSM pretreated group at different time intervals were 0.278 ± 0.044,0.800 ± 0.079,1.056 ± 0.125,0.736 ±0.087 and 0.442 ± 0.047,which were significantly lower than the group at 3,12,24,72 hours (P ＜ 0.05).The relative expression of caspase12 in the control group was 0.983 ± 0.003,and the relative expressions of caspase12 in the sham operation group at different time intervals were 0.974 ± 0.004,0.983 ± 0.005,0.985 ± 0.003,0.981 ± 0.004 and 0.978 ± 0.004,with no significant difference between the 2 groups (P ＞ 0.05).The relative expression of caspase12 in the IR group at 0 hour was increased to 1.018 ± 0.076,while it had no significant difference compared with the control group and the sham operation group (F =1.43,P ＞ 0.05).The relative expressions of caspase12 in the IR group began to rise at 3 hours (2.056 ±0.067),and peaked at the 12 hours (2.804 ± 0.050),still at a relative higher level at 24 hours (1.882 ± 0.037),and began to decrease at 72 hours (1.282 ± 0.066),it had significant difference compared with the control group and the sham operation group (F=1290.69,6490.84,2898.71,103.61,P ＜ 0.05).The relative expressions of caspase12 in the RSM pretreated group at different time intervals were 0.998 ± 0.056,1.442 ± 0.066,1.990 ± 0.068,1.364 ± 0.056and 0.962 ±0.054.The relative expressions of caspase12 in the RSM pretreated group at 3,12,24,72 hours were significantly lower than the IR group (P ＜ 0.05).The results of pathomorphological examination showed that the hepatic lobules were integrated and the nucleuses were large and round in the control group an d the sham operation group; deformation of the hepatic lobule,smaller cell volume,nuclear condensation and necrosis were observed in the IR group; cell swelling and slight spotty necrosis were detected in the RSM pretreated group.Conclusions RSM could protect liver from damages during IR through up-regulating the expression of p-eIF-2α,reducing the apoptosis induced by endoplasmic reticulum stress.
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Objective To study the best extraction method for Radix Salvia Miltiorrhiza effective component. Methods Several extraction methods for Radix Salvia Miltiorrhiza (water decoction, alcohol reflux, ultrasonic extraction and supercritical CO2 extraction) were compared, to optimize the extraction methods by determined of Danshensu and Tanshinone IIA of Radix Salvia Miltiorrhiza. Results Supercritical CO2 extraction had high content of effective component of Danshensu and Tanshinone IIA. Conclusion Supercritical CO2 extraction was the best extraction method for Radix Salvia Miltiorrhiza. This result would be an experimental proof for pre-processing the preparation of Radix Salvia Miltiorrhiza.
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Objective To study the sample quality of 'whitish' Radix Salvia Miltiorrhiza(the root of a grown-environmental-influenced variant sample of Salvia miltiorrhiza,which cortex and transection is white) grown in Luanchuan Region of Henan Province and normal Radix Salvia Miltiorrhiza by the means of TLC and HPLC chromatographic fingerprints;hence to compare the difference of their constituents.Methods TLC Chromatographic condition: Precoated silica gel F_(254) plate;hydrophilic and lipophilic constituents were developed and derivatizated with different solvent systems and visualization reagents,respectively.HPLC Chromatographic condition: Zorbax SB C_(18) column;hydrophilic and lipophilic constituents were gradient eluted,respectively.Results The 'whitish' Radix Salvia Miltiorrhiza contains lower content of lipophilic compounds,the main diterpene quinones was only trace amount,while the content of hydrophilic constituents was relatively higher.Conclusion The 'whitish' phenomenon suggests that the 'micro'-environmental condition can probably influence negatively the bio-synthesis of diterpene-quinones.The factors of influence still need further study.Such phenomenon would be worth concerning seriously for GAP administration.
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Objective To elucidate the ESI-MS performance of lipophilic tanshinones and to establish a fingerprint of lipophilic tanshinones of Radix Salviae Miltiorrhiza.Methods The lipophilic tanshinones were extracted by ultrasonic wave with 95% ethanol from Radix Salviae Miltiorrhiza and the extracts were analyzed directly in positive ion mode by electrospray ion trap mass spectrometry(ESI-MS).Results The lipophilic tanshinones were easily to form molecular ions ~+ and dimeric sodium adduct ions ~+ in positive ion mode,molecular ions of lipophilic tanshinones were fragmentated through lossing H_2O,CO and A-ring cleavages in ESI-MS~2.The ESI-MS fingerprints of lipophilic tanshinone extracts with 14 selected characteristic peaks analyzed with SPSS software were characteristic and stable.Conclusion Lipophilic tanshinones have similar ESI-MS performance;the ESI-MS fingerprint is a useful tool for a rapid and special identification of lipophilic tanshinones in selected traditional Chinese medicine.
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Objective To establish a sensitive HPLC method with UV detection for the determination of tanshinone Ⅱ_A(TSⅡ_A) and cryptotanshinone(CT) in rat plasma and to study pharmacokinetics of Radix Salviae Miltiorrhizae(RSM) liposoluble components in rat in vivo. Methods TS Ⅱ_A and CT in plasma of rat at different sampling time were determined by pretreatment with Gemifibrozil as internal standard,the protein was precipitated in methanol solution after iv administration of alcohol extract of RSM 5 mg/per animal.The mean plasma concentration-time curve was protracted and pharmacokinetic parameters were calculated by 3p87.Results The main pharmacokinetic parameters of TS Ⅱ_A were as follows: t_(1/2?) was(0.088?0.024) h;t_(1/2?) was(2.1?0.7) h;V_C was(0.8?0.6) L;CL was(2.0?1.1) L/h~(-1);AUC_(0-4) and AUC_(0-∞) were(2.2?0.9) mg?h/L and(3.4?1.7) mg?h/L,respectively.Conclusion The concentration-time curve of TS Ⅱ_A can be fitted to two-compartment model after ethanol extract of RSM iv administrated.Concentration of TS Ⅱ_A in rat plasma declines rapidly with large apparent volume of distribution.CT decreases also rapidly and it is difficult to acquire its pharmacokinetic parameters in rat.