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Objective To investigate the effect of Liensinine on lipopolysaccharide ( LPS)-induced acute lung injury (ALI) in mice. Methods BALB/c mice were randomly divided into six group: control group, LPS group, LPS+Liensinine (2 mg/kg, 4 mg/kg, 8 mg/kg) groups, and dexamethasone group. Acute lung injury in mice was induced by nasal instillation of LPS. After 12 h, the pathological changes of lung tissue were observed. The levels of TNF-α, IL-6 and IL-1β in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The number of neutrophils in BALF was detected using Wright-Giemsa staining. Total protein content was detected by BCA protein quantification assay. The pulmonary capillary permeability was examined with Evans blue. The MPO activity, MDA content, SOD activity, and GSH content in lung homogenate supernatant were detected by spectrophotometry. The content of ROS in lung tissue was detected by flow cytometry. Results The LPS group showed inflammatory cell infiltration, thickening of bronchial alveolar wall and pulmonary congestion in the lung tissue, while Liensinine improved the lung injury. In the LPS group, the contents of TNF- α, IL-6 and IL-1β in BALF were significantly increased, the number of neutrophils and the content of total protein were significantly increased, pulmonary capillary permeability, MPO activity and MDA content were increased, SOD activity and GSH content were decreased, the content of ROS was increased; while the Liensinine group reduced the contents of TNF-α, IL-6, IL-1β in BALF, reduced the number of neutrophils and total protein content, decreased the pulmonary capillary permeability, attenuated MPO activity and MDA contents and increased SOD activity and GSH content, and reduced ROS content in the LPS-challenged lung tissue. Conclusions Liensinine protects mice from LPS-induced acute lung injury by its anti-inflammatory and anti-oxidative activities.
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Objective: To optimize the prescription of liensinine solid dispersion osmotic pump controlled release tablet and research its release characteristics in vitro. Methods: The cumulative release percent in 2, 6, and 12 h and the linear correlation coefficient of cumulative release curve were taken as evaluation indexes to select the optimal prescription by using the central composite design- response surface methodology (CCD-RSM). The main factors of influence on drug release, which were the dosage of NaCl and PEG 400, and coating agent thickness. Results: The optimal prescription for liensinine solid dispersion osmotic pump controlled release tablet were as follows: NaCl dose was 166.0 mg, PEG 400 content was 80.5%, and the coating weight gain was 3.5%. Conclusion: The prescription optimization model of liensinine solid dispersion osmotic pump controlled release tablet is optimized by CCD-RSM, and it is proved to follow zero-order release kinetics.
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Objective To optimize the preparation of liensinine HP-β-CD inclusion compound; To investigate its dissolution performance in vitro. Methods The inclusion compound of liensinine was prepared by using saturated water solution method; the cumulative dissolution (45 min) was used as an indicator and Box-Behnken design was adopted to evaluate the influence of feed ratio, mixing time and inclusion temperature on preparation process. Results were analyzed by multiple linear and binomial fitting; response surface methodology was used to screen the optimal inclusion process; predictive parsing and verification experiment were conducted; SEM, DSC, IR, and XRD were applied for the structural characterization of inclusion compound of liensinine. Results The optimal preparation process was: HP-β-CD was 4.5 times the amount of liensinine feeding amount; mixing time was 3.7 h; inclusion temperature was 52 ℃. HP-β-CD inclusion compound of liensinine formed. Conclusion Optimal inclusion process is stable and feasible, which can significantly improve the dissolution of liensinine and increase its bioavailability.
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Objective To study the effects of Liensinine on apoptosis of 5637 cells, and its mechanism thereof. Meth?ods CCK-8 method and the colony formation test were used to detect cell viabilities, and then inhibition rates were calcu?lated. Flow cytometry was used to detect the effects of Liensinine on apoptosis of 5637 cells. Western blot assay was used to detect Caspase-7 protein expression. Results CCK-8 assay and colony formation test indicated that Liensinine inhibited the cell proliferation significantly. Results of flow cytometry indicated that Liensinine induced early apoptosis of 5637 cells. Western blot assay showed that Liensinine improved the expression of Caspase-7 and enhanced the activation of Caspase-7 in 5637 cells. Conclusion Liensinine could inhibit the proliferation of 5637 cells, induce early apoptosis, which may be re?lated with the enhanced expression of Caspase-7 and its activation.
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Objective To observe protective effects of four active liposoluble alkaloids of a Chinese herb, lotusine (Lot), liensinine (Lien), isoliensinine (Iso) and neferine (Nef) of embryo loti (the green embryo), against H2 O2-induced oxidative damage on human umbilical vascular endothelial cell ECV-304.Methods The protective effects of Lot, Lien, Iso and Nef on the survival of normal and oxidatively damaged ECV 304 cells were studied by cell morphology observation and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium (MTT) assay.The levels of nitric oxide (NO) and nitric oxide synthase ( NOS) were measured using colorimetric assay.Results Lot, Lien, Iso and Nef did not affect cell morphology and cell viability of normal ECV 304 cells.The survival of oxidative damaged vascular endothelial cells was rescued by incubating with Lot at 100μmol/L, and Lien, Iso and Nef at 0.1 μmol/L.The proliferative activity of medicated groups increased to 112.8%, 129.3%, 125.6 and 118.2%, respectively (P<0.01 or 0.05), relative to that of the group with H2O2 induced oxidative damage.The four alkaloids restrained oxidative injury of endothelial cells induced by H2 O2 and the protective influences were similar with captopril, which served as a positive control.Each alkaloid except Lot reduced intercellular space and increased the connections of oxidative damaged cells, concomitant with more recognizable cell borders.Lien, Iso and Nef also increased the concentration of NO ( P<0.05 ) .Besides, all of the four alkaloids activated NOS in damaged vascular endothelial cells ( P<0.05 ) . Conclusion The four alkaloids of embryo loti, especially Lien, Iso and Nef, have certain protective effects against H2 O2-induced oxidative damage on vascular endothelial cells.The protective mechanism may be promotion of NO release through the increase of NOS production.
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Objective To explore the effect of liensinine on cortical EEG of epileptic rats induced by lithium chloride-pilocarpine,and investigate the effective spectrum of liensinine on epilepsy.Methods 32 SD rats were randomly divided into four groups:low dose of liensinine group(2.5 mg/mL, 10μL),high dose of liensinine group(5 mg/mL,10μL),the normal saline group(10μL)which was negative control group,levetiracetam group (100 mg/mL,10μL)which was positive control group,8 rats in each group.Electrocorticogram of rats was recorded after chloride lithium-pilocarpine model was induced.The anesthetic rats were fixed on stereotaxic apparatus after the epilepsy model was confirmed by ethology.A trochar was put into the left lateral ventricle.Rats were implanted with epidural recording electrodes.After the cortical EEG was recorded about 30 minutes, liensinine (at concentration of 2.5,5 mg/mL),levetiracetam and 0.9% sodium chloride was injected into lateral ventricle.Electrocorticogram was recorded about 150 minutes again.The frequency of epileptic discharge was observed every 30 minutes.The differences of frequency in the same group and the different change of frequency between groups at the same period were compared.Results The frequency of epileptic discharge decreased in low dose of liensinine group,high dose of liensinine group and levetiracetam group after administration ,there was significantly statistical difference in low dose of liensinine group after administration about 60 minutes(P<0.01 ),there was significant statistics difference in high dose of liensinine group after administration about 30 minutes(P<0.01),and the same change in levetiracetam group within 30 minutes after administration(P<0.01);the change of frequency of epileptic discharge was no significantly statistical difference between pro-and post-administration in the normal saline control group.The difference of the frequency change in epileptic discharge at the same period between liensinine group and levetiracetam group was observed ,there was statistic difference between low dose of liensinine group and levetiracetam group at the period of thirty to sixty minutes after administration,there was no statistic difference at other periods;there was no statistic difference between high dose of liensinine group and levetiracetam group at every period.Conclusion Liensinine could inhibit the epileptic discharges in acute model of epileptic rats induced by chloride lithium-pilocarpine.
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Objective To study the effect of liensinine on the proliferation of human bladder cancer T24 cells.Methods T24 cells were treated with different concentrations of liensinine.Its influence on the cell proliferation was detected by the CCK-8 exper-iment and the clonogenic experiment.After staining of T24 cells,the influence of liensinine on the cell cycle was examined by the flow cytometry.The mRNA change of p2 1 gene was determined by real-time quantitative PCR.Results Compared with the con-trol,liensinine significantly inhibited the proliferation of T24 cells in different doses groups(1.562 5,3.125 0,6.250 0,12.500 0, 25.000 0μg/mL),the differences had statistical significance and showed the dose-dependence;the cell cycle detection results re-vealed that liensinine arrested the T24 cells at the S phase;the real-time quantitative PCR detection results showed that liensinine increased mRNA of p21 gene in T24 cells.Conclusion Liensinine inhibits the proliferation of T24 bladder cancer cells and arrests the T24 cells at S phase,its mechanism may be related with the upregulation of p21 expression.
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Aim To study the influence of Liensinine(LIE) on the oxidative damage of erythrocyte ghost. Method The erythrocyte ghost oxidative injury caused by super oxide anion (O?_2) from the auto-oxidation of pyrogallic acid was used as model. The effect of LIE on oxidated erythrocyte ghost was observed by the measurement of the malonidial aldehyde (MDA)-the degradation product of lipid peroxidation, blocking degree, speed of auto-oxidation and membrane fluidity. Result O?_2 caused lipid peroxidation of erythrocyte ghost, increased the content of MDA and the speed of auto-oxidation, decreased membrane fluidity, and alleviated blocking degree. But LIE(80, 100, 120 mg?L -1 )could decrease the content of MDA and speed of auto-oxidation markedly, improve the membrane fluidity and blocking degree significantly, and inhibit the lipid peroxidation. Conclusion LIE has protective action on the oxidative damage of erythrocyte ghost initiated by O?_2
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OBJECTIVE:To optimize the extracting technology of Nelumbo nucifera.METHODS:The extraction technology of Nelumbo nucifera was optimized by orthogonal experiment with the extraction rate of liensinine as index and with the concentration and amount of alcohol and the extraction time and extraction times as factors.RESULTS:The optimal conditions for extraction technology was as follows:adding four - fold amount of 80%alcohol and refluxing and extracting twice(1h each time).CONCLUSION:The extracting technology optimized under simulated conditions serves as a reference for industrial production.
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Objective To investigate the adsorption function of macroporous adsorption resins for the separation and purification of the effective components in Plumula Nelumbinis. Methods AKS-W, AB-8, and H214 resins were systematically studied for their adsorption capability, and the contents of the liensinine, isoliensinine, and neferine were determined by HPLC. Results AKS-W Resin had a preferable adsorptive and separateive effect for the effective components in Plumula Nelumbinis. The products containing 10.2% liensinine, 6.7% isoliensinine, and 11.9% neferine could be obtained. Conclusion Macro-porous-type and non-polar adsorption resin AKS-W is an ideal adsorbent for the extraction of the effective components in Plumula Nelumbinis.
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Aim To investigate the effects of Liensinine on platelet aggregation and coagulation function in rats,as well as the effect on experimental thrombosis.Methods Inhibition rates of platelet aggregation for Liensinine in vivo were determined by the model of platelet aggregation induced by adenosine diphosphate.Coagulation time of mice was measured by capillary vessel method,and bleeding time of mice was measured by tail-cutting method.The effects of Liensinine were also evaluated on prothrombin time(PT),activated partial thromboplastin time(APTT)and thrombin time(TT).The model of artery-vein bypass thrombosis and Chandler's model were established to observe the effect of Liensinine.Results The result showed that Liensinine 5 and 10 mg?kg-1 had significant effect on inhibition of platelet aggregation and markedly prolonged bleeding time,coagulation time,PT,APTT and TT.Liensinine 5 and 10 mg?kg-1 inhibited the artery-vein bypass and Chandler's thrombus in different degree,reduced the thrombus weight significantly (either wet ordry).Conclusion Liensinine exerts remarkable effect against thrombosis and possesses strong effect against platelet aggregation and coagulation.