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Objective:To investigate the regulatory effect of lumbrukinase (LBK) on the gastric cancer SGC7901cells, and to clarify its mechanism.Methods:The SGC7901cells in the logarithmic growth phase were selected and divided into control group and 2, 4, 8U·mL-1 LBK groups.MTT assay was used to detect the inhibitory rates of proliferation of SGC7901cells in various groups in vitro at different time (24, 48and 72h) .Cell scratch assay was used to detect the migration abilities of the SGC7901cells in vitro in various groups.Flow cytometry was used to determine the apoptotic rates of SGC7901cells and the pencentages of cells at different cell cycles in various groups.The expression levels of Bcl-2, Bax, and caspase-3in the SGC7901cells in various groups were detected by Western blotting method.Results:The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901cells in different doses of LBK groups after treated for 24, 48and 72hwere increased (P＜0.01) .The cell scratch assay results showed that compared with control group, the migration distances of SGC7901cells in4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) .The flow cytometry results showed that compared with control group, the apoptotic rates of SGC7901cells in 4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) ;the percentages of cells in G1and S phases were decreased (P＜0.01) ;the percentages of cells in G2phase were increased (P＜0.01) .The results of Western blotting method showed that compared with control group, the Bcl-2protein expression level in the SGC7901cells in 8U·mL-1 LBK group was decreased (P＜0.05) ;the Bax and caspase-3protein expression levels were increased (P＜0.05) .Conclusion:LBK can inhibit the proliferation and migration abilities of SGC7901cells in vitro and induce the apoptosis;its mechanism is achieved through the regulation of expression levels of Bcl-2, Bax, and caspase-3proteins.
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Objective: To investigate the regulatory effect of lumbrukinase CI.RK) on the gastric cancer SGC7901 Cells, and to clarify its mechanism Methods: The SGC7901 cells in the logarithmic growth phase were selected and divided into control group and 2, 4 , 8 U • ml. I.RK groups. MTT assay was used to detect the inhibitory rates of proliferation of SGC7901 cells in various groups in vitro at different time (24, 48 and 72 h). Cell scratch assay was used to detect the migration abilities of the SGC7901 cells in vitro in various groups. Flow cytometry was used to determine the apoptotic rates of SGC7901 cells and the pencentages of cells at different cell cycles in various groups. The expression levels of Bcl-2, Rax, and caspase-3 in the SGC790I cells in various groups were detected by Western blotting method. Results: The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901 cells in different doses of I.RK groups after treated for 24, 48 and 72 h were increased ( P<0.01). The cell scratch assay results showed that compared with control group, the migration distances of SGC790I cells in 4 and 8 U • mL"1 I.RK groups were increased significantly ( P
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Objective To observe the regulation of lumbrukinase on the expression of α-smooth muscle actin,fibronectin,focal adhesion kinase and Src kinase induced by transforming growth factor β1 in human renal tubular epithelial cells.Methods According to random number table method,the human renal tubular epithelial cells were divided into the normal group,TGF-β1 model group,benazepril group,lumbrukinase low,medium and high dose group.Except the normal group,the other groups were treated with TGF-β1 10 μg/ml.After 30 minutes,the benazepril group added benazepril 10 μmol/L,and the low,medium,high dose groups of lumbrukinase were respectively treated with lumbrokinase 30,60,120 U/ml to intervene.After 24 hours of cultivation.Western blotting and real-time PCR were used to detect the expression of α-SMA,FN,FAK and Src.Results Compared with the model group,the expressions of α-SMA (0.84 ± 0.14,0.72 ± 0.08,0.69 ± 0.05 vs.1.24 ± 0.03) and FN (0.59 ± 0.09,0.55 ± 0.11,0.44 ± 0.08 vs.0.83 ± 0.18) and FAK (0.94 ± 0.04,0.79 ± 0.05,0.70 ± 0.02 vs.1.29 ± 0.07) and Src (0.87 ± 0.20,0.78 ± 0.15,0.71 ± 0.11 vs.1.23 ± 0.01) proteins in the high doses of lumbrical kinase group were significantly lower than those in the model group (P＜0.05),the expressions of α-SMA (3.13 ± 0.62,2.76 ± 0.14,2.15 ± 0.33 vs.4.12 ± 0.32) and FN (3.08 ± 0.34,2.78 ± 0.17,2.49 ± 0.11 vs.4.34 ± 0.06) and FAK (1.73 ± 0.23,1.63 ± 0.36,1.57 ± 0.27 vs.2.61 ± 0.59) and Src (2.11 ± 0.17,1.78 ± 0.25,1.71 ± 0.22 vs.2.78 ± 0.47) mRNA in the high doses of lumbrical kinase group decreased significantly (P＜0.05).Conclusions Lumbrokinase may prevent the development of renal fibrosis by regulating the expression ofFN,FAK and and reducing the production of α-SMA,FN.
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Objective:To explore the clinical efficacy and safety oflumbrokinase in the treatment of acute and moderate risk pulmonary thromboembolism.Methods:The clinical data of 60 patients with acute and moderate risk pulmonary thromboembolism,who were collected from January 2010 to October 2015 in Hunan Provincial People's Hospital,were retrospectively analyzed.According to the different treatments,60 patients were randomly divided into a lumbrokinase group (lumbrokinase in combination with low molecular heparin and sequential warfarin,n=30) and a control group (low molecular heparin and sequential warfarin,n=30).The clinical efficacy and safety were compared between the two groups.Results:Compared with the control group,maximum short axis,ratio of right and left ventricles,systolic pulmonary artery pressure,and the main pulmonary artery diameter in the lumbrokinase group were significant changed after the treatment for 10,20 and 30 d.NT-proBNP level in the lumbrokinase group after the treatment for 10,20 and 30 d was significantly reduced than that in the the control group (P＜0.05).However,the value of PO2 significantly increased after 10,20 and 30 d,and there was no significant difference between 20 d and 30 d (P＞0.05).D-dimer in the two groups was obviously increased after treatment for 10 d,but it was significantly reduced after treatment for 20 d or 30 d (P＜0.05).The clinical efficacy of the lumbrokinase group was better than that in the control group,with significant difference (P＜0.05).Conclusion:Combination of lumbrokinase with low molecular heparin and sequential warfarin is a safe and efficient strategy in treating the patients with acute and moderate risk pulmonary thromboembolism.It is worthy of clinical popularization and application.
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Objective:To establish a fibrin zymography method for identifying the active proteins in lumbrokinase,and investigate the production difference and batch consistency of 5 different manufacturers. Methods:Fibrin zymography was used with the final con-centration of fibrinogen of 5 × 10 - 4 g·ml - 1 ,renature time of 30 min in 2. 5% Triton-x-100,incubation time of 30 min in PBS buffer (pH = 7. 4)at 37℃ and the protein concentration in the sample of 5. 0-37. 5 μg. Results:The sensitivity of the method was high,and the molecular weight distribution of active protein bands for the samples from five manufacturers was between 15KD and 40KD with 6 common active protein bands. The zymography of the samples from the five manufacturers had slight difference,while various batches of the samples from the same manufacturer showed no difference. Conclusion:The method is special. It can reflect molecular weight distribution and species of active protein,batch consistency and production process stability. It is easy to be standardized and suitable for the identification of lumbrokinase,which can lay foundation for the quality consistency evaluation of marketed products.
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Objective To observe the curative effect of Zhengqingfengtongning release tablets and lumbroki-nase enteric -coated capsules and losartan potassium tablets joint for the treatment of chronic nephritis proteinuria, and explore its mechanism.Methods 42 patients with chronic nephritis were randomly divided into two groups. 20 cases in the control group were given losartan potassium tablets,50mg,1 ~2 times a day.22 cases in the treatment group received Zhengqingfengtongning release tablets 120mg,2 times a day,lumbrokinase enteric -coated capsules 600 000u,3 times a day on the basis of the control group.The 24h urine protein,liver and kidney function before and after the treatment were observed.Results 20 days after treatment,24h urinary protein in the treatment group decreased 57.08%,which in the control group decreased 41.59%,the difference was statistically significant (t =2.39,P <0.05).30 days after treatment,24h urinary protein in the treatment group decreased 81.28%,which in the control group decreased 52.34%,the difference was statistically significant (t =3.65,P <0.01 ).Renal function change had no significant difference between the two groups.Conclusion Zhengqingfengtongning release tablets and lumbrokinase enteric -coated capsules combined with losartan potassium tablets in the treatment of chronic nephritis proteinuria work fast,and has good curative effect,less side effects,and is worth to recommend.
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Objective: To investigate the method for study on the effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity and to eliminate the interference of pepsin and trypsin on the detection of crude protein fibrinolytic activity of Armadillidium vulgare (porcellio plasmin) in order to obtain the proteins or peptides which have the smaller molecular weight but higher titer during the pepsin and trypsin degradation. Methods: To study the effect of pepsin and trypsin deactivation on pH value, temperature, metal ions, enzyme inhibitor, surfactant, and responsing fibrinolytic by fiber fibrin plate assay. The better enzyme deactivation process was obtained and used for studying the effect on the fibrinolytic activity of urokinase, lumbrokinase, and porcellio plasmin. Results: All the pH value, temperature, metal ions, enzyme inhibitor, and surfactant have had an impact on pepsin and trypsin fibrinolytic activity. Among them the optimum deactivation of pepsin was pH 6.0-8.0, while the optimum deactivation of trypsin was mixed preparation with TLCK at the concentration of 25 mg/mL and EDTA at the concentration of 1 mmol/L. Conclusion: This study has obtained the better enzyme deactivation process which could be used for the detection of fibrinolytic activity of pepsin and trypsin degradation product by fiber fibrin plate assay, the operation is simple, and the repeatability and stability are good.
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Abstract? AlM: To explore the effect of Danshen injection iontophoresis combined with lumbrukinase on early patients with retinal vein occlusion ( RVO ) , and observe the prognosis to provide objective basis for clinical?METHODS: Eighty-two patients ( 117 eyes ) with early RVO were randomly selected in our hospital from January, 2011 to April, 2014, 41 cases (59 eyes) of control group treated with Lumbrokinase. Forty-one cases ( 58 eyes) of the observation group, treated with Danshen injection iontophoresis combined with lumbrokinase. The main indexes were the treatment effect, visual acuity and retinal circulation time.?RESULTS: The total effective rate of the observation group was 85. 4% ( 35/41 ), more than that of control group 68. 3% ( 28/41 ), the difference was statistically significant (P<0. 05). The invalid rate of the observation group was 14. 6% (6/41), lower than that of the control group 31. 7% ( 13/41 ), the difference was statistically significant (P<0. 05). The vision of the observation group 4. 8 ~5. 0 was 53. 7% (22/41), more than that of control group 41. 5% ( 17/41 ), the difference was statistically significant ( P < 0. 05 ). The restore vision of the observation group 4. 3 ~ 4. 7 was 31. 7% (13/41), lower than that of the control group 41. 5% ( 17/41 ), the difference was statistically significant ( P<0. 05 ). After treatment, retinal circulation time of two groups were lower than before treatment, the difference was statistically significant (P<0. 05). After treatment of retinal vein circulation time of the observation group was 8. 15 ± 1. 30s, 9. 70 ± 1. 28s lower than that of the control group, the difference was statistically significant (P<0. 05).?CONCLUSlON: There is better effective on Danshen injection iontophoresis combined with Lumbrokinase for early retinal vein occlusion, and can improve the patient's visual acuity, promote the rehabilitation of patients.
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Objective To study the effect of lumbrokinase treatment on hemorheology, glucose and lipids of type 2 diabetes mellitus (T2DM). Methods 76 cases with T2DM whose blood glucose was controlled at FPG0.05). Conclusions Lumbrokinase can decrease the levels of the blood rheology and triglyceride in T2DM. It may prevent and cure the complication of diabetes mellitus
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The purpose of the present study is to determine whether lumbrokinase has an in vivo thrombolytic effect in a rabbit cerebral embolism model. In our previous studies, we found that lumbrokinase, an extract from Korean earth worms, has a strong in vitro fibrinolytic effect without the presence of plasminogen and significant in vivo thrombolytic effects of lumbrokinase in a rat human-clot-induced cerebral embolism model. We established the cerebral embolism model in rabbits by injecting a piece of human clot into the internal carotid artery via the external carotid artery and confirmed the occlusion with angiography. Twenty one rabbits were divided into three groups and 5cc of saline, urokinase of 50,000 u/ml, and equipotent LK were injected intraarterially for 30 minutes into each group of 7 animals. Ten minutes after the end of infusion, an angiogram was performed to confirm the recanalization. Clot lysis occurred in one, six, and one animals in the saline, urokinase and lumbrokinase treated groups respectively. With regard to its in vitro effect, lumbrokinase is not as potent in vivo. Further investigation should be performed to determine the cause of its weakened in vivo effect and to develop a method to potentiate it.
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Animals , Rabbits , Endopeptidases/therapeutic use , Fibrinolytic Agents/therapeutic use , Intracranial Embolism and Thrombosis/drug therapy , Thrombolytic Therapy , Urokinase-Type Plasminogen Activator/therapeutic useABSTRACT
AIM: To assess the prophylactic effect of lumbrokinase plus aspirin on recurrence of ischemic stroke in the patients recovering from cerebral infarction. METHODS: 87 patients of cerebral infarction were assigned to two groups:control and treatment group,treated respectively with aspirin in control group and lumbrokinase plus aspirin in the treatment group.The recurrence rate and clinical efficacy were analysed. RESULTS: There were remarkable differences in recurrence rate and clinical efficacy between the two groups(P