Effect of human urinary kallidinogenase on cognitive function of SAMP8 mouse model and its mechanism / 中国药理学通报

Aim To study the effect of human urinary kallidinogenase(HUK)on the cognitive function of SAMP8 mouse model and its mechanism. Methods SAMP8 mice were divided intofive groups:SAMP8 group,treatment group(giving 8.75×10-3,1.75×10-2,3.5×10-2,7.0×10-2 HUK),and the SAMR1 vehicle group was used as blank control. Each group was performed Morris water maze to detect spatial cognition. Afterwards the group with the most obvious cognitive improvement(HUK group)was selected for the follow-up experiments. Immunohistochemical detection of ChAT expression in CA3 area was further verified by RtPCR. Western blot was used to detect the expression of PSD95,SYN,BDNF,and pCREB protein. The activity of MPO and the content of IL-1β and IL-18 were determined. Results The passing times in the SAMP8 group was less than that of the SAMR1 group(P<0.05). The passing times of treatment group increased compared with the SAMP8 group(P<0.05 or P<0.01),and the spatial probe time of the target quadrant was shorter(P<0.05 or P<0.01). We conducted follow-up experiments with group d(HUK group). The expression of ChAT positive cells in CA3 area of SAMP8 group was significantly lower than that of SAMR1 group; the expression of positive cells in HUK group significantly increased; RtPCR showed that ChAT expression in SAMP8 group was lower than that in SAMR1 group,and ChAT expression was significantly higher than that in SAMP8 group after HUK treatment. Compared with the SAMR1 group,the levels of IL-1β,IL-18 and MPO activity in the CA3 area of SAMP8 group significantly increased,and the protein expressions of PSD95,SYN,BNDF and pCREB decreased. After HUK treatment,the content of IL-1β,IL-18 and MPO activity decreased,and the expression of PSD95,SYN,BNDF and pCREB increased. Conclusions HUK can improve the spatial cognition of SAMP8 mice. The mechanism may be achieved by promoting the expression of ChAT in CA3 area,reducing the oxidative stress and increasing synapse-related proteins.

Subproteome of Lachesis muta rhombeata venom and preliminary studies on LmrSP-4, a novel snake venom serine proteinase

Lachesis muta rhombeata is one of the venomous snakes of medical importance in Brazil whose envenoming is characterized by local and systemic effects which may produce even shock and death. Its venom is mainly comprised of serine and metalloproteinases, phospholipases A2 and bradykinin-potentiating peptides. Based on a previously reported fractionation of L. m. rhombeata venom (LmrV), we decided to perform a subproteome analysis of its major fraction and investigated a novel component present in this venom. Methods: LmrV was fractionated through molecular exclusion chromatography and the main fraction (S5) was submitted to fibrinogenolytic activity assay and fractionated by reversed-phase chromatography. The N-terminal sequences of the subfractions eluted from reversed-phase chromatography were determined by automated Edman degradation. Enzyme activity of LmrSP-4 was evaluated upon chromogenic substrates for thrombin (S-2238), plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) and upon fibrinogen. All assays were carried out in the presence or absence of possible inhibitors. The fluorescence resonance energy transfer substrate Abz-KLRSSKQ-EDDnp was used to determine the optimal conditions for LmrSP-4 activity. Molecular mass of LmrSP-4 was determined by MALDI-TOF and digested peptides after trypsin and Glu-C treatments were analyzed by high resolution MS/MS using different fragmentation modes. Results: Fraction S5 showed strong proteolytic activity upon fibrinogen. Its fractionation by reversed-phase chromatography gave rise to 6 main fractions (S5C1-S5C6). S5C1-S5C5 fractions correspond to serine proteinases whereas S5C6 represents a C-type lectin. S5C4 (named LmrSP-4) had its N-terminal determined by Edman degradation up to the 53rd amino acid residue and was chosen for characterization studies. LmrSP-4 is a fibrinogenolytic serine proteinase with high activity against S-2302, being inhibited by PMSF and benzamidine, but not by 1,10-phenantroline. In addition, this enzyme exhibited maximum activity within the pH range from neutral to basic and between 40 and 50 °C. About 68% of the LmrSP-4 primary structure was covered, and its molecular mass is 28,190 Da. Conclusions: Novel serine proteinase isoforms and a lectin were identified in LmrV. Additionally, a kallikrein-like serine proteinase that might be useful as molecular tool for investigating bradykinin-involving process was isolated and partially characterized.(AU)

Pancreatic kininogenase improves erectile function in streptozotocin-induced type 2 diabetic rats with erectile dysfunction / 亚洲男科学杂志(英文版)

Erectile dysfunction (ED) associated with type 2 diabetes is a severe problem that requires effective treatment. Pancreatic kininogenase (PK) has the potential to improve the erectile function of ED patients. This study aims to investigate the effect of PK on erectile function in streptozotocin-induced type 2 diabetic ED rats. To achieve this goal, we divided male Sprague-Dawley rats into five groups. One group was not treated, and the other four groups were treated with saline, sildenafil, PK or sildenafil, and PK, respectively, for 4 weeks after the induction of type 2 diabetic ED. Then, intracavernous pressure under cavernous nerve stimulation was measured, and penile tissue was collected for further study. Endothelial nitric oxide synthase levels, smooth muscle content, endothelium content, cyclic guanosine monophosphate (cGMP) levels in the corpus cavernosum, and neuronal nitric oxide synthase levels in the dorsal penile nerve were measured. Improved erectile function and endothelium and smooth muscle content in the corpus cavernosum were observed in diabetic ED rats. When treating diabetic ED rats with PK and sildenafil at the same time, a better therapeutic effect was achieved. These data demonstrate that intraperitoneal injection of PK can improve erectile function in a rat model of type 2 diabetic ED. With further research on specific mechanisms of erectile function improvement, PK may become a novel treatment for diabetic ED.

Pancreatic kininogenase improves erectile function in streptozotocin-induced type 2 diabetic rats with erectile dysfunction / 亚洲男科学杂志(英文版)

Erectile dysfunction (ED) associated with type 2 diabetes is a severe problem that requires effective treatment. Pancreatic kininogenase (PK) has the potential to improve the erectile function of ED patients. This study aims to investigate the effect of PK on erectile function in streptozotocin-induced type 2 diabetic ED rats. To achieve this goal, we divided male Sprague-Dawley rats into five groups. One group was not treated, and the other four groups were treated with saline, sildenafil, PK or sildenafil, and PK, respectively, for 4 weeks after the induction of type 2 diabetic ED. Then, intracavernous pressure under cavernous nerve stimulation was measured, and penile tissue was collected for further study. Endothelial nitric oxide synthase levels, smooth muscle content, endothelium content, cyclic guanosine monophosphate (cGMP) levels in the corpus cavernosum, and neuronal nitric oxide synthase levels in the dorsal penile nerve were measured. Improved erectile function and endothelium and smooth muscle content in the corpus cavernosum were observed in diabetic ED rats. When treating diabetic ED rats with PK and sildenafil at the same time, a better therapeutic effect was achieved. These data demonstrate that intraperitoneal injection of PK can improve erectile function in a rat model of type 2 diabetic ED. With further research on specific mechanisms of erectile function improvement, PK may become a novel treatment for diabetic ED.

Effect of pancreatic kininogenase for renal function and hemodynamics of patients with clinical diabetic nephropathy / 中国医学装备

Objective:To observe the effect on renal function and hemodynamics of pancreatic kininogenase in patients with clinical diabetic nephropathy(DN,Ⅳ Period).Methods: 145 patients with clinical diabetic nephropathy(DN,Ⅳ Period) were divided into the observation group (75 cases) and the control group (70 cases) as random method. The control group was given conventional treatment, and the observation group was given pancreatic kininogenase based on the conventional treatment. After 8 weeks, to observe and analyze the changes of renal function indexes and hemodynamics indexes of the two groups.Results:After treatment, the serum creatinine(SCr), blood urea nitrogen(BUN), Serum Cys C(CysC) and urinary albumin excretion rate(UAER) of observation group were obviously decreased compared with that before treatment; the Cys C, UAER of control group also were obviously decreased, but SCr, BUN of control group had no significant changes compared with that before treatment; and all indexes of the observation group were lower than those of the control group, with significant differences (t=4.2744, t=3.0832,t=6.8261,t=8.0936;P<0.05). After treatment, the systolic pressure(SBP), diastolic blood pressure(DBP) and resistance index(RI) of two groups were decreased; both of peak systolic blood flow(PSV) and end-diastolic blood flow velocity(EVD) were increased, and the change ranges of every index of the observation group were improved more than those in the control group, with significant difference (t=5.6270,t=6.2009,t=5.0926,t=4.764,t=3.1844;P<0.05). Conclusion: Pancreatic kininogenase has the clear effect on protecting renal function in the treatment of clinical diabetic nephropathy(Ⅳ Period), and it can reduce urinary albumin excretion and improve hemodynamics.

Effects of pancreatic kininogenase enteric-coated tablets on optic discand macular retinal hemodynamics in diabetic retinopathy / 中国生化药物杂志

Objective To explore the pancreatic kininogenase enteric-coated tablets in the treatment of diabetic retinopathy (DR) patients and its effect on the optic disc and macula retinal hemodynamics. Methods 86 cases (140 eyes) of DR patients were randomly divided into pancreatic kininogenase group and normal group with 43 cases in each group, two groups were treated with basic therapy, pancreatic kininogenase group were combined with pancreatic kininogenase treatment. The best corrected visual acuity and the clinical effect of optic disc and macula retinal hemodynamic changes were compared between two groups. Results After treatment, the best corrected visual acuity (BCVA) in pancreatic kininogenase group was greater than the normal group (P<0.05), the retinal neovascularization and fluorescein leakage area in pancreatic kininogenase group was less than the normal group (P<0.05). The disc vascular and macular retinal blood flow volume (VOL), blood flow velocity (FLW)values in pancreatic kininogenase group were larger than those of normal group, and the clinical curative effect of pancreatic kininogenase group was better than that of normal group (P<0.05). Conclusion Pancreatic kininogenase enteric-coated tablets in the treatment of DR patients can improve the optic disc and macular retinal hemodynamic parameters, improve visual acuity and reduce the retinal neovascularization and fluorescein leakage area, so as to improve the clinical treatment effect.

Clinical treatment of erectile dysfunction in type 2 diabeticpatients in the high-altitude area / 中华男科学杂志

Objective@#To investigate the safety and efficacy of pancreatic kininogenase combined with sildenafil in the treatment of erectile dysfunction(ED) in type 2 diabetes mellitus (DM) patients in the high-altitude area.@*METHODS@#This study included 93 ED patients with type 2 DM, all residents of the Xining area 1500 meters above sea level. We randomly divided them into an experimental group (n = 48) and a control group (n = 45), the former treated with pancreatic kininogenase(120 u, tid) and sildenafil (25 mg, qd at bedtime), while the latter with sildenafil only (25 mg, qd at bedtime).After 4 and 8 weeks of medication, we obtained the penile hemodynamic parameters,IIEF-5 scores, and sexual intercourse satisfaction(SIS) scores and compared them between the two groups of patients.@*RESULTS@#There were no statistically significant differences in age or DM course between the two groups of patients (P >0.05).Compared with the baseline, both the experimental and control groups showed remarkably improvement inthe IIEF-5 score (8.81 ± 2.06 vs 11.54 ± 7.72 and 8.29 ± 1.91 vs 9.37± 1.65, P 0.05). Even more remarkable improvement was observed at 8 weeks in the experimental and control groups in the IIEF-5 score (19.29± 1.85 and 15.43± 1.74)(P <0.05), SIS score (11.73 ± 2.57 and 6.55± 2.71) (P <0.05), and penile hemodynamic parameters(P <0.05), all with significant differences between the two groups (P <0.05).@*CONCLUSIONS@#Pancreatic kininogenase combined with sildenafil has a better clinical effect than sildenafil alone on ED in type 2 DM patientsin the high-altitude area.

Analysis of efficacy and safety of mosapride combined with pancreatic kallikrein in the treatment of diabetic neurogenic bladder / 中国基层医药

Objective To explore the efficacy and safety of mosapride combined withpancreatic kallikrein in the treatment of diabetic neurogenic bladder(DNB).Methods According to the digital table,120 patients with DNB were randomly divided into the control group and the observation group,each group 60 cases,All patients received basic treatment,including reducing blood glucose,reducing blood pressure,nutrition nerve and regulating blood lipids,and the observation group was accepted the treatment of original enzyme plus mosapride pancreatic kinin on basis of the above treatment.The treatment time of the two groups was both 3 weeks,The effect,blood sugar,blood hpids,urodynamic,quality of life,and the incidence of adverse reactions were observed.Results The total effective rate of the observation group was 93.33％,which was significantly higher than that of the control group (65.00％) (χ2 =14.602,P ＜ 0.01) ；the adverse reaction rate of the observation group was 8,33％,which was significantly lower than that of the control group(20.00％)(χ2 =11.368,P ＜ 0.01),Conclusion Mosapride combined with pancreatic kallikrein in thetreatment of DNB has better effect,and could significantly improve the clinical symptoms and quality of life of patients.So it should be promoted and applied.

Clinical study on cilostazol combined with pancreatic kininogenase in treatment of LEADDP / 中华内分泌外科杂志

Objective To compare clinical effects between cilostazol(CLT)alone and cilostazol combined with pancreatic kininogenase(PK)on lower extremity arterial disease in diabetic patient(LEADDP).Methods Patients with LEADDP were randomly divided into 2 groups:single medication group or the control group(53 cases)and combination group or the treatment group(53 cases).Sugar-reducing medicines were given before and after treatment to keep a steady blood glucose level.The control group took CLT orally 100 mg each time,twice a day.Besides receiving the same dose of CLT,the treatment group took 120 tablet of PK on an empty stomach,three times a day,with the duration of 3 months.Clinical symptoms,ankle/brachial index(ABI),blood rheology,nailfold microcirculation,intimal medial thickness(IMT)and plaque thickness were compared between the 2 groups after treatment.Results Symptoms in both groups improved after treatment.ABI was elevated significantly compared to that before treatment(P<0.05)and the change was more obvious in combination group.The difference of ABI after treatment between the 2 groups had statistical significance(P<0.05).IMT of the right foot dorsal artery,and left and right posterior tibial artery increased significantly compared to that before treatment(P<0.05).Plaque thickness decreased compared to that before treatment(P<0.01)and the difference between before and after treatment in combination group was significantly greater than that in single medication group(P<0.05).Conclusion Cilostazol combined with pancreatic kininogenase has better clinical effects than cilostazol alone in treatment of patients with LEADDP.

Effect of Human Urinary Kallikrein on Blood Pressure in the Treatment of Acute Cerebral Infarction / 药物流行病学杂志

Objective:To study the reasons and preventive measure of the effect of human urinary kallikrein on blood pressure in the treatment of acute cerebral infarction.Method:Documents about the effect of human urinary kallikrein on blood pressure of pre-clinical study and the phaseⅠ-Ⅲclinical trials were collected and analyzed in combination with the related information of the phaseⅣclinical trial from our hospital.Result:The rate and concentration of medication in combination with ACEI were the reasons for the effect of human urinary kallikrein on blood pressure.Conclusion:Controlling the rate and concentration of medication and forbidding the combined use of ACEI from the 72hr before the medication and the 24hr after it can prevent hypotension caused by human urinary kallikrein.

Effect of Pancreatic Kininogenase on Ventriculer Remodeling in Spontaneously Hypertensive Rats / 中华高血压杂志

Objective To investigate the effect of pancreatic kininogenase on the expression of matrix metalloproteinases-2 2(MMP-2), transfer growth factor-?_ 1 (TGF-?_ 1 ) and ventriculer remodeling in spontaneously hypertensive rats (SHR). Methods Twenty-four male 15 weeks SHR were randomly divided into three groups: SHR group, pancreatic kininogenase treatment group(PK: 7.2 U/kg?d), captopril treatment group(Cap: 10 mg/kg?d)(n=8 in each), 8 Wister Kyoto were served as control. After four weeks, blood pressure were measured througth carotid artery catherization. Myocardial tissue was stained with VG and pathological changes were studied. MMP-2, TGF-?_ 1 were determined by immunohisto-chemical technique(SP method). Results In pancreatic kininogenase treated SHR, SBP(183?12 vs SHR: 234?23)mm Hg, LVMI(2.89?0.15 vs SHR: 3.06?0.18)mg/g, CVF(0.17?0.03 vs SHR: 0.26?0.05)%, PVCA(0.57?0.26 vs SHR: 0.99?0.47)% and expression of MMP-2, TGF-?_ 1 in SHR were significantly improved (P