Predictive value of cathepsin level in peripheral blood on arteriovenous fistula stenosis and therapeutic effect of urokinase combined with argatroban in patients with maintenance hemodialysis / 中国医师进修杂志

Objective:To study the predictive value of peripheral blood cathepsin (Cat) level on arteriovenous fistula stenosis and therapeutic effect of urokinase combined with argatroban in patients with maintenance hemodialysis (MHD).Methods:The clinical data of 120 patients with MHD from January 2017 to January 2021 in the First Affiliated Hospital of Hebei North University were retrospectively analyzed. Among them, 72 patients had arteriovenous fistula stenosis (stenosis group), and 48 patients had not arteriovenous fistula stenosis (non-stenosis group). The patients in stenosis group were treated with urokinase combined with argatroban, and the therapeutic effect was evaluated; the stenosis degree of arteriovenous fistula stenosis was evaluated by digital subtraction angiography (DSA). The levels of Cat K and S in peripheral blood were detected by enzyme linked immunosorbent assay. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of Cat K and S in peripheral blood on arteriovenous fistula stenosis in patients with MHD. The independent risk factor of arteriovenous fistula stenosis in patients with MHD was analyzed by multivariate Logistic regression analysis.Results:The levels of Cat K and S in peripheral blood in stenosis group were significantly higher than those in non-stenosis group: (404.34 ± 12.43) μg/L vs. (344.22 ± 12.09) μg/L and (124.55 ± 13.43) μg/L vs. (84.60 ± 12.45) μg/L, and there were statistical differences ( t = 26.39 and 16.68, P<0.01). The result of DSA showed that mild stenosis of arteriovenous fistula stenosis was in 33 cases, moderate stenosis in 23 cases, and severe stenosis in 16 cases. The levels of Cat K and S in peripheral blood in patients with moderate stenosis and severe stenosis were significantly higher than those in patients with mild stenosis: (399.83 ± 11.79) and (476.27 ± 12.24) μg/L vs. (372.61 ± 12.88) μg/L, (125.77 ± 12.75) and (151.69 ± 11.86) μg/L vs. (110.54 ± 12.07) μg/L, the indexes in patients with severe stenosis were significantly higher than those in patients with moderate stenosis, and there were statistical differences ( P<0.01). After treatment, excellent was in 40 cases, effective in 23 cases, and ineffective in 9 cases. The levels of Cat K and S in peripheral blood in patients with effective and ineffective were significantly higher than those in patients with excellent: (404.78 ± 10.96) and (491.30 ± 10.26) μg/L vs. (384.52 ± 10.36) μg/L, (121.85 ± 10.99) and (232.65 ± 10.61) μg/L vs. (101.78 ± 10.61) μg/L, the indexes in patients with ineffective were significantly higher than those in patients with effective, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the area under the curve of Cat K combined with Cat S in peripheral blood in forecasting arteriovenous fistula stenosis in patients with MHD was larger than that of Cat K and S alone (0.699 vs. 0.635 and 0.611), and the accuracy and specificity were also significantly higher (80.83% vs. 48.33% and 60.00%, 89.58% vs. 76.25% and 81.33%), the optimum cut-off values of Cat K and S in peripheral blood were 401.23 and 123.65 μg/L. Multivariate Logistic regression analysis result showed that the levels of Cat K and S in peripheral blood were the independent risk factor of arteriovenous fistula stenosis in patients with MHD ( OR = 1.02 and 1.63, 95% CI 0.90 to 1.93 and 1.33 to 2.32, P<0.01). Conclusions:The levels of Cat K and S in peripheral blood can predict the occurrence and extent of arteriovenous fistula stenosis in patients with MHD, and could also predict the therapeutic effect of urokinase combined with agatroban.

Correlation between serum cathepsin S level and severity of coronary heart disease in elderly patients / 中华老年医学杂志

Objective To study the expression level of cathepsin S in elderly patients with different degree of coronary heart disease(CHD),and to evaluate the correlation between cathepsin S level and the severity of CHD.Methods A total of 126 elderly patients with CHD were enrolled,including 36 cases with acute myocardial infarction(group AMI),48 cases with unstable angina(group UAP),and 42 cases with stable angina(group SAP).During the same period,40 healthy subjects were selected as controls.Venous blood was collected immediately after admission.Arterial blood was taken through radial/femoral artery sheath in AMI patients who underwent primary percutaneous coronary intervention,and coronary arterial blood was taken in AMI patients undergoing intracoronary thrombus aspiration.SYNTAX score was assessed in patients undergoing coronary angiography.Serum levels of high sensitivity C reactive protein(hs-CRP),matrix metalloproteinase-9 (MMP-9) and cathepsin S were determined in all specimens and compared between groups.Results Serum levels of cathepsin S,MMP-9 and hs-CRP were higher in CHD patients than in the control group(P＜0.05).Among CHD patients,group AMI had the highest serum levels of cathepsin S,MMP-9 and hs-CRP,followed by group UAP and group SAP(F =106.830,197.035 and 310.442,all P =0.000).Spearman's rank correlation test suggested that cathepsin S level was positively correlated with serum levels of MMP-9 (r=0.816,P =0.000)and hs-CRP(r =0.827,P =0.000).SYNTAX score was positively correlated with serum levels of cathepsin S(r=0.581,P=0.000),MMP-9(r=0.511,P=0.000),and hs-CRP (r=0.557,P =0.000).Among the 24 patients with AMI who underwent intracoronary thrombus aspiration,the levels of cathepsin S,MMP-9 and hs-CRP were higher in coronary artery blood than in peripheral artery blood (t =217.288,3.177 and 681.479,all P =0.000).Cathepsin S and hs-CRP levels in peripheral arterial blood had positive correlations with those in coronary arterial blood respectively(r =0.962 and 0.494,P =0.000 and 0.014),but such correlation between in peripheral arterial blood versus in coronary artery blood was not found in MMP-9 levels (r =-0.188,P =0.380).Conclusions Serum cathepsin S level is higher in elderly CHD patients than in healthy people,increases along with the increased severity of CHD,and positively correlates with the degree of coronary stenosis.

The Mechanism of Cathepsins S Inhibitor Affected Ischemia-induced Neovascularization in Experimental Mice / 中国循环杂志

Objectives: To investigate the potential mechanism of cathepsins S inhibitor (CatS-I) affected ischemia-induced neovascularization in experimental mice. Methods: 8 week-old wild type (C57/BL6) mice were randomly divided into 2 groups: Control group, the mice received basic diet with intraperitoneal injection of 5% carboxymethylcellulose sodium and CatS-I group, the mice received basic diet with intraperitoneal injection of CatS-I (1mg/kg·d); all animals were treated for 17 days. n=20 in each group. Hind limb ischemia model was established at 3 days after injection in both groups. Blood flow was measured by laser Doppler blood flow analyzer, the ratio of ischemic area to non-ischemic area was calculated; protein expressions of peroxidase proliferation activated receptors-γ (PPAR-γ), p-Akt, p-eNOS and VEGF were examined by Western blot analysis at day 4 after the operation; frozen section of ischemic skeletal muscle was taken at 7 days after operation to measure capillary density by immunohistochemistry.Results: ① CatS-I inhibited blood flow recovery. Compared with Control group, CatS-I group had slower blood flow recovery in ischemic hind limb, P<0.05. ② CatS-I inhibited capillary formation. At 14 days after operation, capillary formation in non-ischemic skeletal muscles was similar between 2 groups, P>0.05; while in ischemic skeletal muscles, capillary density was lower in CatS-I group than Control group, P<0.01. ③ Compared with Control group, CatS-I group showed decreased protein expressions of PPAR-γ, p-Akt, p-Enos and VEGF, P<0.05. Conclusions: CatS-I regulated ischemia-induced neovascularization might be related to PPAR-γ activation and PI3K/Akt/eNOS signaling pathway in experimental mice.

Advances in Study on Role of Cathepsin S in Irritable Bowel Syndrome / 胃肠病学

Irritable bowel syndrome(IBS)is a common functional intestinal disease,and its etiology and pathogenesis are not completely clear. The pathogenesis of IBS involves disturbed gastrointestinal motility,gut hypersensitivity,intestinal inflammation,immune dysfunction and brain-gut axis abnormality. Cathepsin S(CTSS)is a proteolytic enzyme widely distributed in various cell lysosomes,and participates in a variety of pathophysiological processes. Recent studies have shown that CTSS may be involved in the pathogenesis of IBS. This article reviewed the advances in study on role of CTSS in IBS.

The relationship between cathepsin L and apoptosis cell in rats after cerebral ischemia reperfusion / 中国医师杂志

Objective To investigate the relationship between cathepsin L and apoptosis cell in rats after cerebral ischemia reperfusion.Methods Sixty healthy male Sprague-Dawley Rats (10-12 weeks old,260-300 g) were chosen.Based on the random number table method,the rats were randomly divided into sham-operated control group (Sham group,n =10),ischemia-reperfusion group (model group,n =25),and Z-FY-DMK intervention group (CLI group,n =25).Rats were randomly divided into 6 h,12 h,24 h,and 48 h four subgroups in model group and CLI group,respectively.Modified transient middle cerebral artery occlusion was made as Longa described,the intervention groups were injected intracerebroventricularly Z-FY-DMK (20 μg / 1μ1 ×5 μl) preoperative 30 min prior to surgery,Sham group and schemia reperfusion injury (IRI) group were injected intracerebroventricularly dimethyl sulfoxide (DMSO) 5 μ1 (10ml/L) at the same time.Cell apoptosis was detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) straining.Western blotting was used to detect the expression of cathepsin L and caspase-3.Results In the cortical area of ischemic brain,apoptosis cells of sham operation group were rare,while apoptosis of nerve cells of model group with 6 hours reperfusion were visible,and were gradually increased in the order of 12 hours,24 hours and 48 hours.At the same time point,the apoptosis cells of CL intervention group (6 h,12 h,24 h,48 h) were obviously less than model group (P ＜0.05).Western blotting found little visible cathepsin L protein expression in ischemic cerebral cortex preoptic in the sham group.For model group,the cathepsin L expression initially increased in sub groups with 6 hours reperfusion,reached to a peak in sub groups with 12 hours and 24 hours,and remained a high level in sub groups with 48 hours reperfusion.Compared to model group,the cathepsin L expressions of CL intervention group were obviously decreased at all time points (P ＜ O.05).Conclusions Cathepsin L may be involved in neuronal apoptosis by means of caspases 3 pathway.

Progress on matrix metalloproteinase inhibitors / 华西口腔医学杂志

Continuing advances in dentin bonding technology and adhesives revolutionized bonding of resin-based composite restorations. However, hybrid layers created by contemporary dentin adhesives present imperfect durability, and degradation of collagen matrix by endogenous enzymes is a significant factor causing destruction of hybrid layers. Bond durability can be improved by using enzyme inhibitors to prevent collagen degradation and to preserve integrity of collagen matrix. This review summarizes progress on matrix metalloproteinase inhibitors (including chlorhexidine, ethylenediaminetetraacetic acid, quaternary ammonium salt, tetracycline and its derivatives, hydroxamic acid inhibitors, bisphosphonate derivative, and cross-linking agents) and suggests prospects for these compounds.

Impact of Cathepsins S on Aortic Ring-derived Micro Vascular Cavity Formation in Experimental Mice / 中国循环杂志

Objective: To investigate the impact of cathepsins S (CatS) on aortic ring-derived micro vascular cavity formation in experimental mice. Methods: Hindlimb ischemia model was established in CatS+/+ and CatS-/- mice, n=8 in each group. The blood flow in hindlimb was measured before ischemic surgery; immediately and 1, 4, 7, 14, 21 days after surgery. CatS+/+mice were further divided into 4 subgroups: Normal control subgroup, Selective CatS inhibitor (LHVS) subgroup, Non-selective CatS inhibitor (E64d) subgroup and MMP inhibitor (GM6001) subgroup; n=2 in each subgroup. The mice aortic ring-derived micro vascular cavity formation was observed by FITC-CD31 immunofluorescence method. Results: ① CatS-/- mice had inhibited blood flow recovery after ischemic surgery. Laser Doppler blood flow (LDBF) examination indicated that compared with CatS+/+group, CatS-/- group had slower hindlimb blood flow recovery, P<0.05;② CatS-/-group had the less aortic ring-derived micro vascular cavities, P<0.001. ③ Compared with Normal control subgroup, LHVS subgroup, E64d subgroup and GM6001 subgroup had suppressed micro vascular cavity formation, all P<0.05.④ Aortic ring-derived micro vascular cavity was composed by endothelial cells. Conclusion: CatS plays a beneficial role in ischemic vascular regeneration in experimental mice; it is not only increasing aortic ring-derived micro vascular cavity formation, but also promoting blood flow recovery in ischemic hindlimb. Our finding provides a theoretical basis for new therapeutic target in ischemic vascular regeneration.

Impact of Cathepsins S on Aortic Ring-derived Micro Vascular Cavity Formation in Experimental Mice / 中国循环杂志

Objective: To investigate the impact of cathepsins S (CatS) on aortic ring-derived micro vascular cavity formation in experimental mice. Methods: Hindlimb ischemia model was established in CatS+/+ and CatS-/- mice, n=8 in each group. The blood flow in hindlimb was measured before ischemic surgery; immediately and 1, 4, 7, 14, 21 days after surgery. CatS+/+mice were further divided into 4 subgroups: Normal control subgroup, Selective CatS inhibitor (LHVS) subgroup, Non-selective CatS inhibitor (E64d) subgroup and MMP inhibitor (GM6001) subgroup; n=2 in each subgroup. The mice aortic ring-derived micro vascular cavity formation was observed by FITC-CD31 immunofluorescence method. Results: ① CatS-/- mice had inhibited blood flow recovery after ischemic surgery. Laser Doppler blood flow (LDBF) examination indicated that compared with CatS+/+group, CatS-/- group had slower hindlimb blood flow recovery, P<0.05;② CatS-/-group had the less aortic ring-derived micro vascular cavities, P<0.001. ③ Compared with Normal control subgroup, LHVS subgroup, E64d subgroup and GM6001 subgroup had suppressed micro vascular cavity formation, all P<0.05.④ Aortic ring-derived micro vascular cavity was composed by endothelial cells. Conclusion: CatS plays a beneficial role in ischemic vascular regeneration in experimental mice; it is not only increasing aortic ring-derived micro vascular cavity formation, but also promoting blood flow recovery in ischemic hindlimb. Our finding provides a theoretical basis for new therapeutic target in ischemic vascular regeneration.

Potential targets prediction of apigenin via virtual screening / 重庆医学

Objective To predict the potential targets of apigenin by virtual screening .Methods The targets preliminarily forecast by PharmMapper ,were validated by associating data mining and Autodock Vina in PyRx 0 .8 .Subsequently ,receptor‐ligand interactions were analyzed by Discovery Studio 3 .5 .Results The virtual screening by PharmMapper indicated that apigenin coupled well with the disease‐related targets including insulin receptor ,estradiol 17‐beta‐dehydrogenase 1 ,and cathepsin K .According to the data mining ,insulin receptor was found in related experimental researches ,while the other two had few reports previously .And then ,the interactions between apigenin and the target proteins were analyzed by Autodock Vina and Discovery Studio Visualizer 3 .5 ,involving hydrogen bonds ,electrostatic forces ,van der Waals forces etc .Conclusion The most potential targets of apigenin were insulin receptor ,while 17‐beta‐dehydrogenase 1 and cathepsin K were also possible .

Effect of ultraviolet A radiation on the expression and secretion of cathepsin G by human dermal fibroblasts / 中华皮肤科杂志

Objective To investigate the effect of ultraviolet A (UVA) radiation on the expression and secretion of cathepsin G (CatG) by human dermal fibroblasts.Methods Dermal fibroblasts were isolated from the foreskins of boys,and subjected to primary culture and subculture.After 10 or less passages,the fibroblasts were collected and divided into several groups to be irradiated with 10 J/cm2 UVA followed by 24,48 and 72 hours of additional culture,or be irradiated with 10,20 and 30 J/cm2 UVA followed by 24 hours of additional culture,with those receiving no treatment serving as the control group.Subsequently,cells and culture supernatant were collected,real time PCR and Western blot were performed to detect the expressions of CatG mRNA and protein respectively in these cells,and enzyme-linked immunosorbent assay (ELISA) was conducted to measure the expression of CatG protein in the culture supernatant of these cells.Results Compared with the control group,the fibroblasts irradiated with 10 J/cm2 UVA showed a significant increase at 24,48 and 72 hours in the expressions of CatG mRNA (0.376 ± 0.014 vs.0.183 ± 0.003,0.308 ± 0.022 vs.0.185 ± 0.005,0.296 ± 0.032 vs.0.182 ± 0.004,respectively,all P＜ 0.05) and protein (1.80 ± 0.12 vs.0.96 ± 0.06,1.41 ± 0.17 vs.0.95 ± 0.22,1.27 ± 0.09 vs.1.00 ± 0.14,respectively,all P ＜ 0.05),as well as in the supernatant level of CatG protein ((161.35 ± 7.55) vs.(122.45 ± 6.46) ng/L,(141.76 ± 2.95) vs.(124.17 ± 6.15) ng/L,(139.63 ± 3.04) vs.(121.72 ± 3.17) ng/L respectively,all P ＜0.05),with the strongest increase observed at 24 hours.At 24 hours after 10,20 and 30 J/cm2 of UVA radiation,the expression of CatG mRNA in irradiated fibroblasts was 1.90,2.51 and 3.04 times respectively (all P ＜ 0.05),the expression of CatG protein was 1.88,3.97 and 4.72 times respectively (P ＜ 0.05),and the supernatant level of CatG protein was 1.36,1.50 and 1.66 times respectively (P ＜ 0.05),that in the control group,and there was an increasing trend in all the above three parameters with increasing dose of UVA.Conclusion Acute UVA radiation can promote the expression and secretion of CatG by human dermal fibroblasts.

Progress in Cathepsin K Inhibitor Odanacatib / 天津医药

The prevalence of osteoporosis has increased during recent years. The activation in the function of osteo?clast is the main reason of osteoporosis. Cathepsin is expressed by osteoclast and involved in the degradation of collagen of bone,and causes osteoporosis. Cathepsin K is a kind of most important enzyme in the family of cathepsin. Odanacatib(ODN) is the inhibitor of cathepsin K, and it may be used to anti-osteoporosis thought inhibiting the degradation of bone collagen. It was found that the cortical thickness and bone minerals of cancellous increased after taking ODN in the studies, and then the density and the strength of bone increased. This study reviewed the pharmacological profile of ODN and the progresses of ani?mal study and clinical trials about ODN.

MAPK pathway regulates ultraviolet A-induced cathepsin K expression in human dermal fibroblasts / 中华皮肤科杂志

Objective To investigate whether ultraviolet A UVA)-induced CatK expression is regulated by the mitogen-activated protein kinases (MAPK) signaling pathway in human dermal fibroblasts in vitro.Methods Human dermal fibroblasts were obtained from circumcised foreskin of children,and subjected to primary culture.After several passages of subculture,some fibroblasts were irradiated with UVA at a dose of 10 J/cm2.Western blot was performed to measure the expressions of total and phosphorylated JNK (t-and p-JNK) and P38 (t-and p-P38) at 0.75,1.5,3 and 6 hours after the irradiation.Some fibroblasts were divided into six groups:control group receiving no treatment,SP group treated with SP600125 of 800 nmol/L,SB group treated with SB203580 of 10 μmol/L,UVA group irradiated with UVA at a dose of 10 J/cm2,UVA-SP group treated with SP600125 for 1 hour before and for 1.5 or 48 hours after UVA irradiation at 10 J/cm2,UVA-SB group treated with SB203580 for 1 hour before and for 1.5 or 48 hours after UVA radiation at 10 J/cm2.Subsequently,Western blot was performed to determine the expressions of p-c-Jun and p-MAPKAPK2 in these groups at 1.5 hours after the UVA irradiation,and reverse transcription (RT)-PCR and Western blot to detect the mRNA and protein expressions of CatK at 48 hours after the UVA irradiation,respectively.Statistical analysis was carried out by t test,one way analysis of variance and least significant difference (LSD)-t test.Results The expression levels (gray values) of p-JNK and p-P38 were significantly increased at 0.75 hour (4.77 ± 0.19 and 2.44 ± 0.13 respectively,both P ＜ 0.05) and 1.5 hours (4.68 ± 0.09 and 2.30 ± 0.04 respectively,both P ＜ 0.05),but showed no significant changes at 3 hours (both P ＞ 0.05) and 6 hours (both P ＞ 0.05) after the UVA irradiation compared with those before the irradiation (3.2 ± 0.27 and 1.61 ± 0.08 respectively).A significant decrease was observed in the expression of p-c-Jun in the UVA-SP group and p-MAPKAPK2 in the UVA-SB group compared with the UVA group (p-c-Jun,2.55 ± 0.48 vs.4.85 ±0.96; p-MAPKAPK2,1.16 ± 0.12 vs.2.46 ± 0.09,both P ＜ 0.05).The CatK mRNA and protein expressions were attenuated by 61.1％ and 44.3％ respectively in the UVA-SP group (both P ＜ 0.05),and by 71.3％ and 50.4％ respectively in the UVA-SB group (both P ＜ 0.05) in comparison with the UVA group.The UVA-SP group also showed a significant reduction in CatK mRNA and protein expressions as compared with the UVA-SB group (both P ＜ 0.05).Conclusion Both JNK and P38 signaling pathways,especially the JNK pathway,may contribute to the upregulation of CatK expression in dermal fibroblasts induced by UVA irradiation.

Cathepsin S and atherosclerosis / 国际脑血管病杂志

Cathepsin S is one of the cathepsins that distributes generally at lysosomes of histiocyte in humans body.It has variety of actions when it is unbalancing,including promote inflammation,atherosclerosis,diabetes and obesity,etc.Studies in recent years have demonstrated that there are certain correlations of adiponectin with vascular risk factor and cerebrovascular disease.

Cysteinyl Cathepsins: Multifunctional Enzymes in Cardiovascular Disease / 전남의대학술지

Until recently, the role of lysosomal cysteine protease cathepsins in intracellular protein degradation was believed to be mainly restricted to scavenging. However, recent studies have revealed nontraditional roles for cysteine protease cathepsins in the extracellular space during the development and progression of cardiovascular disease. Although the precise mechanisms are unknown, data from animal studies suggest that members of the cathepsin family, like other extracellular proteases, contribute to extracellular matrix protein remodeling and interstitial matrix degradation, as well as to cell signaling and cell apoptosis in heart disease. Inflammatory cytokines and hormones regulate the expression and secretion of cathepsins in cultured cardiovascular cells and macrophages. Serum levels of cathepsins L, S, and K and their endogenous inhibitor cystatin C may be useful predictive biomarkers in patients with coronary artery disease and cardiac disease. Furthermore, in vivo pharmacological intervention with a synthetic cathepsin inhibitor and cardiovascular drugs (including statins and angiotensin II type 1 receptor antagonists) has the potential for pharmacologic targeting of cathepsins in cardiovascular disease. This review focuses on cathepsin biology (structure, synthesis, processing, activation, secretion, activity regulation, and function) and the involvement of cysteinyl cathepsins in the pathogenesis of several heart and vessel diseases, especially with respect to their potential application as diagnostic and prognostic markers and drug targets to prevent inappropriate proteolysis in cardiovascular disease.

Effect of diphosphonate on biochemistry change in subchondral bone of unstable rabbit knee joints / 中华骨科杂志

Objective To explore the effect of diphosphonate on expression of matrix metalloproteinase-9(MMP-9) and cathepsin K(CK) in subchondral bone of unstable rabbit knee joints.Methods Fifty male New Zealand white rabbits were divided randomly into three groups according to random digits table:the control group (n=10),the model group (n=20).the diphosphonate group (n=20).Hulth model of unstable rabbit knee joint was achieved in the right knee joint.Ten rabbits from the diphosphonate group and 5 rabbits from the control group were sacrificed by aeroembolism at the second and tenth week postoperatively,respectively.Then the medial femoral condyles of the right knee were harvested.Specimens were processed for immunohistochemical analysis of MMP-9 and CK.Results Cells with expression of MMP-9 and CK could be found in the three groups at the second and tenth week after operation.Compared with the control group,there was a significant increase in the number of cells with expression of MMP-9 and CK in model group at the second and tenth week after operation.Diphosphonate could inhibit expression of MMP-9 and CK in cells.Compared with the model group,the number of cells with expression of MMP-9 and CK in diphosphonate group was fewer; there was a statistical significance between them.Conclusion Diphosphonate can inhibit the expression of MMP-9 and CK in subchondral bone of unstable rabbit knee joints,which can resist the bone resorption and protect articular cartilage.

Cysteinyl Cathepsins: Multifunctional Enzymes in Cardiovascular Disease / 전남의대학술지

Until recently, the role of lysosomal cysteine protease cathepsins in intracellular protein degradation was believed to be mainly restricted to scavenging. However, recent studies have revealed nontraditional roles for cysteine protease cathepsins in the extracellular space during the development and progression of cardiovascular disease. Although the precise mechanisms are unknown, data from animal studies suggest that members of the cathepsin family, like other extracellular proteases, contribute to extracellular matrix protein remodeling and interstitial matrix degradation, as well as to cell signaling and cell apoptosis in heart disease. Inflammatory cytokines and hormones regulate the expression and secretion of cathepsins in cultured cardiovascular cells and macrophages. Serum levels of cathepsins L, S, and K and their endogenous inhibitor cystatin C may be useful predictive biomarkers in patients with coronary artery disease and cardiac disease. Furthermore, in vivo pharmacological intervention with a synthetic cathepsin inhibitor and cardiovascular drugs (including statins and angiotensin II type 1 receptor antagonists) has the potential for pharmacologic targeting of cathepsins in cardiovascular disease. This review focuses on cathepsin biology (structure, synthesis, processing, activation, secretion, activity regulation, and function) and the involvement of cysteinyl cathepsins in the pathogenesis of several heart and vessel diseases, especially with respect to their potential application as diagnostic and prognostic markers and drug targets to prevent inappropriate proteolysis in cardiovascular disease.

The expressions of cathepsin D,H and L in hepatocellular carcinoma / 中华消化杂志

Objective To determine the expression of cathepsin D (CD), cathepsin H (CH) and cathepsin L (CL) in human primary hepatocellular carcinoma (HCC), and to investigate their mechanisms. Methods The protein expression of CD, CH and CL in hepatic specimens consisted of control (n = 17), HCC (n = 37) and paracancerous (n = 28) tissues were detected by immunohistochemistry. The relative values of CD, CH and CL protein expression were examined with absorbent density analysis. Data were analyzed using SPSS11. 5 software. The univariate analysis was used to compare the difference among groups. Results The mean absorbance of CD, CH and CL proteins in HCC tissues (1.21± 0.33, 0. 89 ± 0.22 and 1.16± 0. 25, respectively) were significantly higher in comparison with those in control tissues (0. 19 ± 0. 07, 0. 24 ± 0. 12 and 0. 28 ± 0. 14,respectively) and in paracancerous tissues (0.27±0.13,0. 31± 0.14 and 0. 36±0.15)(all P values =0.0001). Whereas there was no difference between control and paracancerous tissues with respect to CD, CH and CL proteins (P ＞0. 05). Three proteins immunohistochemically appeared as a lot of diffused spots and stripes staining in cytoplasma of HCC tissues,but only a few scatted spots staining was found in control and paracancerous tissues. Conclusion The high expression of CD, CH and CL protein in primary HCC may be important markers for carcinogenesis and malignant progress.

The effect of mannose-6-phosphate on adipocyte differentiation and its underlying molecular mechanism / 中华内科杂志

Objective Cathepsin K (CTSK) played an important role in adipocyte differentiation.The activation of CTSK needs to convey by mannose-6-phosphate receptors (M6PR) in osteoclasts. The aim of the present study was to identify the effects of mannose-6-phosphate (M6P) in adipocyte differentiation and its underlying molecular mechanism. Methods Oil red O staining, accumulation of cytoplasmic triglycerides and glycerine release were used to assess its effects on adipocyte differentiation in the 3T3-L1cell line. The enzyme activity of CTSK was observed by laser confocal microscopy. The proliferation of 3T3-L1 preadipocytes was detected by MTT methods. mRNA expression of M6PR was determined by RTPCR. Results M6P could prevent adipocyte differentiation in a dose-dependent manner as evidenced by absence of triglyceride accumulation and glycerol content. Statistical significance was showed when the concentrations of M6P were 5.0 mmol/L and 8. 0 mmol/L respectively(P ＜0. 05). The mRNA expression of M6PR was detected during the whole process of adipocyte differentiation. With the increase of M6Pconcentration, enzyme activity of CTSK was inhibited in a concentration-dependent manner. MTT method showed that the absorbance at 570 nm of 3T3-L1 preadipocytes was 0. 057 ±0. 091, increased about 62. 9%at 10. 0 mmoL/L compared with the control group (P ＜ 0. 05 ). Conclusion M6P inhibits the terminal differentiation of adipocyte, which may be associated with its effect of blocking CTSK activity by competitive binding with M6PR.

Expressions of aspartic proteinase and cysteine proteinase in photoaged fibroblasts / 中华皮肤科杂志

Objective To investigate the expression changes of aspartic proteinase (cathepsin D) and cysteine proteinase (cathepsin K) in photoaged fibroblasts. Methods The senescence of human fibroblasts was induced via culture in the presence of 8-methoxypsralen (MOP) of 50 mg/L in darkness for 24 hours followed by irradiation with UVA of 80 kJ/m~2. Then, aged fibroblasts were confirmed by senescence-associated β-galactosidase (SA-β-gal) staining. Real-time RT-PCR and Western blot were carried out to detect the mRNA and protein expressions of cathepsin D and cathepsin K in photoaged and normal control fibroblasts, respectively. Results Western blot showed a significant difference between photoaged and control fibroblasts in the grey scale of cathepsin D and cathepsin K (3.25 ± 0.33 vs 14.18 ± 2.25, f = 30.61, P < 0.01; 2.39 ± 0.66 vs 29.38 ± 4.62, t = 12.63, P< 0.01). The △Ct values for cathepsin D and cathepsin K mRNA were 2.79 ± 0.17 and -0.92 ± 0.06, respectively, in photoaged fibroblasts, significantly lower than those in the control fibroblasts (4.54 ± 0.34, 2.57 ± 0.13, t = 20.78, 28.50, respectively, both P < 0.01). According to the value of 2~(-△△Ct), the expression of cathepsin D and cathepsin K mRNA decreased 0.24 ± 0.021 and 0.09 ± 0.005 folds, respectively, in photoaged fibroblasts compared with the control fibroblasts. Conclusion The expression of cathepsin D and cathepsin K is decreased in photoaged fibroblasts.

Mechanisms of curcumin protecting endothelial cells against ischemia and reperfusion injury / 中国药理学通报

Aim To investigate the mechanisms in protecting HUVEC against ischemia/reperfusion(I/R) injury directed by curcumin.Methods Hypoxia/reoxgenation(H/R) model was established on HUVEC.MTT colorimetric assay was used to observe the injury degree of hypoxia and reoxygenation at the different time.With preconditioning by different concentration of Cur,the survival rate of HUVEC subjected to H/R was assessed by MTT colorimetric assay.Pretreated with Cur(5 ?mol?L-1),the expression of LC3,cathepsin B,cathepsin L,Bax and Bcl-2 were observed by fluorescent staining and Western blot in HUVEC during H/R process.Results Cur(1.25~5 ?mol?L-1) played a protective role during H/R in HUVEC in a dose-dependent manner.During H/R,the expressions of LC3,cathepsin B and the ratio of Bax/Bcl-2 increased,and the nuclear translocation of cathepsin L was induced;when cur was pretreated,LC3 was furtherstrengthened,at the same time,the up-regulation of cathepsin B,the ratio of Bax/Bcl-2 and the nuclei-location of cathepsin L were inhibited partly by Cur.Conclusions Cur can raise the survival rate of HUVEC in the process of H/R.Cur increases the autophagy activity,depresses cathepsins and Bax/Bcl-2 to protect the endothelial cells.