Coagulation status detection in breast cancer patients by using thrombelastography

Many cancer patients are known to present in a hypercoagulable state, meaning an increased risk of thrombosis. To investigate hypercoagulable state in breast cancer (BC) patients, their coagulation status was compared with a benign disease group (control). The BC patients were divided into earlier stage (stage I and stage Ⅱ ) and later stage (stage Ⅲ and stage Ⅳ ). Thrombelastography (TEG) and other traditional coagulation tests were performed. The results showed that prothrombin time (PT) was significantly shortened and the levels of D-dimer, fibrinogen (Fib) and platelets (PLT) were significantly increased in the traditional BC group test (P< 0.05). According to TEG detection, the average level of blood clot formation time (K) was significantly lower, while the Angle, MA and CI were significantly higher in the BC group than those in benign disease group (P< 0.05). There were 5 cases of lower extremity venous thrombosis in the breast cancer patients, coinciding with hypercoagulable state. The results showed that the BC patients had an increased hypercoagulable state, with hypercoagulability becoming more obvious in advanced stages. This study suggests that BC patients have an increased tendency for clot formation, and TEG monitoring could be a useful tool to predict the risk of thrombosis for clinical prevention and treatment.     

Effects of the monoclonal antibody against porcine 40 kDa adipocyte-specific plasma membrane protein on adipocytes and carcass composition

The effects of the mouse monoclonal antibody against 40 kDa adipocyte-specific plasma membrane protein on porcine adipocytes and carcass composition were investigated in vitro and in vivo. Results revealed that the in vitro complement-mediated cytotoxicity of this monoclonal antibody can lead to adipocyte lysis, remarkable reduction of adipocyte lipid accumulation （P〈0.01）, and significant decrease of well-differentiated fat cells （P〈0.01）. Treatment of adipocytes with this antibody alone in vitro did not induce cell lysis, but could lead to noticeable reduction of well-differentiated cells and lipid accumulation （P〈0.05） at the pre-adipocyte stage. In vivo, pigs injected with 0.5 mg/kg or 1.0 mg/kg of antibody showed smaller adipocyte sizes （P〈0.01） and reduced lipid accumulation of adipocytes （P〈0.01）. Our results also indicated that pigs intraperitoneally or subcutaneously immunized with 0.5 mg/kg of monoclonal antibody at 15 kg or 1.0 mg/kg antibody at 60 kg had a higher lean meat percentage （P〈0.05）, larger loin eye area （P〈0.05）, lower fat meat percentage （P〈0.05）, less backfat thickness （P〈0.05） and smaller leaf fat weight （P〈0.05） than the control pigs, but other carcass traits such as caul fat weight, heart weight, liver weight, spleen weight, kidney weight, lung weight, and dressing percentage were not significantly affected. These results suggested that this monoclonal antibody could be applied to restrain excessive fat deposition in porcine production.     

Increased monoamine oxidase activity and imidazoline binding sites in insulin-resistant adipocytes from obese Zucker rats

BACKGROUND Despite overt insulin resistance,adipocytes of genetically obese Zucker rats accumulate the excess of calorie intake in the form of lipids.AIM To investigate whether factors can replace or reinforce insulin lipogenic action by exploring glucose uptake activation by hydrogen peroxide,since it is produced by monoamine oxidase(MAO)and semicarbazide-sensitive amine oxidase(SSAO)in adipocytes.METHODS 3H-2-deoxyglucose uptake(2-DG)was determined in adipocytes from obese and lean rats in response to insulin or MAO and SSAO substrates such as tyramine and benzylamine.14C-tyramine oxidation and binding of imidazolinic radioligands[3H-Idazoxan,3H-(2-benzofuranyl)-2-imidazoline]were studied in adipocytes,the liver,and muscle.The influence of in vivo administration of tyramine+vanadium on glucose handling was assessed in lean and obese rats.RESULTS 2-DG uptake and lipogenesis stimulation by insulin were dampened in adipocytes from obese rats,when compared to their lean littermates.Tyramine and benzylamine activation of hexose uptake was vanadate-dependent and was also limited,while MAO was increased and SSAO decreased.These changes were adipocyte-specific and accompanied by a greater number of imidazoline I2 binding sites in the obese rat,when compared to the lean.In vitro,tyramine precluded the binding to I2 sites,while in vivo,its administration together with vanadium lowered fasting plasma levels of glucose and triacylglycerols in obese CONCLUSION The adipocytes from obese Zucker rats exhibit increased MAO activity and imidazoline binding site number.However,probably as a consequence of SSAO down-regulation,the glucose transport stimulation by tyramine is decreased as much as that of insulin in these insulin-resistant adipocytes.The adipocyte amine oxidases deserve more studies with respect to their putative contribution to the management of glucose and lipid handling.     

Changes in Carboxypeptidase A,Dipeptidase and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript> ATPase Activities in the Intestine of Rats Orally Exposed to Different Doses of Cadmium

The purpose of the present study was to evaluate the effect of cadmium on some protein digestive and absorption enzymes in rats. Thirty-six rats were grouped into three groups of 12 animals each; one group received deionised water and acted as control. One group received 445 μM Cd and the last group received 890 μM Cd in their drinking water for a period of one month. The results obtained indicate that increasing the level of cadmium from 445 μM to 890 μM in the drinking water of the rats led to 29 and 23 increase in accumulated cadmium in the proximal and distal small intestine respectively. The body weight gain of rats exposed to 445 μM and 890 μMCd was decreased by about 24 and 43 respectively when compared with the control. The activities of carboxypeptidase A, dipeptidase and Na+/K+ ATPase were reduced in the mucosa of the proximal end of the small intestine of cadmium exposed rats. The reduction was dose dependent; with the 890 μM Cd exposed rats displaying the least activities. In the distal small intestine, the activities of these enzymes were restored in the 445 μM Cd exposed rats to levels that were not statistically different (P>0.05) from those observed in the controls. In the 890 μMCd exposed rats, dipeptidase activity improved by about 80 compared with the activity of the enzyme in the proximal small intestine. Likewise, Na+/K+ ATPase activity increased by about 125 compared with the observed level in the proximal small intestine. The study suggests that cadmium given to rats in drinking water compromise protein digestion and absorption of nutrients particularly in the proximal region of small intestine and could account for weight reduction associated with cadmium toxicity. Published online December 2004     

Neuroprotection of aucubin in primary diabetic encephalopathy

Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin, using rats (n=8). Diabetes mellitus was induced in the rats by intraperitoneal (i.p.) injection of streptozotocin (60 mg/kg body weight). After 65 d, half of the DM rats were administered aucubin (5 mg/kg; i.p.) for 15 d, yielding treatment DM A. A third group of rats received no strepto- zotocin or aucibin, and served as controls (CON). Encephalopathy was assessed using Y-maze be- havioral testing. Rats were euthanized on Day 87, and hippocampi were excised for visual (light and transmission electron microscopic) and immunochemical (Western blot; immunohistochemical) as- sessments of the CA1 subfield for apoptosis and expression of regulatory proteins Bcl-2 and Bax. Treatment responses to all the parameters examined (body weight, plasma glucose, Y-maze error rates, pyramidal cell ultrastructure, proportions of apoptotic cells, levels of expression of Bcl-2 and Bax, and survivability of neuronal cells) were identical: there were highly significant differences between DM and CON groups (P<0.001), but the effects were significantly moderated (P<0.01) in DM A compared with DM. These findings confirm the association of apoptosis with the encephalopathic effects of diabetes mellitus, and suggest a major role of the expression levels of Bcl-2 and Bax in the regulation of apop- totic cell death. All of the results suggest that aucubin could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.     

Matrine reduces the proliferation and invasion of colorectal cancer cells via reducing the activity of p38 signaling pathway

Matrine has been used in anti-inflammatory and anti-cancer therapies for a long time. However, the anti-metastatic effect and related mechanism（s） in colorectal cancer （CRC） are still unclear. In this study, we investigated whether the adminis- tration of matrine could inhibit the proliferation, motility, and invasion of human CRC cells via regulating p38 signal- ing pathway. Results showed that matrine inhibited migration and invasion of CRC cells in vitro and in vivo. Additionally, after being treated with matrine for 24 h, the expression levels of matrix metalloproteinase-2 （MMP-2） and MMP-9 as well as proteinase activity in CRC cells were reduced in a dose-dependent manner. Moreover, matrine reduced the phosphorylation level of p38 obviously. Combined treatment with p38 inhibitor （SB203580） and matrine resulted in a syn- ergistic reduction of invasion as well as MMP-2/-9 expression in CRC cells. It was also found that matrine inhibited the pro- liferation and metastasis of CRC tumor in vivo. In conclusion, p38 signaling pathway may involve in matrine＇s inhibitory effects on migration and invasion of CRC cells by reducing the expression of MMP-2/-9, suggesting that matrine may be a potential therapeutic agent for CRC.     

AdipoRon prevents L-thyroxine or isoproterenol-induced cardiac hypertrophy through regulating the AMPK-related pathway

Cardiac hypertrophy is a risk factor which can intrigue heart failure.In the present study,we explored whether AdipoRon attenuates isoprenaline (ISO) or L-thyroxine-induced cardiac hypertrophy in Sprague-Dawley (SD) rats and whether the anti-hypertrophy effect is mediated by AMPK-related pathway.Here,cardiac hypertrophy was induced by injection of L-thyroxine or ISO in SD rats.In the treatment group,AdipoRon was co-administered.We examined the effects of AdipoRon on cardiac hypertrophy and hypertrophy signaling pathway.The weight of SD rats was recorded every day.Rats were killed for collection of blood and heart under anesthesia.The left heart weight and heart weight were weighed.Paraffin-embedded heart tissue regions (4 μm) were stained with hematoxylin and eosin or Masson to detect left heart hypertrophy and myocardial fibrosis.The serum BNP levels were determined by using an enzyme-linked immunosorbent assay.The mRNA levels of ANP,BNP,PGC-1α,and ERRα were evaluated by real-time PCR analysis.The protein expression levels of PGC-1α,ERRα,and pAMPK/AMPK were determined by western blot analysis.The results showed that AdipoRon significantly reversed heart weight (HW)/ body weight (BW) ratio,left ventricular (LV)/BW ratio,serum BNP level and the mRNA level of ANP and BNP induced by ISO or L-thyroxine.ISO or L-thyroxine reduced both the mRNA level and protein level of ERRα and PGC-1α,and also reduced the protein level of pAMPK/AMPK.However,AdipoRon reversed ISO or L-thyroxine-induced changes of pAMPK/AMPK,ERRα,and PGC-1α.Our data indicated that the effects of AdipoRon are mediated partly by activating AMPK-related pathway,and AdipoRon plays a potential role in the prevention of cardiac hypertrophy.     

Alterations of cardiac and lymphocyte β-adrenoceptors in rat with chronic heart failure

The alterations of cardiac and lymphocyte β-adrenoceptors were observed in the rats with chronic heart failure produced by constriction of both abdominal aorta and renal artery. The results showed that β1-adrenocep-tor density and mRNA levels were increased, whereas these levels remained unchanged for β2 The concentration-contractile response curve for isoproterenol was shifted to the right in cardiac atrium, whereas the concentration-cAMP accumulation response curve for isoproterenol in myocardium was not changed. The number of β-adrenoceptors in blood lymphocyte was markedly reduced. Thus in the heart-failure rats the density of cardiac β-adrenoceptor was increased accompanying reduced β-adrenoceptor-mediated positive inotropic response, suggesting a post adenylate cyclase dys-function or impaired contractile components. In contrast, the alteration of β-adrenoceptor in lymphocyte is consistent with the reduced β-adrenoceptor-mediated inotropic response in heart.     

Everolimus and zoledronic acid——a potential synergistic treatment for lung adenocarcinoma bone metastasis

Non-small-cell lung cancer （NSCLC） frequently metasta- sizes to bone. It is known that zoledronic acid is cytostatic to tumors, and everolimus, the inhibitor for mammalian target of the rapamycin, could inhibit many types of cancer. Herein, we evaluated the effect of zoledronic acid alone and in combination with everolimus on treating lung adenocarcinoma bone metastasis in vitro and in vivo. Mice treated with zoledronic acid in combination with everoli- mus had more apoptotic lung cancer cells and more cells were arrested in the G1/G0 phase. The phosphorylation of p70S6K was inhibited in the combination treatment group. Lung cancer cell invasion was also significantly inhibited in the group with combination treatment in vitro. Bone nuclear scans revealed more metastatic lesions in controls compared with those in the combination treatment group. Bone scans and radiographic images indicated that com- bination therapy significantly reduced bone metastasis. The moderate survival rate suggested that the drug com- bination was synergistic, which can delay NSCLC bone metastasis and prolong survival in vivo.     

Pyrrolidine dithiocarbamate protects pancreatic β-cells from oxidative damage through regulation of FoxO1 activity in type 2 diabetes rats

Pyrrolidine dithiocarbamate （PDTC） can lower the bloot glucose level and improve the insulin sensitivity in diabeti, rats. However, the mechanisms underlying this effect o PDTC treatment in diabetic rats remained uncertain, h this study, we evaluated the mechanisms by which PDT（ conferred protection against oxidative damage to pancreat ic islet β-cells in rats with experimental type 2 diabete mellitus （DM）. DM in the rats was elicited by long-tern high-fat diet accompanied with a single intraperitonea （i.p.） injection of a low dose of streptozotocin. After a 7-da1 administration of PDTC （50 mg/kg/day i.p.）, blood glucos levels were measured and pancreatic tissues were collecte / for the determination of various biochemical and enzyma 1 ic activities using immunohistochemistry, immunofluoresI cence, and western blot techniques. The percentage o 1 apoptotic pancreatic islet β-cells was detected by flow cyto metry. The results showed that diabetic rats had elevate blood glucose levels and insulin resistance, accompanieq with an increase in malondialdehyde content, nitrotyrosin production, and inducible nitric oxide synthase expression A decrease in superoxide dismutase and glutathione pero idase activities was also observed in DM rats, culminatin with elevated β-cell apoptosis. PDTC treatment significantl reduced the oxidative damage and the β-cell apoptosi and also increased the insulin production through down-reg lating FoxO1 acetylation and up-regulating nuclear PDX- level. These data suggested that PDTC can protect islet βcells from oxidative damage and improve insulin productio through regulation of PDX-1 and FoxO1 in a DM rat model.     

The influence of Annexin32, a new Ca2+-dependent phospholipid-binding protein, on coagulation time and thrombosis

The pharmacodynamics of Annexin32, a new Ca2+-dependent phospholipid-binding protein, was studied by measuring coagulation time in rabbits and venous thrombosis in rabbits and rats. Rabbits and rats were given Annexin32 by intravenous administration. Then Kaolin partial thromboplastin time (KPTT), thrombosis in vitro and in vivo were assayed. The results showed that KPTT of rabbits was prolonged (p < 0.01), and the length and weight of thrombus in vitro were reduced (p < 0.01) after administration of Annexin32 at 1 mg/kg. It also inhibited thrombosis in vivo and reduced the weight of venous thrombus significantly in rats (p < 0.01). All these results suggested that Annexin32 possesses the characteristic of antithrombotic effect and fewer side effects on coagulation time.     

Antithrombotic effect of L-arginine in hypertensive rats.

The aim of the study was to evaluate the effect of L-arginine (L-Arg) on haemostasis in stasis model of venous thrombosis in renal hypertensive rats. The effect of the single dose (i.v. 300 mg/kg bolus+300 mg/kg/h) and of the 10-day application (p.o. 1 g/kg, once daily) of L-Arg was determined. L-Arg reduced the blood pressure both in the acute and long-term application. The single dose of L-Arg decreased the occurrence rate of the thrombus whereas long-term administration reduced significantly the thrombus weight. There were no differences in prothrombin time and activated partial thromboplastin time while the fibrinogen concentration decreased both in the acute and the long-term experiment. L-Arg shortened euglobulin clot lysis time and bleeding time in the long-term application. The chronic L-Arg treatment also inhibited significantly collagen-induced platelet aggregation. The overall haemostasis and coagulation potentials were inhibited and the fibrinolysis potential was higher in the group receiving this amino-acid. The results show that L-Arg, in a complex way, evokes the antithrombotic effect in the model of venous thrombosis in hypertensive rats.     

Antithrombotic effect of tissue and plasma type angiotensin converting enzyme inhibitors in experimental thrombosis in rats.

This study compared the antithrombotic effect of plasma angiotensin converting enzyme inhibitors (ACE-Is): captopril (CAP), enalapril (ENA) and tissue ACE-Is: perindopril (PER), quinapril (QUIN) in experimental venous and arterial thrombosis. Normotensive Wistar rats were treated p.o. with CAP (75 mg/kg), ENA (20 mg/kg), PER (2 mg/kg) and QUIN (3 mg/kg) for 10 days. The influence of ACE-Is on coagulation and fibrinolytic systems as well as platelet function was evaluated. The hypotensive effect of ACE-Is was equal in all groups. QUIN maintained the final carotid blood flow at the highest value in comparison to PER and plasma ACE-Is. The arterial thrombus weight was reduced in PER and QUIN groups while venous thrombus weight was also reduced after CAP. Tissue and plasma ACE-Is caused the inhibition of platelet adhesion and aggregation. A reduction of fibrin generation, prolongation of prothrombin time (PT), activated partial thromboplastin time (APTT) and shortening of euglobulin clot lysis time (ECLT) were observed after PER and QUIN treatment. In conclusion, given in equipotent hypotensive doses, tissue ACE-Is exerted more pronounced antithrombotic effect than plasma ACE-Is in experimental thrombosis. The differences between tissue and plasma ACE-Is in terms of their more pronounced inhibition of experimental thrombosis may be related to the intensified activation of fibrinolysis and inhibition of coagulation.     

毕赤酵母重组Annexin32的分子特征及其对血凝与血栓形成的影响

采用SDS PAGE、等电聚焦、脱磷酸反应、蛋白质N端氨基酸测序、KPTT及大鼠下腔静脉血栓形成观察 ,发现毕赤酵母重组Annexin32的分子特征较原核产物有了明显变化。有主、次 2条带 ,主带的分子量比原核产物大 ,等电点较理论值低 ,已被磷酸化 ,2条带的N端均携带了来自载体的 4个氨基酸 ,可能对其与活化血小板表面磷脂的结合有所下调 ,导致血凝与血栓形成受到一定的影响 ,但仍具有明显的抗凝及抑制血栓形成作用。     

Sulfatide can markedly enhance thrombogenesis in rat deep vein thrombosis model

Although sulfatide (galactosylceramide I3-sulfate) has been reported to activate blood coagulation factor XII (Hageman factor), it has been administered to animals without subsequent thrombus formation. We recently found that sulfatide binds to fibrinogen and thus disturbs fibrin formation in vitro, suggesting its possible role as an anticoagulant rather than as a coagulant. We therefore examined the in vivo effects of sulfatide on thrombogenesis by using a rat deep vein thrombosis model in which thrombus is induced by ligating the inferior vena cava. Sulfatide and gangliosides were each separately administered to rats 1 min before the vein ligation, and after 3 h, sulfatide but not gangliosides markedly (P < .001) enhanced the thrombogenesis. A kinetic turbidmetric assay of plasma coagulation initiated by CaCl2 in the wells of a microtiter plate revealed that coagulation was also markedly accelerated in the presence of sulfatide but not gangliosides, the results of which seemed to be very consistent with those of the in vivo experiments. Because sulfatide could not induce thrombosis without vein ligation in rats, the enhancement of thrombogenesis by sulfatide in the in vivo model might require endothelial damage and/or venous congestion, both of which could be induced by vein ligation.     

沙棘油对实验性血栓形成及凝血系统的影响

沙棘油能使实验性血栓形成延迟,具有预防血栓形成的作用。沙棘油有一定的抗凝作用,主要参与内源性凝血系统;且有促纤溶作用,明显降低纤维蛋白原含量,使血浆鱼精蛋白副凝试验呈阳性反应。     

Experimental studies on the antithrombotic and haemorrhagic effects of heparin and a low molecular weight heparin derivative

Antithrombotic, haemorrhagic and anticoagulant effects of unfractionated heparin (UH) and the low molecular weight heparin fragment KABI 2165 were studied in rats. In stasis-induced venous thrombosis of the jugular vein intravenous injection of both, UH and KABI 2165, either reduced significantly the size of thrombi or completely prevented thrombus formation in a dose-dependent manner. The dose of KABI 2165 required for prevention of thrombus formation showed a marked anticoagulant activity measured by APTT which was in the same range as that of the equieffective dose of UH. After administration of antithrombotically effective doses only UH caused a significant prolongation of bleeding time after standardized incision of the tail. KABI 2165 produced haemorrhagic effects at about 4-fold higher doses only than those required for the antithrombotic action.     

Inhibition of microsurgical thrombosis by the platelet glycoprotein IIb/IIIa antagonist SR121566A

Despite major improvements in tools and significant refinements of techniques, microsurgical anastomosis still carries a significant risk of failure due to microvascular thrombosis. The key to improving the success of microvascular surgery may lie in the pharmacologic control of thrombus formation. Central to pathologic arterial thrombosis are platelets. Glycoprotein IIb/IIIa is a highly abundant platelet surface receptor that plays a major role in platelet aggregation by binding platelets to each other through the coagulation factor fibrinogen. To explore the ability of antithrombotic agents to prevent microvascular thrombosis, a rabbit ear artery model was used in which a standardized arterial injury results in predictable thrombus formation. This model was used to examine whether SR121566A, a specific and potent glycoprotein IIb/IIIa inhibitor, can successfully prevent microsurgical thrombosis.Using a coded, double-blind experimental design, 20 rabbits (40 arteries) were assigned to four treatment groups: (1) saline injection (n = 10), (2) acetylsalicylic acid 10 mg/kg (n = 10), (3) heparin 0.5 mg/kg bolus with subsequent intermittent boluses of 0.25 mg/kg every 30 minutes (n = 10), and (4) SR121566A 2 mg/kg bolus (n = 10). After vessel damage and clamp release, arteries were assessed for patency at 5, 30, and 120 minutes by the Acland refill test. Coagulation assays, in vivo bleeding times, and ex vivo platelet aggregation studies were also conducted. Scanning electron microscopy was used to examine mural thrombus composition.A significant, fourfold increase in vessel patency following administration of SR121566A over saline control (80 percent versus 20 percent patency, respectively, at 35 minutes after reperfusion, p < 0.01) was noted. This was correlated with marked inhibition of ex vivo platelet aggregation. This antiplatelet treatment did not prolong coagulation assays (mean international normalized ratio: saline, 0.66 +/- 0.04; SR121566A, 0.64 +/- 0.03; mean thromboplastin time: saline, 19.63 +/- 0.67; SR121566A, 17.87 +/- 3.27) and bleeding times (mean bleeding time: saline, 42 +/- 4; SR121566A, 48 +/- 6). Scanning electron microscopy demonstrated extensive platelet and fibrin deposition in control vessel thrombi. In contrast, thrombi from SR121566A-treated vessels demonstrated predominance of fibrin with few platelets when examined under scanning electron microscopy.Administration of SR121566A was associated with a significant increase in vessel patency, without deleterious effects on coagulation assays or bleeding times. The increase in vessel patency was correlated with inhibition of platelet aggregation and decreased platelet deposition, as demonstrated by scanning electron microscopy. Glycoprotein IIb/IIIa antagonists represent a new class of anti-platelet agents that may be suited for inhibiting microsurgical thrombosis. This study supports further investigation into the use of these agents in microsurgery.     

Moderate Intensity Static Magnetic Fields Prevent Thrombus Formation in Rats and Mice

We established three types of thrombosis models to explore the effects of the static magnetic field (SMF) on thrombosis in rats and mice with three different MF intensities. In the carrageenan-induced thrombosis model in rats, the SMF treatments reduced the black tail length of rats, extracorporeal thrombus, and the mass of wet and dry thrombus, and improved the coagulation index value. In FeCl₃-induced arterial thrombosis model in rats, the SMF treatment showed some anti-thrombotic effects. More specifically, the SMF treatment affected rodent blood pressure, plasma plasminogen activator inhibitor, tissue-type plasminogen activator, thrombus mass, and thrombus protein content. In the adrenaline-induced thrombosis model in mice, the SMF treatment had certain effects on the diameter and blood flow velocity of mouse auricle microcirculation in fine veins and arteries. Overall, the highest MF intensities we tested, 20–150 mT, showed a trend of anti-thrombotic effect, indicating that the moderate-intensity SMF might serve as a potential treatment for clot-related diseases in the future. Bioelectromagnetics. 2020;41:52–62 © 2019 Bioelectromagnetics Society.     

Statins Improve the Resolution of Established Murine Venous Thrombosis: Reductions in Thrombus Burden and Vein Wall Scarring

Despite anticoagulation therapy, up to one-half of patients with deep vein thrombosis (DVT) will develop the post-thrombotic syndrome (PTS). Improving the long-term outcome of DVT patients at risk for PTS will therefore require new approaches. Here we investigate the effects of statins—lipid-lowering agents with anti-thrombotic and anti-inflammatory properties—in decreasing thrombus burden and decreasing vein wall injury, mediators of PTS, in established murine stasis and non-stasis chemical-induced venous thrombosis (N = 282 mice). Treatment of mice with daily atorvastatin or rosuvastatin significantly reduced stasis venous thrombus burden by 25% without affecting lipid levels, blood coagulation parameters, or blood cell counts. Statin-driven reductions in VT burden (thrombus mass for stasis thrombi, intravital microscopy thrombus area for non-stasis thrombi) compared similarly to the therapeutic anticoagulant effects of low molecular weight heparin. Blood from statin-treated mice showed significant reductions in platelet aggregation and clot stability. Statins additionally reduced thrombus plasminogen activator inhibitor-1 (PAI-1), tissue factor, neutrophils, myeloperoxidase, neutrophil extracellular traps (NETs), and macrophages, and these effects were most notable in the earlier timepoints after DVT formation. In addition, statins reduced DVT-induced vein wall scarring by 50% durably up to day 21 in stasis VT, as shown by polarized light microscopy of picrosirius red-stained vein wall collagen. The overall results demonstrate that statins improve VT resolution via profibrinolytic, anticoagulant, antiplatelet, and anti-vein wall scarring effects. Statins may therefore offer a new pharmacotherapeutic approach to improve DVT resolution and to reduce the post-thrombotic syndrome, particularly in subjects who are ineligible for anticoagulation therapy.