Vitronectin Regulates the Fibrinolytic System during the Repair of Cerebral Cortex in Stab-Wounded Mice

Abstract

Vitronectin (VN), one of the serum proteins, is known to be involved in the regulation of blood coagulation, fibrinolysis, and cell migration. It has been proposed that the regulation of fibrinolysis by VN promotes the blood-brain barrier (BBB) recovery from brain injuries such as traumatic injury and subarachnoid hemorrhage. The effects of VN on fibrinolysis in the injured brain remain unclear, however. We examined the effects of VN on the fibrinolytic system in the stab-wounded cerebral cortex of VN-knockout (KO) mice. First, hemorrhage and recovery from BBB breakdown in the wounded regions were assessed by serum immunoglobulin G (IgG) extravasation. The level of IgG extravasation increased 3-7 days after the stab wound (D3-7) in the cortex of VN-KO mice, compared with that in wild type mice, indicating that VN deficiency inhibited the recovery from BBB breakdown. The VN deficiency decreased fibrin fiber deposition at D1-3, suggesting that VN deficiency tilts the balance between fibrinogenesis and fibrinolysis toward fibrinolysis. Next, the effects of VN deficiency on the fibrinolytic factors were analyzed in the stab-wounded cortex. The VN deficiency impaired the activity of plasminogen activator inhibitor-1, an inhibitor of the fibrinolytic system, at D3-5. Further, VN deficiency up-regulated the mRNA and protein expression levels of tissue-type plasminogen activator, and urokinase-type plasminogen activator. These results demonstrate that VN contributes to the regulation of the fibrinolytic system and recovery from BBB breakdown in the wounded brain.

2-carba cyclic phosphatidic acid suppresses inflammation via regulation of microglial polarisation in the stab-wounded mouse cerebral cortex

“…Stab wound surgery was performed using a modified version of the procedure previously described by Hashimoto et al . 18 . Mice (6 weeks old) were anaesthetised by a mixture of 0.75 mg/kg Domitor (Nippon Zenyaku Kogyo, Fukushima, Japan), 4 mg/kg midazolam (Sandoz, Holzkirchen, Germany), and 5 mg/kg Vetorphale (Meiji Seika Pharma, Tokyo, Japan).…”

“…After the operation, the Domitor antagonist Antisedan (0.75 mg/kg; Nippon Zenyaku Kogyo) was injected intraperitoneally to facilitate recovery from anaesthesia. A sample size calculation was based on our previous study 18 .…”

“…Immunohistochemistry was performed as described by Hashimoto et al . with modifications 18 . Briefly, naïve and stab-wounded mice at postoperative days 0, 1, 3, 5, and 7 were perfused with PBS and fixed with 4% paraformaldehyde (PFA), and the collected brains were fixed with 4% PFA overnight.…”

Vitronectin deficiency attenuates hepatic fibrosis in a non‐alcoholic steatohepatitis‐induced mouse model

“…Real‐time RT‐PCR analysis was performed to quantify Vn , αv integrin , Srebp , Acc , Fas , Cd11b , F4/80 , Tnf‐α , Il‐1β , α‐Sma and Tgf‐β1 mRNA expression levels. The primer sequences used in the present experiment are listed in Table . The mRNA expression levels were quantified using the KOD SYBR qPCR Mix (Toyobo Co., Ltd.) with an ABI 7300 real‐time PCR machine (Thermo Fisher Scientific, Waltham, MA).…”

“…The primer sequences used in the present experiment are listed in Table 1. [14][15][16][17][18][19][20] The mRNA expression levels were quantified using the KOD SYBR qPCR Mix (Toyobo Co., Ltd.) with an ABI 7300 real-time PCR machine (Thermo Fisher Scientific, Waltham, MA). In all experiments, gene expression levels were determined using the ΔΔCT method after normalization to Gapdh expression levels.…”