3-4-dihydroxyphenyllactic-acid and Brain-Ischemia

Cerebral infraction seriously affects the life quality of patients. Danshensu has been reported to exhibit anti-inflammatory and vascular protective effects. However, the therapeutic function of Danshensu in cerebral vascular injury is still unclear.. Middle cerebral artery occlusion (MCAO) was used to construct the cerebral infraction model. Wound healing and tube formation assays were used to evaluate angiogenesis in vitro. Western blot assay was used to evaluate the activation of the PI3K/Akt/mTOR signaling pathway. The laser Doppler scanner was used to measure the regional cerebral blood flow (rCBF) in the area around the infarction, and the adhesion removal test was used to measure the sensorimotor function. The Modified Neurological Severity Score was performed to evaluate the cognitive functions of mice.. Danshensu promoted the proliferation of bEnd.3 cells and angiogenesis in vitro. Danshensu upregulated the expression of VEGF through PI3K/Akt/mTOR signaling pathway in bEnd.3 cells. Danshensu improved rCBF restoration and attenuated the behavioral deficits in mice post-MCAO/R.. Danshensu enhances angiogenesis through the PI3K/Akt/mTOR/VEGF signaling pathway in a mouse model of cerebral ischemic injury.

Topics: Animals; Brain Ischemia; Endothelial Cells; Infarction, Middle Cerebral Artery; Mice; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases; Vascular Endothelial Growth Factor A

Cerebral ischemia-reperfusion injury (CI/RI) is the main cause of disability and death in stroke without satisfactory therapeutic effect. Inflammation mediated by activation of astrocytes and microglia is the main pathological mechanism of CI/RI. Danshensu (DSS) has been shown to exert anti-inflammatory effects against brain injury. However, limited by its poor cellular permeability and low bioavailability, it is still needed the new DSS preparations with the ability to cross the blood-brain barrier (BBB) and target inflammatory glial cells. In this study, we developed phosphatidylserine (PS) and transferrin (TF) modified liposomes carrying DSS (TF/PS/DSS-LPs) to improve the therapeutic efficacy against ischemic stroke. First, TF molecules targeted transferrin receptor (TfR) that is overexpressed in the BBB. Following the liposomes enter the brain, PS modification allowed the liposomes to target and bind to the overexpressed phosphatidylserine-specific receptors (PSRs) on the surface of astrocytes and microglia. Furthermore, it enhanced the uptake of TF/PS/DSS-LPs by astrocytes and microglia, while polarizing astrocytes from A1 to A2 and microglia from M1 to M2, reducing neuronal inflammation, and ultimately ameliorating cerebral ischemic injury. Thus, TF/PS/DSS-LPs could potentially serve as a promising strategy for the CI/RI treatment.

Topics: Astrocytes; Brain Ischemia; Humans; Inflammation; Lipopolysaccharides; Liposomes; Microglia; Phosphatidylserines; Reperfusion Injury

Ischemic stroke remains a serious threat to human life. There are limited effective therapies for the treatment of stroke. We have previously demonstrated that angiogenesis and neurogenesis in the brain play an important role in functional recovery following ischemic stroke. Recent studies indicate that increased arteriogenesis and collateral circulation are determining factors for restoring reperfusion and outcomes of stroke patients. Danshensu, the Salvia miltiorrhiza root extract, is used in treatments of various human ischemic events in traditional Chinese medicine. Its therapeutic mechanism, however, is not well clarified. Due to its proposed effect on angiogenesis and arteriogenesis, we hypothesized that danshensu could benefit stroke recovery through stimulating neurogenesis and collaterogenesis in the post-ischemia brain. Focal ischemic stroke targeting the right sensorimotor cortex was induced in wild-type C57BL6 mice and transgenic mice expressing green fluorescent protein (GFP) to label smooth muscle cells of brain arteries. Sodium danshensu (SDS, 700 mg/kg) was administered intraperitoneally (i.p.) 10 min after stroke and once daily until animals were sacrificed. To label proliferating cells, 5-bromo-2'-deoxyuridine (BrdU; 50 mg/kg, i.p.) was administered, starting on day 3 after ischemia and continued once daily until sacrifice. At 14 days after stroke, SDS significantly increased the expression of vascular endothelial growth factor (VEGF), stromal-derived factor-1 (SDF-1), brain-derived neurotrophic factor (BDNF), and endothelial nitric oxide synthase (eNOS) in the peri-infarct region. SDS-treated animals showed increased number of doublecortin (DCX)-positive cells. Greater numbers of proliferating endothelial cells and smooth muscle cells were detected in SDS-treated mice 21 days after stroke in comparison with vehicle controls. The number of newly formed neurons labeled by NeuN and BrdU antibodies increased in SDS-treated mice 28 days after stroke. SDS significantly increased the newly formed arteries and the diameter of collateral arteries, leading to enhanced local cerebral blood flow recovery after stroke. These results suggest that systemic sodium danshensu treatment shows significant regenerative effects in the post-ischemic brain, which may benefit long-term functional recovery from ischemic stroke.

Topics: Animals; Brain Ischemia; Cell Proliferation; Cells, Cultured; Cerebrovascular Circulation; Doublecortin Domain Proteins; Doublecortin Protein; Lactates; Male; Mice, Inbred C57BL; Microtubule-Associated Proteins; Neovascularization, Physiologic; Neural Stem Cells; Neurogenesis; Neuropeptides; Recovery of Function

To investigate the protective effect of edaravone and danshensu conjugate (IM-009) on focal cerebral ischemia-reperfusion injury in rats and its underlying mechanisms.
. Rats were randomly assigned into 6 groups, including a sham group, a model group, an edaravone-treated group, a danshensu-treated group, a low dose of IM-009-treated group and a high dose of IM-009-treated group. The focal cerebral ischemia-reperfusion model was established by intraluminal filament. After the drug treatment, the infarct volume and extent of brain edema were measured. The levels of MDA and SOD were determined by the corresponding assay kit. The scavenging effect of IM-009 on hydroxyl radical and superoxide anion was also measured in a cell free system.
. 1) In comparison with the model group, the infarct volume and water content in rat brain after IM-009 treatment were significantly reduced. The protective effect of IM-009 at higher dose was much stronger than that of edaravone or danshensu (all P<0.05). 2) IM-009 significantly reduced the levels of MDA and increased the activity of SOD (all P<0.05). 3) IM-009 demonstrated strong activities in scavenging .OH and .O(2)(-) (all P<0.05).
. IM-009 is able to protect rats from ischemia-reperfusion injury. The protective effect of IM-009 could be due to its radical-scavenging action.. 目的：观察依达拉奉丹参素共轭物(IM-009)对大鼠局灶性缺血再灌注损伤模型的保护作用，对其作用机制进行初步研究。方法：将SD大鼠随机分为假手术组、模型组、依达拉奉治疗组、IM-009低剂量治疗组以及IM-009高剂量治疗组。采用线栓法制作大鼠局灶性脑缺血再灌注损伤模型，药物干预后，观察各组动物的脑梗死体积及其脑水肿程度。用试剂盒检测脑组织中丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活力。采用体外非细胞实验法检测IM-009对羟自由基(.OH)和超氧阴离子(.O2−)的清除能力。结果：1) 与模型组比较，IM-009治疗组大鼠的脑梗死体积明显减小，脑水含量降低，并且IM-009高剂量组大鼠的脑梗死体积小于依达拉奉治疗组(均P<0.05)。2) 与模型组比较，IM-009治疗组大鼠的MDA含量显著降低，SOD活性明显提高(均P<0.05)。3) IM-009对.OH和.O2−具有较强的清除能力(均P<0.05)。结论：IM-009对大鼠局灶性脑缺血再灌注损伤动物模型具有保护作用，其作用机制可能与其清除自由基有关。.

Topics: Animals; Antipyrine; Brain Edema; Brain Ischemia; Cerebral Infarction; Edaravone; Lactates; Malondialdehyde; Rats; Reperfusion Injury; Superoxide Dismutase