Objective:To observe the effect and mechanism of Tanhuo Prescription on regulating the activation of M1 microglia and alleviating brain tissue injury in rats with cerebral ischemia.Methods:Male SD rats were divided into sham-operation group, model group, Tanhuo Prescription high-(3.68 g/kg), medium-(1.84 g/kg), low-dosage(0.92 g/kg) groups, and ginaton group (0.06 g/kg) using random number table method. Except for the sham-operation group, the other groups established cerebral ischemia rat models using the middle cerebral artery occlusion method. The balance beam walking test was used to evaluate the symptoms of neurological deficit. MRI-T2 mapping was used to measure the damage to brain tissue. LFB staining was used to observe the damage to nerve fibers. HE staining was used to observe the damage to nerve cell, and Iba-1 and CD16/Iba-1 immunofluorescence staining were used to observe the condition of microglial activation.Results:Compared with the model group, the scores of balance beam walking ability of rats in Tanhuo Prescription high-dose group and ginaton group at 24 h, 48 h and 72 h after ischemia were significantly improved ( P<0.05, P<0.01). The scores of balance beam walking ability of rats in Tanhuo Prescription low- and medium- dose groups at 72 h after ischemia were improved ( P<0.01). Compared with the model group, the T2 values of the cortex and striatum around the infarct of rats in Tanhuo Prescription high-dose group and ginaton group were significantly reduced ( P<0.05, P<0.01), and the T2 values of the striatum around the infarct of rats in Tanhuo Prescription low- and medium- dose groups were significantly reduced ( P<0.05). Compared with the model group, the LFB IOD of the cortex, striatum and outer capsule around the infarct decreased in the Tanhuo Prescription high-,low-dose group and ginaton group ( P<0.01). The LFB IOD of striatum around infarct decreased in medium- dose Tanhuo Prescription group ( P<0.01). Compared with the model group, the pathological injury degree of the striatum around the infarct of rats in Tanhuo Prescription low- ,medium-, and high-dose groups decreased, and the cell density decreased ( P<0.05, P<0.01). The density of the cortical and striatum cells around the infarct of rats in ginaton group increased ( P<0.01, P<0.05). Compared with the model group, the number of Iba-1 and CD16/Iba-1 positive cells in the cortex and striatum around the infarct decreased in Tanhuo Prescription medium-, high-dose and ginaton groups ( P<0.01). The number of CD16/Iba-1 positive cells in the cortex and striatum around the infarct of rats in Tanhuo Prescription low-dose group decreased ( P<0.01), and the number of Iba-1 positive cells in the striatum around the infarct of rats in Tanhuo Prescription low-dose group decreased ( P<0.01). Conclusion:Tanhuo Prescription can improve the symptoms of neurological deficits in rats with cerebral ischemia, reduce the neuropathological damage in the cerebral area around ischemic infarction, and inhibit the activation of M1 microglia.

Objective:To study the effect of Tanhuo Formula (THW) on the expression of Glial Fibrillary Acidic Protein (GFAP), Caspase-3 and angiogenesis.Methods:Rats were divided into sham group, model group, low-dose THW group, medium-dose THW group, high-dose THW group and Ginaton group according to random number table method. Except the sham group, rats in other groups were subjected to the middle cerebral artery occlusion via a suture method. After 2 hours,rats in the low, medium and high dose of THW groups were gavaged with 0.92, 1.84 and 3.68 g/kg of THW dry extract powder solution respectively, and the Ginaton group were gavaged with 60 mg/kg of Ginaton, once every 24 hours for 3 days. Rats in sham operation group and model group were given equal volume of normal saline by gavage. The limb symmetry score was used to evaluate limb dysfunctions. The immunofluorescence staining of GFAP and Caspase-3 were applied to assess astrocyte activation and neuronal apoptosis, respectively. The double-labeled immunofluorescence staining of platelet-endothelial cell adhesion molecule (CD31) and chondroitinsulphate peoteoglycan (NG2) were performed to detect angiogenesis.Results:Compared with the model group, rats in the high-dose of THW group showed increased limb symmetry score ( P<0.01 or P<0.05), increased number of Caspase-3 (cortex: 765.0±122.4 vs. 1 131.0±392.9; striatum: 895.9±389.8 vs. 1 401.9±453.1) ( P<0.01 or P<0.05) and CD31 +/NG2 + (cortex: 1 355.0±257.9 vs. 825.4±308.1; striatum: 1 290.9±400.9 vs. 675.2±259.7) ( P<0.01) positive cells in the periinfarct cortex and striatum, and attenuated the integrated optical density of GFAP in the perilesional cortex (4 210.00±1 226.38 vs. 7 935.78±2 001.98) ( P<0.01). Conclusions:THW could ameliorate the limb functional disorders, inhibit astroglia activation, down-regulate the expression of Caspase-3, and enhanced angiogenesis in MCAO rats.

Objective To observe the effect of Buyang Huanwu Decoction on hippocampal tissue structure and blood perfusion in rats with focal cerebral ischemia using multi-modal MRI;To analyze the mechanism of Buyang Huanwu Decoction on hippocampal neuroplasticity combined with the expression changes of neuroplasticity proteins and glucose metabolism related transporters.Methods Focal cerebral ischemia rat model induced by right middle cerebral artery occlusion were established.The rats were divided into sham-operation group,sham-operation+Buyang Huanwu Decoction group,model group and model+Buyang Huanwu Decoction group.The rats in the sham-operation group and model group were given normal saline for 30 days,and those in the sham-operation+Buyang Huanwu Decoction group and model+Buyang Huanwu Decoction group were given Buyang Huanwu Decoction for 30 days.T2 mapping imaging was used to detect the changes in hippocampal tissue structure,the changes of cerebral blood perfusion in hippocampus were detected by arterial spin labeling imaging,Western blot was used to detect the protein expressions of SYN,GAP-43,glucose transporters and MCT.Results Compared with the sham-operation group,the T2 relaxation time of the right and left hippocampus in the model group rats significantly increased(P<0.01,P<0.001),the blood flow in the right hippocampus was significantly reduced(P<0.05),the protein expressions of SYN,GAP-43,MCT4 and MCT2 in the right hippocampus were significantly decreased(P<0.01,P<0.001),the protein expressions of GLUT1 and GLUT3 in bilateral hippocappal tissue significantly decreased(P<0.05,P<0.01,P<0.001).Compared with the model group,the T2 relaxation time in the right hippocampus of rats in model+Buyang Huanwu Decoction group was significantly reduced(P<0.05),the blood flow in the right hippocampus significantly increased(P<0.05),the protein expressions of SYN,GAP-43,GLUT1 and GLUT3 in bilateral hippocampal tissues significantly increased(P<0.01,P<0.001),and the protein expressions of MCT4 and MCT2 in right hippocampal tissue significantly increased(P<0.01,P<0.001).Conclusion Buyang Huanwu Decoction can alleviate hippocampal injury in rats with focal cerebral ischemia,which may be related to improving blood perfusion,up-regulating neuroplasticity-related proteins,promoting hippocampal axon regeneration and synaptic remodeling,regulating energy metabolism of nerve cells.

Objective　To explore the clinical value of 3D artistic spin labeling technology in evaluating early perfusion changes of leukaraiosis.Methods　Ninety patients and 30 healthy volunteers with different degrees of LA who were examined by magnetic resonance imaging were collected from January 2018 to April 2020.According to the Fazekas scale,they were divided into LA0,LA1,LA2 and LA3 group,30 cases were included in each group.CBF values of cortical watershed and subcortical watershed were measured according to cerebral blood flow pseudocolor image,and cerebral perfusion differences in anterior,posterior,radial crown and semicircular center of cortical watershed in different LA grades were evaluated,and the characteristics of early hemodynamic changes of LA were analyzed.Results　The CBF values of LA2,LA3 were significantly lower than that of LA0.There was no significant difference between LA1 and LA0 (P>0.05),however,the CBF in subcortical watershed was slightly lower than LA0 (P<0.05).Conclusion　3D ASL perfusion imaging technology can be used to quantify the CBF in the watershed area,and can be used to monitor the state of cerebral perfusion in LA patients.The CBF response in the subcortical watershed area is more sensitive,which can be used to evaluate the early cerebral hemodynamic changes in LA.