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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.
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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.
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Objective:This study aimed to evaluate the neuroprotective effect of Buyang-Huanwu Decoction (BYHWD) against cerebral ischemia by using magnetic T2WI and DTI to observe the infarct volume, formation of cerebral edema and injury of white matter fibers on cerebral ischemic rats. Methods:Rat model of cerebral ischemia was established by middle cerebral artery occlusion (MCAO). The successfullly modeled rats were randomly divided into model group and BYHWD group, with 5 rats in each group, and the other 5 rats were taken as sham operation group. The Rats were intragastrically administrated with BYHWD (16.1 g/kg) once daily for 30 d after MCAO. T2WI and DTI examinations were performed on the 3rd, 7th, 14th, 30th day after the surgery.Results:T2WI showed abnormal hyperintensities in the right hemisphere of the MCAO rats on the 3rd, 7th, 14th, 30th day after the surgery. The infarction percentage reduced with time coursing ( P<0.05). Compared with the model group, the rats treated with BYHWD showed reduced infarction percentage [3 d: (13.9% ± 13.7% vs. 40.1% ± 10.7%); 7 d: (13.9% ± 11.9% vs. 28.2% ± 7.7%); 14 d: (10.2% ± 7.9% vs. 24.5% ± 3.5%); 30d: (6.8% ± 6.5% vs. 24.7% ± 8.7%)] ( P<0.05 or P<0.01). The percentage of edema reached the peak on the 3rd day after MCAO. Due to the cerebral atrophy on 30th day, the percentage of edema exhibited negative growth. The edema percentage of BYHWD group reduced significantly on the 3rd and 7th day compared with model group [3 d: (11.4% ± 6.9% vs. 21.5% ± 3.1%); 7 d: (5.5% ± 3.1% vs. 8.7% ± 1.2%)] ( P<0.05 or P<0.01). DTI showed that the low signals indicating fiber injuries were observed in the infarct areas of the model group rats on the 3rd, 7th, 14th, 30th day after MCAO. The rFA values in the ipsilateral cortex and striatum of MCAO rats were significantly decreased compared with the sham group ( P<0.05 or P<0.01). The rFA values in cortex and striatum in focal cerebral ischemia rats of BYHWD group were higher than those in the model group, while only 14 d showed significance ( P<0.05 or P<0.01). Conclusions:T2WI and DTI clearly provided the informations of the location and morphology of the infarct areas. The ischemic brain showed significant infarction, edema, and white matter injury, which were ameliorated with time going on. BYHWD reduced the infarction percentage, inhibited cerebral edema and stimulated the recovery of neurofibra, suggesting that BYHWD could protect against cerebral ischemia.
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Objective To observe the effects of Buyang-Huanwu decoction (BYHWD) combined with enriched environment (EE) on the abnormal activation of astrocytes and expression of angiogenic factors in rats with cerebral ischemia. Methods The rat model of permanent middle cerebral artery occlusion (pMCAO) was established by using suture method. Male SD rats were randomly divided into sham, model, EE, BYHWD and BYHWD+EE group. Rats in the BYHWD and BYHWD+EE groups were intragastrically adminstratered with BYHWD (16.1g/kg) 24h after MCAO and once daily for consecutive 15 days. Accordingly, rats in the sham, model, and EE groups were adminstratered with the sam volume normal saline. Rats treated with EE and BYHWD+EE were housed in the enriched environment 12 h every day. The EE, BYHWD and BYHWD+EE group rats were given relative treatment continuously for 15 days. Beam walking test was performed to examine the motor function of rats. Next, the HE staining was conducted to detect the pathological alterations of the brain. Immunohistochemical staining with DARPP-32 and GFAP were applied to respectively observe the changes of multiple spinous neurons and activation of astrocytes in the striatum. Subsequently, the mRNA expressions of angiogenic factors including VEGF/VEGFR2 and Ang1/Ang2 were detected by RT-PCR. Results The rats treated with BYHWD+EE revealed markedly elevated beam walking scores on the 7th and 15th day after MCAO compared with model group (P<0.01). Compared with the model, BYHWD+EE treatment significantly increased the number of neurons (31.08 ± 8.06 vs. 19.00 ± 9.12, P<0.01), elevated the number of DARPP-32-positive cells (975.3 ± 589.68 vs. 271.25 ± 164.98) and integral optical density (18621.87 ± 6996.64 vs. 4071.60 ± 2477.27) of DARPP-32 (P<0.01), suppressed the expression of GFAP (6094.07 ± 3211.74 vs. 10002.91 ± 8430.34, P<0.01) and up-regulated the mRNA levels of VEGF (1.59 ± 0.80 vs. 0.77 ± 0.33), VEGFR2 (1.58 ± 0.88 vs. 0.70 ± 0.37), Ang1 (1.58 ± 0.52 vs. 0.84 ± 0.13), and Ang2 (1.25 ± 0.25 vs. 0.90 ± 0.22) in the striatum (P<0.05 or P<0.01). Conclusions The Buyang-Huanwu decoction combined with enriched environment exerted synergistic effect on ischemic rats and promoted motor function by inhibiting the excessive activation of astrocytes and improving the expression of angiogenic factors.
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Objective To investigate the effect of Shenling-Baizhu powder on gastrointestinal hormone,digestive enzyme and inflammatory factors in D-IBS rats.Methods A total of 50 Wistar rats were randomly divided into the normal group,the model group,the Shenling-Baizhu powder large dose group,medium dose group and small dose group,10 rats in each group.D-IBS rat model was induced by high lactose diet and restraint stress except normal group.At the same time,the Shenling-Baizhu powder groups received Shenling-Baizhu powder (large dose group:9 g/kg,medium dose group:3 g/kg,small dose group:1 g/kg),normal group and model group received equal volume of normal saline.Malate dehydrogenase (MDH),lactate dehydrogenase (LDH) and disaccharidase activities were detected from small intestine of rats in each group.Plasma vasoactive intestinal peptide (VIP) and substance P (SP) activities were measured by radioimmunoassay.TNF-α and IL-10 levels in serum were determined by ELISA.Results Compared with the model group,the model group,the ratio of serum TNF-α/IL-10 (0.31 ± 0.15,0.34 ± 0.21,0.41 ± 0.15 vs.0.74 ± 0.19) significantly decreased in large,medium and low dose groups (P＜0.05).The disaccharidase activities in the small intestinal mucosa (38.44 ± 11.30 U/mg,44.77 ± 16.46 U/mg,42.58 ± 13.02 U/mg vs.23.23 ± 6.85 U/mg) significantly increased in the large,medium and low dose groups (P＜0.05).LDH activities in the small intestinal mucosa (1 863.61 ± 1 019.28 U/mg,1 136.13 ± 631.89 U/mg vs.416.50 ± 223.94U/mg) significantly increased in large,medium dose groups (P＜0.05).Plasma SP level (22.88 ± 10.17 pg/mg vs.31.86 ± 7.81 pg/mg) significantly decreased in large dose group (P＜0.05).Conclusions Shenling-Baizhu powder may increase the activities of LDH,MDH and disaccharidase,inhibit the abnormal secretion of SP and regulate of Th1/Th2 immune imbalance to relieve diarrhea.