In this study, we extracted and purified components in the Angong Niuhuang pill. Then we applied transdermal enhancers to Angong Niuhuang stickers by modern technology. The Angong Niuhuang sticker includes extracts fr...In this study, we extracted and purified components in the Angong Niuhuang pill. Then we applied transdermal enhancers to Angong Niuhuang stickers by modern technology. The Angong Niuhuang sticker includes extracts from curcuma, berberine hydrochloride, baicalin, geniposide, borneol, and musk. Angong Niuhuang stickers at different point application doses （1.35, 2.7, and 5.4 g/kg） were administered to Dazhui （DU14）, Qihai （RN6） and Mingmen （DU4）. Rats in the different dose point application and acupuncture groups were continuously administered for 7 days. Then a middle cerebral artery occlusion model was prepared for simulating human cerebral ischemia. Twelve hours later, expressions of Bcl-2, Bax and p53 protein in hippocampal CA1 were detected with immunohistochemistry. The expression level of Bcl-2, an anti-apoptotic protein, significantly increased in the high-, medium- and low-dose point application groups and the acupuncture group, while the expression of the pro-apoptotic proteins, Bax and p53, significantly decreased compared with the middle cerebral artery occlusion rats; and the Bcl-2/Bax ratio was significantly increased. The difference was noticeable for the high-dose point application group, which showed statistical difference compared with the low-dose point application group and the acupuncture group. Our experimental findings indicate that point application with Angong Niuhuang stickers promotes the expression of Bcl-2, and inhibits the expressions of Bax and p53 in the hippocampal CA1 area of rats after focal cerebral ischemia. Thus, point application of Angong Niuhang stickers protects brain tissues from cerebral ischemia.展开更多
Objective: To observe the effect of electroacupuncture (EA) on expression of p53 protein in cerebral cortex of senile rats with global cerebral ischemia/reperfusion (IR) injury and to explore its mechanism. Methods: T...Objective: To observe the effect of electroacupuncture (EA) on expression of p53 protein in cerebral cortex of senile rats with global cerebral ischemia/reperfusion (IR) injury and to explore its mechanism. Methods: The cerebral IR injury rat model was established referring to Pulsinelli 4-vessel occlusion method. Thirty-six SD rats were randomly and evenly divided into the control group, the IR group and the IR plus EA (IR-EA) group. The animals in the control group were subjected to electrocauterization of vertebral arteries in bilateral flank orifice alone with the general carotid arteries unoccluded. To rats in the IR-EA group, immediately and 24h, 48h, 72h after cerebral IR, EA treatment on bilateral acupoint 'Zusanli' (ST36) was applied once a day, lasting for 60 minutes. After the final treatment, all the rats were sacrificed and their brains were taken to examine p53 protein expression by the immunohistochemical method. Results: Cells with positive p53 immunoreactivity in the cerebral cortex of rats in the IR group was significantly higher than that in the control group ( P<0. 05), while that in the IR-EA group was significantly lower than that in the IR group ( P<0. 05). Conclusion: EA could remarkably reduce expression of p53 protein in the cerebral cortex of senile rats with global cerebral IR injury, which might be one of the means for EA to inhibit neuronal ap-optosis after cerebral IR injury.展开更多
Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebr...Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase（MAPK） signaling pathway.We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa＇s method.At 30 minutes before model establishment,p38 MAPK blocker SB20358 was injected into the left lateral ventricles.At 1.5 hours after model establishment,electroacupuncture was administered at acupoints of Chize（LU5）,Hegu（LI4）,Zusanli（ST36）,and Sanyinjiao（SP6） for 20 minutes in the affected side.Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores,but no significant differences were determined among different interventional groups.Hematoxylin-eosin staining also showed reduced brain tissue injuries.Compared with the SB20358 group,the cells were regularly arranged,the structures were complete,and the number of viable neurons was higher in the SB20358 ＋ electroacupuncture group.Terminal deoxynucleotidyl transferase（Td T）-mediated d UTP nick-end labeling assay showed a decreased apoptotic index in each group,with a significant decrease in the SB20358 ＋ electroacupuncture group.Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 ＋ electroacupuncture group compared with the ischemia/reperfusion group.There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group.These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway.A time period of 3 days could promote the repair of ischemic cerebral nerves.展开更多
The tumor suppressor p63 is one of p53 family members and plays a vital role as a regulator of neuronal apoptosis in the development of the nervous system. However, the role of p63 in mature neuronal death has not bee...The tumor suppressor p63 is one of p53 family members and plays a vital role as a regulator of neuronal apoptosis in the development of the nervous system. However, the role of p63 in mature neuronal death has not been addressed yet. In this study, we first compared ischemia-induced effects on p63 expression in the hippocampal regions （CA1-3） between the young and adult gerbils subjected to 5 minutes of transient global cerebral ischemia. Neuronal death in the hippocampal CA1 region of young gerbils was significantly slow compared with that in the adult gerbils after transient global cerebral ischemia, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons in the sham-operated young group was significantly low compared with that in the sham-operated adult group, p63 immunoreactivity was apparently changed in ischemic hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. In the ischemia-operated adult groups, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons was significantly decreased at 4 days post-ischemia; however, p63 immunoreactivity in the ischemia-operated young group was significantly higher than that in the ischemia-operated adult group. At 7 days post-ischemia, p63 immunoreactivity was decreased in the hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. Change patterns of p63 level in the hippocampal CA1 region of adult and young gerbils after ischemic damage were similar to those observed in the immunohistochemical results. These findings indicate that higher and longer-term expression of p63 in the hippocampal CA1 region of the young gerbils after ischemia/reperfusion may be related to more delayed neuronal death compared to that in the adults.展开更多
The fermented Chinese formula Shuan-Tong-Ling is composed of radix puerariae（Gegen）,salvia miltiorrhiza（Danshen）,radix curcuma（Jianghuang）,hawthorn（Shanzha）,salvia chinensis（Shijianchuan）,sinapis alba（Baiji...The fermented Chinese formula Shuan-Tong-Ling is composed of radix puerariae（Gegen）,salvia miltiorrhiza（Danshen）,radix curcuma（Jianghuang）,hawthorn（Shanzha）,salvia chinensis（Shijianchuan）,sinapis alba（Baijiezi）,astragalus（Huangqi）,panax japonicas（Zhujieshen）,atractylodes macrocephala koidz（Baizhu）,radix paeoniae alba（Baishao）,bupleurum（Chaihu）,chrysanthemum（Juhua）,rhizoma cyperi（Xiangfu） and gastrodin（Tianma）,whose aqueous extract was fermented with lactobacillus,bacillus aceticus and saccharomycetes.ShuanTong-Ling is a formula used to treat brain diseases including ischemic stroke,migraine,and vascular dementia.Shuan-Tong-Ling attenuated H_2O_2-induced oxidative stress in rat microvascular endothelial cells.However,the potential mechanism involved in these effects is poorly understood.Rats were intragastrically treated with 5.7 or 17.2 m L/kg Shuan-Tong-Ling for 7 days before middle cerebral artery occlusion was induced.The results indicated Shuan-Tong-Ling had a cerebral protective effect by reducing infarct volume and increasing neurological scores.Shuan-Tong-Ling also decreased tumor necrosis factor-α and interleukin-1β levels in the hippocampus on the ischemic side.In addition,Shuan-Tong-Ling upregulated the expression of SIRT1 and Bcl-2 and downregulated the expression of acetylated-protein 53 and Bax.Injection of 5 mg/kg silent information regulator 1（SIRT1） inhibitor EX527 into the subarachnoid space once every 2 days,four times,reversed the above changes.These results demonstrate that Shuan-Tong-Ling might benefit cerebral ischemia/reperfusion injury by reducing inflammation and apoptosis through activation of the SIRT1 signaling pathway.展开更多
目的探讨^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)在幼龄兔缺氧缺血性脑损伤(HIBD)细胞凋亡显像中的价值。方法制作幼龄兔假手术组、HIBD 4 h 组、HIBD 40 h 组模型,对各组动物行注药后60 min 平面脑显像和脑组织石蜡切片观...目的探讨^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)在幼龄兔缺氧缺血性脑损伤(HIBD)细胞凋亡显像中的价值。方法制作幼龄兔假手术组、HIBD 4 h 组、HIBD 40 h 组模型,对各组动物行注药后60 min 平面脑显像和脑组织石蜡切片观察,对 HIBD 40 h 组还进行注药后5,30,120 min 显像;用 ROI 技术计算各组动物脑患侧与健侧放射性计数比值(T/NT);假手术组和HIBD 4 h 组还进行 MRI、弥散加权成像(DWI)。结果假手术组、HIBD 4 h 组和 HIBD 40 h 组注药后60 min T/NT 分别为0.94±0.14,1.32±0.11,1.81±0.07(F=82.385,P<0.01);HIBD 40 h 组不同时间(5,30,60,120 min)显像获得的 T/NT 分别为1.72±0.04,1.77±0.06,1.81±0.07,1.71±0.03(F=3.994,P>0.01)。切片观察假手术组脑组织未见凋亡细胞,HIBD 4 h 组与 HIBD 40 h 组患侧脑见凋亡细胞,后者较明显;HIBD 4 h 组 MRI、DWI 和表观弥散系数(ADC)图均未见异常。结论^(99)Tc^m-HYNIC-Annexin V 显像可以早期发现神经细胞凋亡,在新生儿 HIBD 细胞凋亡的无创性诊断、抑制凋亡疗效评价和预后估计方面有重要的应用前景。展开更多
基金supported by the Research Project of Administration of TCM of Guangdong Province, No. 20111270
文摘In this study, we extracted and purified components in the Angong Niuhuang pill. Then we applied transdermal enhancers to Angong Niuhuang stickers by modern technology. The Angong Niuhuang sticker includes extracts from curcuma, berberine hydrochloride, baicalin, geniposide, borneol, and musk. Angong Niuhuang stickers at different point application doses （1.35, 2.7, and 5.4 g/kg） were administered to Dazhui （DU14）, Qihai （RN6） and Mingmen （DU4）. Rats in the different dose point application and acupuncture groups were continuously administered for 7 days. Then a middle cerebral artery occlusion model was prepared for simulating human cerebral ischemia. Twelve hours later, expressions of Bcl-2, Bax and p53 protein in hippocampal CA1 were detected with immunohistochemistry. The expression level of Bcl-2, an anti-apoptotic protein, significantly increased in the high-, medium- and low-dose point application groups and the acupuncture group, while the expression of the pro-apoptotic proteins, Bax and p53, significantly decreased compared with the middle cerebral artery occlusion rats; and the Bcl-2/Bax ratio was significantly increased. The difference was noticeable for the high-dose point application group, which showed statistical difference compared with the low-dose point application group and the acupuncture group. Our experimental findings indicate that point application with Angong Niuhuang stickers promotes the expression of Bcl-2, and inhibits the expressions of Bax and p53 in the hippocampal CA1 area of rats after focal cerebral ischemia. Thus, point application of Angong Niuhang stickers protects brain tissues from cerebral ischemia.
基金This item was supported by Fund on Opening Subject of Key Laboratory in Jiangsu Province (No.K9842)
文摘Objective: To observe the effect of electroacupuncture (EA) on expression of p53 protein in cerebral cortex of senile rats with global cerebral ischemia/reperfusion (IR) injury and to explore its mechanism. Methods: The cerebral IR injury rat model was established referring to Pulsinelli 4-vessel occlusion method. Thirty-six SD rats were randomly and evenly divided into the control group, the IR group and the IR plus EA (IR-EA) group. The animals in the control group were subjected to electrocauterization of vertebral arteries in bilateral flank orifice alone with the general carotid arteries unoccluded. To rats in the IR-EA group, immediately and 24h, 48h, 72h after cerebral IR, EA treatment on bilateral acupoint 'Zusanli' (ST36) was applied once a day, lasting for 60 minutes. After the final treatment, all the rats were sacrificed and their brains were taken to examine p53 protein expression by the immunohistochemical method. Results: Cells with positive p53 immunoreactivity in the cerebral cortex of rats in the IR group was significantly higher than that in the control group ( P<0. 05), while that in the IR-EA group was significantly lower than that in the IR group ( P<0. 05). Conclusion: EA could remarkably reduce expression of p53 protein in the cerebral cortex of senile rats with global cerebral IR injury, which might be one of the means for EA to inhibit neuronal ap-optosis after cerebral IR injury.
基金supported by the National Natural Science Foundation of China,No.81173355
文摘Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase（MAPK） signaling pathway.We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa＇s method.At 30 minutes before model establishment,p38 MAPK blocker SB20358 was injected into the left lateral ventricles.At 1.5 hours after model establishment,electroacupuncture was administered at acupoints of Chize（LU5）,Hegu（LI4）,Zusanli（ST36）,and Sanyinjiao（SP6） for 20 minutes in the affected side.Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores,but no significant differences were determined among different interventional groups.Hematoxylin-eosin staining also showed reduced brain tissue injuries.Compared with the SB20358 group,the cells were regularly arranged,the structures were complete,and the number of viable neurons was higher in the SB20358 ＋ electroacupuncture group.Terminal deoxynucleotidyl transferase（Td T）-mediated d UTP nick-end labeling assay showed a decreased apoptotic index in each group,with a significant decrease in the SB20358 ＋ electroacupuncture group.Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 ＋ electroacupuncture group compared with the ischemia/reperfusion group.There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group.These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway.A time period of 3 days could promote the repair of ischemic cerebral nerves.
基金supported by 2013 Research Grant from Kangwon National University(120131480)Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2014R1A6A3A01056005)
文摘The tumor suppressor p63 is one of p53 family members and plays a vital role as a regulator of neuronal apoptosis in the development of the nervous system. However, the role of p63 in mature neuronal death has not been addressed yet. In this study, we first compared ischemia-induced effects on p63 expression in the hippocampal regions （CA1-3） between the young and adult gerbils subjected to 5 minutes of transient global cerebral ischemia. Neuronal death in the hippocampal CA1 region of young gerbils was significantly slow compared with that in the adult gerbils after transient global cerebral ischemia, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons in the sham-operated young group was significantly low compared with that in the sham-operated adult group, p63 immunoreactivity was apparently changed in ischemic hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. In the ischemia-operated adult groups, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons was significantly decreased at 4 days post-ischemia; however, p63 immunoreactivity in the ischemia-operated young group was significantly higher than that in the ischemia-operated adult group. At 7 days post-ischemia, p63 immunoreactivity was decreased in the hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. Change patterns of p63 level in the hippocampal CA1 region of adult and young gerbils after ischemic damage were similar to those observed in the immunohistochemical results. These findings indicate that higher and longer-term expression of p63 in the hippocampal CA1 region of the young gerbils after ischemia/reperfusion may be related to more delayed neuronal death compared to that in the adults.
基金supported by the National Natural Science Foundation of China,No.81202625Open Fund of Key Laboratory of Cardiovascular and Cerebrovascular Diseases Translational Medicine of China Three Gorges University of China,No.2016xnxg101
文摘The fermented Chinese formula Shuan-Tong-Ling is composed of radix puerariae（Gegen）,salvia miltiorrhiza（Danshen）,radix curcuma（Jianghuang）,hawthorn（Shanzha）,salvia chinensis（Shijianchuan）,sinapis alba（Baijiezi）,astragalus（Huangqi）,panax japonicas（Zhujieshen）,atractylodes macrocephala koidz（Baizhu）,radix paeoniae alba（Baishao）,bupleurum（Chaihu）,chrysanthemum（Juhua）,rhizoma cyperi（Xiangfu） and gastrodin（Tianma）,whose aqueous extract was fermented with lactobacillus,bacillus aceticus and saccharomycetes.ShuanTong-Ling is a formula used to treat brain diseases including ischemic stroke,migraine,and vascular dementia.Shuan-Tong-Ling attenuated H_2O_2-induced oxidative stress in rat microvascular endothelial cells.However,the potential mechanism involved in these effects is poorly understood.Rats were intragastrically treated with 5.7 or 17.2 m L/kg Shuan-Tong-Ling for 7 days before middle cerebral artery occlusion was induced.The results indicated Shuan-Tong-Ling had a cerebral protective effect by reducing infarct volume and increasing neurological scores.Shuan-Tong-Ling also decreased tumor necrosis factor-α and interleukin-1β levels in the hippocampus on the ischemic side.In addition,Shuan-Tong-Ling upregulated the expression of SIRT1 and Bcl-2 and downregulated the expression of acetylated-protein 53 and Bax.Injection of 5 mg/kg silent information regulator 1（SIRT1） inhibitor EX527 into the subarachnoid space once every 2 days,four times,reversed the above changes.These results demonstrate that Shuan-Tong-Ling might benefit cerebral ischemia/reperfusion injury by reducing inflammation and apoptosis through activation of the SIRT1 signaling pathway.
文摘目的探讨^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)在幼龄兔缺氧缺血性脑损伤(HIBD)细胞凋亡显像中的价值。方法制作幼龄兔假手术组、HIBD 4 h 组、HIBD 40 h 组模型,对各组动物行注药后60 min 平面脑显像和脑组织石蜡切片观察,对 HIBD 40 h 组还进行注药后5,30,120 min 显像;用 ROI 技术计算各组动物脑患侧与健侧放射性计数比值(T/NT);假手术组和HIBD 4 h 组还进行 MRI、弥散加权成像(DWI)。结果假手术组、HIBD 4 h 组和 HIBD 40 h 组注药后60 min T/NT 分别为0.94±0.14,1.32±0.11,1.81±0.07(F=82.385,P<0.01);HIBD 40 h 组不同时间(5,30,60,120 min)显像获得的 T/NT 分别为1.72±0.04,1.77±0.06,1.81±0.07,1.71±0.03(F=3.994,P>0.01)。切片观察假手术组脑组织未见凋亡细胞,HIBD 4 h 组与 HIBD 40 h 组患侧脑见凋亡细胞,后者较明显;HIBD 4 h 组 MRI、DWI 和表观弥散系数(ADC)图均未见异常。结论^(99)Tc^m-HYNIC-Annexin V 显像可以早期发现神经细胞凋亡,在新生儿 HIBD 细胞凋亡的无创性诊断、抑制凋亡疗效评价和预后估计方面有重要的应用前景。
