Effects of Echinacoside on Histio-central Levels of Active Mass in Middle Cerebral Artery Occlusion Rats

Objective To investigate the effects of echinacoside on the extracellular striatal levels of norepinephrine（NE）,dopamine（DA）,homovanillic acid（HVA）,3,4-dihydroxyphenylethanoid acid（DOPAC）,5-hydroxyindoleacetic acid（HIAA）,and 5-hydroxytryptamine（5-HT） in middle cerebral artery occlusion（MCAO rats.Methods The middle cerebral artery was occluded in male Sprague-Dawley rats.Three days later microdialysis probes were placed into the right striatum of MCAO rat brains and the brains were perfused with Ringer＇s solution at a rate of 1.5 μL/min.Cerebral microdialysates were collected every 30 minutes from awake and freely moving rats before assaying for NE,DA,HVA,DOPAC,HIAA,and 5-HT levels by reverse phase HPLC with electrochemistry.Results Three days after MCAO,the extracellular striatal levels of NE,DA,DOPAC,HIAA,HVA,and 5-HT of the MCAO rats increased significantly（at least P0.05 vs.control）.However,simultaneous treatment with echinacoside（30.0 or 15.0 mg/kg） attenuated these increases（at least P0.05 vs.non-treated model rats）.Conclusion These results imply that echinacoside may protect striatal dopa minergic neurons from the injury induced by MCAO and may help prevent and treat cerebral ischemic diseases.

Effects of preventive acupuncture or moxibustion at the Guanyuan acupoint on expression of interleukin-2 and its receptors in hypothalamus, serum, and uterus of menopausal rats

BACKGROUND： Previous studies have indicated preventive acupuncture and moxibustion at the Guanyuan point have positive effects on the immune system of menopausal rats. OBJECTIVE： To observe the effects of preventive acupuncture or moxibustion at the Guanyuan point on interleukin-2 （IL-2） and its receptor （IL-2R mRNA） levels in hypothalamus, serum, and uterus of menopausal rats. DESIGN： Randomized control animal experiment. SETTING： School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine. MATERIALS： This study was performed at the Laboratory of Beijing University of Chinese Medicine from November 2004 to July 2005. The vaginal smear method was used to determine the estrus cycle of 9.5-month-old Sprague Dawley （SD） female rats. Three continuous estrus cycles were measured, and 160 rats with estrus cycle disorder, weighing （300 _＋ 20） g, were included in this study. The rats were randomly divided into four groups： preventive acupuncture group （n =48）, preventive moxibustion group （n =48）, model groups （n =48）, and 10-month-old group （n =16）. Sixteen 3.5-month-old healthy SD female rats, weighing （240 ± 20） g, were also assigned to the young group. All rats were clean grade and were purchased from the Laboratory Animal Center, Beijing University of Chinese Medicine. The experiment was in accordance with animal ethics standards. METHODS： Acupuncture was applied to the preventive acupuncture group at 10 months of age. The needle was inserted upward and perpendicularly two fen to a point that was two fen below the Guanyuan point. The rats freely moved around during needle retention （20 minutes）. Acupuncture was administered twice a week and continued for eight weeks. A moxa cone （small） moxibustion was applied to the preventive moxibustion group at 10 months of age. The rats were caught gently and kept at the same position as the preventive acupuncture group. The moxa cone was ignited and held to the Guanyuan point directly. Only one moxa cone was used each time. After moxibustion, the local skin turned slightly red. Moxibustion was used twice a week and continued for eight weeks. Nothing except gently catching and lying was administered to the model group and the young group. MAIN OUTCOME MEASURES： Radioimmunoassay, in situ hybridization, and similar methods were employed to detect IL-2 in serum and the uterus, as well as IL-2R mRNA expression in the hypothalamus and uterus arcuate nuclei, of the preventive acupuncture group, preventive moxibustion group, model groups （at 12, 14, and 16 months）, 10 month old group （at 10 months）, and the young group （at 4 months）. RESULTS： All 176 rats were included in the final analysis. （1） IL-2R mRNA： IL-2R mRNA expression decreased significantly in the uterus of 12-, 14-, and 16-month-old model groups and in the hypothalamus of 14- and 16-month-old model groups （P 〈 0.05-0.01）. Compared to the same month-aged model groups, the expression of IL-2R mRNA increased significantly in the hypothalamus and uterus of 16-month-old preventive acupuncture or moxibustion groups （P 〈 0.05-0.01）.（2） IL-2：IL-2 in serum of 12- and 14-month-old model groups and in the uterus of 12-, 14-, and 16-month-old model groups decreased significantly compared to the young group （P 〈 0.05-0.01）. IL-2 serum levels increased significantly in the 16-month-old group with preventive moxibustion; in addition, IL-2 expression levels increased significantly in the uterus of the 12-month-old group with preventive moxibustion, as well as the 12- and 16-month-old groups with preventive acupuncture （P 〈 0.05-0.01）. CONCLUSION： As a rat ages and becomes menopausal, IL-2R mRNA expression in the hypothalamus and uterus, as well as IL-2 levels in the serum and uterus, decrease. However, preventive acupuncture and preventive moxibustion can repress the decrease to some extent. This effect is even greater in the late stages of menopause （16-month-old rats）.

Increasing doses of diminazene aceturate:adverse reproductive effects in female Wistar rats

Objective:To investigate the effects of comparatively high doses of diminazene aceturate on the reproductive performance of female rats in the early stage of pregnancy.Methods:After oestrus synchronisation and successful mating,20 pregnant female rats were randomly divided into four groups(A-D).Group A rats served as the control and were given single intraperitoneal injection of 0.5 mL sterile water(vehicle only) while groups B.C and D rats were given single intraperitoneal doses of 7,14 and 21 mg/kg body weight diminazene aceturate respectively,on day 7 of pregnancy.The gestation length,litter size and weight at birth,and areas of foetal resorption in the uterus were determined post partum.The post-implantation survival index(%) and the gestation index(group%) were also evaluated for rats in all the groups.Results:There was a graded increase in the number of observed resorbed foetuses as the dose of diminazene aceturate was increased,although only groups C(14 mg/kg) and D(21 mg/kg) revealed a significant decrease(P<0.01,ANOVA) in the post implantation survival index of rat embryos.There was also a significant decrease(P<0.05) in the litter weights of rats in groups C and D.Conclusions: Although the pregnant rats showed no overt signs of systemic toxicity even at the highest dose of 21 mg/kg body weight diminazene aceturate in this study,it was concluded that the use of high doses of diminazene aceturate in an effort to combat resistant trypanosomes could have adverse reproductive effects on female animals in the early period of pregnancy.

电针神庭、百会穴对MCAO/R大鼠γ-氨基丁酸受体Gabbr1表达的影响

目的通过观察电针神庭、百会穴对大脑中动脉缺血再灌注模型(middle cerebral artery occlusion and reperfusion,MCAO/R)认知功能障碍大鼠海马区γ-氨基丁酸B受体gabbr1表达的影响,探讨电针改善MCAO/R大鼠学习记忆能力的可能机制。方法27只雄性健康SD大鼠随机分为假手术组、模型组、电针组;模型组、电针组采用longa线栓法制备MCAO/R模型,假手术组仅分离颈总、颈内及颈外动脉,不插入线栓,造模后通过longa五分法验证模型。电针组取神庭、百会穴在造模后24 h进行干预,治疗7 d;干预第3 d采用Morris水迷宫实验检测各组大鼠学习记忆能力,为期4 d;干预第7 d断头取大鼠完整海马组织,采用实时荧光定量PCR(qRT-PCR)检测各组大鼠海马组织中gabbr1 mRNA的表达。结果与假手术组相比,模型组水迷宫实验的第三象限距离比显著减小(P<0.05),穿越平台次数减少(P<0.01);与模型组相比,电针组经治疗后第三象限距离比扩大(P<0.05),穿越平台次数增加(P<0.05)。与假手术组相比,模型组海马区gabbr1 mRNA表达量显著增高(P<0.01);与模型组相比,电针组海马区gabbr1 mRNA表达降低(P<0.01)。结论电针神庭、百会穴能改善脑缺血再灌注大鼠的学习记忆能力,其机制可能与调节gabbr1 mRNA在海马中的表达有关。

Neuroprotective effect of Phyllanthus urinaria extract on ischemic brain injury in rats

Objective:To investigate the neuroprotective effect of Phyllanthus urinaria on ischemic stroke and its primary mechanism.Methods:Adult SD rats were selected as the research object,and the right middle cerebral artery infarction rat model was established by the modified suture method(MCAO).Observe the neurological deficit score at 24h,48h and 72h after the model is successfully prepared,then TTC staining method to detect the area of cerebral infarction,the content of superoxide dismutase(SOD),nitric oxide(NO)and endothelial NOS(eNOS)in brain tissue;Immunofluorescence method was used to detect the expression of Caspase-3 positive cells in brain tissue;Western blot method was used to detect the expression of PI3K and AKT protein in brain tissue.72 experimental animals were randomly divided into 4 groups,sham operation group,model group(MCAO),extract of Phyllanthus urinaria low dose group(PuL5 g/kg),high dose group(PuL10 g/kg),and make MCAO model after 7 days of continuous administration,and continue to infuse the medicine once/d until the material is obtained.Results:No neurological deficits in the sham operation group,The 24h,48h and 72h of the modelling showed that the neurological impairment of the two doses of extract of Phyllanthus urinaria and the MCAO group was more severe than that of the sham operation group(P<0.01),however,with the prolongation of the modeling time,the neurological function scores of the two doses of extract of Phyllanthus urinaria were lower than those of the MCAO group,the most significant at 72h(P<0.01);The infarct size of the rats in the two dose groups of extract of Phyllanthus urinaria was lower than that of the MCAO group(P<0.01),and there was no dose dependence between the two groups,the content of SOD in the MCAO group was reduced,and the content of NO and eNOS was increased than the sham operation group(P<0.05),Compared with the MCAO group,the two administration groups significantly increased the content of SOD,decreased the content of NO and eNOS(P<0.05);Although the expression of Caspase-3 positive cells in the two administration groups was higher than that in the sham operation group,it was significantly lower than that in the MCAO group(Plow<0.05,Phigh<0.01);The expression of PI3K and AKT protein in the brain tissue of the MCAO group was significantly lower than that of the sham operation group(P<0.05),but the expression of PI3K and AKT protein in brain tissue of extract of Phyllanthus urinaria in high and low dose groups was significantly higher than that in MCAO group(P<0.05).Conclusion:Extract of Phyllanthus urinaria can improve neurological damage and cerebral infarction area in rats,and reduce the expression of Caspase-3 positive cells in MCAO rats,and then increase the expression levels of PI3K and AKT proteins to protect ischemic brain injury.

Effect of calcitonin gene-related peptide and nerve growth factor on spatial learning and memory abilities of rats following focal cerebral ischemia/reperfusion

BACKGROUND: Calcitonin gene-related peptide (CGRP) and nerve growth actor (NGF) cam improve spatial learning and memory abilities of rats with cerebral ischemia/reperfusion; however, the effect of combination of them on relieving learning and memory injury following cerebral ischemia/reperfusion should be further studied. OBJECTIVE: To study the effects of exogenous CGRP and NGF on learning and memory abilities of rats with focal cerebral ischemia/reperfusion. DESIGN: Randomized controlled animal study. SETTING: Department of Neurosurgery, the Second Hospital of Xiamen; Department of Neurosurgery, the Second Affiliated Hospital of China Medical University; Department of Neurobiology, Basic Medical College of China Medical University. MATERIALS: A total of 30 healthy male SD rats, aged 8 weeks, of clean grade, weighing 250-300 g, were provided by Experimental Animal Department of China Medical University. All rats were randomly divided into sham-operation group, ischemia/reperfusion group and treatment group with 10 in each group. The main reagents were detailed as the follows: 100 g/L chloral hydrate, 0.5 mL CGRP (2 mg/L, Sigma Company, USA), and NGF (1× 106 U/L, 0.5 mL, Siweite Company, Dalian). METHODS: The experiment was carried out in the Department of Neurobiology, Basic Medical College of China Medical University from February to July 2005. Rat models of middle cerebral artery occlusion were established by method of occlusion, 2 hours after that rats were anesthetized and the thread was slightly drawn out for 10 mm under direct staring to perform reperfusion. Rats in the ischemia/reperfusion group received intraperitoneal injection of 1 mL saline via the abdomen at two hours later, while rats in the treatment group at 2 hours later received intraperitoneal injection of 2 mg/L CGRP (0.5 mL) and 1×106 U/L NGF (0.5 mL) once a day for 10 successive days. First administration was accomplished within 15 minutes after ischemia/reperfusion. Rats in the sham-operation group were separated of the vessels without occlusion or administration. The neural function was evaluated with Zea Longa 5-grade scale. Animals with the score of one, two and three points received Morris water-maze test to measure searching time on platform (omitting platform-escaping latency). Meanwhile, leaning and memory abilities of animals were reflected through testing times of passing through platform per minute. MAIN OUTCOME MEASURES: Experimental results of omitting platform-escaping latency and spatial probe. RESULTS: Three and two rats in the ischemia/reperfusion group and treatment group respectively were not in accordance with the criteria in the process, and the rest were involved in the final analysis. ① Comparisons of platform-escaping latency during Morris water-maze test in all the three groups: Ten days after modeling, the platform-escaping latency in the ischemia/reperfusion group was obviously longer than that in sham-operation group (P < 0.01), and was significantly shorter than that in the treatment group (P < 0.01). ② Comparisons of times of passing through platform in all the three groups: Times of passing through platform were remarkably less in the ischemia/reperfusion group than those in the sham-operation group [(1.79±0.39), (4.30±0.73) times/minute, P < 0.01], and those were markedly more in the treatment group than the ischemia/reperfusion group [(3.16±1.03), (1.79±0.39) times/minute, P < 0.01]. CONCLUSION: CGRP and NGF are capable of ameliorating the abilities of spatial learning and memory in MCAO rats, which indicates that CGRP and NGF can protect ischemic neurons.

Distribution and excretion of N-Ile1 Thr2-63-desulfatohirudin in rats

Objective To investigate distribution and excretion of N-Ile1Thr2-63-desulfatohirudin(rH)a recombinant hirudin newly developed in China,in rats for its development as a novel anticoagulant agent.Methods ELISA was used to determine the rH concentration in related tissues and body fluids.Tissues were collected at 15,60 and 180min respectively,after iv administration of rH 1.0 mg·kg-1 to 3 groups of 5 rats,and homogenized.Urine,bile and feces were collected at pre-selected intervals of time after iv dosing 1.0 mg·kg-1 to 3 groups of 5 rats and assayed.Results rH following iv dosing was distributed rapidly,the rH levels in all tissues being found to be the highest at 15 min post-injection,afterwards gradually reduced.The highest concentration of rH was found in blood,the next in lung and heart,the lowest in brain.With 15 min post dose as an example,the rH contents in tissues were ranked in order of plasma>lung>heart >adipose>skeletal muscles>kidney>liver>spleen>brain.The 12 h-cumulative excretion amount of rH in urine and feces accounted for 0.03%and 0.001% of administered dose,respectively;the 6 h-cumulative excretion amount in bile was 0.02%of the dose.Conclusions The rH is distributed mainly in blood circulation system with very low content in other tissues.The drug is excreted from urine,feces and bile of rats in extremely minute amount(only 0.051% dose),suggesting that rH undergoes extensive metabolic elimination in rat body.

Evaluation of the effect of hydrogen sulfide postconditioning by p-v loop against myocardial i/r injury in rats

Objectives To evaluate the effect of hydrogen sulfide(H2S) postconditioning on myocardial ischemia-reperfusion (I/R) by pressure-volume loop(P-V loop). Methods The I/R model of rat in vivo was established by ligating the left anterior descending coronary artery for 30min and reperfusing for 120 min.Wistar rats(n=32) were ran- domly divided into 4 groups;Sham operation,ischemia-reperfusion (I/R),Ischemic postconditioning(IPO) and H2S postconditioning.In sham operation,there was no ligation.In IPO,at the start of reperfusion,three cycles of 30s reperfusion and 30s LAD reocclusion preceded the 3h of reperfusion. In H2S postconditioning,NaHS(15μmol/kg,Sodium hydrosulfide)was administrated before coronary artery reperfusion. The heart rate(HR),I/R arrhythmia,the left ventricular end-systolic pressure(LVESP),left ventricular enddiastolic pressure(LVEDP),the slope of the end- systolic P-V relation(ESPVR) and the slope of the end-diastolic P-V relation(EDPVR) were detected.Infarct size was determined by scanning the images of the rat heart ventricular sections stained with Evans blue and TTC.Results Compared with I/R group,the I/R arrhythmia and the infarct size were decreased significantly(PPP2S postconditioning group.Conclusions Myocardial I/R injury was decreased by H2S post-conditioning, and it was sensitive and accurate to evaluate the heart function by P-V loop.

额尔敦-乌日勒对MCAO/R大鼠脑前额叶皮质BDNF及NGF表达的影响

目的:探讨额尔敦-乌日勒对大脑中动脉阻塞/再灌注损伤(Middle Cerebral Artery Occlusion/Reperfusion,MCAO/R)大鼠皮质脑源性神经营养因子(Brain-Derived Neurotrophic Factor,BDNF)及神经生长因子(Nerve Growth Factor,NGF)的影响。方法:取清洁级健康SD大鼠60只,随机分为假手术组、模型组、尼莫地平片组、额尔顿-乌日勒组,采用改良Zea-Longa法,制备MCAO/R模型。实验结束后,各组大鼠麻醉、取脑,采用HE染色、SP等免疫组化方法评价脑组织病理形态学的改变情况,并且采用双抗夹心ELISA法和荧光定量RT-PCR法检测大鼠脑前额叶皮质BDNF m RNA、NGF m RNA相对表达量及其蛋白含量。结果:与模型组相比,额尔敦-乌日勒组大鼠脑前额叶皮质坏死细胞数显著减少(P<0.05),BDNF、NGF蛋白含量及m RNA表达明显增加(P<0.05)。结论:额尔敦-乌日勒可上调MCAO/R大鼠脑前额叶皮质BDNF、NGF表达,促进星形胶质细胞活化,从而保护神经元,进而起到脑保护作用,这可能是额尔敦-乌日勒治疗"白脉"病的传统功效机制之一。

VEGF提高MCAO/R小鼠的EPCs表达及促进脑梗死的康复

目的：观察局灶性脑缺血再灌注模型（MCAO/R）小鼠外周循环中血管内皮祖细胞（EPCs）的数量变化,并应用血管内皮生长因子（VEGF）动员小鼠自体骨髓中的EPCs,达到治疗脑梗死的目的。方法：以MCAO/R小鼠为研究对象,通过腹腔注射VEGF动员体内的EPCs,（每日一次,持续一周）,在动员过程中的第1d、4d、7d、14d从内眦静脉采血,使用流式细胞仪检测MCAO/R组及MCAO/R＋VEGF组小鼠外周血中EPCs的数量;然后断头取脑,用免疫组化法检测新生血管的密度,并对脑组织TTC染色后计算并比较小鼠脑梗死体积的变化。结果：MCAO/R＋VEGF组在动员过程中,外周血中的EPCs在给药第1d开始增加,第4d达到高峰并持续到第7d,第14d高峰持续存在;并且在这四个时间点上,MCAO/R＋VEGF组与其它两组比较均具有统计学意义（P〈0.01）。新生血管密度：MCAO/R＋VEGF组1d开始有新生血管生成,4d血管新生明显,14d达高峰,且各时间点MCAO/R＋VEGF组血管新生数量明显多于MCAO/R组。TTC染色后通过图像分析系统计算脑梗死体积：MCAO/R＋VEGF组在给药第14d较第1d、4d、7d梗死灶体积明显缩小（P〈0.01）,且MCAO/R＋VEGF组在给药第4d,7d、14d的梗死灶体积较同时间点的MCAO/R组有显著缩小（P〈0.01）。结论：在急性脑缺血/再灌注损伤的情况下,应用VEGF能显著动员小鼠骨髓中的EPCs,增强出生后的血管新生,达到缩小梗死灶,治疗脑梗死的目的。

蒙成药额日敦-乌日勒对大脑中动脉阻塞再灌注模型(MCAO/R)大鼠血清及脑组织IL-6、IL-1β的影响

目的:探讨额日敦-乌日勒对大脑中动脉阻塞再灌注模型(MCAO/R)大鼠血清及脑内IL-6、IL-1β的影响。方法:选取144只wistar大鼠随机分为假手术组、模型组、对照组、额日敦-乌日勒低、中、高剂量组共6组。除假手术组外其他组采用改良Zea-longa法,建立MCAO/R模型。手术造模成功后24h开始灌胃给药,治疗组(额日敦-乌日勒125mg/kg、额日敦-乌日勒250mg/kg、额日敦-乌日勒500mg/kg),对照组(血塞通236.25mg/kg)。假手术组、模型组wistar大鼠每日一次灌胃纯净水。分别再灌注1、3、7天给药后行神经功能评分、心脏取血、断头取脑组织。采用ELISA法检测血清、脑组织IL-6、IL-1β的含量进行统计学分析。结果:wistar大鼠再灌注1、3、7天模型组与假手术组相比血清、脑组织IL-6、IL-1β含量均明显升高,具有统计学意义(P<0.01)。再灌注1天额日敦-乌日勒各组和血塞通组与模型组相比血清、脑组织IL-6、IL-1β含量无明显差异,无统计学意义(P>0.05)。再灌注3、7天额日敦-乌日勒各组和血塞通组与模型组相比血清、脑组织IL-6、IL-1β含量明显降低,具有统计学意义(P<0.01)。再灌注3、7天额日敦-乌日勒高剂量组与血塞通组相比明显降低,具有统计学意义(P<0.01,P<0.05)。结论:额日敦-乌日勒治疗缺血型脑卒中有一定的疗效,其作用途径可能是通过抑制IL-6、IL-1β的表达而起到神经功能恢复的作用。

间断低压缺氧预处理对MCAO大鼠脑保护作用及对TLR4表达影响

目的探讨间断低压缺氧预处理对MCAO大鼠的神经保护作用及对脑组织TLR4、NF-κB表达的影响。方法实验大鼠分为:正常对照(C)组30只、假手术(S)组26只、单纯低氧处理(H)组35只,低压氧仓内间断低压处理、单纯大脑中动脉缺血(M)组43只,行单侧大脑中动脉栓塞处理、预处理后脑梗死(HM)组37只;Longa评分,TTC染色及尼氏染色观察神经损伤程度,免疫组织化学法观察TLR4的细胞表达情况,Western blot法测定TLR4表达量,ELISA法测定TLR4及NF-κB表达变化。结果 HM组大鼠神经功能评分及梗死体积低于M组(P<0.05),残存神经元增多,TLR4表达下调(P<0.05),NF-κB表达下调(P<0.05)。结论低压缺氧预处理可有效减轻神经损伤,降低TLR4及NF-κB的表达。

Middle cerebral artery occlusion methods in rat versus mouse model of transient focal cerebral ischemic stroke

Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.

Intervention effect and mechanism of Jisheng Shenqi Decoction plus Panax notoginseng and Bionjia on CCl4-induced hepatic fibrosis rat model

Objective:To investigate the intervention effect and specific mechanism of Jisheng Shenqi Decoction plus Panax notoginseng and Turmeric on carbon tetrachloride(CCl4)-induced liver fibrosis in rats.Methods:SPF SD rats were randomly divided into control group,model group,and Chinese medicine treatment(low,medium and high dose)groups.The model group and Chinese medicine treatment group were given intraperitoneal injection of 40%CCl4 olive oil solution twice a week.A rat model of liver fibrosis was replicated by the method for 8 weeks.After successful modeling,each drug-administered group was gavaged with corresponding drugs,and the control group and model group were gavaged with equal volume of distilled water,once a day,for 4 weeks.After the last intragastric administration,HE staining and Masson staining were used to observe the pathological morphology of liver tissue,and liver function(ALT,AST)was detected;ELISA method was used to determine transforming growth factorβ(TGF-β),angiotensin II(Ang-Ⅱ),chitinase 3-like protein 1(CHI3L1),interleukin-1(IL-1),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)content;The expression levels of type I,type III collagen(Col-Ⅰ,Col-Ⅲ)and TGF-βmRNA were determined by RT-PCR.Results:①HE and Masson staining showed that a large number of liver cells in the model group were inflamed and necrotic,the collagen fibers in the liver tissue were extensively proliferated,the fibrous septa were interconnected,and the pseudolobules were formed.Compared with the model group,the levels of ALT,AST,TGF-β,Ang-Ⅱ,CHI3L1,IL-1,IL-6 and TNF-αin the traditional Chinese medicine treatment group were decreased,while the levels of Col-Ⅰ,Col-ⅢThe expressions ofⅢand TGF-βmRNA were decreased,and the effect was most obvious in the middle and high dose groups of traditional Chinese medicine(P<0.01).Conclusion:Jisheng Shenqi Decoction combined with Panax notoginseng and Biejia can significantly improve liver fibrosis induced by CCl4 in rats,and its mechanism may be related to the down-regulation of angiotensin-converting enzyme(ACE)-angiotensin II(Ang-II)-angiotensin II receptor 1(AT1R)signaling pathway related gene expression,reducing the level of related cytokines,promoting the degradation of extracellular matrix and so on.

Different Concentrations of Notoginsenoside Rg1 Attenuate Hypoxic and Hypercapnia Pulmonary Hypertension by Reducing the Expression of ERK in Rat PASMCs

Pulmonary arterial hypertension (PAH) is a serious disease which is characterized by increased vascular resistance and pressure. We have previously hypothesized that panax notoginseng saponins (PNS) might attenuate pulmonary vasoconstriction under hypoxia and hypercapnia condition. This study aims to investigate the effect of notoginsenoside Rg1, a main ingredient of PNS, with various concentrations (8, 40, 100 mg/L, respectively) on extracellular signal regulated kinase (ERK1/2) signaling pathway in pulmonary arterial smooth muscle cells (PASMCs). In addition, PASMCs were randomly divided into six groups: SD rat under normoxic condition as control group (N group), hypoxia hypercapnia group (H group), DMSO control group (HD group), Rg1-treatment groups (RgLRgM and RgH group). Western-blot and RT-PCR were used to test the expression of p-ERK protein and the expression of ERK1 mRNA and ERK2 mRNA. This study provided the evidence that the expression of p-ERK protein and the expression of ERK1 mRNA and ERK2 mRNA in HD group and H group were obviously higher than that in N group (P < 0.01), Whereas the level of ERK1/2 mRNA in Rg1-treatment groups was significantly lower than that in HD group and H group (P < 0.01), and the proper concentration of Rg1 is 40 mg/L. These results suggested that notoginsenoside Rg1 can attenuate pulmonary vasoconstriction which may lead to HHPV through reducing the expression of ERK1/2.

人参皂苷Rd通过SHH信号通路促进MCAO模型大鼠血管新生机制研究

目的探讨人参皂苷Rd对大脑中动脉闭塞(MCAO)模型大鼠血管新生的影响及相关机制。方法40只SD大鼠随机数字表法分为假手术组、模型组、人参皂苷Rd组(Rd组)、人参皂苷Rd+环靶明(CYC)组(Rd+CYC组),每组10只。采用线栓塞法建立MCAO模型大鼠,假手术组仅分离左侧颈总动脉、颈外动脉和颈内动脉。造模成功后,Rd组大鼠按照50 mg/kg给予人参皂苷Rd,Rd+CYC组按50μg/kg给予CYC腹腔注射,假手术组和模型组给予同等剂量的生理盐水,每天给药1次,连续给药2周。各组大鼠神经功能评分,TTC方法检测脑梗死体积,显微镜下观察脑组织毛细血管密度,蛋白质免疫印记检测血管生成相关分子和声波刺猬信号蛋白(SHH)、血管内皮生长因子(VEGF)、血管紧张素-1(ANG-1)、血管紧张素-2(ANG-2)蛋白表达水平。结果与模型组比较,Rd组大鼠神经功能评分显著降低[(1.000±0.471)分比(3.100±0.738)分,P<0.01];脑梗死体积显著减少[(18.120±3.735)%比(33.920±2.954)%,P<0.01];脑梗死区域毛细血管数量显著增加[(389.60±23.34)条比(54.37±11.21)条,P<0.01];血管生成相关分子SHH、VEGF、ANG-1、ANG-2蛋白表达水平显著升高[SHH:(1.267±0.038)比(0.533±0.043);VEGF:(0.866±0.035)比(0.596±0.055);ANG-1:(0.880±0.036)比(0.510±0.056);ANG-2:(0.870±0.040)比(0.386±0.045),P均<0.01]。与Rd组比较,Rd+CYC组大鼠神经功能评分显著升高[(2.700±0.675)分比(1.000±0.471)分,P<0.01];脑梗死体积显著增加[(28.910±4.491)%比(18.120±3.735)%,P<0.01];脑梗死区域毛细血管数量显著减少[(180.90±37.37)条比(389.60±23.34)条,P<0.01];血管生成相关分子VEGF、ANG-1、ANG-2蛋白表达水平显著下降[VEGF:(0.600±0.007)比(0.866±0.035);ANG-1:(0.570±0.027)比(0.880±0.036);ANG-2:(0.476±0.038)比(0.87±0.04),P均<0.01]。结论人参皂苷Rd可能通过SHH信号通路促进脑梗死区域血管生成,改善MCAO模型大鼠预后并发挥保护作用。

线栓法大鼠局灶性脑缺血模型(PMCAO)的实验研究

目的:探讨改进后的线栓法大鼠永久性大脑中动脉梗塞(PMCAO)局灶性脑缺血模型的使用价值。方法:分离颈总动脉、颈内动脉、颈外动脉,结扎颈外动脉,经由颈总动脉剪一小口,将制备好的栓线插入颈总动脉,并推进送入颈内动脉内,阻断大脑中动脉,制备成PMCAO模型,并观察大鼠脑组织的病理改变。结果:成功率可高达95%,模型符合脑梗死病理过程。结论:改进后的方法(PMCAO)简便、易重复,降低了动物死亡率,提高了成功率,可做为缺血性脑血管病研究的理想动物模型。

三七皂苷Rg_1对脑缺血再灌注损伤的保护作用

目的探讨三七皂苷Rg1对大鼠局灶性脑缺血再灌注损伤后大脑皮质中BDNFmRNA表达的影响。方法♂SD大鼠36只随机分为脑缺血再灌注模型组、三七皂苷Rg1组和尼莫地平组,采用线栓法制作模型,各组分别于术后1、3、5 d随机取4只大鼠处死后,取出脑组织,经石蜡包埋切片,片厚5μm。用地高辛标记的寡聚核苷酸探针进行原位杂交实验,用HPIAS-100高清晰度彩色病理图文报告分析系统测量分析大鼠大脑皮质的BDNFmRNA的含量和阳性神经元数目的变化。结果在脑缺血再灌注后不同的时间段上,与模型组比较,三七皂苷Rg1均能增加大脑皮质的BDNFmRNA含量及其阳性神经元的数量。用药3 d时,三七皂苷Rg1的作用达到高峰,且其作用在各时间段上均强于尼莫地平;5 d后,虽然BDNFmRNA的含量及阳性神经元的数量有一定程度的下降,但仍与三七皂苷Rg1组1 d时的表达水平相当,且三七皂苷Rg1组5 d时的表达水平仍强于模型组1 d和3 d时表达的水平及尼莫地平组1 d时的水平。结论三七皂苷Rg1通过上调大鼠脑缺血再灌注损伤后,大脑皮质BDNFmRNA的含量和阳性神经元的数量,可发挥其治疗作用,疗效强于阳性对照药尼莫地平。

高胆固醇膳食对大鼠和仓鼠肝脏HMG-CoA-R、LDL-R蛋白和基因表达的影响

大鼠和仓鼠作为常用的降胆固醇研究模型而被广泛研究。通过喂饲大鼠和仓鼠不同含量膳食胆固醇（0%~0.9%）,对大鼠和仓鼠高胆固醇模型进行比较。结果显示,喂饲胆固醇膳食,仓鼠和大鼠肝脏3-羟基-3甲基-戊二酰CoA还原酶（HMG-CoA-R）的蛋白和基因表达均受到明显的抑制。随着膳食胆固醇添加量的增加,大鼠肝脏低密度脂蛋白受体（LDL-R）表达增加,但是仓鼠和大鼠的LDL-R基因表达均受到抑制。

针刺水沟穴抑制MCAO大鼠海马区神经细胞坏死的作用研究

目的:对比针刺水沟穴与水沟旁非穴对MCAO大鼠海马区坏死神经元作用。方法:将72只健康雄性Wistar大鼠按照随机数字表分为正常组、假手术组、模型对照组、非针刺组、水沟旁组、水沟组6组,每组12只。模型对照组、水沟旁组、水沟组参照Zea-Longa线栓法复制大鼠大脑中动脉缺血模型(MCAO)。水沟旁组、水沟组以频率180次/min、持续5s进行针刺干预。观察大鼠神经行为学,血流量,脑梗死面积比率,以及海马区神经细胞坏死率。结果:水沟组分别与非针刺组和水沟旁组比较差异都具有显著差异性P<0.05,而增加血流量方面水沟组也明显优于水沟旁组P<0.01。水沟旁组与非针刺组无显著性差异(P>0.05)。结论:针刺水沟穴抑制MACO大鼠神经细胞坏死,增加血流量从而降低梗死面积,而水沟旁无此作用.