黄芩苷对干酵母致热大鼠的解热作用及血清TNF-α、IL-1β、IL-6、PGE_(2)、cAMP和脑组织NF-κB表达的影响

目的:观察黄芩苷对干酵母致热大鼠的解热作用并探讨其作用机制。方法:采用背部皮下注射干酵母构建大鼠发热模型,SD雄性大鼠随机分为正常对照,模型组,阳性组(阿司匹林,0.1 g/kg),黄芩苷高、中、低剂量组(160、80、40 mg/kg),连续给药3 d,测定各组大鼠肛温的变化;酶联免疫法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白细胞介素-6(IL-6)、前列腺素E_(2)(PGE_(2))与环磷酸腺苷(cAMP)水平;Western Blot检测各组大鼠脑组织NF-κB p65(核转录因子-κB p65)蛋白表达。结果:黄芩苷高剂量组有显著解热效果(P<0.01),黄芩苷各剂量组均可不同程度降低大鼠血清TNF-α、IL-1β、IL-6、PGE 2和cAMP含量;与正常组比较,模型组脑组织NF-κB p65蛋白表达增多,黄芩苷高剂量组可明显降低大鼠脑组织NF-κB p65表达(P<0.05)。结论:黄芩苷可显著性降低干酵母引起的体温升高,解热机制可能与抑制TNF-α、IL-1β、IL-6、PGE_(2)与cAMP的分泌和减少脑组织NF-κB p65蛋白表达有关。

Expressions of MMP-2,-9,TIMP-1,-2,-3 mRNA in Rat Uterus during Estrous Cycle

Zymography and in situ hybridization were used to investigate matrixmetalloproteinase -2, -9 (MMP -2, MMP-9) activities and expressions of MMP -2, -9 and TIMP1, -2, -3 (tissue inhibitors of matrix metallo-proteinases) mRNA in the rat uterus during estrouscycle. The relative activity was semiquanted by using densitometric analysis. The MMP-2(67 kDa) activity in every stage during estrpus cycle was detected by zymography. MMP-2activity was highest at proestrus; higher at estrus and metaestrus; lowest at diestrus. Throughin situ hybridization, MMP -2, -9, TIMP -1~ -3 mRNA mainly in hasal stroma cells of uterineendometrium were detected. The positive signals of MMP -2 and -9 mRNAs in hasal stromacells were shown stronger at proestrus, estrus and metaestrus while they showed the weakest atdiestrus. The expression of MMP -2 mRNA coincided with MMP -2 activity change. MMP-2and -9 mRNAs were also highly expressed in uterine circular muscle at estrus. Weak signals ofMMP -9 mRNA were detected in uterine luminal and glandular epithelial cells at estrus.TIMP -1 mRNA in hasal stroma cells was shown as the strongest expression at estrus andmetaestrus; stronger at proestrus and the weakest at diestrus. TIMP-2 mRNA in basal stromacells was stronger at estrus and diestrus; weaker at proestrus and metaestrus. TIMP -1 and -2mRNAs were also highly expressed in uterine luminal and glandular epithelial cells at estrus.TIMP -3 mRNA in hasal stroma cells revealed the strongest expression at estrus; stronger atdiestrus and metaestrus and showed the weakest at proestrus. The mRNA was also highlyexpressed in uterine circular muscle at estrus. In short, our present results provide evidencethat MMP -2, -9 and TIMP -1~ -3 were involved in rat uterine endometrium reconstructionduring estrous cycle.

Effects of V2O5 on Proliferation and Differentiation of Limb Bud Cells of Rat

The micromass culture was used to determine the effects of vanadium pentoxide (V2O5 ) on the proliferation and differentiation of limb bud cells of rat. In the in vitro test, the results showed that V2O5 had obvious inhibiting effects on both proliferation and differentiation of limb bud cells with a dosedependent response, its proliferating and differentiating IC50 being 13.64 and 4.77μmol/L, respectively. In the in vivo/in vitro test, the results showed that V2O5 had no obvious effect on cell proliferation but had obvious inhibiting effect on cell differentiation. These results indicated that V2O5 might have a specific inhibiting effect on the differentiation of limb bud cells.

尼莫地平对脊髓损伤大鼠脊髓组织Ca^(2+)及细胞焦亡的影响

目的研究尼莫地平(Nimodipine)对脊髓损伤(SCI)大鼠及脊髓组织Ca^(2+)及细胞焦亡的影响。方法将36只SD雄性大鼠按照随机数表法分为假手术组(Sham组)、脊髓损伤组(SCI组)和SCI+Ca^(2+)抑制剂尼莫地平组(Nimodipine组)各12只。脊髓撞击法制备大鼠SCI模型,BBB评分评估大鼠运动功能,荧光探针检测脊髓组织Ca^(2+)含量,酶联免疫吸附法检测白介素1β(IL-1β)及白介素18(IL-18)含量,蛋白免疫印迹检测细胞焦亡相关蛋白NOD样受体蛋白3(NLRP3)及半胱氨酸/天冬氨酸蛋白酶1(Caspase-1)的表达水平。结果与Sham组比较,SCI组大鼠BBB评分降低,差异有统计学意义(P<0.05);与Sham组比较,SCI组大鼠Ca^(2+)(18.92±3.60)、IL-1β[(383.66±45.42)pg/mL]、IL-18[(364.21±38.23)pg/mL]含量和NLRP3(0.93±0.19)、Caspase-1(0.98±0.08)的表达水平明显增加,差异均有统计学意义(P<0.05);与SCI组比较,Nimodipine组大鼠1 d、3 d、7 d的BBB评分稍增高,但差异均无统计学意义(P>0.05),而14 d、21 d及28 d大鼠BBB评分明显增高,差异均有统计学意义(P<0.05);与SCI组比较,Nimodipine组大鼠Ca^(2+)(11.73±4.31)、IL-1β[(292.93±28.48)pg/mL]、IL-18[(279.81±22.52)pg/mL]含量和NLRP3(0.79±0.18)及Caspase-1(0.63±0.10)的表达水平明显减少,差异均有统计学意义(P<0.05)。结论尼莫地平可能通过抑制Ca^(2+)来改善SCI大鼠的细胞焦亡及运动功能。

运动联合益生菌干预2型糖尿病大鼠糖脂代谢、氧化应激及骨骼肌卫星细胞成肌分化水平的变化

目的观察运动联合益生菌干预2型糖尿病大鼠糖脂代谢、氧化应激及骨骼肌卫星细胞表达的影响。方法8周龄SPF级SD雄性大鼠60只,随机选择10只大鼠为正常对照组(NC组),造模组大鼠腹腔注射链脲佐菌素,建立2型糖尿病大鼠模型,随机分为模型组(TN组)、运动+糖尿病组(YTN组)、益生菌+糖尿病组(GTN组)、运动+益生菌+糖尿病组(YGTN组),6周的递增负荷有氧跑台运动(第1周跑台速度为15 m/min,持续30 min;第2周跑台速度为15 m/min,持续60 min;第3~6周跑台速度为19.3 m/min,运动时间持续60 min,持续训练到第6周,每周5 d);GTN组和YGTN组在训练前1 h灌胃10.0 mL/(kg·d)的Lactibiane Iki益生菌溶液(浓度为10~7 CFU/mL)。最后,测定大鼠糖脂代谢指标、氧化应激指标及骨骼肌卫星细胞蛋白的表达。结果(1)TN组、YTN组、GTN组和YGTN组大鼠空腹血糖(fasting plasma glucose,FBG)、血清胰岛素(fasting serum insulin,FINS)、糖化血红蛋白(hemoglobin A1c,HBA1C)和胰岛素抵抗指数(homeostasis model assessment-insulin resistance index,HOMA-IR)水平均高于NC组(P<0.01),其中,YGTN组大鼠FBG水平均低于TN组、YTN组、GTN组(P<0.05);YTN组、GTN组和YGTN组大鼠血清FINS水平均与TN组比较均有所下降(P<0.01);YGTN组大鼠血清HBA1C水平均低于TN组、YTN组和GTN组水平(P<0.01);YGTN组大鼠HOMA-IR指数与TN组、YTN组和GTN组比较下降较显著(P<0.01)。结果表明有氧运动联合益生菌干预组大鼠FBG、FINS、HBA1C和HOMA-IR含量下降较明显。(2)TN组大鼠血清总胆固醇(total cholesterol,T-CHO)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL)和游离脂肪酸(free fatty acid,FFA)水平均高于NC组(P<0.05),YGTN组大鼠血清T-CHO、TG、LDL和FFA水平均低于TN组(P<0.05),基本上恢复到NC组的水平。TN组大鼠血清高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL)水平低于NC组(P<0.05),YGTN组大鼠血清HDL水平高于TN组(P<0.05),基本上恢复到NC组水平。结果表明有氧运动联合益生菌干预糖尿病大鼠,可降低糖尿病大鼠血清T-CHO、TG、LDL、FFA含量,提高HDL水平较明显。(3)TN组大鼠血清丙二醛(malondialdehyde,MDA)和游离8-异前列腺素F2α(8-iso-prostaglandin F2α,8-isoPGF2α)水平均高于NC组(P<0.05或P<0.01),YGTN组大鼠血清MDA和8-isoPGF2α水平均低于TN组(P<0.05或P<0.01);TN组大鼠血清过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和总抗氧化能力(total antioxidant capacity,T-AOC)水平均低于NC组(P<0.01),YGTN组大鼠血清CAT、SOD、GSH-Px和T-AOC水平均高于NC组(P<0.01),但基本上恢复到NC组的水平。(4)TN组大鼠骨骼肌卫星细胞配对盒基因7(paired box gene 7,Pax7)、成肌分化因子(myogenic determination gene,MyoD)、肌细胞生成素(myogenin,MyoG)和生肌因子5(myogenic factor 5,Myf5)蛋白表达均低于NC组(P<0.05或P<0.01),YGTN组大鼠骨骼肌Pax7、MyoD、MyoG和Myf5蛋白表达均高于TN组(P<0.05或P<0.01);基本上恢复到NC组水平。TN组大鼠骨骼肌生成抑制素(myostatin,MSTN)蛋白表达高于NC组(P<0.05),YGTN组大鼠骨骼肌MSTN蛋白表达低于TN组(P<0.05),基本上恢复到NC组水平。结论益生菌和有氧运动具有减轻胰岛素抵抗,改善血脂和氧化激的作用,在改善2型糖尿病大鼠血糖和胰岛素水平具有协同作用。此外,有氧运动联合益生菌干预促进了肌细胞的分化,可能对防止T2DM引起肌肉质量的流失和力量下降以及肌肉组织的并发症发挥一定的作用。

桑黄素通过抑制ERK1/2-p38信号通路介导对老年大鼠骨骼保护作用

【目的】探讨桑黄素(SSS)治疗对老年大鼠骨代谢及骨量影响,并阐明其可能的作用机制。【方法】10只3月龄年轻雌性Sprague-Dawley(SD)大鼠和20只24月龄老年雌性SD大鼠随机分为3组,对照组(CON,10只年轻大鼠)、模型组(MOD,10只老年大鼠)和桑黄素组(SSS,10只老年大鼠)。在实验过程中,SSS组每日接受腹腔注射桑黄素(10 mg/kg)治疗。治疗为期12周,待治疗结束后使用Micro-CT、HE染色切片、血清学检测以及蛋白质印迹观察治疗效果以及可能的机制。【结果】治疗12周后,与MOD组相比,SSS组的大鼠骨小梁数量和密度得到明显的改善。SSS组左侧股骨BMD、Conn.D、Tb.N、Tb.Th和Tb.Sp较MOD组明显改善(P<0.05)。治疗12周时,SSS组CTX-1、骨钙素、TRACP-5b和PINP水平较MOD显著降低(P<0.05)。和MOD组比较,SSS组的大鼠ERK1/2-p38信号通路显著抑制,ERK1/2和p38水平显著降低,比较有统计学差异(P<0.05)。【结论】桑黄素通过抑制ERK1/2-p38信号通路和降低骨转换来介导对老年大鼠骨骼保护作用。

常压高浓度氧对新生大鼠脑微血管内皮细胞损伤与Nrf2/HO-1信号通路的影响分析

目的探究常压高浓度氧(NBO)对新生大鼠脑微血管内皮细胞损伤及核因子E2相关因子2/血红素氧合酶-1(Nrf2/HO-1)信号通路的影响。方法取新生SD大鼠45只,采用随机数字表法分为常氧组、NBO组和NBO+Nrf2激活剂组,每组各15只。常氧组大鼠置于普通空气(21%氧气)中饲养,NBO组和NBO+Nrf2激活剂组大鼠置于90%常压氧气饲养,NBO+Nrf2激活剂组每日灌胃5 mg/kg Nrf2激动剂莱菔硫烷。测定脑组织伊文思蓝(EB)含量,采用酶联免疫法检测血管内皮生长因子(VEGF)和基质金属蛋白酶9(MMP-9)含量,干湿重法检测脑组织含水量,HE染色和TUNEL染色观察脑组织病理变化,蛋白质印迹法检测海马组织Nrf2/HO-1信号通路蛋白表达,水迷宫检测大鼠认知功能。结果与常氧组比较,NBO组脑组织EB、VEGF、MMP-9含量及脑组织含水量升高,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组脑组织EB、VEGF、MMP-9含量及脑组织含水量降低,差异均有统计学意义(P<0.05)。病理染色结果显示,常氧组大鼠神经细胞形态及结构完整,未见明显病理变化和细胞凋亡;NBO组神经细胞形态及结构不规则,出现明显的水肿和空泡,并伴有大量的凋亡细胞;NBO+Nrf2激活剂组脑组织病理损伤较NBO组明显减轻。与常氧组比较,NBO组脑组织Nrf2、HO-1蛋白相对表达量降低,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组Nrf2、HO-1蛋白相对表达量升高,差异均有统计学意义(P<0.05)。与常氧组比较,NBO组第2~4天逃避潜伏期延长,穿越平台次数减少,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组第2~4天逃避潜伏期缩短,穿越平台次数增多,差异均有统计学意义(P<0.05)。结论NBO可诱导新生大鼠脑微血管内皮细胞损伤,导致远期认知功能障碍,可能与下调Nrf2/HO-1信号通路表达有关。

羟基红花黄色素A通过调控环氧合酶2/前列腺素E_(2)信号通路减轻大鼠脑缺血再灌注损伤

目的观察羟基红花黄色素A(HSYA)对环氧合酶2(COX-2)/前列腺素E_(2)(PGE_(2))信号通路的影响,探讨HSYA对脑缺血再灌注损伤(CIRI)的保护机制。方法90只雄性SD大鼠随机分为假手术组(S组)、手术组(CIRI组)、HSYA组和塞来昔布组(C组),HSYA组进一步分为HSYA低剂量组(HSYA-L组)、HSYA中剂量组(HSYA-M组)和HSYA高剂量组(HSYA-H组),每组15只。线栓法制备脑缺血再灌注损伤模型。各组大鼠于术前30 min腹腔注射给药,HSYA各组分别给予HSYA 10、15、25 mg/kg,C组给予塞来昔布40 mg/kg,S组和CIRI组给予等量的生理盐水。各组大鼠模型制作苏醒后立刻进行神经功能学评分,再灌注24 h时进行脑梗死体积检测,同时Nissl染色观察神经细胞损伤,Real-time PCR和Western blotting检测COX-2 mRNA和蛋白的变化,ELISA检测PGE_(2)、肿瘤坏死因子α(TNF-α)和白细胞介素(IL)-1β的变化。结果与S组比较,CIRI组神经功能学评分显著升高(P<0.05),脑梗死体积显著增加(P<0.05),神经细胞损伤较重,数目显著降低(P<0.05),COX-2 mRNA和蛋白的表达显著增多,同时PGE_(2)、TNF-α和IL-1β的表达也显著增多(P<0.05);与CIRI组比较,HSYA组及C组神经功能学评分明显降低(P<0.05),脑梗死体积明显减少(P<0.05),神经细胞损伤减轻,数目明显增加(P<0.05),COX-2 mRNA和蛋白及PGE_(2)、TNF-α和IL-1β的表达均明显下降(P<0.05),且HSYA各组之间及HSYA-L组和HSYA-M组与C组比较差异均较显著(P<0.05),而HSYA-H组与C组比较差异无显著性(P>0.05)。结论HSYA减轻缺血性脑卒中再灌注损伤可能与抑制COX-2/PGE_(2)信号通路有关。

Effects of genistein on neuronal apoptosis,and expression of Bcl-2 and Bax proteins in the hippocampus of ovariectomized rats

Genistein is one of several isoflavones that has a structure similar to 17β-estradiol, has a strong antioxidant effect, and a high affinity to estrogen receptors. At 15 weeks after ovariectomy, the expression of Bcl-2 in the hippocampus of rats decreased and Bax expression increased, with an obvious upregulation of apoptosis. However, intraperitoneal injection of genistein or 17β-estradiol for 15 consecutive weeks from the second day after operation upregulated Bcl-2 protein expression downregulated Bax protein expression, and attenuated hippocampal neuron apoptosis. Our experimental findings indicate that long-term intervention with genistein can lead to a decrease in apoptosis in hippocampal neurons following ovadectomy, upregulate the expression of Bcl-2, and downregulate the expression of Bax. In addition, genistein and 17β-estradiol play equal anti-apoptotic and neuroprotective roles.

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）.

Emblica officinalis improves glycemic status and oxidative stress in STZ induced type 2 diabetic model rats

Objective:To evaluate the antidiabetic and antioxidant potential of Emblica officinalis(E.officinalis)fruit on normal and type 2 diabetic rats.Methods:Type 2 diabetes was induced into the male Long-Evans rats.The rats were divided into nine groups including control groups receiving water,type 2 diabetic controls,type 2 diabetic rats treated with glibenclamide(T2GT)and type 2diabetic rats treated with aqueous extract of fruit pulp of E.officinalis.They were fed orally for8 weeks with a single feeding.Blood was collected by cutting the tail tip on 0 and 28 days and by decapitation on 56 day.Packed red blood cells and serum were used for evaluating different biochemical parameters.Results:Four weeks administration of aqueous extract of E.officinalis improved oral glucose tolerance in type 2 rats and after 8 weeks it caused significant(P<0.007)reduction in fasting serum glucose level compared to 0 day.Triglycerides decreased by 14%but there was no significant change in serum ALT,creatinine,cholesterol and insulin level in any group.Furthermore,reduced erythrocyte malondialdehyde level showed no significant change(P<0.07)but reduced glutathione content was found to be increased significantly(P<0.05).Conclusions:The aqueous extract of E.officinalis has a promising antidiabetic and antioxidant properties and may be considered for further clinical studies in drug development.

柴胡疏肝汤对癫痫大鼠海马组织中Bcl-2、NLRP1、Caspase-3、Caspase-1表达的影响

目的探讨柴胡疏肝汤对癫痫大鼠海马组织中B细胞淋巴瘤2(Bcl-2)、含NLR家族pyrin结构域1因子(NLRP1)、Caspase-3、Caspase-1表达的影响。方法基于生物信息学分析筛选柴胡疏肝汤治疗癫痫的核心活性成分及核心靶点。将100只SD大鼠随机分为癫痫组(n=80)与空白组(n=20)。对空白组大鼠不作任何处理,对癫痫组大鼠建立癫痫模型,成功建模后将其随机分为模型组、柴胡疏肝汤低剂量组、柴胡疏肝汤中剂量组、柴胡疏肝汤高剂量组,每组20只。给予柴胡疏肝汤低、中、高剂量组大鼠柴胡疏肝汤(2.5 mg/100 g、5 mg/100 g、10 mg/100 g)灌胃,给予模型组及空白组大鼠生理盐水灌胃。给药4周后,比较模型组与柴胡疏肝汤低、中、高剂量组的大鼠癫痫发作等级,然后处死5组大鼠并获取其海马组织,分别采用实时荧光定量PCR和Western blot检测Bcl-2、NLRP1、Caspase-3、Caspase-1的mRNA及蛋白表达水平。结果(1)柴胡疏肝汤治疗癫痫的核心活性成分包括槲皮素、山柰酚、柚皮素、β-谷甾醇、川陈皮素、异鼠李素、木犀草素、豆甾醇、7-甲氧基-2-甲基异黄酮、罗通定,核心靶点包括Bcl-2、NLRP1、Caspase-3、Caspase-1。槲皮素、山柰酚、柚皮素、β-谷甾醇、川陈皮素和木犀草素与核心靶点表现出较好的结合活性。(2)干预后,与模型组相比,柴胡疏肝汤中、高剂量组的癫痫发作等级降低(P<0.05);与柴胡疏肝汤低剂量组相比,柴胡疏肝汤中、高剂量组的癫痫发作等级降低(P<0.05)。与空白组比较,模型组Bcl-2的mRNA和蛋白表达水平降低,NLRP1、Caspase-3、Caspase-1的mRNA和蛋白表达水平升高(P<0.05);与模型组比较,柴胡疏肝汤各剂量组Bcl-2的m RNA和蛋白表达水平升高,NLRP1、Caspase-3、Caspase-1的mRNA和蛋白表达水平降低(P<0.05);柴胡疏肝汤低剂量组、柴胡疏肝汤高剂量组、柴胡疏肝汤中剂量组Bcl-2的mRNA和蛋白表达水平依次升高,柴胡疏肝汤中剂量组、柴胡疏肝汤低剂量组、柴胡疏肝汤高剂量组NLRP1的m RNA和蛋白表达水平依次升高,柴胡疏肝汤高剂量组、柴胡疏肝汤低剂量组、柴胡疏肝汤中剂量组Caspase-3、Caspase-1的mRNA和蛋白表达水平依次升高(P<0.05)。结论柴胡疏肝汤可能通过上调大鼠海马组织中Bcl-2的表达,抑制NLRP1、Caspase-3、Caspase-1的表达,发挥抗癫痫作用。

Morin enhances hepatic Nrf2 expression in a liver fibrosis rat model

AIM To investigate whether morin can reduce hepatic fibrosis by activating the NF-E2-related factor 2(Nrf2) signaling pathway.METHODS Twenty male Sprague-Dawley rats were randomly divided into four groups: control group, morin group, carbon tetrachloride(CCl4) group, and morin + CCl4 group. Rats in both the CCl4 and morin + CCl4 groups were injected intraperitoneally with CCl4 at a dose of 2 mL/kg twice a week. Rats in both the morin and morin + CCl4 groups were treated orally with morin at a dose of 50 mg/kg twice a week. Control rats were treated with vehicle only twice a week. At the end-point of the 8 wk of the experimental period, serum AST, ALT, and ALP were measured, and the liver specimenswere obtained for pathological assessment. Real-time PCR and Western blot methods were used to analyze the expression of α-smooth muscle actin(α-SMA), collagen Ⅰ, collagen Ⅲ, Nrf2, heme oxygenase(HO-1), and quinone oxidoreductase 1(NQO1) using frozen liver specimens.RESULTS Morin-treated rats in the morin + CCl4 group had less hyperplasia of fiber tissue, minimal inflammatory cells, and less body weight loss with favorable liver enzyme measurements compared to rats treated with CCl4 only. Additionally, morin-treated rats had significantly lower m RNA and protein expression of α-SMA, collagen Ⅰ, and collagen Ⅲ, but significantly higher m RNA and protein expression of Nrf2, HO-1, and NQO1 compared to rats treated with CCl4 only(P < 0.05).CONCLUSION Morin could play a protective role by inducing the expression of Nrf2 and its downstream antioxidant factors(HO-1 and NQO1) and reducing the expression of α-SMA, collagen Ⅰ, and collagen Ⅲ in CCl4-induced liver fibrosis rats.

Expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic stellate cells during rat hepatic fibrosis and its intervention by IL-10

AIM: To investigate the expression of matrix metallopr-oteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10). METHODS: Hepatic fibrosis was induced by CCI4 administration and 60 male Sprague-Dawley rats were randomly divided into normal control group (group N, 8 rats), CCI4-induced group (group C, 28 rats) and IL-10-treated group (group I, 24 rats). At the beginning of the 7th and 11th wk, rats in each group were routinely perfused with pronase E and type IV collagenase through portal vein catheter and the suspension was centrifuged by 11% Nycodenz density gradient to isolate hepatic stellate cells (HSCs). RT-PCR was used to analyze mRNA of MMP-2 and TIMP-1 from freshly isolated cells. Densitometric data were standardized with β-actin signals. Immunocytochemistry was performed to detect MMP-2 and TIMP-1 expression in HSC cultured for 72 h. RESULTS: Compared to group N in the 7th wk, MMP-2 and TIMP-1 mRNA increased in group C (P= 0.001/0.001) and group I (P= 0.001/0.009). The level of MMP-2 and TIMP-1 mRNA in group I was significantly lower than that in group C (P= 0.001/0.001). In the 11th wk, MMP-2 mRNA in group I was still lower than that in group C (P = 0.005), but both dropped compared with that in the 7th week (P = 0.001/0.004). TIMP-1 mRNA in group I was still lower than that in group C (P= 0.001), and increased in group C (P= 0.001) while decreased in group I (P = 0.042) compared with that in the 7th wk. Same results were found by immunocytochemistry. CONCLUSION: Expression of MMP-2 and TIMP-1 is increased in hepatic fibrosis. IL-10 exhibits an antifibrogenic effect by suppressing MMP-2 and TIMP-1 expression.

Induction of Endoplasmic Reticulum Stress by Cadmium and Its Regulation on Nrf2 Signaling Pathway in Kidneys of Rats

Objective This study was conducted to investigate the regulation of endoplasmic reticulum stress on Nrf2 signaling pathway in the kidneys of rats. Methods Rats were divided into twelve groups of six animals each. Some groups were pre-administered with bacitracin or tauroursodeoxycholic acid(TUDCA), and all of them were treated with 5-20 μmol/kg cadmium(Cd) for 48 h. The oxidative stress levels were analyzed using kits. The mRNA and protein expression levels of endoplasmic reticulum stress-related factors and Nrf2 signaling pathway-related factors were determined using RT-PCR and western blot. Results Cd exposure resulted in oxidative stress in the kidneys of rats and upregulated the expression of endoplasmic reticulum stress(ERS)-related factors and Nrf2 signaling pathway-related factors, especially at doses of 10 and 20 μmol/kg Cd, and the expression changes were particularly obvious. Moreover, after pretreatment with bacitracin, Cd upregulated the expression of ERS-related factors to a certain extent and, at higher doses, increased the mRNA expression of Nrf2. After pretreatment with TUDCA, Cd reduced the level of ERS to a certain extent; however, at these doses, there were no significant changes in the expression of Nrf2. Conclusion Cadmium can result in ERS and oxidative stress in the kidneys of rats, activate Nrf2, and upregulate the transcriptional expression of phase II detoxification enzymes under these experimental conditions. ERS has a positive regulation effect on Nrf2 signaling pathway but has little effect on the negative regulation of Nrf2 signaling pathway in cadmium toxicity.

THE EXPERIMENTAL STUDY ON THE CELL APOPTOSIS AND EXPRESSION OF BCL-2 PROTEIN IN INTRACEREBRAL HEMORRHAGE IN MODEL OF RATS

Objective To study whether there is the apoptosis of neural cells and the expression of Bcl-2 protein in intracerebral hemorrhage (ICH) in model of rats, for the further understanding the mechanism of the delayed damage of the neural cells around the hematoma after ICH. Methods Fifty SD rats were randomly divided into 5 groups, ten in each. With the Group A as the control, the rest 40 were used to set up intracerebral hemorrhage model. The brains were taken out at 12 th , 24 th , 48 th and 72 th hours, respectively. Apoptosis cells were detected with terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL), and the expression of Bcl-2 protein was detected with immunochemical stainging methed (SP). Results In the control group, no apoptosis cells and Bc1-2 protein were detected. In rest groups, the apoptosis cells and Bc1-2 protein were expressed in different degree. Apoptosis rates verified and corresponded with the time after ICH, with the peak at 48 th -72 th hour after hemorrhage. The peak rate of apoptosis cells was (24.50±2.69)% and Bcl-2 protein expression was (20.76±1.97)% . There was significant difference between the experimental groups and control (P<0.05), and no linear relationship between the apoptosis rate and the expression of Bcl-2 protein. Conclusion Apoptosis may be an important factor in the secondary trauma of ICH. There is a time leg after hemorrhage. All this is instructive to clinical treatment in time. Bcl-2 protein keeps increasing in a certain time after hemorrhage, but not synchronize with the cell apoptosis. This indicates that bcl-2 has the effect to reduce the apoptosis of neural cells.

Effect of Shoutai Pills on Th1/Th2 Cytokines in Serum and Endometrium of Rats with Stimulated Ovulation

In our previous study,we found that Shoutai pills could improve the embryo implantation rate as well as the levels of estrogen,progesterone and estrogen receptor in rats with stimulated ovulation.However,the mechanism is not clear.This study was designed to investigate the effect of Shoutai pills on the levels of Th1 and Th2 cytokines in rats with stimulated ovulation and the mechanism.The rat model of stimulated ovulation was established by combined injection of pregnant mare serum gonadotropin(PMSG)and human chorionic gonadotropin(HCG).Then the rats were randomly divided into model group(M),Shoutai pills group(S),progesterone group(P)and normal group(N).All the pregnant rats were treated from the first day.The S and P groups were administrated with gavage of Shoutai pills and injection of progesterone respectively,and N and M groups were given the same volume of normal saline and distilled water respectively.After treatment for 7 days,the animals were executed for serum and uterine tissues.The ELISA method was adopted to detect the contents of Thl cytokines[interferon-γ(INF-γ),interleukin-2(IL-2)]and Th2 cytokines(IL-4,IL-6,IL-10).The expression of leukemia inhibitory factor(L1F)and leukemia inhibitory factor receptor(LIFR)was detected by Western blotting and real-time PCR.As compared with N group,the expression levels of IFN-y and IL-2 in M group were significantly increased,and those of IL-4,IL-6,IL-10.LIF and LIFR were significantly decreased(P<0.05).As compared with M group,the levels of IL-4,IL-6,IL-10,LIF and LIFR in S group were significantly increased(P<0.05),and those of IFN-γand IL-2 were significantly decreased(P<0.05).It was suggested that Shoutai pills can increase the levels of IL-4.1L-6,IL-10,LIF and LIFR as well as reduce the levels of INF-γand IL-2 in rats with stimulated ovulation.The Shoutai pills may improve endometrial receptivity and promote embryo implantation by maintaining the balanee of Th1/Th2 cytokines.

Effects of melatonin on the expression of iNOS and COX-2 in rat models of colitis

AIM: To investigate the effects of melatonin (MT) on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in rat models of colitis.METHODS: Healthy adult Sprague-Dawlay (SD) rats of both sexes, weighing 280±30 g, were employed in the present study. The rat models of colitis were induced by either acetic acid or 2,4,6-trinitrobenzene sulfonic acid (TNBS) enemas. The experimental animals were randomly divided into melatonin treatment and model control group that were intracolicly treated daily with melatonin at doses of 2.5, 5.0, 10.0 mg.kg-1 and equal amount of saline respectively from 24 h following induction of colitis in rats inflicted with acetic acid enema and the seventh day in rats with TNBS to the end of study. A normal control group of rats treated with neither acetic acid nor TNBS but saline enema was also included in the study. On the 28th day of the experiment, the rat colon mucosal damage index (CDMI) was calculated, and the colonic prostaglandin E2(PGE2), nitric oxide (NO), as well as the iNOS and COX-2expression were also determined biochemically or immunohistochemically.RESULTS: CDMI increased to 2.87±0.64 and 3.12±1.12respectively in rats treated with acetic acid and TNBS enema,which was in accordance with the significantly elevated colonic NO and PGE2 contents, as well as the up-regulated colonic iNOS and COX-2 expression in both of the two rat models of colitis. With treatment by melatonin at the doses of 5.0 and 10.0 mg@kg-1, CDMI in both models of rat colitis was significantly decreased (P＜0.05-0.01), which accorded synchronously and unanimously with the reduced colonic NO and PGE2 content, as well as the down-regulated expression of colonic iNOS and COX-2.CONCLUSION: Melatonin has a protective effect on colonic injury induced by both acetic acid and TNBS enemas, which is probably via a mechanism of local inhibition of iNOS and COX-2 expression in colonic mucosa.

Comparison of T-2 Toxin and HT-2 Toxin Distributed in the Skeletal System with That in Other Tissues of Rats by Acute Toxicity Test

Twelve healthy rats were divided into the T-2 toxin group receiving gavage of 1 mg/kg T-2 toxin and the control group receiving gavage of normal saline. Total relative concentrations of T-2 toxin and HT-2 toxin in the skeletal system（thighbone, knee joints, and costal cartilage） were significantly higher than those in the heart, liver, and kidneys（P 〈 0.05）. The relative concentrations of T-2 toxin and HT-2 toxin in the skeletal system（thighbone and costal cartilage） were also significantly higher than those in the heart, liver, and kidneys. The rats administered T-2 toxin showed rapid metabolism compared with that in rats administered HT-2 toxin, and the metabolic conversion rates in the different tissues were 68.20%-90.70%.

Polysaccharide Nucleic Acid of Bacillus Calmette Guerin Modulates Th1/Th2 Cytokine Gene Expression in Lipopolysaccharide-Induced Mastitis in Rats

The aim of this study was to evaluate, in rats, the changes in the T helper type 1（Th1）/Th2 radio in mammary glands after an intramammary infusion of lipopolysaccharide （LPS） and to characterize the moderating effects of the polysaccharide nucleic acid of Bacillus Calmette Guerin （BCG-PSN） on the mammary gland. In the control group, the levels of IL-2 and INF-7 mRNA expression increased, whereas IL-4 mRNA expression decreased after LPS challenge. As a consequence, the INF-γ/IL-4 mRNA ratio was significantly higher at 3, 6, and 9 h post-infusion （PI） compared to the control value （0 h; P〈0.01）. BCG-PSN increased mRNA expression of both INF-γ and IL-4 before infusion of LPS. LPS challenge significantly the reduced Th1/Th2 cytokine ratio due to Thl cytokine IFN-γ suppression and Th2 cytokine IL-4 upregulation compared with the control group. A significant reduction of N-acetyl-β-D-glucosaminidase （NAGase） was observed at 24 h PI in the BCG-PSN treatment group compared to the control group （P〈 0.05）. Thus, it was demonstrated that level of BCG-PSN might change the Th1/Th2 ratio mainly by enhancing the Th2 immune response. This is the first report of a Th1/Th2 change induced by coliform mastitis and characterization of the effect of BCG-PSN on mammary gland inflammation. This study makes a better understanding of the mechanisms of coliform mastitis and provides a putative novel strategy for the prevention and/or treatment of mastitis.