眼针对腹泻型肠易激综合征模型大鼠结肠AQP8调节机制的研究

[目的]研究眼针对腹泻型肠易激综合征（D-IBS）模型大鼠结肠组织水通道蛋白8（AQP8）的表达的影响,探讨眼针对D-IBS模型大鼠结肠组织水液代谢的调控及其机制.[方法]32只雄性Wistar大鼠随机分成对照组、D-IBS模型组、眼针组、眼针＋VIP受体拮抗组（针抗组）,采用慢性应激与束缚相结合的方法建立D-IBS模型,免疫组织化学SABC法检测大鼠结肠组织AQP8的蛋白表达.[结果]与正常组相比,模型组AQP8的蛋白表达明显降低（P＜0.01）;与模型组相比,眼针组AQP8的蛋白表达显著升高（P＜0.01）;与眼针组相比,针抗组AQP8的蛋白表达显著下降（P＜0.01）.[结论]眼针治疗D-IBS的机制之一可能是通过上调结肠组织AQP8的蛋白表达,进而调控了结肠的水液代谢,其中AQP8的调控可能是通过VIP通路实现的.     

眼针疗法对D-IBS模型大鼠结肠VIPR1表达的影响

目的 观测眼针对腹泻型肠易激综合征(D-IBS)模型大鼠血清、结肠组织中血管活性肠肽(VIP)含量及结肠组织VIP受体1(VIP-Rl)表达的变化,探讨眼针治疗D-IBS的作用机制.方法 30只雄性SPF级大鼠,分为对照组、IBS模型组和眼针组,采用慢性应激与束缚相结合的方法复制D-IBS模型后,进行眼针治疗7d.应用ELISA方法检测VIP含量；采用免疫组化、RT-PCR方法检测VIP-R1表达.结果 与对照组比较,模型组血清、结肠组织中VIP含量及VIP-R1表达明显增高和上调(P ＜0.01,P＜0.05)；与模型组比较,眼针组血清、结肠组织中VIP含量及VIP-R1表达明显下降和下调.结论 眼针治疗D-IBS的机制之一可能与抑制血清、结肠组织VIP释放、下调VIP-R1表达有关.     

眼针对腹泻型肠易激综合征模型大鼠结肠水通道蛋白3表达的影响

目的:研究眼针对腹泻型肠易激综合征(D-IBS)模型大鼠结肠血管活性肠肽(VIP)和水通道蛋白3(AQP3)表达的影响,探讨眼针对D-IBS模型大鼠结肠水液代谢的调控作用和机制.方法:SPF级Wistar大鼠30只,随机分为对照组、D-IBS模型组和眼针组,每组10只.采用慢性应激结合束缚方法建立D-IBS大鼠模型.采...     

健脾化湿颗粒对D-IBS模型大鼠脑-肠轴中CRF和CRFR1的影响

目的:从结肠、海马及下丘脑中促肾上腺皮质激素释放因子(corticotropin releasing factor,CRF)和CRF受体1(CRF receptor 1,CRFR1)角度探讨健脾化湿颗粒改善腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)模型大鼠结肠运动和内脏敏感性的作用机制.方法:采用番泻叶灌胃结合束缚应激法建立D-I B S大鼠模型,应用健脾化湿颗粒进行干预,采用酶联免疫法(ELISA)检测大鼠结肠中CRF含量,采用免疫组织化学法检测结肠中CRFR1及海马、下丘脑中CRF,CRFR1阳性表达,采用RT-PCR法检测结肠、海马中CRF m RNA和CRFR1 m RNA的表达水平.结果:与正常组相比,模型组结肠中CRF含量(67.1±3.8 vs 36.0±3.0),海马、下丘脑中CRF阳性表达(0.23±0.02 vs 0.09±0.01,0.17±0.02 v s 0.09±0.01)明显升高(P0.01);结肠、海马、下丘脑中C R F R1阳性表达(0.17±0.01 vs 0.03±0.01,0.20±0.02 vs 0.09±0.01,0.19±0.02 vs 0.07±0.01)明显升高(P0.01);结肠、海马中C R F m RNA和CRFR1 m RNA的表达(结肠:0.89±0.04 vs 0.09±0.01,1.09±0.09 vs 0.21±0.04;海马:0.56±0.01 vs 0.15±0.05,1.26±0.14 vs 0.23±0.06)显著升高(P0.01).与模型组相比,各治疗组结肠、海马中C R F(51.0±3.4,54.6±4.1,45.1±4.7,43.3±3.9 vs 67.1±3.8;0.18±0.02,0.19±0.02,0.15±0.02,0.11±0.01 vs 0.23±0.02)显著下降(P0.01),阳性对照组、中、高剂量组下丘脑中CRF(0.15±0.02,0.13±0.01,0.12±0.01 vs 0.17±0.02)下降显著(P0.05,P0.0 1);阳性对照组、中、高剂量组结肠、海马、下丘脑中C R F R1表达(结肠:0.10±0.01,0.08±0.01,0.05±0.01 vs 0.17±0.01;海马:0.16±0.01,0.14±0.02,0.13±0.01 vs 0.20±0.02;下丘脑:0.15±0.02,0.13±0.01,0.11±0.01 vs 0.19±0.02)下降显著(P0.05,P0.01);结肠中CRF m RNA表达(0.63±0.04,0.76±0.06,0.32±0.06,0.13±0.03 v s 0.89±0.04)及中、高剂量组海马中CRF m RNA表达(0.76±0.11,0.67±0.10 v s 1.09±0.09)显著降低(P0.01);阳性对照组、中、高剂量组结肠中C R F R1m RNA表达(0.47±0.03,0.40±0.06,0.24±0.06 vs 0.56±0.01)及中、高剂量组海马中CRFR1 m RNA表达(0.62±0.06,0.60±0.07vs 1.26±0.14)显著降低(P0.05,P0.01).结论:健脾化湿颗粒可能通过下调结肠、海马及下丘脑中CRF、CRFR1表达来改善D-IBS模型大鼠结肠运动和内脏敏感性.     

眼针与体针对急性脑缺血再灌注损伤大鼠脑皮质脑源性神经营养因子表达的影响

目的分别观察眼针与体针对急性脑缺血再灌注损伤大鼠脑源性神经营养因子(BDNF)表达的影响,探讨眼针疗法与体针疗法的效应差异性,为眼针疗法自成诊疗体系提供相关依据。方法线栓法制备SD大鼠大脑中动脉梗死(MCAO)再灌注模型。将SD大鼠随机分为六组,即正常对照组、假手术组、模型组、眼针穴区组、眼针穴区外组与体针组。采用Zea Longa评分法进行大鼠神经功能评分;实时定量PCR方法检测缺血再灌注3 h后脑皮质BDNF mRNA的表达;Western印迹法检测缺血再灌注3 h后脑皮质BDNF蛋白的表达。结果脑缺血再灌注3 h后,眼针穴区组和体针组大鼠神经功能缺损评分较模型组大鼠降低(P0.01),眼针穴区组与体针组相比差异无统计学意义(P0.05);眼针穴区组、体针组和眼针穴区外组较模型组大鼠脑皮质BDNF mRNA和蛋白质含量均升高(P0.01),但以眼针穴区组和体针组升高较明显,眼针穴区组与体针组比较无统计学差异(P0.05)。结论眼针与体针在改善急性脑缺血再灌注损伤中的效应相当,且两种疗法的作用机制可能与上调脑皮质BDNF表达有关。     

肠易激综合征模型大鼠血清胰高糖素样肽-1及结肠组织中其受体的变化

目的 观察肠易激综合征( IBS)不同亚型模型大鼠血清胰高糖素样肽(GLP)-1及结肠组织中GLP-1受体的变化,初步探讨GLP-1及其受体在IBS发病中的作用.方法 40只雄性SD大鼠均分为腹泻型IBS(D-IBS)模型组、灌肠对照组、便秘型IBS(C-IBS)模型组、灌胃对照组及空白对照组.乙酸加束缚应激法制备D-IBS模型,冰水灌胃法制备C-IBS模型.观察大鼠粪便变化,检测粪便重量、粪便含水量及大鼠小肠推进率,给予结直肠扩张(CRD)刺激,记录腹外斜肌放电活动(EMG),评价模型大鼠的内脏敏感性.酶联免疫法测定各组大鼠血清中活性GLP-1的含量.免疫组织化学法、实时定量PCR法及Western印迹法检测各组大鼠近端结肠及远端结肠组织中GLP-1 受体的分布和表达.结果 与各自的对照组及空白对照组相比,D-IBS模型组大鼠粪便湿重、粪便含水量及小肠推进率均上升(P＜0.05);C-IBS模型组粪便湿重、粪便含水量及小肠推进率均降低(P＜0.05).在压力为20、40及60 mm Hg(1 mm Hg=0.133 kPa)的结直肠扩张刺激下各模型组大鼠腹外斜肌放电幅值均较各对照组明显增加,且D-IBS模型组高于C-IBS模型组(P＜0.05).C-IBS模型组血清中活性GLP-1的水平高于D-IBS模型组(P＜0.05),IBS模型组和对照组之间差异无统计学意义.GLP-1受体主要分布在结肠黏膜组织、环肌层及肌间神经丛中.C-IBS模型组结肠组织中GLP-1受体mRNA及蛋白表达量显著高于灌胃对照组,D-IBS模型组结肠组织中表达量低于灌肠对照组(P＜0.05).结论 不同亚型IBS结肠组织中GLP-1受体的表达水平不同,血清GLP-1水平也不同,提示GLP-1及其受体的改变可能与IBS不同亚型的发生有关.     

美沙拉嗪对溃疡性结肠炎大鼠结肠组织TLR4表达的影响

目的探讨美沙拉嗪对溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠组织TLR4表达的影响。方法选取30只SD大鼠随机分为正常对照组、UC组及治疗组。后两组均采用三硝基苯磺酸(TNBS)诱导制备UC大鼠模型。成功建模2 d后,治疗组采用美沙拉嗪+蒸馏水混悬液3 ml灌胃,正常对照组、UC组仅用单纯蒸馏水3 ml灌胃,连续灌注14 d后,处死三组大鼠,对三组大鼠的疾病活动指数、肠黏膜及组织学损伤进行评估,RT-PCR反应测定结肠组织TLR4 m RNA的表达;蛋白质印迹法测定TLR4蛋白的表达。结果 UC组大鼠疾病活动指数、肠黏膜及组织学损伤评分均显著高于正常对照组和治疗组(P0.05)。UC组及治疗组大鼠结肠组织TLR4 m RNA及蛋白表达水平显著高于正常对照组(P0.05)。与UC组比较,治疗组大鼠结肠组织TLR4 m RNA及蛋白表达均显著降低,差异有统计学意义(P0.05)。结论美沙拉嗪对UC大鼠结肠组织TLR4表达有抑制作用,可有效减少肠组织TLR4 m RNA的表达,缓解UC大鼠结肠组织损伤,有效保护UC大鼠结肠组织黏膜。     

辣椒素对水浸-束缚应激大鼠胃动力的作用及机制研究

目的探讨辣椒素(capsaicin,CAP)对水浸-束缚应激(water immersion restraint stress,WIRS)大鼠胃动力的作用及机制。方法雄性SD大鼠30只,随机分为3组:正常对照组、WIRS模型组、CAP干预组。其中正常对照组自由摄食进水;WIRS模型组每日WIRS 1 h后自由摄食进水;CAP干预组每日WIRS 1 h后喂食CAP饲料。4周后所有大鼠给予酚红溶液灌胃后麻醉处死,测定胃酚红排空率;采用ELISA法检测血浆胃动素(motilin,MTL)水平;采用免疫组化方法检测胃窦辣椒素受体(transient receptor potential vanilloid1,TRPV1)、P物质(substance P,SP)的表达;用RT-PCR方法检测c-kit m RNA、SCF m RNA的转录水平。结果大鼠胃酚红排空率:WIRS模型组胃酚红排空率显著低于其余两组(P0.05);正常对照组胃酚红排空率与CAP干预组相比,差异无统计学意义(P0.05)。血浆MTL水平:WIRS模型组血浆MTL水平显著低于其余两组(P0.05);正常对照组血浆MTL水平显著低于CAP干预组(P0.05)。免疫组化结果:大鼠胃窦组织TRPV1表达积分:CAP干预组TRPV1表达水平显著高于其余两组(P0.05);正常对照组TRPV1表达水平与WIRS模型组相比,差异无统计学意义(P0.05)。大鼠胃窦组织SP表达积分:WIRS模型组SP表达水平显著低于其余两组(P0.05);正常对照组SP表达水平显著低于CAP干预组(P0.05)。RT-PCR方法检测结果:大鼠胃窦组织c-kit m RNA表达结果:WIRS模型组c-kit m RNA转录水平显著低于正常对照组(P0.05);正常对照组、WIRS模型组c-kit m RNA转录水平与CAP干预组相比,差异无统计学意义(P0.05)。大鼠胃窦组织SCF m RNA表达结果:WIRS模型组SCF m RNA转录水平显著低于其余两组(P0.05);正常对照组SCF m RNA转录水平显著低于CAP干预组(P0.05)。结论 CAP可能对WIRS大鼠胃动力具有改善作用,其机制可能与CAP调节TRPV1、SP的表达及MTL的释放有关,是否与Cajal间质细胞相关尚不明确。     

化瘀通便汤对慢传输型便秘大鼠结肠P物质和血管活性肽的影响

[目的]探讨化瘀通便汤对慢传输型便秘(STC)大鼠结肠P物质(SP)和血管活性肽(VIP)表达的影响。[方法]72只雄性大鼠随机分为空白对照组、模型对照组、化瘀通便汤低、中、高剂量组、西药对照组,每组12只,采用大黄粉灌胃制造慢传输型便秘模型,RT-PCR法检测化瘀通便汤对慢传输型便秘大鼠结肠SP和VIP mRNA表达情况。[结果]STC大鼠结肠SP表达显著增高、VIP表达显著降低(P0.01),化瘀通便汤高、中、低剂量组不同程度下调SP表达、上调VIP表达(P0.01)。[结论]STC大鼠便秘的发生可能与结肠组织中SP表达增高、VIP表达降低有关,化瘀通便汤治疗STC的机制可能与下调结肠组织中SP,上调结肠组织中VIP的表达有关。     

疏肝健脾方对腹泻型肠易激综合征大鼠的治疗作用及对肥大细胞、5-HT通路的影响

[目的]探讨疏肝健脾方对腹泻型肠易激综合征(diarrhea-predominant pattern irritable bowel syndrome,D-IBS)大鼠的治疗作用及肥大细胞、5-HT通路的影响。[方法]1日龄SD雄性大鼠随机分为正常组、模型组、疏肝健脾方组。采用母婴分离联合束缚应激方法建立D-IBS大鼠模型。造模成功后,疏肝健脾方组给予疏肝健脾方灌胃(2.33g·100g~(-1)·d~(-1)),正常组和模型组则给予等体积的生理盐水灌胃。灌胃结束后,测定各组大鼠粪便含水量、直结肠球囊扩张刺激时腹壁撤退反射(abdominal withdrawal reflex,AWR)评分及腹外斜肌肌电(electromyography,EMG)积分变化率。并分别采用甲苯胺蓝染色检测结肠肥大细胞表达,酶联免疫法(ELISA)测定结肠5-HT水平,蛋白免疫印记法(Western blot,WB)检测结肠5-HT3R(5-hydroxytryptamine receptor 3,5-HT3R)、5-HT4R(5-hydroxytryptamine receptor 4,5-HT4R)蛋白表达。[结果]疏肝健脾方组大鼠粪便含水量较模型组降低(P0.05);直结肠内球囊压力在40、60、80mmHg(1mmHg=0.133kPa)时,AWR评分及EMG积分变化率较模型组均降低(P0.05);疏肝健脾方组结肠肥大细胞数目、5-HT含量、5-HT3AR及5-HT3BR蛋白表达较模型组均降低(P0.05),5-HT4R蛋白表达较模型组升高(P0.05)。[结论]疏肝健脾方对D-IBS大鼠有较好治疗作用,其作用机制可能与降低结肠肥大细胞数目、5-HT的含量、5-HT3AR及5-HT3BR的表达,升高5-HT4R的表达有关。     

Anti-malarial efficacy of pyronaridine and artesunate in combination in vitro and in vivo

Pyronaridine is a Mannich base anti-malarial with demonstrated efficacy against drug resistant Plasmodium falciparum, P. vivax, P. ovale and P. malariae. However, resistance to pyronaridine can develop quickly when it is used alone but can be considerably delayed when it is administered with artesunate in rodent malaria models. The aim of this study was to evaluate the efficacy of pyronaridine in combination with artesunate against P. falciparum in vitro and in rodent malaria models in vivo to support its clinical application. Pyronaridine showed consistently high levels of in vitro activity against a panel of six P. falciparum drug-sensitive and resistant strains (Geometric Mean IC50=2.24 nM, 95% CI=1.20-3.27). In vitro interactions between pyronaridine and artesunate showed a slight antagonistic trend, but in vivo compared to pyronaridine and artesunate administered alone, the 3:1 ratio of the combination, reduced the ED90 of artesunate by approximately 15.6-fold in a pyronaridine-resistant P. berghei line and by approximately 200-fold in an artesunate-resistant line of P. berghei. Complete cure rates were achieved with doses of the combination above or equal to 8 mg/kg per day against P. chabaudi AS. These results indicate that the combination had an enhanced effect over monotherapy and lower daily doses of artesunate could be used to obtain a curative effect. The data suggest that the combination of pyronaridine and artesunate should have potential in areas of multi-drug resistant malaria.     

Modulation of enterotoxin binding and function in vitro and in vivo

The use of the nontoxic B subunits of cholera and Escherichia coli enterotoxins in vitro and in vivo led to a decrease in toxin binding to target cells and a decrease in toxin-induced effects (i.e., morphological effects, adenylate cyclase activation, and fluid secretion). The reduction in toxin binding involves a process of down-regulation of cellular receptors for the toxin and not toxin occupancy of receptors. The extent of inhibition was dependent on the amount of B subunit used and on the duration of time after its use. Thus, in vivo exposure to a single bolus of B subunit was sufficient to block toxin binding and activity for up to 18 h. Because the B subunit binds extensively to the esophagus and the stomach, peroral administration will require a preparation that allows the subunit to reach the small bowel in a protected form. Our data provide a rationale for using B subunit therapy for short-term protection against the effects of enterotoxins, before the development of an immune response.     

我国高血压防治现状和策略

<正>我国现患高血压2亿人,由于人群高血压患病率的不断升高和防控力度不够,我国高血压人群的知晓率、治疗率、控制率仍处于较低水平。对于像高血压这样的群体性慢性病,应当采取全人     

Cooperation of B- and T-lymphocytes of the human in vitro and in vivo

It is reported on the experimental proofs for the existence of a cooperation of different populations of lymphocytes in man. Regulatory lymphocytes play a part in the regulation of the synthesis of immunoglobulins by polyclonally stimulated B-lymphocytes, in the generation of killer-T-cells and in the regulation of the DNA-synthesis by mitogenically stimulated T- and B-cells. Typical helper- and suppressor-effects may be proved. Disturbances of lymphocytic interactions may be a cause for the development of immune deficiency diseases. It is very probable that also in several chronic infections a dysfunction of regulatory T-lymphocytes is present.     

Uptake of labeled alloxan in mouse organs and mitochondria in vivo and in vitro

[14C]2-Alloxan was administered in vivo and in vitro for study of the uptake of alloxan in different organs and their mitochondia of mice. After in vivo administration, radioactivity was demonstrated in all organs investigated, with quantitative differences: endocrine pancreas greater than liver greater than exocrine pancreas and heart. No significant difference was found between the iv and ip routes of injection. An in vivo uptake of alloxan was also found in mitochondria, with significant quantitative differences as to the origin of the organelles: endocrine pancreas greater than liver greater than exocrine pancreas and heart. Pretreatment with D-glucose caused significantly decreased uptake in liver, exocrine pancreas, and heart, but significantly increased uptake in endocrine pancreas, whereas the uptake was significantly decreased in the mitochondria from all of these organs. In vitro uptake was observed in all kinds of mitochondria studied. This uptake was higher than the in vivo uptake in mitochondria from liver, exocrine pancreas, and heart, whereas the uptake in vivo was higher than the in vitro uptake in islet mitochondria. The presence of D-glucose did not affect the in vitro uptake of alloxan in mitochondria. The findings show that in vivo, alloxan passes across plasma membranes and is taken up by mitochondria, and data obtained with mitochondrial subfractions may also indicate a passage across mitochondrial membranes. D-Glucose protection against alloxan diabetogenicity may be associated with prevention of mitochondrial uptake of alloxan. This prevention seems to be dependent on the metabolism of glucose.