摘要:目的:观察杏仁中央核(CeA)γ-氨基丁酸(GABA)能神经元及下丘脑室旁核(PVN)在胃俞募配穴针刺调节胃功能中的作用,基于CeA(GABA)-PVN环路探讨胃俞募配穴针刺调节胃功能的中枢机制。方法:实验分3部分进行:第1部分选取C57BL/6小鼠,随机分为正常组和电针组(电针"中脘"和右侧"胃俞"20 min),每组6只,用免疫荧光染色法检测CeA、PVN区c-fos表达;第2部分用GAD2-Cre小鼠4只,在CeA区注射顺行示踪病毒(rAAV-EF1α-DIO-mcherry-WPRE-pA),观察CeA中GABA能神经元是否投射到PVN区;第3部分选取GAD2-Cre小鼠,随机分为空载病毒组(CeA注射rAAV-EF1α-DIO-mcherry-WPRE-pA)、化学遗传激活病毒组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA]、化学遗传激活病毒+电针组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA+电针"中脘"和右侧"胃俞"20 min],每组6只,记录各组小鼠的进食量,检测胃排空率,同时用高效液相色谱法测定PVN区GABA含量。结果:1)与正常组比较,电针组CeA、PVN区c-fos表达均明显增加(P<0.01)。2)小鼠脑区切片可见病毒在CeA区GABA能神经元胞体大量表达,在PVN区可见大量有病毒表达的投射纤维,表明CeA中GABA能神经元可投射到PVN区,即CeA到PVN有直接的纤维联系。3)与空载病毒组比较,化学遗传激活病毒组小鼠进食量和胃排空率明显下降(P<0.01),PVN区GABA含量明显升高(P<0.01);与化学遗传激活病毒组比较,化学遗传激活病毒+电针组进食量和胃排空率明显增加(P<0.01),PVN区GABA含量明显下降(P<0.01)。结论:电针胃俞募配穴对胃功能的调控可通过CeA(GABA)-PVN环路探讨胃俞募配穴针刺调节胃功能的中枢机制。方法:实验分3部分进行:第1部分选取C57BL/6小鼠,随机分为正常组和电针组(电针"中脘"和右侧"胃俞"20 min),每组6只,用免疫荧光染色法检测CeA、PVN区c-fos表达;第2部分用GAD2-Cre小鼠4只,在CeA区注射顺行示踪病毒(rAAV-EF1α-DIO-mcherry-WPRE-pA),观察CeA中GABA能神经元是否投射到PVN区;第3部分选取GAD2-Cre小鼠,随机分为空载病毒组(CeA注射rAAV-EF1α-DIO-mcherry-WPRE-pA)、化学遗传激活病毒组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA]、化学遗传激活病毒+电针组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA+电针"中脘"和右侧"胃俞"20 min],每组6只,记录各组小鼠的进食量,检测胃排空率,同时用高效液相色谱法测定PVN区GABA含量。结果:1)与正常组比较,电针组CeA、PVN区c-fos表达均明显增加(P<0.01)。2)小鼠脑区切片可见病毒在CeA区GABA能神经元胞体大量表达,在PVN区可见大量有病毒表达的投射纤维,表明CeA中GABA能神经元可投射到PVN区,即CeA到PVN有直接的纤维联系。3)与空载病毒组比较,化学遗传激活病毒组小鼠进食量和胃排空率明显下降(P<0.01),PVN区GABA含量明显升高(P<0.01);与化学遗传激活病毒组比较,化学遗传激活病毒+电针组进食量和胃排空率明显增加(P<0.01),PVN区GABA含量明显下降(P<0.01)。结论:电针胃俞募配穴对胃功能的调控可通过CeA(GABA)-PVN环路实现。Objective To explore the effect of γ-aminobutyric acid(GABA)ergic neuronal circuit of the central amygdaloid nucleus(CeA) and the paraventricular nucleus of hypothalamus(PVN) on electroacupuncture(EA)-induced regulation of gastric function by way of CeA-PVN projection. Methods The present study included 3 parts: 1) C57 BL/6 mice were randomly divided into control and EA groups(n=6 in each group). EA was applied to right "Weishu"(BL21, Back-shu point) and "Zhongwan"(CV12, Front-mu point) for 20 min, followed by detecting the expression of c-fos in the CeA and PVN by using immunofluorescence staining; 2) Microinjection of anterograde tracer(rAAV-EF1α-DIO-mcherry-WPRE-pA) into the CeA was conducted in GAD2-Cre mice for confirming the projection of GABAergic neurons from CeA to PVN; 3) GAD2-Cre mice were randomly divided into rAAV-DIO-mcherry(intra-CeA injection of rAAV-EF1α-DIO-mcherry-WPRE-pA), rAAV-DIO-hM3 D(Gq)-mcherry(intra-CeA injection of rAAV-EF1α-DIO-hM3 D(Gq)-mcherry-WPRE-pA) and rAAV-DIO-hM3 D(Gq)-mcherry+EA groups(n=6 in each group). The food intake and gastric empty were detected, and the concentration of GABA in the PVN was assayed by using high performance liquid chromatography on the 28(GABA)-PVN环路实现。
摘要:目的:观察电针对腹泻型肠易激综合征(IBS-D)大鼠肠上皮黏膜屏障功能的影响,探讨电针干预IBS-D的可能机制。方法:将SD大鼠随机分为空白组、模型组、电针组和药物组,每组10只,雌雄各半。采用慢性不可预知性温和应激联合番泻叶浸剂灌胃法建立IBS-D大鼠模型。电针组交替干预单侧"足三里""天枢""三阴交""太冲"穴,15 min/d,药物组予2.7 mg/mL匹维溴胺灌胃(10 mL·kg(-1)·d(-1)·d(-1)),均治疗14 d。采用内脏疼痛阈值和腹泻指数评价各组大鼠肠道症状;免疫组织化学法检测各组大鼠结肠组织紧密连接蛋白Claudin-1、Occludin表达水平;分光光度法检测各组大鼠血浆二胺氧化酶(DAO)活性。结果:与空白组比较,模型组大鼠腹泻指数及血浆DAO活性显著升高(P<0.01),内脏疼痛阈值、结肠Claudin-1及Occludin表达显著降低(P<0.01)。与模型组比较,电针组和药物组腹泻指数及血浆DAO活性显著降低(P<0.01),内脏疼痛阈值、结肠Claudin-1及Occludin表达显著升高(P<0.05,P<0.01)。结论:电针可以明显改善IBS-D大鼠腹痛、腹泻等肠道症状,其机制可能与增加肠上皮紧密连接蛋白Claudin-1和Occludin表达水平,降低肠道通透性,恢复肠上皮黏膜屏障功能有关。Objective To observe the effect of electroacupuncture(EA) on intestinal epithelial mucosal barrier function in diarrhea-predominant irritable bowel syndrome(IBS-D) rats, so as to explore its mechanisms underlying improvement of IBS-D.Methods Forty SD rats(half male and half female) were randomly divided into control, model, EA and medication(Pinaverium Bromide, PB) groups, with 10 rats in each group. The IBS-D model was established by chronic unpredictable mild stress combined with gavage of Senna-leaf solution. EA(2 Hz/15 Hz,0.1-1 mA) was applied to unilateral "Zusanli"(ST36),"Tianshu"(ST25), "Sanyinjiao"(SP6) and "Taichong"(LR3) alternatively for 15 min, once daily for 14 days. Rats of the medication group was treated by gavage of PB(10 mL·kg(-1)),均治疗14 d。采用内脏疼痛阈值和腹泻指数评价各组大鼠肠道症状;免疫组织化学法检测各组大鼠结肠组织紧密连接蛋白Claudin-1、Occludin表达水平;分光光度法检测各组大鼠血浆二胺氧化酶(DAO)活性。结果:与空白组比较,模型组大鼠腹泻指数及血浆DAO活性显著升高(P<0.01),内脏疼痛阈值、结肠Claudin-1及Occludin表达显著降低(P<0.01)。与模型组比较,电针组和药物组腹泻指数及血浆DAO活性显著降低(P<0.01),内脏疼痛阈值、结肠Claudin-1及Occludin表达显著升高(P<0.05,P<0.01)。结论:电针可以明显改善IBS-D大鼠腹痛、腹泻等肠道症状,其机制可能与增加肠上皮紧密连接蛋白Claudin-1和Occludin表达水平,降低肠道通透性,恢复肠上皮黏膜屏障功能有关。
摘要:目的:观察慢性萎缩性胃炎(CAG)大鼠胃黏膜淤血与缺氧诱导因子(HIF)-1α、血管内皮生长因子(VEGF)表达的关系,以及穴位埋线对两者的影响,探讨穴位埋线改善CAG的机制。方法:将健康成年雄性SD大鼠分为空白组10只、模型组9只和埋线组10只。采用自由饮用100μg/mL N-甲基-N’-硝基-N-亚硝基胍溶液配合饥饱失常法制备CAG大鼠模型。埋线组大鼠予"中脘"及双侧"足三里""脾俞"穴埋线治疗,10 d 1次,共治疗6次。采用HE染色法观察各组大鼠胃黏膜病理情况,采用Western blot法检测各组大鼠胃黏膜HIF-1α、VEGF蛋白表达水平。结果:HE染色显示,空白组大鼠胃黏膜结构清晰,胃腺丰富;模型组大鼠黏膜固有层局部腺体发生萎缩、减少和变性坏死,部分伴有黏膜淤血;埋线组可见部分胃黏膜上皮细胞脱落、胞质淡染,腺体扩张、坏死。与空白组相比,模型组腺体坏死、萎缩与黏膜淤血大鼠数量显著增多(P<0.001,P<0.05);与模型组相比,埋线组腺体坏死、萎缩与黏膜淤血大鼠数量显著减少(P<0.01,P<0.05)。模型组共有5只大鼠存在胃黏膜淤血情况,设为淤血组,另4只设为非淤血组;与淤血组相比,非淤血组胃黏膜HIF-1α、VEGF蛋白表达降低(P<0.01)。与空白组相比,模型组胃黏膜HIF-1α、VEGF蛋白表达升高(P<0.01);与模型组相比,埋线组胃黏膜HIF-1α、VEGF蛋白表达降低(P<0.05,P<0.01)。结论:胃黏膜淤血与HIF-1α、VEGF蛋白的异常表达密切相关,穴位埋线可能通过降低HIF-1α、VEGF蛋白表达,从而发挥治疗CAG的作用。
摘要:目的:基于磁共振体素内不相干运动成像(IVIM)技术观察针刺联合高压氧(HBO)对一氧化碳中毒迟发性脑病(DEACMP)患者脑血流灌注的影响,从而间接评价其疗效。方法:将28例DEACMP患者随机分为对照组和针刺组,每组14例。对照组给予常规治疗及HBO治疗,针刺组在对照组基础上联合醒脑开窍针刺法治疗,每日1次,5 d为1个疗程,疗程间隔2 d,共治疗6个疗程。治疗前、治疗后行常规头颅磁共振功能成像(MRI)平扫、IVIM扫描,测量双侧半卵圆中心伪扩散系数(D*)、灌注分数(f)值,获得血流相关参数fD*)、灌注分数(f)值,获得血流相关参数fD*值;采用Barthel指数评分评估患者日常生活能力;采用Pearson法评价参数值间的相关性。结果:与治疗前比较,治疗后两组DEACMP患者f、fD*值;采用Barthel指数评分评估患者日常生活能力;采用Pearson法评价参数值间的相关性。结果:与治疗前比较,治疗后两组DEACMP患者f、fD*值与Barthel指数评分均升高(P<0.05)。治疗后,与对照组比较,针刺组f、fD*值与Barthel指数评分均升高(P<0.05)。治疗后,与对照组比较,针刺组f、fD*值与Barthel指数评分升高更显著(P<0.05)。f、fD*值与Barthel指数评分升高更显著(P<0.05)。f、fD*值与Barthel指数评分两两之间均呈正相关(P<0.05),其中f与fD*值与Barthel指数评分两两之间均呈正相关(P<0.05),其中f与fD*值相关系数r=0.822,f值与Barthel指数评分相关系数r=0.636,fD*值相关系数r=0.822,f值与Barthel指数评分相关系数r=0.636,fD*值与Barthel指数评分相关系数r=0.061。结论:IVIM技术可以对脑血流灌注作定量评估,针刺联合HBO疗法可明显改善早期DEACMP患者日常生活能力和脑血流灌注情况,其疗效优于单纯HBO治疗。Objective To observe the effect of acupuncture plus hyperbaric oxygen(HBO) on cerebral blood perfusion in patients with delayed encephalopathy after carbon monoxide poisoning(DEACMP).Methods Twenty-eight patients with DEACMP were randomly divided into acupuncture group(n=14 cases) and control group(n=14 cases). Patients of the acupuncture group were treated by using "Xingnao Kaiqiao" needling technique(specific set of acupuncture points as Yintang [EX-HN3], Shangxing [GV23], Baihui [GV20], Sishenchong [EX-HN1], Fengchi [GB20], Taichong [LR3], etc., and strong stimulation) combined with HBO in an air pressurized tank, and those of the control group treated by simple HBO. The treatment was conducted once a day, 5 days a week for 6 weeks. All the patients underwent head routine magnetic resonance imaging(MRI) and intravoxel incoherent motion imaging(IVIM) scan before and after the treatment. The values of pseudo-diffusion coefficient D(D*值与Barthel指数评分相关系数r=0.061。结论:IVIM技术可以对脑血流灌注作定量评估,针刺联合HBO疗法可明显改善早期DEACMP患者日常生活能力和脑血流灌注情况,其疗效优于单纯HBO治疗。