重症急性胰腺炎外周血中性粒细胞凋亡检测的实验研究

目的初步探讨重症急性胰腺炎（SAP）外周血中性粒细胞凋亡的动态变化。方法采用胰胆管逆行注射4％牛磺胆酸钠建立大鼠SAP模型；观察血浆淀粉酶水平变化及胰腺组织病理评分；TUNEL荧光标记法检测外周血中性粒细胞凋亡。结果SAP组血浆淀粉酶及胰腺病理评分均较假手术组（SO）明显升高（P〈0．05）。SAP组中性粒细胞凋亡率均较SO组明显降低（P〈0．05），随时间延长更明显。结论重症急性胰腺炎外周血中性粒细胞凋亡明显延迟，在SAP的发病机制中可能具有重要作用。     

Effect of electroacupuncture on the bladder function in rats with bladder hyperactivity

目的：观察电针对排尿频率、骶髓背角血管活性肠肽（VIP）表达的影响,探讨针刺抑制大鼠左旋多巴诱发的膀胱机能亢进的神经生物学机制。方法：将48只SD雌性大鼠按随机数字表分成正常组6只,对照组14只,模型组14只,电针组14只。全部动物均施行膀胱插管术,正常组不做腹腔注射;对照组腹腔注射生理盐水;模型组和电针组动物腹腔内先后注射卡比多巴和左旋多巴;应用连续膀胱测压技术动态观察电针中膂俞对大鼠排尿频率的影响;应用免疫组织化学的方法,观察电针后骶髓背角的vIP动态变化,并用图像分析技术进行定量分析。结果：模型组在腹腔注射左旋多巴后排尿频率较处理前迅速增加（P〈0．05）,电针组在腹腔注射左旋多巴后15～75min时段排尿频率较处理前增加（P〈0．05）,75～105min时段排尿频率接近处理前水平。在腹腔注射左旋多巴后3h时电针组骶髓背角的VIP阳性纤维和终末比模型组、正常组显著增加（P〈0．05）。在腹腔注射左旋多巴后8h时模型组骶髓背角的VIP阳性纤维和终末较正常组显著增加（P〈0．05）,而电针组与正常组则无统计学差异。结论：电针可促进左旋多巴引起的膀胱机能亢进大鼠的躯体传入神经释放VIP,从而降低排尿频率。     

攻下清热活血中药对重症胰腺炎大鼠血清TNF-αIL-1β的影响

目的：观察攻下清热活血中药对重症胰腺炎大鼠血清TNF-α、IL-1β的影响。方法：胰胆管逆行注射建立大鼠模型后,分别以不同剂量的攻下清热活血中药灌胃治疗,以奥曲肽皮下注射为对照。取胰腺组织HE染色观察大鼠胰腺组织学改变,酶法测定大鼠血清淀粉酶含量,酶联免疫吸附法（ELISA法）检测血清中TNF-α、IL-1β的含量。结果：模型组大鼠胰腺组织病理评分及血清淀粉酶明显高于正常对照组（P〈0.01）;应用大、小剂量组中药和奥曲肽治疗后大鼠胰腺组织病理评分及血清淀粉酶较模型组明显下降（P〈0.05）。模型组大鼠血清TNF-α、IL-1β含量明显高于正常对照组（P〈0.05）;应用中药治疗后大、小剂量组和奥曲肽后大鼠血清TNF-α、IL-1β含量较模型组明显下降（P〈0.05）。结论：攻下清热活血中药能明显改善重症胰腺炎大鼠胰腺组织病理和血清淀粉酶,抑制大鼠血清TNF-α、IL-1β含量可能是其作用机理之一。     

益气养血法对小鼠胃肠动力影响的实验研究

目的：观察以益气养血法组方药物对正常及便秘模型小鼠胃肠动力的影响。方法：用酚红法检测胃排空功能；墨水推进法检测小肠、结肠运动功能；用NADPH．黄递酶（NDP）组化技术和分层铺片方法检测结肠肌间神经丛一氧化氮合酶（NOS）阳性神经元的灰度值。用复方地芬诺酯制备便秘动物模型。结果：益气养血法组方药物3个剂量组的小鼠胃排空率，小肠和结肠推进率与正常对照组相比都有显著性差异（P〈0．001～0．05）。便秘状态下3个剂量组的小鼠胃排空率，结肠推进率和结肠肌间神经丛NOS阳性神经元的灰度值与模型组相比都有显著性差异（P〈0．001～0．05）。结论：以益气养血法组方药物对小鼠胃肠动力具有促进作用，可调节便秘小鼠结肠肌间神经丛NOS阳性神经元的兴奋性。     

神经降压素介导的脑-肠轴在电针治疗功能性消化不良大鼠中的作用

目的:探讨神经降压素(neurotensin,NT)介导的脑-肠轴在电针治疗功能性消化不良(FD)大鼠中的作用。方法:SD大鼠48只按照随机数字表法分为空白组、模型组、电针组,每组16只。采用夹尾刺激法配合隔日进食制备FD大鼠模型。电针组电针右侧"足三里"和"太冲"穴,每次30min,每日1次,共14d。检测胃排空率和小肠推进率,运用ELISA法检测血浆中NT含量,应用免疫组织化学法检测下丘脑、胃窦及回肠组织中NT的表达。结果:模型组较空白组大鼠胃排空率明显降低(P0.01),小肠推进率显著下降(P0.01);电针组较模型组胃排空率增加,小肠推进率升高(P0.05)。模型组血浆NT含量较空白组增加(P0.05),电针组较模型组明显减少(P0.05)。模型组下丘脑、胃窦及回肠组织中NT表达较空白组升高(P0.05),电针组较模型组降低(P0.05)。结论:电针可明显降低FD大鼠下丘脑、胃窦、回肠及血浆中NT的表达,通过中枢及外周两种途径介导脑-肠轴作用,加快胃排空及小肠推进,改善胃肠动力障碍,该作用可能是电针治疗FD的重要机制之一。     

电针足三里对牛磺胆酸钠诱导的重症急性胰腺炎大鼠炎症反应及小肠SIgA表达的影响

目的 探讨电针足三里对牛黄胆酸钠诱导的重症急性胰腺炎（SAP）大鼠炎症反应及肠道免疫功能的影响。方法 将72只SD雄性大鼠随机分为电针组、模型组、假手术组,每组24只。通过逆行胰胆管注射3.5%牛磺胆酸钠制作SAP模型。电针组于模型制作成功后和处死前,固定在操作台上给予双侧“足三里”穴电针治疗各30 min。模型组及假手术组仅固定30 min不针刺。各组大鼠分别于造模成功后3、6、12 h分批处死,用酶联免疫吸附法测定大鼠血清白细胞介素-4(IL-4)、白细胞介素-10(IL-10)、转化生长因子-β1(TGF-β1)和干扰素-γ（IFN-γ）的水平,苏木精和伊红染色观察各组胰腺组织、小肠组织病理变化并进行评分,并用免疫组化法检测小肠分泌型免疫球蛋白A(SIgA)的表达水平。结果 电针组术后3、6、12 h血清IL-4、IL-10、IFN-γ含量均低于模型组（P <0.05）,血清TGF-β1含量均高于模型组（P <0.05）;假手术组及电针组各时间点胰腺病理评分均低于模型组（P <0.05）;假手术组各时点小肠病理评分明显低于模型组（P <0.05）,电针组小...     

四磨汤口服液对胃肠运动功能障碍模型大鼠胃残留、小肠推进率及结肠P物质的影响

目的：观察四磨汤口服液对胃肠运动功能障碍模型大鼠胃残留率、小肠推进率及结肠P物质（SP）的影响．方法：6f）只大鼠随机分为空白组,模型组,四磨汤口服液高、中、低剂量组以及西药组,按夹尾法配合饥饱失常法造模后给予相应药物灌胃,检测半固体营养糊在体内的胃残留率和小肠推进率,按免疫组化要求染色并观察sP的阳性面积。结果：与空白组比较,模型组大鼠胃残留率明显升高以及小肠推进率显著降低（P〈0．05）;四磨汤口服液高剂量组较模型组大鼠胃残留率降低、小肠推进率增加（P〈0．05）;模型组结肠SP表达水平较空白组显著降低（P〈0.05）;四磨汤口服液高剂量组与西药组结肠SP阳性表达均较模型组增高（P〈0．05）。结论：夹尾法配合不规则喂养可导致大鼠胃肠运动紊乱,四磨汤口服液能促进胃肠运动障碍模型大鼠胃肠蠕动功能的恢复,能够增高胃肠功能障碍大鼠兴奋性神经递质SP的表达,可能是其调整胃肠运动功能障碍的作用机制之一。     

电针“足三里”穴对重症急性胰腺炎大鼠小肠闭锁蛋白和核因子-κB表达的影响

目的:探讨电针"足三里"穴调控急性胰腺炎大鼠肠道炎性反应的作用机制。方法:将54只雄性SD大鼠随机分为重症急性胰腺炎(SAP)模型组、电针组、假手术组,每组18只。通过逆行胆胰管注射3.5%牛磺胆酸钠制作SAP模型。模型制作成功后电针组在双侧"足三里"穴给予电针治疗30min。假手术组及模型组大鼠在同一时间固定30min不予治疗。各组大鼠均于造模后3h、6h、12h分批处死,观察胰腺以及小肠病理学改变,并用免疫组化SP法检测小肠上皮组织闭锁蛋白(occludin protein)以及核因子-κB(NF-κB p65)表达水平。结果:各时间点胰腺病理评分假手术组、电针组均低于模型组(P0.05);小肠上皮组织occludin蛋白表达假手术组与电针组均高于模型组(P0.05);小肠上皮组织NF-κB p65表达假手术组与电针组均低于模型组(P0.05)。结论:电针"足三里"穴可以降低SAP模型大鼠小肠上皮组织NF-κB p65表达,提高小肠上皮组织occludin蛋白表达,从而减轻胰腺损伤。     

点刺与电针委中穴对家兔腰椎间盘突出症影响的对照研究

目的：观察点刺与电针委中穴对实验性腰椎间盘突出症的影响。方法：50只健康新西兰家兔随机分为5组，即空白组、LIDP模型组、点刺委中组、电针委中组和点刺非穴对照点组。自制的LIDP动物病理模型造模器造实验性LIDP病理模型，采用Siegal记录触觉和步态功能，酶联免疫吸附（ELISA）法与放射免疫方法测定腰椎间盘髓核组织中PLA2与IL-1α的含量。结果：点刺委中组与电针委中组可改善家兔的触觉与步态功能，降低LIDP家兔髓核组织中PLA2与IL-1α含量，且点刺委中穴LIDP家兔髓核组织中PLA2的含量与电针委中穴组比较有显著性差异（P〈0．05）。结论：点刺与电针委中穴治疗LIDP的机制之一可能是抑制髓核组织中炎症介质尤其是PLA2的含量，其点刺放血的作用优于电针。     

胰腺穴对急性胰腺炎的诊断与治疗之临床研究

目的：观察胰腺穴对急性胰腺炎的诊断、治疗的临床意义。方法：按压及针刺胰腺炎，有于诊断、治疗急性胰腺炎，设立针刺组及西药对照组；系统观察分析胰腺穴在诊断急性胰腺炎、消除腺痛、镇痛持续时间及降低血清淀粉酶作用的比较。结果：胰腺穴组在诊断和治疗方面较对照组有明显的优势，经统计学处理，P＜0．05。在治愈时间上，二者比较无显著性意义，P＞0．05。结论：胰腺穴是诊断、治疗急性胰腺炎的特效腧穴。     

Effect of point-moxibustion on ghrelin and GHSR-1a expressions in lateral septal nucleus of rats with diabetic gastroparesis

Objective

To observe the effect of point-moxibustion on gastrointestinal motility, mRNA and protein expressions of ghrelin and growth hormone secretagogue receptor 1a (GHSR-1a) in lateral septal nucleus of rats with diabetic gastroparesis (DGP), and to investigate the central regulatory mechanism of DGP treatment with point-moxibustion.

Methods

Forty SPF-grade Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, an electroacupuncture (EA) group and a point-moxibustion group, with 10 rats in each group. A DGP rat model was established by intraperitoneal injection of 2% streptozotocin (STZ) with 8-week irregular high-sugar and high-fat diet in the model group, the EA group and the point-moxibustion group; and rats in the blank group were injected intraperitoneally with 0.1 mmoL/L (pH 4.5) citric acid-sodium citrate buffer with 8-week normal diet. Eight weeks later, rats in the EA group were treated by EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6); while rats in the point-moxibustion group were treated by point-moxibustion at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) for successive 15 d. Rats in the blank group and the model group were fixed as the control without intervention. After treatment, intestinal propulsion rate and gastric emptying rate were measured. The mRNA and protein expressions of ghrelin and GHSR-1a in the lateral septal nucleus were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB).

Results

Compared with the blank group, the intestinal propulsion rate and gastric emptying rate of the model group were significantly lower (both P<0.01); compared with the model group, the intestinal propulsion rate and gastric emptying rate of the EA group and the point-moxibustion group increased significantly (all P<0.05). The mRNA and protein expressions of ghrelin and GHSR-1a were lower in the model group than those in the blank group (all P<0.01). The mRNA and protein expressions of ghrelin and GHSR-1a were significantly higher in the EA group and the point-moxibustion group than those in the model group (all P<0.05). There were no statistically significant differences between the EA group and the point-moxibustion group (all P>0.05).

Conclusion

Point-moxibustion at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) can increase the intestinal propulsion rate and gastric emptying rate of DGP rats, and promote the mRNA and protein expressions of ghrelin and GHSR-1a in the central nervous system. The mechanism may be related to the activation of ghrelin pathway in hypothalamic arcuate nucleus-lateral septal nucleus.

     

EFFECT OF ELECTROACUPUNCTURE OF "ZUSANLI" ON VIP CONTENTS OF THE PERIPHERAL BLOOD, GASTRIC MUCOSAL AND BRAIN TISSUES IN GASTRIC MUCOSAL INJURY RATS

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针刺对胶原诱导性关节炎大鼠胃肠动力的影响

目的：利用制备的胶原诱导性关节炎(CIA)大鼠模型,探讨针刺足三里对CIA大鼠胃肠动力的影响。方法：将60只12周龄雄性大鼠随机分为4组：正常组、模型组、电针组、手针组,每组15只,建立牛Ⅱ型胶原与不完全弗氏佐剂诱导CIA大鼠模型。电针组和手针组于造模完成后第1天取足三里穴治疗,15 min/次,1次/d,连续治疗14 d。在3 d, 7 d, 14 d时,采用跖围法测大鼠跖围长度,并在治疗14 d后,用苏木精-伊红(HE)染色观察膝关节的病理学变化,计算大鼠的胃排空率和肠推进率,结肠蠕动波测定方法检测大鼠的肠蠕动波次数,通过蛋白质免疫印迹法检测胃黏膜中促肾上腺皮质激素释放因子(CRF)和血管活性肠肽(VIP)的表达。结果：与正常组比较,模型组大鼠跖关节肿胀明显(P<0.01),软骨组织破坏严重并有炎症细胞浸润,胃排空率和肠推进率均降低(P<0.01),胃黏膜中CRF和VIP表达明显增多(P<0.01)。与模型组比较,针刺后大鼠跖关节肿胀程度明显降低(P<0.05),软骨破坏程度减轻和炎症细胞浸润明显减少,胃排空率和肠推进率提升(P<0.05),肠蠕动波次...     

Effect of electroacupuncture on electrogastrogram and gastric antrum ghrelin in rats with diabetic gastroparesis

Objective

To observe the effect of electroacupuncture (EA) on the electrogastrogram and gastric antrum ghrelin in rats with diabetic gastroparesis (DGP).

Methods

Fifty Sprague-Dawley (SD) rats were randomly divided into group A, group B, group C, group D and group E, 10 rats in each group. Group A was the blank control group without intervention. Group B, Group C, Group D and Group E were treated with single dose intraperitoneal injection of 2% streptozotocin (STZ), combined with 8-week high glucose and high fat diet to establish DGP rat models. Group B was the model group without treatment. Group C was the EA at acupoint group, was treated with EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Group D was the EA at non-acupoint group, was treated by EA at the control points of Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Rats in the metoclopramide control group received 1.7% metoclopramide solution [10 mL/(kg·bw)] by gavage. Rat’s blood glucose level was measured by blood glucose meter; gastric emptying rate was detected using phenol red as a marker; the electrogastrogram was detected by BL-420F biological function system; the protein level of ghrelin was detected by enzyme-linked immunosorbent assay (ELISA); the expression of ghrelin mRNA was detected by real-time polymerase chain reaction (RT-PCR).

Results

Compared with group A, the blood glucose of group B, C, D and E were significantly increased before and after the treatment (all P<0.01); after treatment, the gastric emptying rate of group B was significantly decreased (P<0.01), the migration rates of small intestine in group B, C, D and E were significantly lower (all P<0.01), and the protein content of ghrelin in group C was significantly decreased (P<0.01); the expressions of ghrelin mRNA were significantly increased in group B, C, D and E (all P<0.01), the mean amplitudes of electrogastrogram in group B and D were significantly decreased (both P<0.01). After treatment, compared with group B, the blood glucose of group C was significantly decreased (P<0.05), the gastric emptying rate and small intestine migration rate were significantly increased in group C and E (P<0.05, P<0.01), the small intestinal migration rate was significantly increased in group D (P<0.05), the expression of ghrelin in protein and mRNA in group C was significantly lower (P<0.01), the expression of ghrelin mRNA in group E was significantly lower (P<0.05), and the mean amplitude of electrogastrogram in group C was significantly increased (P<0.05). After treatment, compared with group D, the protein and mRNA expressions of ghrelin in group C were significantly decreased (P<0.01). After treatment, compared with group E, the protein expression of ghrelin in group C was significantly decreased (P<0.01).

Conclusion

EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) could regulate the blood glucose level of DGP model rats, enhance electrogastrogram activity, promote gastric emptying, and regulate ghrelin expression in protein and mRNA.

     

电针足阳明经穴对家兔胃粘膜损伤细胞保护作用的研究

本实验采用无水乙醇灌胃造成胃粘膜损伤后 ,分别电针足阳明经“四白”、“梁门”、“足三里”三个不同段代表穴 ,及“足三里”外 2cm的对照点 ,观察其对胃粘膜损伤后的细胞保护作用 ,以证实足阳明经与胃的相关性。结果发现 ,分别电针“四白”、“梁门”、“足三里”7日后 ,均能使胃粘膜损伤指数显著降低。电针“足三里”后 ,胃液及胃粘膜PGE2 与其他各组比较均显著增高 (P <0 0 5或 0 0 1 ) ,血清NO与模型组、电针“足三里”外 2cm组比较亦见明显增高 (P <0 0 5) ;电针“四白”、“梁门”、“足三里”以及空白组胃粘膜EGF与未经电针的模型组比较均有显著差异 (P <0 0 5或0 0 1 )。提示 ,电针家兔足阳明经不同节段的腧穴均对胃粘膜损伤细胞有保护作用 ,其中以“足三里”最满意 ,说明足阳明经与胃具有相关性 ,与此同时 ,同一经脉的穴位对相关脏腑的作用亦有着相对的特异性     

电针治疗肠黏膜损伤大鼠基本腧穴配伍“肠病方”的筛选

目的:选用临床治疗肠病最常用的3个腧穴即中脘、天枢、上巨虚,通过观察不同组合配伍的效应,选出最优的穴位组合。方法:SD大鼠126只,按随机数字表分为空白组、模型组、中脘组、天枢组、上巨虚组、中脘+上巨虚组、天枢+上巨虚组、中脘+天枢组、中脘+天枢+上巨虚组。采用8%冰乙酸灌肠法造模。各治疗组在相应穴位施以电针,每日1次,共治疗3次。治疗结束后,分别观察各组大鼠的结肠黏膜损伤指数、病理学损伤积分及超微结构变化。结果:与空白组相比,模型组结肠黏膜损伤指数显著降低(P<0.05),病理学损伤积分显著升高(P<0.05);7个电针组与模型组比较,结肠黏膜损伤指数显著升高(P<0.05),病理学损伤积分显著降低(P<0.05);天枢+中脘+上巨虚组与其它6个电针组相比,两项指标变化更显著(P<0.05)。超微结构观察表明,各电针组结肠黏膜损伤程度较模型组减轻,天枢+中脘+上巨虚组更加明显。结论:同时电针"中脘"天枢"上巨虚"减轻结肠黏膜损伤的作用优于其它单穴或双穴使用,因而可作为治疗肠病的基本处方。     

电针“足三里”对急性胰腺炎大鼠血清促炎因子及胰腺核因子-κB活性的影响

目的:探讨电针"足三里"穴治疗重症急性胰腺炎(SAP)的作用机制。方法:66只雄性SD大鼠随机分为假手术组、模型组、电针组,每组22只。通过胆胰管注射3.5%牛磺胆酸钠制作SAP模型。电针组在模型制作成功后及处死前给予电针"足三里"穴治疗各30 min。3组大鼠均于造模后3 h(n=7)、6 h(n=7)、12 h(n=8)分批处死,检测各时间点腹水量,观察胰腺病理学评分变化,并用ELISA方法检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6浓度的变化,运用免疫组化法检测胰腺组织核因子(NF)-κB P 65表达水平。结果:各时间点模型组的胰腺组织病理评分、腹水量、血清TNF-α和IL-6浓度及胰腺NF-κB P65表达水平均较假手术组增高(均P<0.05);电针组各时间点上述各项指标均较模型组明显降低(均P<0.05)。结论:电针"足三里"穴可以减轻牛磺胆酸钠诱导的SAP大鼠胰腺病理损伤,其机制可能与抑制NF-κB的活性、降低血清促炎因子TNF-α、IL-6浓度有关。     

大柴胡汤对急性坏死性胰腺炎大鼠胰腺水通道蛋白1的作用

目的:研究大柴胡汤对急性坏死性胰腺炎(ANP)大鼠胰腺水通道蛋白1(AQP1)的作用。方法:72只雄性SD大鼠随机分为对照组、ANP组、0.9%氯化钠溶液(NS)组和大柴胡汤组,造模后3h、6h、12h时间点每组分别处死6只。测定血清淀粉酶、血清AQP1含量;胰腺组织HE染色并按Schmidt方法对其进行严重度评分;伊文思蓝(EB)染料血管外渗法检测组织毛细血管通透性;免疫组化及Westen Blot检测胰腺组织AQP1蛋白表达。结果:大柴胡汤组腹水量明显低于ANP组(P<0.05);各时间点ANP组血清淀粉酶水平高于对照组(P<0.01);ANP组胰腺组织EB含量显著高于对照组(P<0.05);ANP组胰腺免疫组化及WB检测AQP1蛋白含量均低于对照组(P<0.05);大柴胡汤组血清淀粉酶水平较ANP组显著降低(P<0.05);大柴胡汤组胰腺组织病理改变较ANP组减轻;胰腺组织EB含量低于ANP组(P<0.05),免疫组化及WB检测AQP1蛋白表达均显著低于ANP组(P<0.05)。结论:大柴胡汤预处理可以改善ANP大鼠模型的疾病严重程度,降低毛细血管通透性,其机制可能与上调ANP大鼠胰腺AQP1的表达有关。