Objective:To conduct a glucagon like peptide-1(GLP-1)controllability model rat by chemical genetics, and observe the impact of GLP-1 neuron excitability on appetite.Methods:Fifteen rats were evenly divided into Green fluorescent protein(GFP)group, HM3D group, and HM4D group. Various combinations of adeno-associated virus(rAAV)were injected into the nucleus tractus solitarius(NTS). rAAV-GLP-1-cre and rAAV-GFP-dio were administered in rats of GFP group. The rats of HM3D group were injected with rAAV-GLP-1-cre and rAAV-HM3D-mCherry-dio while rAAV-GLP-1-cre and rAAV-HM4D-mCherry-dio were injected in rats of HM4D group . The optimal dose of clozapine N-oxide(CNO)was selected based on feeding behavior and body weight changes of rats after intraperitoneal injection of different doses of CNO. The controllability of GLP-1 neurons was confirmed by comparing with intraperitoneal injection of saline. The number of activated GLP-1 neurons in the NTS area and the expression of POMC neurons in the hypothalamus were detected 30 minutes after CNO injection.Results:GLP-1 neurons in the NTS area of rats were successfully labeled. The rat of HM3D group revealed a decrease in food intake( P=0.021)while the rat of HM4D group showed an increase( P=0.002), when given 1 mg/kg of CNO, no changes at the dose of 0.5 mg/kg and 3.0 mg/kg. Immunofluorescence showed that the activity of GLP-1 neurons in NTS of GFP group was lower than that of HM3D group( P=0.022), and higher compared with that of the HM4D group( P=0.049). The expression of GLP-1 neurons in NTS and POMC neurons in the hypothalamus of the HM3D group after intraperitoneal injection of CNO was also higher than that in the HM4D group( P=0.003). Conclusion:Using chemical genetics technology, GLP-1 controllability model rat could be successfully established via injecting varying combinations of rAAV into the NTS area of rat. Injection of 1 mg/kg CNO can effectively activate or inhibit the neuron to regulate appetite.

目的:观察电针对肥胖大鼠胰岛素敏感性及脂联素的影响,探讨电针调节脂联素的可能机制.方法:正常组、模型组、电针组大鼠各10只,正常组给予普通饲料喂养,模型组给予高脂饲料喂养,电针组在高脂饲料喂养的基础上给予电针治疗,观察各组大鼠观察治疗前后体质量、空腹及餐后血糖、血清胰岛素、血清脂联素水平的变化,观察干预7周后各组大鼠的腹腔糖耐量和腹腔胰岛素耐量,分别检测各组大鼠白色脂肪组织S1RT1、FoxO1、Adiponectin的蛋白和基因表达水平.结果:电针组的体质量上升幅度小于模型组,肥胖大鼠接受电针后胰岛素敏感性及脂联素水平上升;电针组中SIRT1、FoxO1、Adiponectin蛋白表达高于模型组,低于正常组;正常组SIRT1、FoxO1的基因表达高于模型组和电针组,模型组和电针组之间比较差异无统计学意义,但电针组的脂联素基因表达水平高于模型组.结论:电针能够明显改善高脂饮食诱导肥胖大鼠的胰岛素抵抗状态,其作用机制可能与电针调控白色脂肪组织中SIRT1/FoxO1通路进而影响脂联素的基因表达有关.

目的 观察电针对单纯性肥胖大鼠体重及脂质代谢的影响.方法 20只单纯性肥胖模型大鼠随机分为肥胖组和电针组,两组均给予高脂喂养,电针组在高脂喂养的基础上,取穴中脘、丰隆、足三里给予电针治疗,疗程为8周.检测两组0、2、4、6、8周的体重及血清血清游离脂肪酸(NEFA)、甘油三酯(TG)、胆固醇(TC)水平.结果 两组在8周干预过程中体重仍呈上升趋势,但电针组体重增长在第6周时被抑制(P＜0.05).8周后,电针组NEFA、TG、TC水平明显低于肥胖组(P＜0.05).结论 电针能够通过调节脂质代谢,改善大鼠的单纯性肥胖状态.

Objective To investigate the effectiveness of three different anesthetic techniques in intraventricular catheterization and its effect on the survival rate of rats. Methods Thirty Wistar rats were equally allocated into 3 groups:chloral hydrate group,pentobarbital sodium group and isoflurane group. Intraventricular catheterization was performed in the rats after anesthesia with i. p. injection of chloral hydrate and pentobarbital sodium, and isoflurane inhalation, respectively. Levels of blood glucose were detected before and at 15 and 30 minutes and 1, 3, 7, 14, 28 days after anesthesia. Body mass and 24-hour food intake were recorded before and at 1, 3, 7 days after anesthesia. The onset time and effective time of anesthesia, operation time and the survival rates on 30 days of the rats were compared and analyzed. Results The onset time and effective time of anesthesia, and the operation time in the isoflurane group were shorter than that in the chloral hydrate group, while these parameters in this group were shorter than that in the pentobarbital sodium group. Blood glucose in the chloral hydrate group was apparently increased during the surgical operation, while the body mass, 24-hour food intake and blood glucose were decreasing since one day after operation, and all the rats in this group died during the 30-day observation, mainly, due to enteroplegia. Blood glucose in the pentobarbital sodium group was mildly increased after anesthesia, while the body mass, 24-hour food intake and blood glucose were mildly decreased at one day after operation and recovered within one week. In this group, 3 rats died of respiratory distress due to overdose anesthesia and one rat died during the 30 day-observation. The blood glucose in the isoflurane group was mildly increased after operation, while the 24-hour food intake and blood glucose did not markedly changed, the body mass was stably increased, and no rat died during the 30-day-observation. Conclusions Intraperitoneal injection of chloral hydrate is not suitable for intraventricular catheterization in rats. Intraperitoneal injection of pentobarbital sodium can be only carefully applied for intraventricular catheterization under poorly-limited conditions. Isoflurane inhalation anesthesia is recommended for intraventricular catheterization in rats.

目的 观察不同剂量黄芪对羟基脲致排卵障碍大鼠血清性激素水平、卵巢和子宫指数、血清抗苗勒激素的影响,探讨黄芪改善大鼠排卵的可能机制.方法 除正常组外,运用羟基脲灌胃法对未交配的成熟雌性SD大鼠造模,将造模成功排卵障碍大鼠随机分为模型组、克罗米芬组和黄芪高、中、低剂量组.用药干预28d后,于动情期间处死大鼠.观察大鼠一般情况,测定治疗前后体重;心尖取血测定血清FSH、LH、E2及抗苗勒激素(AMH)水平;剥离子宫和卵巢称湿重,计算卵巢和子宫指数.结果 造模后大鼠出现不同程度倦怠、活动度下降,体重低于正常组(P＜0.01);克罗米芬组治疗后体重仍低于正常组(P＜0.01);黄芪治疗组大鼠的活动度不同程度好转,体重显著高于克罗米芬组(P＜0.05,P＜0.01).模型组卵巢指数显著低于正常组(P＜0.01),克罗米芬组和黄芪高、中剂量组卵巢指数均高于模型组(P＜0.05);模型组子宫指数低于正常组(P＜0.05),黄芪高剂量组和克罗米芬组子宫指数均高于模型组(P＜0.05).模型组大鼠血清E2低于正常组(P＜0.05),黄芪高、中剂量组和克罗米芬组E2水平高于模型组(P＜0.05).模型组血清AMH水平低于正常组(P＜0.05),治疗后仅黄芪高剂量组AMH水平高于模型组(P＜0.05),其它组无显著性变化.结论 高剂量黄芪能够更好地改善排卵障碍大鼠的肾虚症状和性激素水平,提高卵巢储备能力,综合效应优于克罗米芬.

目的:观察电针调控肝脏FoxO1磷酸化及其对肝脏脂质代谢的影响,探讨电针改善肥胖调控脂质代谢的可能机制.方法:将20只高脂饲料诱导的肥胖大鼠和10只普通饲料喂养的大鼠分为正常组(n=10)、模型组(n=10)、电针组(n=10).正常组给予普通饲料喂养,模型组给予高脂饲料喂养,电针组在高脂饲料喂养的基础上给予电针治疗.取穴为中脘、关元、足三里、丰隆,电针频率2 Hz,强度0.5～1 mA,每周治疗3次,共治疗8周.观察各组大鼠治疗前后的体质量、lee's指数的变化;观察各组大鼠治疗后的血清TC、TG、VLDL的水平;通过Western Blotting检测各组大鼠肝脏FoxO1、p-FoxO1的蛋白表达水平;通过Realtime-PCR法检测肝脏MTP、ApoCⅢ的基因表达水平.结果:所有大鼠的体质量均较干预前上升,但电针组的体质量上升幅度小于模型组(P＜0.05);正常组与模型组的lee's指数较干预前无明显变化,电针组Lee's指数较治疗前下降(P＜0.05),亦明显低于模型组(P＜0.05).模型组大鼠血清TC、TG、VLDL水平显著高于正常组,电针组TC、TG、VLDL水平低于模型组(P＜0.05).模型组大鼠的FoxO1和p-FoxO1水平显著低于正常组(P＜0.05),电针组FoxO1水平高于模型组、低于正常组(P＜0.05),p-FoxO1水平高于模型组(P＜0.05),与正常组无显著性差异(P＞0.05).正常组大鼠的MTP、ApoCⅢ基因表达水平高于模型组和正常组(P＜0.05),电针组MTP、ApoCⅢ基因表达水平高于模型组(P＜0.05).结论:电针能够上调肝脏FoxO1的蛋白表达,特别是能够明显上调FoxO1的磷酸化水平,FoxO1磷化后从细胞核内转移至胞浆,进而介导下游载脂蛋白MTP、ApoCⅢ的基因表达水平被下调,VLDL蛋白合成减少,最终导致TC、TG向细胞外的转运减少,VLDL的构建被抑制,达到调控指质代谢的目的.