电针胆下合穴阳陵泉等对AC模型豚鼠α7nAchR及TNF-α的影响

发布时间：2018-01-06 13:12

本文关键词：电针胆下合穴阳陵泉等对AC模型豚鼠α7nAchR及TNF-α的影响　出处：《湖南中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:对比观察电针急性胆囊炎(AC)模型豚鼠阳陵泉、上巨虚、下巨虚、足三里穴对胆囊组织中肿瘤坏死因子-α(TNF-α)、烟碱型乙酰胆碱受体α7(α7nAchR)及血清中的高迁移率族蛋白B1(HMGB1)表达水平的影响,探讨胆之下合穴阳陵泉治疗胆腑病是否存在相对特异性。方法:将72只健康SD豚鼠随机分为空白、模型、阳陵泉、上巨虚、下巨虚、足三里组共六组,每组12只,雌雄各半。除空白组外均胆囊注射大肠杆菌建立豚鼠急性胆囊炎模型,造模成功并治疗5天后,取胆囊组织进行HE染色,观察各组豚鼠胆囊形态学变化,运用Western blot法检测豚鼠胆囊组织α7 nAchR受体的表达,免疫组化法检测胆囊组织中TNF-α的表达,ELISA法检测豚鼠血清中HMGB1水平。结果:1.光镜下胆囊形态学的观察(10x40倍),结果显示:空白组:粘膜上皮细胞为单层柱状上皮,排列整,固有膜较薄,含部分小血管和淋巴管,固有膜及粘膜下层无淤血扩张,基层排列不规整,浆膜完整,无炎症反应。模型组:有大量粘膜上皮坏死脱落,充血水肿,固有层有大量炎性细胞浸润。阳陵泉组:粘膜上皮大部分完好,固有层松弛,有充血水肿,部分淋巴细胞浸润,有较少中性粒浸润,肌层和浆膜层充血。其他穴位组:粘膜上皮有部分脱落,固有层及粘膜下层及肌层有中性粒细胞浸润,血管明显扩张淤血。2.各组豚鼠组织α7nAch R含量检测,结果显示:与空白组比,余组胆囊组织中α7nAchR表达水平均明显偏低(P0.01),差异有显著的统计学意义(P0.01)。电针治疗后与模型组比较,各电针治疗组胆囊组织中α7nAchR表达均明显偏高(P0.01),差异有显著的统计学意义(P0.01)。与阳陵泉比较,上巨虚、下巨虚、足三里组豚鼠胆囊组织中α7nAchR表达均偏低,差异有统计学意义(P0.01或P0.05)。3.各组豚鼠血清HMGB1含量检测,结果显示:与空白组比,血清中HMGB1含量表达均明显偏高,差异有显著的统计学意义(P0.01)。与模型组比较,各电针治疗组血清中HMGB1含量表达均明显偏低,差异有显著的统计学意义(P0.01)。与阳陵泉比较,上巨虚、下巨虚、足三里组血清中HMGB1含量表达均偏高,差异有显著的统计学意义(P0.01)。4.各组组织中TNF-α的表达检测,结果显示:与空白组比,胆囊组织中TNF-α表达均明显偏高,差异有显著的统计学意义(P0.01)。电针治疗后与模型组比,胆囊组织中TNF-α表达均明显偏低,差异有显著的统计学意义(P0.01)。与阳陵泉组比,上巨虚组、下巨虚组、足三里组胆囊组织中TNF-α表达均偏高,差异有统计学意义(P0.01或P0.05)。结论1.电针消化系相关的内腑下合穴对AC豚鼠均可产生不同程度的治疗作用,其机制可能是通过调节α7nAchR、TNF-α表达及HMGB1含量而达到抑制炎症因子、减轻炎症反应、保护AC模型豚鼠胆囊粘膜实现的2.对比可知电针阳陵泉对AC豚鼠炎症干预效果优于上巨虚、下巨虚及足三里穴,表明电针阳陵泉穴治疗AC具有一定的相对特异性。
[Abstract]:Objective: to observe the effect of electroacupuncture (EA) on tumor necrosis factor- 伪 (TNF- 伪) in gallbladder tissue of guinea pig (Yanglingquan, Shangjuxu, Xijuxu, Zusanli). Effects of nicotinic acetylcholine receptor 伪 7 (伪 7 nAchR1) and high mobility group protein B1 HMGB1 in serum. Methods: 72 healthy Sprague-Dawley guinea pigs were randomly divided into six groups: blank model, Yanglingquan, Shangjuxu, Xijuxu and Zusanli groups. The guinea pig model of acute cholecystitis was established by injecting Escherichia coli into the gallbladder except the blank group. The model was successfully established and treated for 5 days. The gallbladder tissue was stained with HE. The changes of gallbladder morphology in each group were observed, and the expression of 伪 7 nAchR receptor in gallbladder tissue of guinea pigs was detected by Western blot method. The expression of TNF- 伪 in gallbladder tissue was detected by immunohistochemical method and HMGB1 level in serum of guinea pigs was detected by Elisa. Results: the morphology of gallbladder was observed 10 x 40 times under light microscope). The results showed that in the blank group, the mucosal epithelial cells were monolayer columnar epithelium, arranged neatly, thin lamina propria, including some small blood vessels and lymphatic vessels, the lamina propria and submucous layer had no congestion and dilatation, and the basic layer was irregular. Model group: a large number of mucosal epithelial necrosis, hyperemia and edema, a large number of inflammatory cells infiltration in the lamina propria. Yanglingquan group: most of the mucosal epithelium intact, lamina propria relaxation. There were hyperemia and edema, some lymphocytic infiltration, less neutrophil infiltration, hyperemia in myometrium and serous layer. Other acupoints group: mucosal epithelium had partial exfoliation, neutrophil infiltration in lamina propria and submucous layer and myometrium. The content of 伪 7nAch R in guinea pig tissues of each group was detected. The results showed that compared with the blank group, the content of 伪 7nAch R was higher than that of the control group. The expression of 伪 7nAchR in gallbladder tissues in the remaining group was significantly lower than that in the model group (P 0.01), and the difference was statistically significant (P 0.01). After electroacupuncture treatment, the expression of 伪 7nAchR was compared with that of the model group. The expression of 伪 7nAchR in gallbladder tissues of each electroacupuncture treatment group was significantly higher than that of P0.01a, and the difference was statistically significant (P 0.01). Compared with Yangling Quan, the expression of 伪 7nAchR in the gallbladder tissue was significantly higher than that in the upper giant asthenia group and the lower giant asthenia group. The expression of 伪 7nAchR in gallbladder tissue of Zusanli group was lower than that of Zusanli group, and the difference was significant (P0.01 or P0.05). The serum HMGB1 content of each group was detected. The results showed that the expression of HMGB1 in serum was significantly higher than that in the blank group, and the difference was statistically significant (P 0.01), compared with the model group. The expression of HMGB1 in serum of each electroacupuncture treatment group was significantly lower than that of Yanglingquan group, and the difference was statistically significant (P 0.01). The expression of TNF- 伪 in serum of Zusanli group was higher than that of control group, and the difference was statistically significant (P 0.01). The results showed that the expression of TNF- 伪 was higher than that of blank group. The expression of TNF- 伪 in gallbladder tissue was significantly higher than that in the model group, and the difference was statistically significant (P 0.01). Compared with the model group, the expression of TNF- 伪 in gallbladder tissue was significantly lower than that in the model group. Compared with Yanglingquan group, the expression of TNF- 伪 in gallbladder tissues of upper giant asthenia group, lower giant asthenia group and Zusanli group was higher than that in Yanglingquan group. The difference is statistically significant P0.01 or P0.050.Conclusion 1. Electroacupuncture of the digestive system related to the internal fu-fu points can produce different degrees of treatment for AC guinea pigs. 2. The mechanism may be to inhibit the inflammatory factors and alleviate the inflammatory response by regulating the expression of 伪 7nAchR- 伪 TNF- 伪 and the content of HMGB1. The effect of electroacupuncture on inflammation of AC guinea pigs was better than that of upper giant asthenia, lower giant asthenia and Zusanli point. The results showed that the treatment of AC by electroacupuncture at Yanglingquan point had certain relative specificity.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245;R-332

【相似文献】