基于胃Cajal间质细胞自噬探讨柴胡疏肝散促进功能性消化不良大鼠胃动力的作用机制

发布时间：2018-03-01 01:13

本文关键词： 柴胡疏肝散功能性消化不良 Cajal间质细胞自噬　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:基于胃Cajal间质细胞(interstitial cells of cajal,ICC)自噬探讨柴胡疏肝散促进功能性消化不良(functional dyspepsia,FD)大鼠胃动力的作用机制。方法:将60只成年健康SD大鼠按随机分组法分为正常对照组、模型组、柴胡疏肝散低剂量(0.16 g/ml)组(柴低组)、柴胡疏肝散中剂量(0.32g/ml)组(柴中组)、柴胡疏肝散高剂量(0.64 g/ml)组(柴高组),每组12只。除外正常对照组,余各组均采取改良郭氏夹尾刺激法构建FD大鼠模型并予相应药物灌胃,共连续4w。半固体糊灌胃法测算各组大鼠胃排空率;正常光线下肉眼观察大鼠胃组织;对大鼠胃组织行苏木精-伊红染色(hematoxylin-eosin staining,HE)并在光学显微镜观察胃组织结构以完成病理检查;透射电镜观察胃ICC超微结构;免疫荧光法定位胃ICC;免疫荧光法及原位杂交法分别检测大鼠胃ICC内Beclin1、微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3B,LC3B)蛋白及m RNA的表达。结果:1.与正常对照组比较,模型组大鼠胃排空率降低(P0.01);与模型组比较,柴高组、柴中组大鼠胃排空率升高(P0.01);与柴中组比较,柴高组胃排空率升高(P0.05),柴低组胃排空率下降(P0.05)。2.各组大鼠胃组织肉眼观察均未见明显糜烂及溃疡,病理检查各组大鼠胃组织均未见明显炎性细胞浸润、结构异常。3.正常对照组ICC内线粒体结构正常,粗面内质网丰富,偶见自噬泡;模型组、柴低组ICC内细胞结构出现异常,线粒体肿胀明显,粗面内质网颗粒脱落,可见大量自噬泡;柴高组、柴中组ICC内细胞内结构未见明显异常,线粒体肿胀不明显,粗面内质网少量脱颗粒,仅见少量自噬泡。4.与正常对照组比较,模型组ICC内Beclin1、LC3B蛋白及m RNA的表达均升高(P0.01);与模型组比较,柴高组、柴中组ICC内Beclin1、LC3B蛋白及m RNA的表达均下降(P0.01);与柴中组比较,柴高组ICC内Beclin1、LC3B蛋白及m RNA的表达均下降(P0.05),柴低组ICC内Beclin1、LC3B蛋白及m RNA的表达均升高(P0.05)。结论:1.FD大鼠胃动力障碍的机制可能与胃ICC内发生过度自噬有关。2.柴胡疏肝散可能是通过抑制胃ICC过度自噬进而促进FD大鼠胃动力,其机制可能与下调Beclin1蛋白及m RNA的表达从而抑制细胞自噬活性并减少LC3B蛋白及m RNA以及自噬泡的表达有关。
[Abstract]:Objective: to explore the mechanism of Chaihu Shugan Powder promoting gastric motility in rats with functional dyspepsia based on interstitial cells of cajalicum. Methods: sixty adult healthy SD rats were randomly divided into normal control group and model group. The low dose of Chaihu Shugan San 0.16 g / ml (Chaihu Shugan Powder 0.32 g / ml) group (Chaihu Shugan Powder 0.32 g / ml) group (Chaihu Shugan Powder 0.64 g / ml) group (Chaigao group, 12 rats in each group), except the normal control group, The FD rat model was established by modified Guo's clip tail stimulation method, and the gastric emptying rate of each group was measured by semi-solid paste method for 4 weeks, and the gastric tissue was observed with naked eyes under normal light. The gastric tissues of rats were stained with hematoxylin-eosin stinging-hehe (hematoxylin-eosin stinging-hehe), and the histological structure of the stomach was observed under optical microscope to complete the pathological examination, and the ultrastructure of gastric ICC was observed by transmission electron microscope (TEM). The expression of Beclin1, microtubule-associated protein 1, microtubule-associated protein 1 light chain 3BHLC3B and m RNA in gastric ICC of rats were detected by immunofluorescence and in situ hybridization respectively. Compared with the model group, the gastric emptying rate of the model group decreased P0.01A; compared with the model group, the gastric emptying rate increased in the Chaigao group and the Chaizhong group; and compared with the Caizhong group, the gastric emptying rate increased. The gastric emptying rate in Chaigao group increased P0.05A, and the gastric emptying rate decreased P0.05.2.There was no obvious erosion and ulcer in gastric tissue of rats in each group, and no inflammatory cell infiltration was found in gastric tissue of rats in each group by pathological examination. In the normal control group, the mitochondrial structure was normal, the rough endoplasmic reticulum was abundant, and the autophagy was occasionally seen in the normal control group, while in the model group, the cellular structure in ICC was abnormal, the mitochondria swelling was obvious, and the coarse endoplasmic reticulum granules were falling off in the model group. A large number of autophagy was observed in Chaigao group and Chaizhong group. The intracellular structure of ICC was not abnormal, mitochondria swelling was not obvious, coarse endoplasmic reticulum was degranulated a little, and only a few autophagy vesicles were observed in Chaigao group, compared with normal control group. The expression of Beclin1 ICC LC3B protein and m RNA increased in model group, and the expression of Beclin1 + LC3B protein and m RNA in ICC group decreased compared with model group, and compared with model group, the expression of m RNA and Beclin1 + LC3B protein decreased significantly in model group, and compared with that in model group, the expression of m RNA in ICC group decreased significantly than that in model group. The expression of Beclin1 ICC LC3B protein and m RNA decreased in Chaigao group, and the expression of Beclin1hl LC3B protein and m RNA increased in ICC of chaigao group. Conclusion 1. The mechanism of gastric motility disorder in FD rats may be related to excessive autophagy in gastric ICC. Chaihu Shugan San. It can promote gastric motility of FD rats by inhibiting gastric ICC hyperautophagy. The mechanism may be related to the down-regulation of the expression of Beclin1 protein and m RNA, thereby inhibiting the autophagy activity and reducing the expression of LC3B protein, m RNA and autophagy.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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