本文關鍵詞： 柴胡疏肝散 功能性消化不良 Cajal間質(zhì)細胞 自噬　出處：《廣西醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:基于胃Cajal間質(zhì)細胞(interstitial cells of cajal,ICC)自噬探討柴胡疏肝散促進功能性消化不良(functional dyspepsia,FD)大鼠胃動力的作用機制。方法:將60只成年健康SD大鼠按隨機分組法分為正常對照組、模型組、柴胡疏肝散低劑量(0.16 g/ml)組(柴低組)、柴胡疏肝散中劑量(0.32g/ml)組(柴中組)、柴胡疏肝散高劑量(0.64 g/ml)組(柴高組),每組12只。除外正常對照組,余各組均采取改良郭氏夾尾刺激法構建FD大鼠模型并予相應藥物灌胃,共連續(xù)4w。半固體糊灌胃法測算各組大鼠胃排空率;正常光線下肉眼觀察大鼠胃組織;對大鼠胃組織行蘇木精-伊紅染色(hematoxylin-eosin staining,HE)并在光學顯微鏡觀察胃組織結(jié)構以完成病理檢查;透射電鏡觀察胃ICC超微結(jié)構;免疫熒光法定位胃ICC;免疫熒光法及原位雜交法分別檢測大鼠胃ICC內(nèi)Beclin1、微管相關蛋白1輕鏈3B(microtubule-associated protein 1 light chain 3B,LC3B)蛋白及m RNA的表達。結(jié)果:1.與正常對照組比較,模型組大鼠胃排空率降低(P0.01);與模型組比較,柴高組、柴中組大鼠胃排空率升高(P0.01);與柴中組比較,柴高組胃排空率升高(P0.05),柴低組胃排空率下降(P0.05)。2.各組大鼠胃組織肉眼觀察均未見明顯糜爛及潰瘍,病理檢查各組大鼠胃組織均未見明顯炎性細胞浸潤、結(jié)構異常。3.正常對照組ICC內(nèi)線粒體結(jié)構正常,粗面內(nèi)質(zhì)網(wǎng)豐富,偶見自噬泡;模型組、柴低組ICC內(nèi)細胞結(jié)構出現(xiàn)異常,線粒體腫脹明顯,粗面內(nèi)質(zhì)網(wǎng)顆粒脫落,可見大量自噬泡;柴高組、柴中組ICC內(nèi)細胞內(nèi)結(jié)構未見明顯異常,線粒體腫脹不明顯,粗面內(nèi)質(zhì)網(wǎng)少量脫顆粒,僅見少量自噬泡。4.與正常對照組比較,模型組ICC內(nèi)Beclin1、LC3B蛋白及m RNA的表達均升高(P0.01);與模型組比較,柴高組、柴中組ICC內(nèi)Beclin1、LC3B蛋白及m RNA的表達均下降(P0.01);與柴中組比較,柴高組ICC內(nèi)Beclin1、LC3B蛋白及m RNA的表達均下降(P0.05),柴低組ICC內(nèi)Beclin1、LC3B蛋白及m RNA的表達均升高(P0.05)。結(jié)論:1.FD大鼠胃動力障礙的機制可能與胃ICC內(nèi)發(fā)生過度自噬有關。2.柴胡疏肝散可能是通過抑制胃ICC過度自噬進而促進FD大鼠胃動力,其機制可能與下調(diào)Beclin1蛋白及m RNA的表達從而抑制細胞自噬活性并減少LC3B蛋白及m RNA以及自噬泡的表達有關。
[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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5

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