本文選題：動物模型 + 功能性消化不良��； 參考：《北京中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的探討功能性消化不良中醫(yī)核心證型—脾虛證的生物學(xué)機制,進一步闡釋并完善中醫(yī)脾虛證相關(guān)理論。方法理論方面,首先對功能性消化不良臨床文獻進行整理,探討功能性消化不良中醫(yī)證型的分布規(guī)律,并對出現(xiàn)頻次較高的中醫(yī)證型的癥狀表現(xiàn)進行分析。挖掘疾病的核心癥狀。在此基礎(chǔ)上,對疾病主癥—“脹”,“滿”相關(guān)文獻進行中醫(yī)古籍整理,梳理“腹脹”的源流及中醫(yī)病機。同時,對“腹脹”西醫(yī)文獻進行整理,探討“腹脹”產(chǎn)生的信號通路,尋找其可能的生物學(xué)機制。研究還梳理了以“腹脹”,“腹?jié)M”為主癥的中醫(yī)疾病源流,尋找最符合腹脹型功能性消化不良的中醫(yī)病名,并研究此疾病的中醫(yī)學(xué)病機。研究另外對“痞滿”相關(guān)的方劑及中草藥文獻進行整理,探討治療“痞滿”的中醫(yī)用藥規(guī)律。進而指導(dǎo)功能性消化不良脾虛證的中醫(yī)用藥。以理論研究為基礎(chǔ),實驗方面,首先建立功能性消化不良脾虛證動物模型。運用碘乙酰胺復(fù)合小平臺勞倦因素造模法,制作功能性消化不良脾虛證動物模型,并觀察動物一般狀態(tài),體重,飲食情況及胃病理改變,運用糖水偏好試驗,球囊試驗對功能性消化不良脾虛證進行評價。在模型建立的基礎(chǔ)上,運用免疫組化,Elisa, Real-time PCR對大鼠胃,下丘腦及血清中的腦腸肽(Ghrelin, CCK, VIP, SS, SP, CGRP)進行檢測。此外,運用中藥香砂六君子加減方對模型進行干預(yù),西藥多潘立酮作為陽性藥物對照。研究期間,記錄大鼠飲食情況,并運用糖水偏好試驗,球囊試驗對藥物干預(yù)療效進行評價。同時也運用免疫組化,Elisa, Real-time PCR對各組大鼠胃,下丘腦及血清中的腦腸肽(Ghrelin, CCK, VIP, SS, SP, CGRP)進行檢測。結(jié)果理論方面,通過對功能性消化不良臨床文獻進行整理發(fā)現(xiàn),功能性消化不良中醫(yī)證型出現(xiàn)頻次較高的依次為：肝胃不和,脾胃虛弱,寒熱錯雜,飲食積滯,肝郁脾虛,肝郁氣滯,脾胃虛寒,脾胃濕熱證,且各中醫(yī)證型的共有癥狀為“上腹脹”。以此為切入點,對“脹”,“滿”相關(guān)文獻進行中醫(yī)古籍整理,研究發(fā)現(xiàn),中醫(yī)“腹脹”主要由氣虛不運,寒,熱,濕,痰,食,血積聚腹內(nèi)而成,病位涉及脾,胃,肝,腎等臟。其形成的生物學(xué)機制可與腦腸肽類激素的改變有關(guān)。從疾病角度,中醫(yī)“痞滿”的腹脹部位與起病時間與腹脹型功能性消化不良最相似,可以作為此病的中醫(yī)病名。而“痞滿”的病機以脾氣虛為本,食欲的減少與否是鑒別“痞滿”虛實的標準。而“痞滿”治療方劑中,以理氣,益氣健脾,燥濕化痰類中藥出現(xiàn)頻次較高。出現(xiàn)頻次最高的藥物,與中藥成方香砂六君子湯的組成具有高度一致性,因此香砂六君子湯可以作為本病的主方。實驗方面,建立的碘乙酰胺及復(fù)合小平臺模型大鼠,均出現(xiàn)飲食減少,糖水偏好降低,胃敏感性增高。但碘乙酰胺復(fù)合小平臺組大鼠體重下降明顯,并伴有倦怠等精神癥狀。因此,兩模型符合功能性消化不良脾虛證標準,但碘乙酰胺復(fù)合小平臺組脾虛證表征更加明顯。生物學(xué)指標方面,Ghrelin, CCK, VIP在兩種功能性消化不良脾虛證大鼠胃中分泌減少,循環(huán)血及下丘腦中含量亦減少,但下丘腦中mRNA表達水平相反,其原因,可能與循環(huán)血中進入下丘腦肽類激素含量減少,引起腦內(nèi)負反饋調(diào)節(jié),從而使模型組中的mRNA表達含量升高有關(guān)。另外,SS在兩種功能性消化不良脾虛證大鼠胃及下丘腦中分泌增加,循環(huán)血中含量增加,并且以單純施加碘乙酰胺組大鼠增加程度大。研究也發(fā)現(xiàn),復(fù)合小平臺組大鼠胃內(nèi)SP及CGRP蛋白升高較為明顯,血清中CGRP明顯降低,其余無顯著性差異。因此,碘乙酰胺及復(fù)合小平臺大鼠共同表現(xiàn)為具有促進食欲及降低胃壁緊張度作用的腦腸肽Ghrelin, CCK, VIP分泌及循環(huán)減少,體內(nèi)抑制性腦腸肽SS含量增加,這可能是功能性消化不良脾虛證的病機。兩脾虛證模型在胃內(nèi)SP與CGRP蛋白表達的差異,以及SS的表達含量的差異可能是兩種脾虛亞型的機制所在。不能食而瘦的脾虛證,以痛閾降低,敏感性相對偏高更顯著,不能食而肥的脾虛證,則主要表現(xiàn)為胃腸激素紊亂。對模型施加藥物治療。研究發(fā)現(xiàn),健脾方劑香砂六君子加減方,可增加模型大鼠飲食量。同時,健脾中藥還可顯著提高組內(nèi)大鼠糖水消耗率75%所占百分比。且呈現(xiàn)量效關(guān)系。胃腸敏感性方面,健脾中藥香砂六君子湯可顯著降低模型組大鼠胃高敏感性,呈量效相關(guān)性。研究顯示,健脾方劑香砂六君子加減方有效改善功能性消化不良脾虛證食少,納呆,腹脹三大主癥。另外,對大鼠毛色不澤,神情倦怠脾虛次癥也有改善作用。因此,健脾方劑香砂六君子加減方對功能性消化不良脾虛證有治療作用,同時也驗證了本次研究復(fù)制的碘乙酰胺模型以及碘乙酰胺復(fù)合小平臺模型與功能性消化不良脾虛證動物模型高度一致。在生物學(xué)指標方面,健脾方劑香砂六君子加減方可增加Ghrelin, CCK, VIP的分泌,進而升高循環(huán)血,及下丘腦中的含量。同時可以降低胃黏膜,血清及下丘腦中的SS蛋白含量。對于SP與CGRP,藥物治療組與碘乙酰胺組之間比較,無顯著性差異。而對于以食少而瘦的脾虛證模型一復(fù)合小平臺模型組,香砂六君子加減方組胃黏膜中SP與CGRP蛋白較模型組降低,其中以SP變化較為明顯,但香砂六君子加減方對血清及下丘腦中SP與CGRP改變不明顯。研究發(fā)現(xiàn),健脾方劑香砂六君子加減方對脾虛型功能性消化不良具有治療作用,其作用靶點集中的胃黏膜上,通過調(diào)節(jié)腦腸肽的生成,進而改善脾虛型功能性消化不良的癥狀。研究也證明了健脾中藥治療功能性消化不良脾虛證的作用機制與調(diào)節(jié)腦腸肽的生成有關(guān)。結(jié)論綜合以上研究發(fā)現(xiàn),中醫(yī)學(xué)認為的功能性消化不良以“上腹脹”作為主要癥狀,而以“上腹脹”作為主要癥狀的中醫(yī)病名為“痞滿”�！捌M”的病機以脾虛為本,飲食的減少,食欲的減退與否是判定邪實盛衰的標準。根據(jù)這一特點,我們建立了功能性消化不良脾虛證動物模型,進而探討了消化不良型脾虛證的生物學(xué)機制。研究發(fā)現(xiàn),碘乙酰胺及復(fù)合小平臺站立法符合功能性消化不良脾虛證標準,而腦腸肽類激素含量的變化與消化不良型脾虛證密切相關(guān)。運用健脾中藥對模型反證,發(fā)現(xiàn)健脾中藥可調(diào)整腦腸肽類激素的生成,進而改善脾虛證癥狀。因此,本研究證明了功能性消化不良脾虛證可能的生物學(xué)機制是腦腸肽的改變,表現(xiàn)為具有胃腸促進及抑制作用的腦腸肽失衡。研究成果一定程度上拓寬了傳統(tǒng)中醫(yī)“脾虛”的理論。
[Abstract]:Objective to explore the biological mechanism of the core syndrome of functional dyspepsia, the core syndrome of TCM - spleen deficiency syndrome, and to further explain and improve the theory of the syndrome of spleen deficiency in Chinese medicine. On the basis of the analysis of the symptoms of the disease, the core symptoms of the disease were excavated. On this basis, the literature of "bloating", "bloating" and the pathogenesis of traditional Chinese medicine were sorted out in the literature of "bloating" and "full", and the literature of "abdominal distention" was sorted out to explore the possible signal pathways of "abdominal distention". The study also combed the origin of TCM disease with abdominal distention and abdominal fullness as the main disease, looking for the name of traditional Chinese medicine which most conforms to abdominal distention and functional dyspepsia and studying the pathogenesis of traditional Chinese medicine. On the basis of theoretical research and experiment, the animal model of functional dyspepsia spleen deficiency syndrome was established, and the animal model of functional dyspepsia spleen deficiency syndrome was made by using the compound iiacetate compound small platform fatigue factor, and the general state of the animal was observed and the body was observed. Weight, diet and gastric pathological changes, using sugar water preference test and balloon test to evaluate the spleen deficiency of functional dyspepsia. On the basis of the establishment of the model, the brain gut peptide (Ghrelin, CCK, VIP, SS, SP, CGRP) in rats' stomach, hypothalamus and serum were detected by immunohistochemistry, Elisa and Real-time PCR. At the time of the study, the diet of rats was recorded, and the diet of rats was recorded during the study, and the sugar water preference test was used to evaluate the effect of the drug intervention on the effect of the drug intervention. At the same time, the Elisa and Real-time PCR were used in the stomach, the hypothalamus and the serum of the rats in each group. Ghrelin, CCK, VIP, SS, SP, CGRP). Results theoretically, by sorting out the clinical literature of functional dyspepsia, it was found that the frequency of TCM syndrome of functional dyspepsia was higher in sequence: liver and stomach disharmony, spleen and stomach weakness, cold and heat, stagnation of liver depression, stagnation of liver depression, stagnation of liver depression, deficiency of spleen and stomach, damp heat syndrome of spleen and stomach, The common symptoms of TCM syndrome type are "upper abdominal distention". As a breakthrough point, the literature of "bloating" and "full" related literature of traditional Chinese medicine is arranged. It is found that "abdominal distention" mainly consists of qi deficiency, cold, heat, dampness, phlegm, food and the accumulation of blood in the abdomen. The biological mechanism of the disease is related to the spleen, stomach, liver and kidney. The biological mechanism of its formation can be related to the brain intestine. From the point of view of disease, the abdominal distention parts of the Chinese medicine "ruffian" and the onset time and abdominal distension type functional dyspepsia are the most similar, which can be used as the name of the disease of traditional Chinese medicine. In addition, the frequency of traditional Chinese medicine, which has the highest frequency, is highly consistent with the composition of the decoction of six Jun Zi Decoction of Chinese herbal medicine. Therefore, the fragrant sand six gentleman soup can be used as the main recipe of this disease. The preference of sugar and water decreased and the sensitivity of stomach increased. However, the weight decline of rats in the IIb compound small platform group was obvious and accompanied by mental symptoms such as burnout. Therefore, the two model was in line with the standard of functional dyspepsia, but the characterization of spleen deficiency syndrome in the IIB complex small platform group was more obvious. The biological indexes, Ghrelin, CCK and VIP were two kinds of work. In the rats with dyspeptic dyspepsia, the secretion of the stomach in the stomach was reduced, and the content of circulating blood and hypothalamus decreased, but the expression level of mRNA in the hypothalamus was opposite, which may be related to the decrease in the content of peptide hormone in the hypothalamus and the adjustment of the negative feedback in the brain, which is related to the increase of mRNA expression in the model group. In addition, SS is two The secretion of gastric and hypothalamus increased in the rats with functional dyspepsia and increased in the gastric and hypothalamus, and increased in circulating blood. The increase of SP and CGRP protein in the stomach of the compound small platform group was more obvious, and the CGRP in the serum was obviously decreased, and the rest had no significant difference. Therefore, iodine B was not significant. Ghrelin, CCK, VIP secretion and circulation decreased, and the content of inhibitory brain gut peptide SS in the body increased, which may be the pathogenesis of functional dyspepsia spleen deficiency syndrome. The difference between the expression of SP and CGRP protein in the two spleen deficiency syndrome model in the stomach, as well as the difference in the expression of SP and CGRP protein in the stomach, and The difference in the expression content of SS may be the mechanism of the two types of spleen asthenia. The deficiency of the spleen deficiency syndrome can not be eaten and thin, the pain threshold is reduced, the sensitivity is relatively high, and the spleen deficiency syndrome can not be fed and fat, and the main manifestation is the gastrointestinal hormone disorder. At the same time, the traditional Chinese medicine can also significantly increase the percentage of the sugar water consumption rate of 75% of the rats in the group. And there is a quantitative effect relationship. In the case of gastrointestinal sensitivity, the spleen six gentleman soup of Jianpi traditional Chinese medicine can significantly reduce the high sensitivity of the stomach in the model group, and it has a quantitative effect correlation. The study shows that the prescription of Six Gentlemen of Jianpi prescription is effectively modified. Good functional dyspepsia spleen deficiency syndrome has three major main symptoms, such as less food, Na, and abdominal distention. In addition, it also improves the color of the rats and the symptoms of burnout and spleen deficiency. Therefore, the prescription of Six Gentlemen of Jianpi prescription has a therapeutic effect on the spleen deficiency syndrome of functional dyspepsia, and the model of iodine acetamide and iodine B copied in this study are also verified. Amido compound small platform model is highly consistent with the animal model of functional dyspepsia with spleen deficiency. In biological indexes, the addition and subtraction of Six Gentlemen of Jianpi prescription fragrant sand can increase the secretion of Ghrelin, CCK, VIP, and then increase circulating blood and the content of hypothalamus. Meanwhile, it can lower the content of SS protein in the gastric mucosa, serum and hypothalamus. There was no significant difference between SP and CGRP, and there was no significant difference between the drug treatment group and the IIb group. The SP and CGRP protein in the gastric mucosa of the addition and subtraction group of the Six Gentlemen of the fragrant sand was lower than that in the model group, and the changes of the SP in the gastric mucosa were significantly lower than those in the model group, but the sera and hypothalamus of the addition and subtraction of the six monarch of the fragrant sand were found in the hypothalamus. The changes of SP and CGRP were not obvious. The study found that the prescription of Jianpi Fang Decoction Six Gentlemen added and subtraction had a therapeutic effect on the spleen deficiency functional dyspepsia. Its target focused on the gastric mucosa, by regulating the formation of the brain gut peptide and then improving the symptoms of the functional dyspepsia of the spleen deficiency type. The mechanism of the dysfunction of the spleen deficiency syndrome is related to the regulation of the formation of the brain gut peptide. Conclusion the comprehensive study found that the main symptoms of functional dyspepsia in Chinese medicine are "upper abdominal distention", while "upper abdominal distention" as the main symptom of TCM is "ruffian". The loss of appetite is the criterion for determining the prosperity and decline of the pathogenic factors. According to this characteristic, we establish the animal model of functional dyspepsia and spleen deficiency syndrome, and then discuss the biological mechanism of the spleen deficiency syndrome of indigestion. The study found that the IIb and the compound small platform stand method conforms to the standard of the spleen deficiency of the dyspepsia, and the brain gut peptide. The change of hormone content is closely related to the spleen deficiency syndrome of indigestion type. Using Chinese medicine Jianpi to prove the model, it is found that the Chinese herbal medicine can adjust the formation of the peptide hormone of the brain and then improve the symptoms of the spleen deficiency syndrome. Therefore, this study proves that the possible biological mechanism of the functional dyspepsia is that the possible biological mechanism of the spleen deficiency syndrome is the change of the brain gut peptide, which is manifested as the stomach with the stomach. The gut gut peptide imbalance in intestinal promotion and inhibition has broadened the theory of "Spleen Deficiency" in traditional Chinese medicine to a certain extent.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R259
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本文編號：1984186