本文選題：高脂血癥 + 健脾消痰方�。� 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:觀察健脾消痰方對(duì)高脂血癥大鼠血清中甘油三酯(TG)、膽固醇(TC)、高密度脂蛋白膽固醇(HDL-C)、低密度脂蛋白膽固醇(LDL-C)的含量及結(jié)腸組織中水通道蛋白3(AQP3)、水通道蛋白4(AQP4)表達(dá)的影響,探討其對(duì)高脂血癥的治療作用及其可能的作用機(jī)制。方法:選用60只健康的SD大鼠,體重在160g~180g,雄性,適應(yīng)性喂養(yǎng)一周后,按照體重分層的原則,隨機(jī)分為6組,分別是:正常對(duì)照組(簡(jiǎn)稱正常組)、模型對(duì)照組(簡(jiǎn)稱模型組)、辛伐他汀對(duì)照組(簡(jiǎn)稱對(duì)照組)、健脾消痰方低劑量組(簡(jiǎn)稱低劑量組)、健脾消痰方中劑量組(簡(jiǎn)稱中劑量組)、健脾消痰方高劑量組(簡(jiǎn)稱高劑量組)。采用喂飼高脂飼料的方法復(fù)制高脂血癥模型,除正常組喂飼普通飼料外,其余各組均喂飼高脂飼料。與此同時(shí),正常組與模型組灌服適量蒸餾水,其余各組灌服治療藥或?qū)φ账?用藥量均按照1ml/100g計(jì)算。第5周末,于最后一次給藥后,將大鼠禁食不禁水24小時(shí),麻醉狀態(tài)下,股動(dòng)脈取血,將血樣離心,分離血清,檢測(cè)血清中TG、TC、HDL-C、LDL-C的含量,并迅速剖取肝臟,將肝臟最大葉部分肝組織放入4%的多聚甲醛溶液中,用于病理觀察。同時(shí)剖取結(jié)腸,取位置相對(duì)固定的3塊結(jié)腸組織,剪開(kāi)沖洗干凈后,將其中1塊放入4%多聚甲醛溶液中,運(yùn)用免疫組織化學(xué)法觀察AQP3及AQP4的表達(dá),其余2塊放入凍存管內(nèi),運(yùn)用RT-PCR及Western Blot法觀察AQP3mRNA、AQP4mRNA、AQP3及AQP4的表達(dá)。結(jié)果:1各組大鼠血清中TG、TC、HDL-C、LDL-C含量的變化與正常組相比,模型組大鼠血清中TC、TG、LDL-C的含量均明顯升高(P0.01),HDL-C含量明顯降低(P0.01或P0.05),提示大鼠造模成功。藥物干預(yù)后,各用藥組TG、TC、LDL-C的含量均較模型組降低,HDL-C含量均升高;與對(duì)照組相比,高劑量組血脂無(wú)明顯變化(P0.05),中、低劑量組TG、TC、LDL-C的含量升高,HDL-C含量降低(P0.01或P0.05);健脾消痰方各劑量組之間比較,高劑量組TG、TC、LDL-C的含量低于中、低劑量組,hdl-c的含量高于中、低劑量組(p0.01或p0.05);與低劑量組相比,中劑量組tc含量降低(p0.05),但tg、hdl-c、ldl-c的含量無(wú)顯著差異(p0.05)。2各組大鼠的肝組織形態(tài)學(xué)的變化正常組大鼠肝組織肝細(xì)胞排列整齊,形狀為規(guī)則六邊形,圍繞中央靜脈呈放射狀分布,肝血竇結(jié)構(gòu)清晰,無(wú)脂肪滴及炎性細(xì)胞浸潤(rùn)。模型組大鼠肝組織中肝細(xì)胞排列混亂,且出現(xiàn)大量脂肪滴,細(xì)胞核位置被擠壓偏離,肝血竇充血水腫。對(duì)照組肝細(xì)胞結(jié)構(gòu)清晰,脂肪滴明顯減少,肝血竇充血水腫減輕,但存在細(xì)胞壞死及炎性細(xì)胞浸潤(rùn)。健脾消痰方低中高劑量組較模型組結(jié)構(gòu)排列清晰,脂肪滴也不同程度減少,肝血竇形態(tài)趨于正常,尤以高劑量組效果顯著。3各組大鼠結(jié)腸組織中aqp3表達(dá)的變化免疫組織化學(xué)法:正常組大鼠陽(yáng)性細(xì)胞表達(dá)相對(duì)較少,著色較淺,模型組大鼠結(jié)腸上皮細(xì)胞的細(xì)胞核、細(xì)胞漿及腸腺上皮細(xì)胞陽(yáng)性表達(dá)相對(duì)較多,著色較深,結(jié)締組織有少量陽(yáng)性表達(dá),提示模型組大鼠結(jié)腸組織aqp3的表達(dá)較正常組明顯升高(p0.01)。對(duì)照組、低劑量組、中劑量組、高劑量組陽(yáng)性細(xì)胞的表達(dá)及著色程度依次降低;藥物干預(yù)后,各用藥組aqp3的表達(dá)均低于模型組(p0.01);其中,高、中、低劑量組aqp3的表達(dá)明顯低于對(duì)照組(p0.01);三個(gè)劑量組中,高劑量組aqp3明顯低于中、低劑量組(p0.01);中劑量組aqp3的表達(dá)又低于低劑量組(p0.05)。rt-pcr法:模型組大鼠結(jié)腸組織aqp3mrna的表達(dá)較正常組明顯升高(p0.01);藥物干預(yù)后,各用藥組aqp3mrna的表達(dá)均低于模型組(p0.01);其中,高、中劑量組aqp3mrna的表達(dá)低于對(duì)照組(p0.01),而低劑量組與對(duì)照組相比無(wú)差異(p0.05);三個(gè)劑量組中,高劑量組aqp3mrna低于中、低劑量組(p0.01),中劑量組aqp3mrna的表達(dá)有低于低劑量組(p0.01)。westernblot法:研究結(jié)果顯示,模型組大鼠結(jié)腸組織aqp3的表達(dá)較正常組明顯升高(p0.01);藥物干預(yù)后,各用藥組aqp3的表達(dá)均低于模型組(p0.01);其中,高、中劑量組aqp3的表達(dá)明顯低于對(duì)照組(p0.01),而低劑量組與對(duì)照組相比無(wú)差異(p0.05);三個(gè)劑量組中,高劑量組aqp3的表達(dá)明顯低于中、低劑量組(p0.01),中劑量組aqp3的表達(dá)又低于低劑量組(p0.01)。4各組大鼠結(jié)腸組織中aqp4表達(dá)的變化免疫組織化學(xué)法:正常組大鼠陽(yáng)性細(xì)胞表達(dá)相對(duì)較少,著色較淺,模型組大鼠結(jié)腸上皮細(xì)胞的細(xì)胞核、細(xì)胞漿及腸腺上皮細(xì)胞陽(yáng)性表達(dá)相對(duì)較多,著色較深,結(jié)締組織有少量陽(yáng)性表達(dá),提示模型組大鼠結(jié)腸組織中aqp4的表達(dá)較正常組明顯升高(p0.01)。對(duì)照組、低劑量組、中劑量組、高劑量組陽(yáng)性細(xì)胞的表達(dá)及著色程度依次降低;藥物干預(yù)后,各用藥組aqp4的表達(dá)均低于模型組(p0.01);其中,高、中、低劑量aqp4的表達(dá)低于對(duì)照組(p0.01或p0.05);三個(gè)劑量組中,高劑量組aqp4的表達(dá)低于中、低劑量組(p0.01),中劑量組aqp4的表達(dá)與低劑量組無(wú)顯著差異(p0.05)。rt-pcr法:模型組大鼠結(jié)腸中aqp4mrna的表達(dá)較正常組明顯升高,(p0.01);藥物干預(yù)后,各用藥組aqp4mrna的表達(dá)均低于模型組(p0.01);其中,高、中劑量aqp4mrna的表達(dá)明顯低于對(duì)照組(p0.01),而低劑量組與對(duì)照組相比無(wú)顯著性差異(p0.05)。三個(gè)劑量組中,高劑量組aqp4mrna的表達(dá)低于中、低劑量組(p0.01),中劑量組aqp4mrna的表達(dá)又低于低劑量組(p0.05)。westernblot法:模型組大鼠結(jié)腸中aqp4的表達(dá)較正常組明顯升高(p0.01);藥物干預(yù)后,各用藥組aqp4的表達(dá)均低于模型組(p0.05);其中,對(duì)照組aqp4的表達(dá)明顯高于低、中、高劑量(p0.01或p0.05)。三個(gè)劑量組中,高劑量組aqp4的表達(dá)低于中、低劑量組(p0.05或p0.01),中劑量組aqp4的表達(dá)又低于低劑量組(p0.05)。結(jié)論:1健脾消痰方能夠降低血清中tc、tg、ldl-c水平,升高h(yuǎn)dl-c水平,減輕肝細(xì)胞的脂變程度,具有良好地調(diào)節(jié)血脂作用。2健脾消痰方通過(guò)調(diào)控結(jié)腸組織中aqp3、aqp4、aqp3mrna、aqp4mrna的表達(dá),可有效地改善結(jié)腸因過(guò)度重吸收水液導(dǎo)致的水液停聚,減少痰濕的生成,消除高脂血癥的病因,從而達(dá)到防治高脂血癥的目的。
[Abstract]:Objective: To observe the effect of Jianpi Xiao Fu Fang on the content of triglyceride (TG), cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and the expression of water channel protein 3 (AQP3) and water channel protein 4 (AQP4) in the colon tissue of hyperlipidemia rats, and to explore the therapeutic effect and possibility of the hyperlipidemia on hyperlipidemia. Methods: 60 healthy SD rats were selected and the body weight was 160g~180g, male, after adaptive feeding for one week, according to the principle of weight stratification, they were randomly divided into 6 groups: normal control group (normal group), model control group (abbreviated as model group), simvastatin control group (abbreviated as control group), low dose group of Jianpi eliminating phlegm prescription (abbreviated for short) The low dose group), the middle dose group of Jianpi eliminating phlegm prescription (medium dose group), the high dose group of Jianpi eliminating phlegm prescription (high dose group). The hyperlipidemia model was replicated by feeding high fat diet. Besides the normal diet, the other groups were fed with high fat diet. At the same time, the normal group and the model group were given proper amount of distilled water. Each group was given the treatment or control, and the dosage was calculated according to 1ml/100g. At the end of the fifth week, after the last administration, the rats were fasted for 24 hours. Under the anesthetic state, the femoral artery was taken blood, the blood samples were centrifuged, the serum was separated and the content of TG, TC, HDL-C, LDL-C in the serum was detected, and the liver was quickly dissected and the liver tissue of the largest lobe of the liver was dissected. In 4% of the 4% polycondensation Formaldehyde Solution, it was used for pathological observation. At the same time, the colon was dissected and 3 colonic tissues with relative fixed position were taken. After sciscent and rinse, 1 of them were put into 4% polycondensation of Formaldehyde Solution. The expression of AQP3 and AQP4 was observed by immunohistochemistry. The remaining 2 pieces were placed in the cryopreservation tube, and AQP3mR was used to observe AQP3mR by RT-PCR and Western Blot. The expression of NA, AQP4mRNA, AQP3 and AQP4. Results: 1 the content of TG, TC, HDL-C, LDL-C in serum of rats in each group was significantly higher than that in the normal group. The content of TC, TG, LDL-C in the serum of the model rats was significantly increased (P0.01), which suggested that the rat model was successful. Compared with the control group, the level of HDL-C in the high dose group had no obvious change (P0.05), and the content of TG, TC, LDL-C in the low dose group increased and the content of HDL-C decreased (P0.01 or P0.05) in the low dose group; the content of TG, TC, LDL-C in the high dose group was lower than in the high dose group, and the low dose group was higher than in the low dose group, and the low dose was higher than the low dose. Compared with low dose group (P0.01 or P0.05), the content of TC in middle dose group decreased (P0.05), but the content of TG, HDL-C, LDL-C had no significant difference (P0.05) the change of liver histomorphology in each group of rats (P0.05).2 group, the liver cells in the normal group were arranged orderly, the shape was regular hexagon, the central vein was radially distributed, and the structure of hepatic sinusoid was clear. In the model group, the liver cells in the model group were arranged in confusion and a large number of fat drops, the nucleus position was squeezed off and the hepatic sinusoid congestion and edema. The control group had clear liver cell structure, the fat drops obviously decreased, the hepatic sinusoid congestion and edema lightened, but there was cell necrosis and inflammatory cell infiltration. Invigorating the spleen and eliminating phlegm. In the middle and high dose group, the structure of the model group was clearer than the model group, the fat droplets were reduced in different degrees and the morphology of the hepatic sinusoids tended to be normal. The effect of the high dose group on the expression of AQP3 in the colon tissue of the.3 rats was significant. The expression of the positive cells in the normal group was less, the coloring was shallow, and the colon epithelium of the model group rats. The positive expression of the cell nucleus, cytoplasm and intestinal gland epithelial cells was relatively more, the coloring was deeper, and the connective tissue had a small number of positive expressions, suggesting that the expression of AQP3 in the colon tissue of the model group was significantly higher than that of the normal group (P0.01). The expression and coloring degree of the positive cells in the low dose group, middle dose group and high dose group decreased in turn; The expression of AQP3 in each group was lower than that of the model group (P0.01), and the expression of AQP3 in the high, middle and low dose groups was significantly lower than that of the control group (P0.01); in the three dose group, the AQP3 in the high dose group was significantly lower than the low dose group (P0.01); the expression of AQP3 in the middle dose group was lower than the low dose group (P0.05).Rt-pcr method: the colon tissue of the model group was AQP3. The expression of mRNA was significantly higher than that in the normal group (P0.01), and the expression of aqp3mrna in each drug group was lower than that in the model group (P0.01), and the expression of aqp3mrna in the middle dose group was lower than that of the control group (P0.01), while the low dose group had no difference compared with the control group (P0.05); in the three dose group, the high dose group aqp3mrna was lower than the low dose group (P0.01), The expression of aqp3mrna in the medium dose group was lower than the low dose group (P0.01).Westernblot method. The results of the study showed that the expression of AQP3 in the colon tissue of the model group was significantly higher than that of the normal group (P0.01). The expression of AQP3 in each drug group was lower than that of the model group (P0.01), and the expression of AQP3 in the middle dose group was significantly lower than that of the control group (P0.01), but the expression of the medium dose group was significantly lower than that of the control group (P0.01). There was no difference between the low dose group and the control group (P0.05); in the three dose group, the expression of AQP3 in the high dose group was significantly lower than that in the middle, low dose group (P0.01), and the expression of AQP3 in the middle dose group was lower than that in the colon tissue of the low dose group (P0.01).4. The expression of AQP4 in the colon tissues of the rats of each group was less than that in the normal group. In the model group, the nucleus of the colonic epithelial cells in the model group, the cytoplasm and the intestinal gland epithelial cells were more positive, the coloring was more deep, and the connective tissue had a small amount of positive expression. It suggested that the expression of AQP4 in the colon tissue of the model group was significantly higher than that of the normal group (P0.01). The control group, the low dose group, the middle dose group, the high dose group positive cells. The expression and coloring degree decreased in turn, and the expression of AQP4 in each drug group was lower than that in the model group (P0.01), and the expression of high, medium and low dose AQP4 was lower than that of the control group (P0.01 or P0.05); in the three dose group, the expression of AQP4 in the high dose group was lower than that in the low dose group (P0.01), and the expression of AQP4 in the middle dose group was not significantly worse than that in the low dose group. P0.05.Rt-pcr method: the expression of AQP4mRNA in the colon of the model rats was significantly higher than that in the normal group, (P0.01), and the expression of AQP4mRNA in each drug group was lower than that of the model group (P0.01). The expression of AQP4mRNA in the middle dose group was significantly lower than that of the control group (P0.01), but there was no significant difference between the low dose group and the control group (P0.05). Three In the dose group, the expression of AQP4mRNA in the high dose group was lower than that in the low dose group (P0.01), and the expression of AQP4mRNA in the middle dose group was lower than that of the low dose group (P0.05).Westernblot method. The expression of AQP4 in the colon of the model group was significantly higher than that in the normal group (P0.01). The expression of AQP4 in each drug group was lower than that of the model group (P0.05), and the control group was AQ (P0.05); and the control group AQ was aq. The expression of P4 was significantly higher than low, middle and high dose (P0.01 or P0.05). The expression of AQP4 in the high dose group was lower than that of the middle, low dose group (P0.05 or P0.01), and the expression of AQP4 in the middle dose group was lower than that of the low dose group (P0.05). Conclusion: 1 the prescription of strengthening spleen and eliminating phlegm can reduce TC, TG, LDL-C level, increase HDL-C level, and reduce the lipid change of liver cells. By regulating the expression of AQP3, AQP4, aqp3mrna and AQP4mRNA in colon tissue, the expression of.2 Jianpi eliminating phlegm can effectively improve the accumulation of water caused by excessive heavy absorption of water in colon, reduce the formation of phlegm dampness, eliminate the cause of hyperlipidemia, and thus achieve the purpose of preventing hyperlipidemia.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉萍,張靜生;冠心康對(duì)高脂血癥大鼠血脂影響的研究[J];中成藥;2003年09期

2 祝麗娣;，

本文編號(hào)：1931973