本文選題：丹皮酚 + 潰瘍性結(jié)腸炎�。� 參考：《鄭州大學(xué)》2017年博士論文

【摘要】：目的:潰瘍性結(jié)腸炎是一類臨床常見疾病,目前缺乏較好的治療方法,本研究在臨床實踐中發(fā)現(xiàn)了丹皮酚(Pae)注射液治療潰瘍性結(jié)腸炎的療效顯著。本文觀察其臨床效果,并對其作用機(jī)制進(jìn)行研究,為潰瘍性結(jié)腸炎的臨床治療提供可參考的新方向。方法:采用臨床實驗結(jié)合動物實驗,通過丹皮酚注射液對潰瘍性結(jié)腸炎患者的臨床治療,觀察其臨床治療效果;通過動物實驗,探究其可能存在的作用機(jī)理。全文實驗共分為五部分:(1)丹皮酚對潰瘍性結(jié)腸炎的臨床療效觀察。選取2015年1月-2016年9月鄭州大學(xué)第二附屬醫(yī)院消化內(nèi)科收治的80例潰瘍性結(jié)腸炎患者作為研究對象,分為對照組(40例)和治療組(40例)。對照組采用常規(guī)藥物(美沙拉嗪或潑尼松片)治療;治療組在常規(guī)藥物治療的基礎(chǔ)上給予肌內(nèi)注射丹皮酚注射液。治療周期14天,觀察指標(biāo)包括血小板水平、血紅蛋白水平、白蛋白水平、C反應(yīng)蛋白水平、紅細(xì)胞沉降速率、機(jī)體氧化應(yīng)激水平、炎性因子含量變化情況及ICAM-1、MMP-9蛋白表達(dá)水平,評價療效;(2)丹皮酚對葡聚糖硫酸鈉(DSS)誘導(dǎo)潰瘍性結(jié)腸炎大鼠模型的保護(hù)作用。利用DSS誘導(dǎo)制備潰瘍性結(jié)腸炎大鼠模型,隨機(jī)分為對照組、模型組、丹皮酚(低、高)劑量組及陽性對照組,觀察大鼠一般狀態(tài)、體重變化情況及疾病活動指數(shù)(DAI),ELISA法檢測血清炎性因子水平,血漿MDA和SOD水平,RT-PCR法檢測結(jié)腸組織炎性因子mRNA表達(dá)水平,Western Blot法檢測結(jié)腸粘膜組織ICAM-1、MMP-9蛋白表達(dá)水平,并評價組織損傷程度;(3)丹皮酚對DSS誘導(dǎo)所致腸炎的抗炎效果及機(jī)制研究。利用DSS誘導(dǎo)制備潰瘍性結(jié)腸炎大鼠模型,隨機(jī)分為對照組、模型組、丹皮酚(低、中、高劑量)組及陽性對照組,ELISA法測定大鼠血清炎性因子水平,RT-PCR法測定大鼠血清炎性因子mRNA表達(dá)水平,Western Blot法測定NF-κB和MAPK信號通路蛋白表達(dá)水平;(4)丹皮酚對大鼠CD4+T細(xì)胞亞群及其相關(guān)細(xì)胞因子表達(dá)的影響及機(jī)制研究。利用DSS誘導(dǎo)制備潰瘍性結(jié)腸炎大鼠模型,隨機(jī)分為正常對照組、模型組、丹皮酚治療組,流式細(xì)胞儀檢測CD4+T細(xì)胞亞群Thl、Th2、Thl7和Treg細(xì)胞的比例,RT-PCR法檢測CD4+T細(xì)胞各亞型轉(zhuǎn)錄因子的表達(dá),抗體芯片檢測CD4+T各細(xì)胞因子的表達(dá),免疫組織化學(xué)染色法檢測大鼠結(jié)腸粘膜組織Thl、Th2、Thl7和Treg細(xì)胞的表達(dá),Western Blot法測定IL-23R/Jak2/Stat3信號通路蛋白表達(dá)水平;(5)丹皮酚對模型大鼠腸黏膜相關(guān)生長因子的影響。利用DSS誘導(dǎo)制備潰瘍性結(jié)腸炎大鼠模型,隨機(jī)分為對照組、模型組、丹皮酚(低、高劑量)組及陽性對照組,SABC免疫組化染色法觀察大鼠腸黏膜平均血管密度,MASSON染色法觀察腸黏膜血管微血栓形成,ELISA法檢測血漿及結(jié)腸黏膜血管內(nèi)皮生長因子(VEGF)、成纖維細(xì)胞生長因子(bFGF)、血小板衍生因子(PDGF)水平,放射免疫分析法測定血漿及結(jié)腸一氧化氮(NO)水平,Western Blot法測定血漿及結(jié)腸缺氧誘導(dǎo)因子(HIF-1)水平。結(jié)果:(1)丹皮酚可有效改善患者的臨床癥狀,改善腸黏膜病變,消除并發(fā)癥,降低血小板、C反應(yīng)蛋白濃度,降低紅細(xì)胞沉降速率,降低ICAM-1、MMP-9蛋白表達(dá)水平,提高血紅蛋白、白蛋白水平,同時不會引起嚴(yán)重的不良反應(yīng);(2)丹皮酚注射液可有效抑制大鼠體重下降趨勢,改善模型大鼠DAI指數(shù),降低體內(nèi)IL-1、IL-4、INF-γ、TNF-α及氧化應(yīng)激水平,提高IL-10水平,同時對炎性因子mRNA表達(dá)具有調(diào)節(jié)作用,降低ICAM-1、MMP-9蛋白的表達(dá),改善大鼠腸黏膜平均血管密度,抑制微血栓形成;(3)丹皮酚可以顯著抑制DSS誘導(dǎo)的實驗性腸炎大鼠促炎癥因子IL-1β、IL-4、IL-6、IFN-γ、TNF-α和MCP-1及其mRNA的表達(dá),免疫印跡結(jié)果顯示,丹皮酚可抑制NF-κB通路p-IκB、p-p65、p-IKKα/β水平和MAPK通路中p-Erk1/2、p-JNK、p-p38水平,從而抑制NF-κB通路和MAPK通路活性;(4)丹皮酚可提高CD4+T細(xì)胞亞型Treg細(xì)胞的比例,下調(diào)Thl、Th2和Thl7細(xì)胞的比例,提高FoxP3的表達(dá)水平,下調(diào)GATA-3、RORc及T-bet表達(dá)水平,降低細(xì)胞因子IL-2、IL-4、IL-21和IL-17表達(dá)水平。抑制IL-23R和JAK-2蛋白的表達(dá),并抑制stat3磷酸化,促進(jìn)smad磷酸化,由此抑制IL-23R/Jak2/Stat3信號通路活性;(5)丹皮酚注射液可有效改善模型大鼠腸黏膜平均血管密度及腸黏膜血管微血栓形成個數(shù),降低血漿及結(jié)腸組織VEGF、NO、bFGF及HIF-1水平。結(jié)論:(1)丹皮酚注射液對潰瘍性結(jié)腸炎具有較好的治療效果,治療機(jī)制可能與減低體內(nèi)氧化應(yīng)激、炎性因子水平及血管細(xì)胞因子的表達(dá)有關(guān);(2)丹皮酚對模型大鼠的保護(hù)作用可能是通過降低氧化應(yīng)激水平及炎性因子mRNA的表達(dá),消除炎癥反應(yīng),抑制ICAM-1、MMP-9蛋白的表達(dá),改善腸道及血管功能,從而起到對模型大鼠的保護(hù)作用;(3)丹皮酚可降低腸炎模型大鼠炎性水平,其作用機(jī)制可能與抑制NF-κB通路和MAPK通路活性有關(guān);(4)丹皮酚通過抑制CD+T細(xì)胞各亞型失衡對慢性潰瘍性結(jié)腸炎產(chǎn)生治療效果,該治療效果與丹皮酚可以抑制IL-23R/Jak2/Stat3信號通路活性有關(guān);(5)丹皮酚對潰瘍性結(jié)腸炎大鼠腸黏膜的保護(hù)作用機(jī)制可能為改變腸黏膜相關(guān)生長因子的水平,影響相關(guān)的代謝通路。
[Abstract]:Objective: ulcerative colitis is a kind of common clinical disease and lack of good treatment. This study found the curative effect of Pae injection in the treatment of ulcerative colitis in clinical practice. The clinical effect of this study was observed and the mechanism of action was studied to provide reference for the clinical treatment of ulcerative colitis. New directions. Methods: the clinical treatment of ulcerative colitis was treated by clinical experiment combined with animal experiment, and the clinical therapeutic effect of ulcerative colitis was observed by Paeonol Injection. The possible mechanism of action was explored through animal experiments. The full text was divided into five parts: (1) the clinical effect of Paeonol on ulcerative colitis. 80 cases of ulcerative colitis were treated in the digestive department of the Second Affiliated Hospital of Zhengzhou University, January 2015 -2016 in September. The patients were divided into control group (40 cases) and treatment group (40 cases). The control group was treated with conventional drugs (mesalazine or prednisone), and the treatment group was given intramuscular injection of Paeonol on the basis of conventional drug treatment. The treatment period was 14 days. The indexes included platelet level, hemoglobin level, albumin level, C reaction protein level, erythrocyte sedimentation rate, oxidative stress level, inflammatory factor content change and ICAM-1, MMP-9 protein expression level, evaluation of therapeutic effect, and (2) paeonol induced ulcerative junction with dextran sodium sulfate (DSS). The rat model of ulcerative colitis was induced by DSS. The rat model of ulcerative colitis was induced and divided into control group, model group, paeonol (low, high) dose group and positive control group. The general state, body weight change and disease activity index (DAI) were observed in rats. The level of serum inflammatory factors was detected by ELISA method, plasma MDA and SOD levels, RT-P CR assay was used to detect the expression level of inflammatory factor mRNA in colon tissue, Western Blot method was used to detect the expression of ICAM-1 and MMP-9 protein in colonic mucosa, and the degree of tissue damage was evaluated. (3) the anti-inflammatory effect and mechanism of Paeonol on DSS induced enteritis. The rat model of ulcers induced colitis was induced by DSS, and the model was randomly divided into the control group and the model was randomly divided into the control group. Group, paeonol (low, medium, high dose) group and positive control group, ELISA assay was used to determine the level of inflammatory factors in rat serum, RT-PCR assay was used to determine the expression level of inflammatory factor mRNA in rat serum. Western Blot method was used to determine the expression level of NF- kappa B and MAPK signaling pathway protein; (4) the effect of Paeonol on the expression of CD4+T lymphocyte subsets and related cytokines in rats The rat model of ulcerative colitis was prepared by DSS. The model was randomly divided into normal control group, model group, paeonol treatment group, flow cytometry to detect the proportion of Thl, Th2, Thl7 and Treg cells in CD4+T cell subsets. RT-PCR method was used to detect the expression of transcription factors of each subtype of CD4+T cells, and the antibody chip was used to detect the various cytokines of CD4+T. The expression of Thl, Th2, Thl7 and Treg cells in the colon mucosa of rats was detected by immunohistochemical staining. The expression level of IL-23R/Jak2/Stat3 signaling protein was measured by Western Blot method; (5) the effect of Paeonol on the intestinal mucosa related growth factors in the model rats. The rat model of ulcerative colitis was prepared by DSS inducement, and was randomly divided into control. Group, model group, paeonol (low, high dose) group and positive control group, SABC immunohistochemical staining method was used to observe the average vascular density of intestinal mucosa of rats, MASSON staining method was used to observe the formation of vascular microthrombus in intestinal mucosa, and ELISA method was used to detect vascular endothelial growth factor (VEGF), fibroblast growth factor (bFGF) and platelet derived factor (bFGF). PDGF) level, radioimmunoassay to determine plasma and colon nitric oxide (NO) levels, Western Blot method for the determination of plasma and colon hypoxia inducible factor (HIF-1). Results: (1) paeonol can effectively improve the clinical symptoms of patients, improve intestinal mucosal lesions, eliminate complications, reduce platelet, C reaction protein concentration, and decrease the rate of erythrocyte sedimentation Rate, decrease ICAM-1, MMP-9 protein expression level, improve hemoglobin and albumin level, and not cause serious adverse reactions. (2) Paeonol Injection can effectively inhibit the trend of weight loss in rats, improve the DAI index of model rats, decrease the level of IL-1, IL-4, INF- gamma, TNF- A and oxidative stress in the body, improve the level of IL-10, and also cause inflammatory causes. The expression of submRNA can reduce the expression of ICAM-1 and MMP-9 protein, improve the mean blood vessel density and inhibit the formation of microthrombus in the intestinal mucosa of rats. (3) paeonol can significantly inhibit the expression of IL-1 beta, IL-4, IL-6, IFN- gamma, TNF- A and MCP-1 and its mRNA in the DSS induced experimental enteritis rats, and the results of Western blot show that the Western blot results show paeonol. Inhibition of NF- kappa B pathway p-I kappa B, p-p65, p-IKK alpha / beta level and MAPK pathway p-Erk1/2, p-JNK, p-p38 levels, thus inhibiting the NF- B pathway and the activity of the pathway; (4) paeonol can improve the proportion of subtype cells. Low cytokine IL-2, IL-4, IL-21 and IL-17 expression levels, inhibit the expression of IL-23R and JAK-2 protein, inhibit STAT3 phosphorylation, promote Smad phosphorylation, and inhibit the activity of IL-23R/Jak2/Stat3 signaling pathway. (5) Paeonol Injection can effectively improve the average vascular density of intestinal mucosa and the number of intestinal mucosal microthrombus formation in the model rats, and reduce the number of intestinal mucosal microthrombus formation. Low plasma and colonic tissue VEGF, NO, bFGF and HIF-1 levels. Conclusions: (1) Paeonol Injection has a good therapeutic effect on ulcerative colitis. The mechanism of treatment may be related to reducing oxidative stress in the body, the level of inflammatory factors and the expression of vascular cell factors; (2) the protective effect of Dan paeonol on model rats may be by reducing oxidation. Stress level and inflammatory factor mRNA expression, eliminate inflammatory reaction, inhibit the expression of ICAM-1, MMP-9 protein, improve intestinal and vascular function, and thus protect the model rats; (3) paeonol can reduce the inflammatory level of rats with enteritis model, its mechanism can be related to the inhibition of NF- kappa B pathway and MAPK pathway activity; (4) paeonol The effect of inhibition of CD+T cell subtypes on chronic ulcerative colitis, the effect of the treatment and paeonol can inhibit the activity of IL-23R/Jak2/Stat3 signaling pathway; (5) the protective mechanism of Paeonol on the intestinal mucosa of ulcerative colitis rats may be related to the level of altered intestinal mucosa related growth factors. Metabolic pathways.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R574.62

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 徐剛;余林蔓;何興元;;潰瘍性結(jié)腸炎感染巨細(xì)胞病毒的危險因素、臨床特征、臨床結(jié)局分析[J];胃腸病學(xué)和肝病學(xué)雜志;2016年10期

2 Adil Coskun;Erdogan Malatyali;Hatice Ertabaklar;Mustafa B Yasar;Ali O Karaoglu;Sema Ertug;;Blastocystis in ulcerative colitis patients:Genetic diversity and analysis of laboratory findings[J];Asian Pacific Journal of Tropical Medicine;2016年09期

3 沈浩;;潰瘍性結(jié)腸炎患者免疫功能狀態(tài)的臨床研究[J];現(xiàn)代消化及介入診療;2016年04期

4 黎莉;楊衛(wèi)文;譚松;劉正勇;何天蘭;戴振媛;;潰瘍性結(jié)腸炎患者外周血及結(jié)腸黏膜TNF-a、IL-6和IL-2變化的臨床意義[J];中國現(xiàn)代醫(yī)學(xué)雜志;2016年14期

5 陳晶;李巧霞;李瑋;李宏;湯菲;王縛鯤;;Allicin對DSS誘導(dǎo)的小鼠結(jié)腸炎的治療作用及可能機(jī)制[J];中國免疫學(xué)雜志;2016年07期

6 孫文佳;王曦;;炎癥小體在潰瘍性結(jié)腸炎發(fā)病機(jī)制中的作用[J];中華醫(yī)學(xué)雜志;2016年24期

7 熊真;;不同嚴(yán)重程度潰瘍性結(jié)腸炎炎癥因子表達(dá)水平的臨床研究[J];現(xiàn)代消化及介入診療;2016年03期

8 王鳳;吳秀美;甘麗萍;龍宇;李燕舞;巫燕莉;劉梓峰;杜群;;潰結(jié)靈對潰瘍性結(jié)腸炎大鼠Th17分化的調(diào)節(jié)機(jī)制研究[J];中藥藥理與臨床;2016年02期

9 黃循銣;王承黨;王瑞幸;焦海霞;;潰瘍性結(jié)腸炎小鼠腸道通透性改變與TNF-α及NF-κB P65的關(guān)系[J];中國應(yīng)用生理學(xué)雜志;2016年02期

10 胡流芳;王迎;任汝靜;霍海如;孫建輝;李洪梅;朱亞英;譚余慶;;Keap1-Nrf2/ARE信號通路的抗氧化應(yīng)激作用及其調(diào)控機(jī)制[J];國際藥學(xué)研究雜志;2016年01期

相關(guān)博士學(xué)位論文 前3條

1 武超;腸內(nèi)營養(yǎng)對大鼠缺血/再灌注損傷后腸黏膜屏障功能的保護(hù)及機(jī)制研究[D];南京大學(xué);2016年

2 盛益華;潰瘍性結(jié)腸炎大鼠肺損傷細(xì)胞凋亡機(jī)制及中藥單體干預(yù)作用機(jī)制研究[D];北京中醫(yī)藥大學(xué);2013年

3 王文生;腸缺血再灌注條件下IFN-γ通過調(diào)控HIF-1α在腸上皮屏障功能損害中的作用及機(jī)制研究[D];第三軍醫(yī)大學(xué);2013年

相關(guān)碩士學(xué)位論文 前1條

1 馬y曃，

本文編號：1979402