自拟活血祛湿方治疗非酒精性脂肪性肝炎临床和实验研究

发布时间：2018-01-23 22:29

本文关键词： 非酒精性脂肪性肝炎自拟活血祛湿方 NF-κB信号转导通路　出处：《辽宁中医药大学》2016年博士论文　论文类型：学位论文

【摘要】：目的:根据导师卢秉久教授治疗非酒精性脂肝性肝炎的临床经验,确立活血祛湿为治疗大法。临床研究旨在观察自拟活血祛湿方的临床疗效,并对其安全性作出评价;实验研究旨在探讨自拟活血祛湿方对非酒精性脂肪性肝炎大鼠模型的疗效及机制。材料与方法:临床研究:将60例NASH患者随机分成试验组和对照组,两组患者除基础治疗外,对照组给予水林佳胶囊治疗,试验组给予自拟活血祛湿方水煎剂和水林佳胶囊治疗。比较治疗前后两组患者中医症状体征量化积分,体重指数,肝功能,血脂,血清SOD、MDA、TNF-ɑ水平以及肝脏B超结果观察加用自拟活血祛湿方治疗NASH的疗效,并对其安全性作出评价。实验研究:纯化高脂饲料建立大鼠NASH模型,随机分为空白组、模型组、自拟活血祛湿方高、中、低剂量组以及水林佳组,采用相应药物干预,观察相应药物对模型大鼠肝细胞保护、血脂调节、抗氧化反应以及NF-κB信号转导通路的作用,并探讨自拟活血祛湿方对大鼠NASH模型肝脏保护作用的可能机制。结果:实验(一)自拟活血祛湿方治疗非酒精性脂肪性肝炎的临床试验研究1.中医症状体征量化积分试验组治疗后总有效率为86.67%,对照组治疗后总有效率为70.00%,差异有统计学意义(p0.05)。2.体重指数、肝功能、血脂两组患者治疗后BMI、肝功能、血脂(TG、TC)较治疗前均下降明显(p0.05),试验组优于对照组(p0.05)。试验组治疗前后VLDL下降明显(p0.05);对照组治疗前后VLDL差异无统计学意义(p0.05)。3.血清SOD、MDA、TNF-ɑ两组患者治疗后血清TNF-ɑ、MDA较治疗前下降明显(p0.05),试验组优于对照组(p0.05)。两组患者治疗后血清SOD水平较治疗前上升明显(p0.05),试验组优于对照组(p0.05)。4.肝脏B超对照组B超总有效率为46.67%,试验组B超总有效率为70.00%,差异无统计学意义(p0.05)。在改善肝脏形态方面,试验组并不优于对照组。5.综合疗效评定对照组总有效率46.67%,试验组总有效率73.33%,差异有统计学意义(p0.05)。试验组在总体疗效方面优于对照组(p0.05)。6.安全性分析两组患者治疗过程中未出现毒、副作用和不良反应事件。实验(二)自拟活血祛湿方对大鼠非酒精性脂肪性肝炎模型肝细胞保护以及血脂调节作用的实验研究1.一般情况及病理形态学与空白组比较,模型组大鼠体质量及肝脏湿重明显增加,肉眼观察肝脏颜色变黄,体积增大;光镜下观察肝组织,胞浆内可见脂肪空泡及炎性细胞浸润,NAS组织学病理评分5分,证明NASH造模成功;各干预组较模型组相比,大鼠的体重以及肝脏湿重明显下降,差异有统计学意义(P0.05),高剂量组以及水林佳组在脂肪变性、炎症、气球样变性以及总分均较其他组下降明显(P0.05或P0.01)。2.肝功能及血脂与空白组比较,模型组大鼠血清ALT、AST、TG、VLDL显著升高;各干预组大鼠较模型组大鼠血清ALT、AST、TG、VLDL均明显降低(P0.05或P0.01),其中高剂量组下降更为明显(P0.05或P0.01)。各干预组大鼠血清TC水平有下降趋势,但无统计学意义。实验(三)自拟活血祛湿方对大鼠非酒精性脂肪性肝炎模型抗氧化作用的实验研究1.肝组织SOD、MDA水平与空白组比较,模型组大鼠肝脏SOD酶活力显著降低,MDA含量显著增加(P0.01)。各干预组大鼠较模型组大鼠肝脏SOD酶活力显著升高,MDA含量显著降低(P0.01);高剂量组效果最为显著(p0.01或p0.05)。2.肝组织中CYP2E1 mRNA以及蛋白表达及分布与空白组比较,模型组大鼠肝组织CYP2E1 m RNA及蛋白表达均明显升高(p0.01或p0.05),自拟活血祛湿方高、中剂量组以及水林佳组大鼠肝组织CYP2E1 m RNA及蛋白表达、分布明显降低(p0.01或p0.05);高剂量组以及水林佳组效果最为显著(p0.01或p0.05)。实验(四)自拟活血祛湿方对大鼠非酒精性脂肪性肝炎模型NF-κB信号转导通路的干预作用与空白组比较,模型组大鼠肝组织中NF-κB p65、TNFR1、TNF-ɑm RNA以及蛋白的表达明显增多(p0.01或p0.05),各干预组大鼠肝组织中NF-κB p65、TNFR1、TNF-ɑm RNA以及蛋白的表达明显降低(p0.01或p0.05);自拟活血祛湿方高剂量组以及水林佳组效果最为显著(p0.01或p0.05)。结论:1自拟活血祛湿方联合水林佳可通过改善患者BMI指数、肝功能、血脂水平、肝脏影像学等方面治疗NASH,未出现毒副作用以及不良反应,且可明显改善患者不适症状和体征。减轻内脏脂肪堆积,促进自由基清除,增加抗氧化能力,抑制炎症因子可能是其治疗NASH的作用机理。2自拟活血祛湿方通过降低大鼠体重以及肝脏湿重,保护肝功能,调节血脂,改善大鼠肝组织病理等方面治疗NASH。3自拟活血祛湿方能够增加肝细胞抗氧化能力,具体体现在:增强机体以及肝脏局部SOD酶活力,促进排除血清以及肝脏中脂质氧化反应的产物MDA,抑制肝脏中CYP2E1基因以及蛋白的表达,减少CYP2E1蛋白分布。4自拟活血祛湿方增强肝组织抗炎症反应能力,抑制肝脏炎性反应。可能机制有二:(1)下调肝脏中TNF-ɑ以及NF-κB p65炎症因子的表达水平;(2)调节NF-κB信号转导通路。5.根据各项指标比较结果,自拟活血祛湿方高剂量较中、低剂量效果更佳。
[Abstract]:Objective: according to the clinical experience of Professor Lu Bingjiu in Treating Nonalcoholic fatty liver hepatitis, establish the blood dampness treatment method. The clinical study is to observe the curative effect of Huoxue Qushi Decoction ", and to evaluate its safety; experimental study aimed to investigate the effect and mechanism of Huoxue Qushi Decoction on nonalcoholic steatohepatitis in rats model. Materials and methods: clinical research: 60 cases of NASH patients were randomly divided into experimental group and control group, two patients in addition to the basic treatment, the control group was given Shui Linjia capsule treatment, experimental group treated with live blood dampness Decoction and silybinin capsules treatment. Comparison of two groups before and after treatment of TCM symptoms and signs quantitative score, body mass index, liver function, blood lipid, serum SOD, MDA, TNF- alpha level and liver ultrasonography results observed with curative effect of Huoxue Qushi Decoction in the treatment of NASH, and the safety of Evaluation. Experimental study on the purification of high fat diet rats NASH model, were randomly divided into control group, model group, Huoxue Qushi, high, low dose group and silybinin group, with the corresponding drug intervention, observe the corresponding drugs on liver cell protection, rat model of regulating blood lipid, antioxidant response and NF- B signal transduction pathway, and to explore the possible mechanism of the protective effect of NASH model in rat liver by self-made dampness. Results: (a) the experiment of Huoxue Qushi Decoction "in treatment of nonalcoholic steatohepatitis in the treatment of 1. medical symptoms quantify integral test group clinical trials after the total efficiency was 86.67% the control group after treatment, the total effective rate was 70%, the difference was statistically significant (P0.05).2. body weight index, liver function, serum BMI in two groups after treatment, liver function, blood lipid (TG, TC) than before treatment decreased significantly (P0.05), the test group was better than that of The control group (P0.05). The experimental group before and after the treatment of VLDL decreased significantly (P0.05); VLDL control group before and after treatment there was no significant difference of serum SOD (P0.05).3., MDA, TNF-. Two groups of patients after treatment. Serum TNF-, MDA decreased significantly (P0.05), the experimental group than in control group (P0.05). Two groups of patients after treatment, the serum level of SOD increased significantly than before treatment (P0.05), the experimental group than the control group (P0.05).4. liver ultrasound control group B the total effective rate was 46.67%, the experimental group B the total effective rate was 70%, the difference was not statistically significant (P0.05). The improvement of liver morphology, test group.5. is not better than that of the control group curative effect was 46.67% in control group, the total efficiency of experimental group was 73.33%, the difference was statistically significant (P0.05). The experimental group than the control group in terms of overall efficacy and safety analysis of.6. (P0.05) virus does not appear in the two groups during treatment, and no side effects Adverse events (two). The experimental study of Huoxue Qushi Decoction on rats with non alcoholic steatohepatitis model to protect liver cells and blood lipids of 1. in general and pathological changes compared with the control group, rats body weight and liver weight increased significantly, visual observation of liver color yellow, volume increased; liver tissues were observed under light microscope, the infiltration of inflammatory cells and visible fat vacuoles in the cytoplasm, NAS histological score was 5, NASH model was successful; the intervention group compared with the model group, the rat body weight and liver weight were significantly decreased, the difference was statistically significant (P0.05), high dose group and silybinin group in fatty degeneration, inflammation, ballooning degeneration and the total score was significantly higher than the other group significantly decreased (P0.05 or P0.01).2. liver function and blood lipid compared with the control group, serum ALT, AST in the model group, TG, VLDL increased significantly in the intervention group; Compared with the model group rats serum ALT, rat AST, TG, VLDL were significantly decreased (P0.05 or P0.01), the high dose group significantly decreased (P0.05 or P0.01). The serum level of TC in the intervention group had a downward trend, but no statistical significance. The experiment (three) experimental study of Huoxue Qushi in rats with non alcoholic fatty liver antioxidant model 1. liver SOD, MDA levels compared with the control group, the enzyme activity of liver SOD in rats of model group was significantly decreased, MDA content increased significantly (P0.01). The rats of each intervention group compared with the model of SOD activity in liver of rats significantly increased, MDA the content was significantly decreased (P0.01); the effect of high dose groups (P0.01 or P0.05).2. in the liver tissue of CYP2E1 mRNA and protein expression and distribution compared with the blank group, the expression model of CYP2E1 m RNA and protein in liver tissue of rats were significantly increased (P0.01 or P0.05), Huoxue Qushi party high dose Group CYP2E1 and m RNA and protein in liver tissue of silybinin group rats, distribution decreased significantly (P0.01 or P0.05); high dose group and silybinin group was the most significant (P0.01 or P0.05). (four) experimental intervention compared with control group, Huoxue Qushi Decoction on rats with non alcoholic Nash model of NF- B signal transduction pathway in the liver tissue of rats in model group in NF- B p65 TNFR1 TNF-, kappa, alpha m and RNA protein expression increased significantly (P0.01 or P0.05), the liver tissue of rats in each intervention group NF- kappa B p65, TNFR1 TNF-, m RNA and protein. The expression was significantly lower (P0.01 or P0.05); Huoxue Qushi recipe high dose group and silybinin group was the most significant (P0.01 or P0.05). Conclusion: 1 Huoxue dehygrosis recipe combined with silybinin can improve BMI index, liver function, blood lipids, liver imaging and other aspects of the treatment of NASH, no adverse effects on And adverse reactions, and can significantly improve patients with symptoms and signs. Reducing visceral fat accumulation, promote the increase of free radical scavenging, antioxidant capacity, inhibition of inflammatory cytokines may be the therapeutic mechanism of NASH..2 Huoxue Qushi party by reducing the body weight of rats and liver wet weight, protect liver function, regulate blood lipids, improve pathology the liver tissue of rats with NASH.3 and other aspects of the treatment of Huoxue Qushi could increase liver cell antioxidant ability, embodied in: enhance the body and liver SOD enzyme activity, promote lipid oxidation and elimination of serum liver product MDA, inhibition of liver CYP2E1 gene and protein expression, reduced CYP2E1 protein distribution of.4 Huoxue Qushi we enhance the liver tissue inflammatory response, inhibit liver inflammatory reaction. There are two possible mechanisms: (1) downregulation of TNF- in liver and the expression of NF- alpha kappa B p65 inflammatory factor The level of NF-; (2) regulating B signal transduction pathway of.5. according to the comparison results of the indicators, Huoxue Qushi party high dose than in low dose, better effect.

【学位授予单位】：辽宁中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R259

【参考文献】