醒脑静对A型肝性脑病大鼠肝功能、内毒素及GS、NOS表达影响的研究

发布时间：2018-09-18 19:42

【摘要】：研究背景与目的：目前肝性脑病发生机理尚未完全明了，治疗也无特效疗法，危害极大，长期以来一直是国内外中西医研究的对象。A型肝性脑(Aepatic Encephalopathy ofType A，AHE)病死率高达80%以上，为临床较为常见的急危重症。醒脑静注射液能缩短肝性脑病昏迷时间，促进清醒，有明确的临床疗效。但现有临床疗效观察资料非常有限，且入选病例较少，缺乏有力的临床数据支持醒脑静用于治疗AHE。本研究探讨300mg·kg-1·d-1的硫代乙酰胺（Thioacetamide，TAA）诱导大鼠A型肝性脑病的造模最适时间；探讨醒脑静对A型肝性脑病大鼠肝功能、内毒素及谷氨酰胺合成酶（Glutamine Synthetase，GS）、一氧化氮合酶（Nitric OxideSynthase，NOS）表达的影响。为醒脑静治疗A型肝性脑病提供科学依据。方法: 1探讨TAA（300mg·kg-1·d-1）诱导大鼠A型肝性脑病的造模最适时间： (1)将60只大鼠分为A、B、C、D四组，其中A组为正常对照组，用同实验组用药量相等的生理盐水灌胃4天。B、C、D三组为实验组，分别用TAA（300mg·kg-1·d-1）连续灌胃2天、3天和4天。 (2)比较各组大鼠行为学变化、大鼠AHE的诱导率和致死率；分析各组给药结束24小时后血氨、谷草转氨酶(Glutamic Oxalacetic Transaminase，AST)、谷丙转氨酶(Glutamic-Pyruvic Transaminase，ALT)、总胆红素(Total Bilirubin，TBIL)的差异；比较各组肝组织形态学变化。 2探讨醒脑静对A型肝性脑病大鼠肝功能、内毒素及GS、NOS表达的影响： (1)将60只大鼠分为A、B、C、D、E、F六组，其中A组为正常对照组；B组为A型肝性脑病模型组；C、D、E、F组为治疗组，即分别给予已建立A型肝性脑病模型的大鼠醒脑静（2.5ml·kg-1·d-1）、醒脑静（5ml·kg-1·d-1）、醒脑静（10ml·kg-1·d-1）、门冬氨酸鸟氨酸（2g·kg-1·d-1）治疗5天。 (2)比较各组大鼠行为学变化、AST、ALT、TBIL的差异；比较各组大鼠肝组织形态学变化；检测分析各组INF-α、内毒素及GS、NOS表达量的差异。结果： 1TAA（300mg·kg-1·d-1）诱导大鼠A型肝性脑病的造模最适时间： (1) B、C、D实验组较A组正常组大鼠脑功能分级高（P0.0083)，差异有统计学意义；C、D组较B组大鼠脑功能评分高（P0.0083)，差异有统计学意义。 (2) A组正常组，无AHE诱导大鼠也无死亡大鼠；C、D组较B组诱导率高（P0.0083），差异有统计学意义；D组较B、C组的大鼠致死率高（P0.0083)，差异有统计学意义。 (3) B、C、D实验组大鼠血氨、AST、ALT、TBIL均显著高于A组正常组（P0.05)，差异有统计学意义；C、D组血氨、AST、ALT、TBIL均较B组高（P0.05)，差异有统计学意义。 (4) A组大鼠肝组织形态正常；C、D组肝组织学观察炎症侵润、坏死、纤维化等损害较明显。 2探讨醒脑静对A型肝性脑病大鼠肝功能、内毒素及GS、NOS表达的影响： (1) A组正常组大鼠无行为学改变；C、D、E、F用药组大鼠行为学改变和脑功能分级均较B组低（P0.0033)，差异有统计学意义。 (2) B、C、 D、E、F组大鼠血氨、AST、ALT、TBIL均较A组正常组高（P0.05)，差异有统计学意义；C、 D、E、F组大鼠血氨、AST、ALT、TBIL均较B组有不同程度的降低（P0.05），差异有统计学意义；E组血氨较C组低（P0.05），差异有统计学意义。 (3) A组大鼠肝组织形态正常；E组肝组织形态变化较C、D、F组轻，为少量炎症侵润，有点状及少量灶状坏死，部分肝细胞肿胀.。 (4) B、C、D、E、F组均较A组正常组大鼠INF-α和内毒素浓度高（P0.05），差异有统计学意义；E组INF-α和内毒素浓度均低于C、 D组（P0.05），差异有统计学意义。 (5) B、C、 D、E、F组均较A组正常组大鼠GS、NOS表达量低（P0.05），差异有统计学意义；E组较C、D组GS表达量较低（P0.05），差异有统计学意义；C、D、E、F组GS、NOS表达量均较B组低（P0.05），差异有统计学意义；D、E、F组较C组NOS表达量低（P0.05），差异有统计学意义。结论： 1300mg·kg-1·d-1的TAA连续灌胃3天为诱导大鼠急性肝性脑病适宜时间。大鼠行为学改变显著、肝功能损害大、肝组织坏死较严重、有较高诱导率及较低致死率。 2A型肝性脑病大鼠通过醒脑静注射液或门冬氨酸鸟氨酸注射液治疗，可改善大鼠行为学变化及肝功能，，减少肝细胞炎症侵润及坏死，降低细胞炎症，降低血液中内毒素和INF-α水平，降低脑内GS和NOS的表达量。在不同剂量醒脑静治疗中，以10ml·kg-1·d-1的醒脑静治疗综合效果较好。
[Abstract]:Background and purpose:
At present, the pathogenesis of hepatic encephalopathy has not been fully understood, and there is no specific therapy for it. It is harmful for a long time. It has been studied by Chinese and Western medicine both at home and abroad. However, the available clinical observation data are very limited, and fewer cases are enrolled. There is a lack of strong clinical data to support Xingnaojing in the treatment of AHE.
The aim of this study was to investigate the optimal time for the establishment of hepatic encephalopathy model induced by thioacetamide (TAA) of 300mg 65 To provide scientific basis for the treatment of type A hepatic encephalopathy.
Method:
1 to explore the optimal time of TAA (300mg. Kg-1. D-1) to induce A type hepatic encephalopathy in rats.
(1) Sixty rats were divided into four groups: A, B, C and D. Group A was the normal control group. Group B, C and D were given saline of the same dosage for 4 days. Group B, C and D were given TAA for 2, 3 and 4 days respectively.
(2) To compare the behavioral changes, the induction rate and lethality of AHE, the blood ammonia, glutamic oxalate transaminase (AST), glutamic-Pyruvic transaminase (ALT) and total bilirubin (TBIL) levels 24 hours after the end of administration in each group. Change.
2 to explore the effects of Xingnaojing on liver function, endotoxin, GS and NOS expression in rats with A hepatic encephalopathy.
(1) Sixty rats were divided into six groups: A, B, C, D, E and F, in which group A was the normal control group, group B was the hepatic encephalopathy model group, and group C, D, E and F was the treatment group, which were given Xingnaojing (2.5ml.kg-1.d-1), Xingnaojing (5ml.kg-1.d-1), Xingnaojing (10ml.kg-1.d-1), ornithine aspartate (2g.kg-1.d-1.d-1), respectively. -1) for 5 days.
(2) Comparing the behavioral changes, the differences of AST, ALT and TBIL, the morphological changes of liver tissues, and the expression of INF-alpha, endotoxin, GS and NOS in each group.
Result:
The optimal time for 1TAA (300mg. Kg-1. D-1) to induce A type hepatic encephalopathy in rats:
(1) The brain function grading of B, C, D experimental group was higher than that of A normal group (P 0.0083), the difference was statistically significant; the brain function score of C, D experimental group was higher than that of B experimental group (P 0.0083), the difference was statistically significant.
(2) Normal group A, no AHE-induced rats and no death rats; C, D group than B group induced rate higher (P 0.0083), the difference was statistically significant; D group than B, C group rats mortality higher (P 0.0083), the difference was statistically significant.
(3) The serum ammonia, AST, ALT and TBIL of rats in group B, C and D were significantly higher than those in group A (P 0.05), and the differences were statistically significant.
(4) The morphology of liver tissue in group A was normal, and the liver histology in group C and D showed obvious damage of inflammation, invasion, necrosis and fibrosis.
2 to explore the effects of Xingnaojing on liver function, endotoxin, GS and NOS expression in rats with A hepatic encephalopathy.
(1) There were no behavioral changes in normal rats in group A, and the behavioral changes and brain function grading of rats in group C, D, E and F were lower than those in group B (P 0.0033).
(2) The levels of serum ammonia, AST, ALT and TBIL in group B, C, D, E and F were higher than those in group A (P 0.05), and the difference was statistically significant; the levels of serum ammonia, AST, ALT and TBIL in group C, D, E and F were lower than those in group B (P 0.05), and the difference was statistically significant.
(3) The morphology of liver tissue in group A was normal; the morphological changes of liver tissue in group E were slighter than those in group C, D and F, with a small amount of inflammation and invasion, a small amount of focal necrosis, and some hepatocytes swelling.
(4) The concentrations of INF-a and endotoxin in group B, C, D, E and F were higher than those in group A (P 0.05), and the differences were statistically significant. The concentrations of INF-a and endotoxin in group E were lower than those in group C and D (P 0.05).
(5) The expression of GS and NOS in group B, C, D, E and F was lower than that in group A (P 0.05), and the difference was statistically significant; the expression of GS and NOS in group E was lower than that in group C and D (P 0.05), and the difference was statistically significant; the expression of GS and NOS in group C, D, E, F was lower than that in group B (P 0.05), and the expression of NOS in group D, E, F was lower than that in group C (P 0.05).
Conclusion:
The suitable time for inducing acute hepatic encephalopathy was 3 days of continuous administration of TAA at 1300 mg kg 1 D 1. The behavior of the rats was significantly changed, the liver function was damaged, the necrosis of liver tissue was serious, the inducing rate was high and the mortality was low.
Xingnaojing injection or ornithine Aspartate Injection can improve the behavioral changes and liver function of rats with hepatic encephalopathy type 2A, reduce inflammation, invasion and necrosis of hepatocytes, reduce inflammation of hepatocytes, reduce the levels of endotoxin and INF-alpha in blood, and decrease the expression of GS and NOS in brain. Xingnaojing treatment of L. Kg-1 D-1 has a good comprehensive effect.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R747.9

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