本文選題：失血性休克　切入點(diǎn)：腸淋巴液　出處：《河北北方學(xué)院》2013年碩士論文

【摘要】：大量研究表明,腸淋巴管結(jié)扎或腸淋巴液引流可減輕失血性休克后多器官的功能障礙與結(jié)構(gòu)損傷,經(jīng)淋巴途徑的腸源性感染是重癥休克后器官損傷的重要因素；氣體信號分子硫化氫(hydrogen sulfide, H2S)加劇了失血性休克后的炎癥反應(yīng),抑制H2S合成可降低器官損傷的程度。但在休克腸淋巴液介導(dǎo)器官損傷的這一過程中,H2S是否發(fā)揮作用,其具體作用機(jī)制如何?尚不清楚。 為此,本研究應(yīng)用胱硫醚-γ-裂解酶(cystathionine-y-lyase, CSE)抑制劑D,L-炔丙基甘氨酸(D, L-propargylglycine, PPG)和H2S供體硫氫化鈉(sodium hydrosulfide hydrate, NaHS)作用于行腸淋巴液引流的大鼠,觀察H2S對腸淋巴液引流改善器官功能與結(jié)構(gòu)以及Toll樣受體4(Toll-like receptor4, TLR4)、白介素10(interleukin10, IL-10)、IL-12、腫瘤壞死因子α(tumor necrosis factor a, TNFa)含量的影響,探討H2S在腸淋巴液引流減輕休克大鼠器官損傷中的作用與機(jī)制。 雄性Wistar大鼠30只隨機(jī)均分為：假手術(shù)組(Sham)、休克組(Shock)、休克+引流組(Shock+drainage)、休克+引流+PPG組(Shock+drainage+PPG)、休克+引流+NaHS組(Shock+drainage+NaHS)。放血前30min,休克+引流十PPG組、休克+引流+NaHS組腹腔分別注射PPG (45ml.kg-1)、NaHS (28μmol.kg-1);其它三組大鼠腹腔注射等量的生理鹽水(1ml.kg-1)。所有大鼠經(jīng)戊巴比妥鈉肌肉注射麻醉后,行股部手術(shù),右股靜脈注射肝素鈉全身抗凝,右股動脈插管連接生物信號采集系統(tǒng),連續(xù)監(jiān)測平均動脈血壓(mean artery pressure, MAP);左股側(cè)動脈插管后連接注射器固定于抽注機(jī)上備放血用；腹部手術(shù),剝離腸系膜淋巴管。待所有手術(shù)完成、穩(wěn)定30min后,休克組、休克+引流組、休克+引流+PPG組、休克+引流+NaHS組大鼠經(jīng)左股動脈勻速放血,10min內(nèi)將MAP降至40mmHg,復(fù)制失血性休克模型。維持低血壓60min后,將放出的全血與林格氏液按1：1混勻,經(jīng)右側(cè)股靜脈輸液復(fù)蘇,30min完成；觀察至輸液結(jié)束后3h。假手術(shù)組進(jìn)行相同的手術(shù)操作,但不放血、不輸液。休克+引流組、休克+引流+PPG組、休克+引流+NaHS組在輸液結(jié)束后,行腸系膜淋巴管插管,引流休克腸淋巴液至休克3h。 各組大鼠在相當(dāng)于輸液結(jié)束后3h這一時(shí)間點(diǎn),經(jīng)腹主動脈取血,制備血漿,檢測反映心肌細(xì)胞損傷的指標(biāo)肌酸激酶同工酶(creatine phosphokinase isoenzyme, CK-MB)與乳酸脫氫酶-1(lactic acid dehydrogenase-1, LDH-1)、反映肝功能的指標(biāo)天門冬氨酸氨基轉(zhuǎn)移酶(aspartate aminotransferase, AST)、丙氨酸氨基轉(zhuǎn)移酶(alanine aminotransferase, ALT)與總膽汁酸(total bile acid, TBA)、反映腎功能的指標(biāo)尿素(Urea)、肌酐(Cre),以及總蛋白、H2S含量。取肺左葉,制備支氣管肺泡灌洗液(bronchoalveolar lavage fluid, BALF),進(jìn)行細(xì)胞計(jì)數(shù)以及蛋白檢測,計(jì)算肺通透性指數(shù)(lung permeability index, LPI),同時(shí)取右肺上葉,測定肺干／濕(D/W)比。留取固定位置的肺、心肌、肝、腎組織,部分經(jīng)多聚甲醛固定用于觀察組織形態(tài)學(xué)變化；另一部分制備組織勻漿用于檢測H2S含量、CSE活性以及TLR4、IL-10、IL-12、TNFα的含量。 結(jié)果發(fā)現(xiàn),腸淋巴引流可提高失血性休克大鼠液體復(fù)蘇后一些時(shí)間點(diǎn)的MAP,盡管PPG沒有明顯的作用,但NaHS降低了腸淋巴引流對MAP的提升作用。組織形態(tài)學(xué)觀察表明,假手術(shù)組大鼠肺、心肌、肝、腎組織結(jié)構(gòu)基本正常,休克組大鼠肺、心肌、肝組織出現(xiàn)了形態(tài)學(xué)損傷,休克+引流組與休克+引流+PPG組大鼠肺、心肌、肝、腎的組織損傷較輕,NaHS加重了各器官的損傷程度。休克組大鼠BALF中細(xì)胞總數(shù)、LPI顯著高于假手術(shù)組,休克+引流組BALF中細(xì)胞總數(shù)、LPI顯著低于休克組,休克+引流+PPG組BALF中細(xì)胞總數(shù)、LPI顯著低于休克+引流組,休克+引流+NaHS組BALF細(xì)胞總數(shù)、LPI顯著高于休克+引流組；而大鼠肺組織的D/W比出現(xiàn)了相反的變化。同時(shí)可見,休克組大鼠血漿AST、ALT、TBA、LDH-1、CK-MB、Urea、Cre均顯著高于假手術(shù)組,休克+引流組大鼠血漿的AST、ALT、TBA、 Urea、Cre含量顯著低于休克組,休克+引流+PPG組血漿AST、ALT、TBA、Cre水平顯著低于休克+引流組,休克+引流+NaHS組血漿AST、ALT、Urea、Cre含量顯著高于休克+引流組。 同時(shí),研究發(fā)現(xiàn),休克組血漿以及肺、肝、腎、心肌組織的H2S含量均顯著升高,腸淋巴液引流顯著降低了休克大鼠血漿以及肺、肝、腎、心肌組織H2S的含量,PPG進(jìn)一步降低了休克+引流組大鼠這些組織H2S的含量,H2S供體NaHS則提高了休克+引流組大鼠的血漿以及肺、肝、腎、心肌組織H2S的含量。進(jìn)一步的研究結(jié)果顯示,休克組大鼠肺、肝、腎、心肌組織的TLR4、IL-10、IL-12、TNFα含量均顯著升高,腸淋巴液引流則降低了肝勻漿IL-10、肺、肝勻漿IL-12含量以及肺、肝、腎、心肌組織的TNFα含量；休克+引流組大鼠給予CSE抑制劑PPG后,進(jìn)一步降低了各器官組織的TLR4含量,肺、腎、心肌IL-10含量、肺組織IL-12含量、肺、肝、腎、心肌的TNFα含量均顯著下降；H2S供體NaHS在增加休克+引流組大鼠肺、肝、腎、心肌的TLR4含量的同時(shí),增加了肺、肝、腎組織IL-10、IL.12以及肺、肝、腎、心肌的TNFα含量。 研究結(jié)果表明,H2S在休克腸淋巴液引流減輕肺、肝、心、腎等重要器官的功能障礙及結(jié)構(gòu)損傷中起負(fù)面作用,其作用機(jī)制與TLR4引起的炎癥反應(yīng)加重有關(guān)。研究結(jié)果補(bǔ)充完善了“多器官損傷的腸淋巴途徑學(xué)說”,為以淋巴為靶點(diǎn),針對H2S的調(diào)節(jié),進(jìn)一步應(yīng)用于危重病的臨床防治奠定了實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:A large number of studies show that intestinal lymph duct ligation or lymph drainage can reduce the structural damage and multiple organ dysfunction after hemorrhagic shock, the intestinal lymph infection is an important factor in organ injury after severe hemorrhagic shock; HYDROGENSULFIDE (hydrogen sulfide H2S) exacerbated inflammation after hemorrhagic shock. Inhibition of H2S synthesis can reduce the extent of organ damage. But in shock lymph mediated the process of organ damage, whether H2S play a role in how the specific mechanism is not clear.?
Therefore, the research and application of cystathionine gamma lyase (cystathionine-y-lyase, CSE) inhibitor D, L- propargylglycine (D, L-propargylglycine, PPG) and H2S donor sodium hydrosulfide (sodium hydrosulfide, hydrate, NaHS) on mesenteric lymph drainage in rats, the effects of H2S on intestinal lymph drainage to improve organ function with the structure and Toll like receptor 4 (Toll-like, receptor4, TLR4), interleukin 10 (Interleukin10, IL-10), IL-12, tumor necrosis factor alpha (tumor necrosis factor A, TNFa) on the content of H2S in intestinal lymph drainage to reduce the role and mechanism of organ injury in shock rats.
30 male Wistar rats were randomly divided into sham operation group (Sham), shock group (Shock), the shock + drainage group (Shock+drainage), the shock + drainage group +PPG (Shock+drainage+PPG), the shock + drainage group +NaHS (Shock+drainage+NaHS). Blood 30min before the shock + drainage group ten PPG, the shock + drainage +NaHS intraperitoneal injection of PPG group respectively (45ml.kg-1), NaHS (28 mol.kg-1); the other three groups of normal saline was injected to rats (1ml.kg-1). All rats were anesthetized with pentobarbital sodium after intramuscular injection, femoral surgery, right femoral vein injection of heparin sodium anticoagulation, the right femoral artery cannula connected with biological the signal acquisition system, continuous monitoring of mean arterial pressure (mean, artery pressure, MAP); the left femoral artery after intubation side connected to the injector is fixed on the pump injection machine bloodletting; abdominal surgery, dissection of mesenteric lymph duct. After all the operation is complete, stable 30min, shock group, Hugh G + drainage group, the shock + drainage group +PPG, the shock + drainage group +NaHS rats via left femoral artery at MAP to 40mmHg 10min of the blood, the rat model of hemorrhagic shock. Maintain hypotension after 60min, will release the whole blood and Ringer's solution according to 1:1 mix, the right femoral vein infusion recovery. 30min; to observe 3h. infusion after the sham operation group were same operation, but no bleeding, no infusion. The shock + drainage group, the shock + drainage group +PPG, the shock + drainage in group +NaHS after infusion for mesenteric lymph duct intubation, drainage of shock lymph to shock 3h.
The rats in the equivalent of 3H infusion after this point in time, abdominal aortic blood plasma preparation, detection index of creatine kinase myocardial cell injury (creatine phosphokinase isoenzyme CK-MB -1 (lactic) and lactate dehydrogenase acid dehydrogenase-1, LDH-1), index of aspartate amino liver metastasis enzyme (aspartate aminotransferase, AST), alanine aminotransferase (alanine, aminotransferase, ALT) and total bile acid (total bile, acid, TBA), renal function indexes of urea (Urea), creatinine (Cre), and the total protein content of H2S. The left lung lobe preparation, bronchoalveolar lavage fluid (bronchoalveolar lavage fluid, BALF), detect cell count and protein, calculate the lung permeability index (lung permeability, index, LPI), at the same time, the upper lobe of the right lung, lung wet / dry (D/W) ratio. Take a fixed position The lung, myocardium, liver and kidney tissue were partly fixed by paraformaldehyde to observe histomorphology. The other part was prepared tissue homogenate to detect H2S content, CSE activity and TLR4, IL-10, IL-12 and TNF alpha content.
The results showed that the intestinal lymph drainage can improve the MAP some time points of hemorrhagic shock rats after resuscitation, while PPG had no obvious effect, but NaHS reduced the intestinal lymph drainage to increase MAP. It shows that the histomorphology of the sham rats myocardium, liver, lung, kidney tissue structure was normal, shock rats lungs, myocardium, liver tissue morphological damage, the shock + drainage group and shock + drainage group +PPG rat lungs, myocardium, liver, renal tissue damage was lower, NaHS increased the damage degree of various organs. The total number of cells in BALF shock group rats, LPI was significantly higher than that of sham operation group the total number of cells, the shock + drainage group BALF, LPI group was significantly lower than the shock, the shock + drainage group +PPG the total number of cells in BALF, LPI was significantly lower than the shock + drainage group, the total number of BALF cells in +NaHS shock + drainage group, LPI was significantly higher than that of the shock + drainage group; while the rat lung tissue than D/W Now the opposite change. At the same time visible, plasma AST, rats in group TBA, shock ALT, LDH-1, CK-MB, Urea, Cre were significantly higher than those in sham group, shock + drainage group rat plasma AST, ALT, TBA, Urea, Cre were significantly lower than in the shock group, shock + drainage of plasma AST, +PPG group ALT, TBA, Cre level was significantly lower than the shock + drainage group, the shock + drainage group +NaHS plasma AST, ALT, Urea, Cre were significantly higher than those in shock + drainage group.
At the same time, the study found that shock plasma and lung, liver, kidney, myocardial H2S content increased significantly and the intestinal lymph drainage significantly decreased the plasma shock rats, lung, liver, kidney, myocardial H2S content, PPG further decreased the content of H2S of the drainage group shock +. H2S donor NaHS can improve the shock + drainage group rat plasma and lung, liver, kidney, myocardial H2S content. Further research results show that the shock group rat lung, liver, kidney, myocardium TLR4, IL-10, IL-12, TNF were all significantly increased, decreased intestinal lymph drainage the liver homogenate IL-10, lung, liver IL-12 content and lung, liver, kidney, TNF alpha in myocardial tissue; the shock + drainage group rats were treated with CSE inhibitor PPG, to further reduce the content of TLR4, the organs and tissues of lung, kidney, myocardial IL-10 content, IL-12 content, lung and liver. Kidney, myocardium The level of TNF alpha decreased significantly. H2S donor NaHS increased the TLR4 content of lung, liver, kidney and myocardium in shock + draining group, and increased TNF, IL-10 and IL.12 in lung, liver and kidney as well as TNF, alpha in lung, liver, kidney and myocardium.
The results show that the H2S in shock lymph drainage reduce the lung, liver, heart, dysfunction and structural damage play a negative role in kidney and other important organs in the inflammatory reaction and its mechanism caused by TLR4 increases. The results of the multiple organ injury of intestinal lymphatic pathway theory ", as for the lymph the target, the regulation of H2S, and further applied to clinical prevention and treatment of critically ill lay the foundation.

【學(xué)位授予單位】：河北北方學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R459.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 牛春雨;趙自剛;陳瑞華;張靜;張玉平;劉艷凱;;休克淋巴液對大鼠肺微血管內(nèi)皮細(xì)胞凋亡相關(guān)基因表達(dá)的影響[J];中國病理生理雜志;2008年02期

2 趙自剛;牛春雨;陳瑞華;張玉平;張靜;劉艷凱;李繼承;;腸淋巴途徑對休克大鼠心肌自由基、炎性介質(zhì)的影響[J];中國應(yīng)用生理學(xué)雜志;2007年04期

3 牛春雨;李繼承;趙自剛;陳瑞華;張靜;劉艷凱;張玉平;姜華;;腸系膜淋巴管結(jié)扎對失血性休克大鼠肺組織一氧化氮及其表達(dá)的影響[J];中國危重病急救醫(yī)學(xué);2006年09期

4 趙自剛;牛春雨;李志鵬;張敏;許幗杰;姜華;張靜;;腸淋巴液引流對失血性休克大鼠紅細(xì)胞流變性的影響[J];中國應(yīng)用生理學(xué)雜志;2012年02期

5 趙克森;;重癥難治性休克發(fā)生機(jī)制之進(jìn)展[J];中國病理生理雜志;2010年10期

，

本文編號：1688091