盐酸法舒地尔对脂多糖诱导的血管组织中ROCK1、Cx43、Cav1表达变化的影响

发布时间：2018-03-10 12:08

本文选题：内皮　切入点：血管　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨LPS诱导大鼠血管组织急性损伤后,其主动脉组织中Rho相关的卷曲蛋白激酶1(ROCK1)、缝隙连接蛋白(Cx)43和小窝蛋白(Cav)1表达的变化。Rho激酶抑制剂盐酸法舒地尔(HF)对脂多糖(lipopolysaccharide,LPS)诱导的大鼠主动脉组织中ROCK1、Cx43、Cav1表达变化的影响,阐明HF的抗炎作用及机制。方法:采用随机数字表法将24只雄性Sprague Dawley大鼠分为对照组(不做特殊处理)、HF组(腹腔注射HF 30 mg/kg)、脂多糖组(尾静脉注射脂多糖1 mg/kg)和脂多糖+HF组(腹腔注射HF 30 mg/kg半小时后,尾静脉注射脂多糖1 mg/kg),每组大鼠均为6只。LPS作用8 h后处死,提取主动脉组织。分别采用实时荧光定量PCR、Western blot和免疫组织化学法检测主动脉组织中ROCK1、Cx43和Cav1的m RNA和蛋白表达水平。结果:(1)实时荧光定量PCR显示,脂多糖组ROCK1(2.67±0.03)、Cx43(1.73±0.03)和Cav1(1.85±0.04)的m RNA表达水平均高于对照组中ROCK1(1.0±0.04)、Cx43(1.00±0.08)和Cav1(1.0±0.03)的m RNA表达水平及HF组ROCK1(0.77±0.04)、Cx43(0.91±0.01)和Cav1(0.82±0.03)的m RNA表达水平(P均0.05);而脂多糖+HF组ROCK1(0.38±0.02)、Cx43(0.58±0.02)和Cav1(0.27±0.01)的m RNA表达水平均低于脂多糖组(P均0.05)。(2)Western blot显示,脂多糖组中ROCK1(3.46±0.82)、Cx43(0.33±0.09)和Cav1(3.45±0.74)的蛋白表达水平均高于对照组中ROCK1(2.19±0.56)、Cx43(0.21±0.09)和Cav1(2.25±0.91)的蛋白表达水平及HF组中ROCK1(1.57±0.38)、Cx43(0.18±0.07)和Cav1(2.06±0.40)(P均0.05);而脂多糖+HF组ROCK1(1.09±0.52)、Cx43(0.11±0.06)和Cav1(2.06±0.40)的蛋白表达水平均低于脂多糖组(P均0.05)。(3)免疫组织化学法显示,脂多糖组ROCK1(84.1±0.9)、Cx43(99.1±2.1)和Cav1(167.0±6.4)的蛋白表达水平均高于对照组中ROCK1(53.7±2.9)、Cx43(46.2±0.8)和Cav1(84.9±1.0)的蛋白表达水平及HF组中ROCK1(40.1±0.9)、Cx43(35.1±0.6)和Cav1(74.4±0.5)的蛋白表达水平(P均0.05);而脂多糖+HF组ROCK1(30.4±0.6)、Cx43(21.4±1.3)和Cav1(55.8±2.8)的蛋白表达水平均低于脂多糖组(P均0.05)。结论:ROCK1、Cx43和Cav1均与LPS导致的血管内皮功能障碍相关。LPS可以通过上调ROCK1、Cx43和Cav1的表达,而发挥其致炎作用。HF可以通过直接抑制Rho A/ROCK1信号通路,降低ROCK1、Cx43的表达。同时也可能通过抑制Cav1的表达而调控Rho A/ROCK1信号通路的活性,改善LPS诱导的血管内皮的炎性损伤,从而发挥其抗炎作用。
[Abstract]:Objective: to investigate the acute vascular injury induced by LPS in rats. Changes in the expression of Rho associated convoluted protein kinase 1 (ROCK1), creating-junction protein (CxF43) and fossa protein (Cav1) in aorta. Effects of Rho kinase inhibitor, fasudil hydrochloride, on the expression of ROCK1Cx43 Cav1 in rat aorta induced by lipopolysaccharide polysaccharide (LPS). Methods: 24 male Sprague Dawley rats were randomly divided into control group (30 mg / kg intraperitoneal injection of HF 30 mg / kg), lipopolysaccharide group (1 mg / kg lipopolysaccharide). Lipopolysaccharide HF group (30 mg/kg after intraperitoneal injection of HF), Lipopolysaccharide (LPS) 1 mg / kg was injected into caudal vein. Six rats in each group were killed after exposure to LPS for 8 h. The expression levels of m RNA and protein of Rock1Cx43 and Cav1 in aortic tissue were detected by real-time fluorescence quantitative PCR Western blot and immunohistochemistry, respectively. The results showed that the expression of m RNA and protein in aortic tissue was detected by real-time fluorescence quantitative PCR. The expression level of m RNA in lipopolysaccharide group (ROCK1(2.67 卤0.03Cx43C 1.73 卤0.03) and Cav1(1.85 卤0.04) was higher than that in control group (ROCK1(1.0 卤0.04Cx43C 1.00 卤0.08) and Cav1(1.0 卤0.03), and the expression level of m RNA in HF group (ROCK1(0.77 卤0.04Cx430.91 卤0.01) and Cav1(0.82 卤0.03) was lower than that in lipopolysaccharide group (ROCK1(0.38 卤0.02Cx43C 0.58 卤0.02) and Cav1(0.27 卤0.01). In group A, P was 0.05 and P was 0. 05 and 0. 05, respectively. Western blot was used to display the results. The expression levels of ROCK1(3.46 卤0.82Cx43C 0.33 卤0.09 and Cav1(3.45 卤0.74) in the lipopolysaccharide group were higher than those in the control group (ROCK1(2.19 卤0.56Cx43C 0.21 卤0.09) and Cav1(2.25 卤0.91), ROCK1(1.57 卤0.38Cx430.18 卤0.07) and Cav1(2.06 卤0.40P in the HF group, while the expression levels of ROCK1(1.09 卤0.52Cx43C 0.11 卤0.06 and Cav1(2.06 卤0.40) in the lipopolysaccharide group were lower than those in the lipopolysaccharide group (P < 0.05). Histochemistry shows that. The expression levels of ROCK1(84.1 卤0.9Cx4399.1 卤2.1) and Cav1(167.0 卤6.4) in lipopolysaccharide group were higher than those in control group (46.2 卤0.8) and Cav1(84.9 卤1.0), ROCK1(40.1 卤0.9Cx43C 35.1 卤0.6.1 and Cav1(74.4 卤0.5) protein expression levels in HF group were lower than those in lipopolysaccharide HF group (ROCK1(30.4 卤0.6Cx4321.4 卤1.3) and Cav1(55.8 卤2.8). Conclusion both Cav1 and Cx43 are related to vascular endothelial dysfunction induced by LPS. LPs can up-regulate the expression of Cx43 and Cav1. By directly inhibiting Rho A / ROCK1 signaling pathway and decreasing the expression of ROCK1Cx43, HF could also regulate the activity of Rho A / ROCK1 signaling pathway by inhibiting the expression of Cav1, and improve the inflammatory injury of vascular endothelium induced by LPS. In order to play its anti-inflammatory effect.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R54

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