【摘要】：目的:探討Apelin-13對大鼠急性壞死性胰腺炎(Acute necrotic pancreatitis,ANP)自噬相關(guān)蛋白LC3-II、Beclin-1的影響。方法:取健康雄性SD大鼠54只,將它們隨機(jī)分為C組(對照組)、P組(ANP組)、S組(ANP+Apelin-13治療組),各組18只。C組為假手術(shù)組,P組SD大鼠以5%的牛黃膽酸鈉膽胰管逆行推注制造急性壞死性胰腺炎模型,S組是造模后5min給予Apelin-13尾靜脈注射干預(yù)組。P組和S組在建立急性壞死性胰腺炎模型后5min,分別尾靜脈注射等量生理鹽水和Apelin-13(0.1ug/g)。每組大鼠按不同的時(shí)相點(diǎn)分別于術(shù)后第3,6,12h麻醉后腹主動脈采血,采取血清檢測淀粉酶;并取一部分胰腺組織用4%的甲醛固定,做常規(guī)病理切片和HE染色;另一部分用Western-Blot檢測自噬有關(guān)蛋白LC3-II、Beclin-1的表達(dá)情況。結(jié)果:1.胰腺病理學(xué)評分:(1)P組、S組和C組之間各時(shí)間點(diǎn)的病理損傷評分差異有著顯著性(P0.05);(2)P組損傷評分明顯增加,隨時(shí)間增加損傷程度增加,各時(shí)間點(diǎn)差異有顯著性(P0.05):(3)S組較P組各時(shí)間點(diǎn)的病理損傷評分有所減輕(P0.05)。2.血清淀粉酶的變化:(1)C組的各個(gè)時(shí)間點(diǎn)大鼠血清淀粉酶未見明顯差異(P0.05)。(2)各個(gè)時(shí)點(diǎn)P組大鼠血清淀粉酶較C組(對照組)顯著升高(P0.05);與P組相比較,S組大鼠各時(shí)間點(diǎn)的血清淀粉酶有明顯降低(P0.05)。3.Western-Blot檢測LC3-II,Beclin-1蛋白的表達(dá)結(jié)果:與C組相比,在大鼠發(fā)生ANP 3h,6h,12h后P組自噬相關(guān)蛋白LC3-II、Beclin-1的蛋白表達(dá)均有著顯著增加(P0.05);然而與P組相比,S組各時(shí)間點(diǎn)自噬相關(guān)蛋白LC3-II和Beclin-1的蛋白表達(dá)水平出現(xiàn)明顯降低(P0.05)。結(jié)論:1、急性壞死性胰腺炎時(shí)自噬的表達(dá)是增加的,且這種自噬的表達(dá)會隨著病理損傷程度加重而增加。2、Apelin-13可以明顯抑制急性壞死性胰腺炎后大鼠胰腺組織細(xì)胞自噬相關(guān)蛋白Beclin-1,LC3-II的表達(dá)。
[Abstract]:Aim: to investigate the effect of Apelin-13 on autophagy associated protein (LC3-II,Beclin-1) in (Acute necrotic pancreatitis,ANP of rats with acute necrotizing pancreatitis. Methods: Fifty-four healthy male SD rats were randomly divided into group C (control group,), P group (ANP group), S group (ANP Apelin-13 treatment group), 18 rats in each group. Acute necrotizing pancreatitis was induced by retrograde injection of 5% sodium taurocholate into the biliary and pancreatic duct of SD rats in group P. the rats in group S were treated with Apelin-13 caudal vein injection after model making, and group P and group S were treated with Apelin-13 for 5 min after the establishment of acute necrotizing pancreatitis. The same amount of saline and Apelin-13 (0.1ug/g) were injected into caudal vein respectively. The blood samples of abdominal aorta were collected from each group according to different time phase points at 6h after anesthesia, and serum amylase was detected, and some pancreatic tissues were fixed with 4% formaldehyde for routine pathological sections and HE staining. In the other part, the expression of autophagy related protein LC3-II,Beclin-1 was detected by Western-Blot. Results: 1. Pancreatic pathological score: (1) there was significant difference in pathological injury score between P group, S group and C group at each time point (P0.05); (2) the injury score of group P increased significantly, and the degree of injury increased with the increase of time. There was significant difference at each time point (P0.05): (3). The pathological injury score of group S was lighter than that of group P (P0.05). The changes of serum amylase: (1) there was no significant difference in serum amylase in group C (P0.05). (2). The serum amylase in group P was significantly higher than that in group C (P0.05). Compared with group P, serum amylase in group S was significantly lower than that in group P (P0.05). The expression of LC3-II,Beclin-1 protein was detected by 3.Western-Blot: compared with group C, ANP occurred in rats for 3 h or 6 h. After 12 hours, the protein expression of autophagy associated protein LC3-II,Beclin-1 was significantly increased in P group (P0.05). However, compared with P group, the expression levels of autophagy associated protein LC3-II and Beclin-1 in S group were significantly decreased at each time point (P0.05). Conclusion: 1. The expression of autophagy was increased in acute necrotizing pancreatitis, and the expression of autophagy increased with the severity of pathological injury. Apelin-13 could significantly inhibit the expression of autophagy associated protein (Beclin-1,LC3-II) in pancreatic tissue of rats after acute necrotizing pancreatitis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R576

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