右美托咪定通过HMGB1/TLR4/NF-κB抑制肠缺血再灌注诱导的自噬性细胞凋亡-现代医学

右美托咪定通过HMGB1/TLR4/NF-κB抑制肠缺血再灌注诱导的自噬性细胞凋亡

单位：广东医科大学附属医院麻醉科, 广东湛江 524001

分类号：R574

出版年·卷·期（页码）：2021·49·第二期（123-128）

摘要：

目的：探讨右美托咪定通过HMGB1/TLR4/NF-κB抑制肠缺血再灌注诱导的自噬性细胞凋亡的作用机制。方法：按选取40只健康雄性SD大鼠按随机数字表法分为对照组、肠缺血再灌注组（I/R组）、小剂量右美托咪定+肠缺血再灌注组（D1组）、中剂量右美托咪定+肠缺血再灌注组（D2组）、大剂量右美托咪定+肠缺血再灌注组（D3组），每组8只。5组大鼠均给予1%戊巴比妥钠50 mg/kg腹腔内注射，麻醉满意后经腹部正中切口，钝性分离暴露肠系膜上动脉，夹闭肠系膜上动脉60 min后松夹再灌注60 min制备肠缺血再灌注模型，对照组只开腹120 min，不夹闭肠系膜上动脉。D1、D2及D3组于夹闭前10 min通过尾静脉分别给予右美托咪定1 μg/kg，5 μg/kg及10 μg/kg，对照组及I/R组给予等量生理盐水。采用HE染色观察肠组织病理变化，ELISA法检测血清炎症因子TNF-α、IL-1及HMGBI水平，Western blotting法检测肠组织中Beclin-1、HMGB1、TLR4、NF-κB蛋白表达水平，TUNEL法检测肠组织细胞凋亡情况。结果：与对照组比较，I/R组肠组织可见大量中性粒细胞浸润，肺泡壁增厚，而D1、D2、D3组仅少量炎症细胞浸润；与对照组比较，I/R、D1、D2及D3组血清炎症因子TNF-α、IL-1及HMGBI水平升高，与I/R组比较，D1、D2及D3组TNF-α、IL-1及HMGBI水平降低；与对照组比较，I/R、D1、D2及D3组肠组织Beclin-1、HMGB1、TLR4、NF-κB蛋白表达升高，与I/R组比较，D1、D2及D3组Beclin-1、HMGB1、TLR4、NF-κB蛋白表达降低；与对照组比较，I/R组TUNEL阳性细胞[（62±10）个／视野]显著增加，D1、D2、D3组中TUNEL阳性细胞分别是[（36±7）个/视野]、[（35±6）个/视野]、[（32±5）个/视野]，与I/R组比较明显下降（P<0.05）。结论：肠缺血再灌注损伤激活细胞自噬性凋亡，右美托咪定通过HMGB1/TLR4/NF-κB抑制肠缺血再灌注诱导的自噬性细胞凋亡，从而发挥对肠缺血再灌注损伤的保护作用。

Objective:To observe the inhibitory effect of dexmedetomidine on autophagic apoptosis induced by intestinal ischemia-reperfusion through HMGB1/TLR4/NF-κB. Methods: Forty healthy male SD rats were randomly divided into control group, ischemia-reperfusion group (I/R group), low-dose dexmedetomidine+ischemia-reperfusion group (D1 group), medium-dose dexmedetomidine+ischemia-reperfusion group (D2 group) and high-dose dexmedetomidine+ischemia-reperfusion group (D3 group), 8 rats in each group. Rats in all 5 groups were given intraperitoneal injection of 1% pentobarbital sodium for 50 mg/kg. After satisfactory anesthesia, the superior mesenteric artery was exposed by blunt separation through a median abdominal incision, and the superior mesenteric artery was clamped for 60 min and then reperfused for 60 min to establish intestinal ischemia-reperfusion model. The control group only underwent laparotomy for 120 min, without clamping the superior mesenteric artery. Group D1, D2 and D3 were given dexmetomidine 1 μg/kg,5 μg/kg and 10 μg/kg via caudal vein 10 min before clamping, respectively, while the control group and I/R group were given the same amount of normal saline. The pathological changes of intestinal tissue were observed by HE staining, and the levels of serum inflammatory factors TNF-α, IL-1 and HMGB1 were detected by ELISA. The protein expression levels of Beclin-1, HMGB1, TLR4 and NF-κB in intestinal tissue were detected by Western blotting, and the apoptosis of intestinal tissue was detected by TUNEL. Results: Compared with the control group, there was neutrophils infiltration in the intestinal tissue of I/R group, but only a few inflammatory cells infiltrated in group D1, D2 and D3. Compared with the control group, the levels of serum inflammatory cytokines TNF-α, IL-1 and HMGBI ingroups I/R,D1,D2 and D3 were higher, and the levels of TNF-α, IL-1 and HMGB1 in group D1, D2 and D3 were lower than those in group I/R. Compared with the control group, the expression of Beclin-1, HMGB1, TLR4 and NF-κB protein in intestinal tissue of group I/R,D1, D2 and D3 increased, while the expression of Beclin-1, HMGB1, TLR4 and NF-κB protein decreased in group D1, D2 and D3 as compared with that of group I/R. Compared with the control group, the number of TUNEL positive cells in group I/R was significantly increased, and the number of TUNEL positive cells in group D1, D2 and D3 were significantly lower than that in group I/R (P<0.05). Conclusion: Intestinal ischemia-reperfusion injury activates autophagic apoptosis. Dexmetomidine plays a protective role in intestinal ischemia-reperfusion injured by inhibiting autophagic apoptosis induced by intestinal ischemia-reperfusion through HMGB1/TLR4/NF-κB.

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