大黄牡丹汤调控HMGB1/RAGE/NF-κB信号通路干预急性胰腺炎模型大鼠的机制

doi:10.11843/j.issn.0366-6964.2021.011.027

摘要/Abstract

摘要： 旨在研究大黄牡丹汤对急性胰腺炎模型大鼠的干预作用及可能的分子机制。将96只大鼠随机分为6组：假手术组、模型组、阳性药物对照组和大黄牡丹汤高、中、低3个剂量试验组（n=16）。采用胰胆管逆行注射牛磺胆酸钠溶液复制急性胰腺炎模型，模型组及假手术组均采用灌胃方式给予生理盐水；对照组皮下注射奥曲肽；高、中、低3个剂量试验组分别灌胃不同剂量大黄牡丹汤水煎液，1次·d^-1，连续6 d。常规麻醉后脱颈椎处死，心脏取血，开腹剖取胰腺组织。在光学显微镜下对胰腺的病理学改变进行观察，采用IHC和Western blot法检测胰腺组织HMGB1、RAGE、p-NF-κBp65、p-IκBα蛋白表达水平，采用RT-PCR法检测胰腺组织HMGB1、RAGE基因表达水平，ELISA检测胰腺组织中IL-1β、TNF-α、IL-6含量。结果发现：1）与假手术组相比，模型组大鼠一般生存状况较差，镜下可见胰腺组织明显肿胀以及扩张充血，胰腺组织中HMGB1、RAGE基因和蛋白相对表达量，下游蛋白NF-κBp65、IκBα磷酸化水平，炎性因子TNF-α、IL-1β、IL-6含量显著升高（P<0.05）；2）与模型组相比，治疗组大鼠一般生存状况不同程度改善，镜下间质性水肿以及坏死灶明显改善，HMGB1、RAGE基因和蛋白相对表达量，下游蛋白NF-κBp65、IκBα磷酸化水平，炎性因子TNF-α、IL-1β、IL-6含量显著下降（P<0.05），其中，尤以大黄牡丹汤高剂量组最为明显（P<0.05）。本研究表明，大黄牡丹汤能够显著缓解大鼠急性胰腺炎疾病进展，其机制与其能够抑制HMGB1/RAGE/NF-κB通路激活，进而阻断炎性级联反应有关。

关键词: 大黄牡丹汤, 急性胰腺炎, HMGB1/RAGE/NF-κB信号通路

Abstract: To investigate the intervention effect of Dahuang Mudan decoction on acute pancreatitis model rats and its possible molecular mechanism. Ninty-six rats were selected and randomly divided into 6 groups: sham operation group, model group, positive drug control group and Dahuang Mudan decoction high, medium and low dose groups (n=16). The acute pancreatitis model was established by retrograde injection of sodium taurocholate into the pancreatic duct. The model group and sham operation group were given normal saline by gavage; The positive drug control group was subcutaneously injected with octreotide; the with high, medium and low dose experimental group were given Dahuang Mudan decoction for 6 days. Blood was taken from the heart and pancreatic tissue was dissected by laparotomy. The expression levels of HMGB1, RAGE, p-NF-κBp65 and p-IκBα in pancreatic tissue were detected by IHC and Western blot methods. The expression levels of HMGB1 and RAGE in pancreatic tissue were detected by RT-PCR. The contents of IL-1β, TNF-α and IL-6 in pancreatic tissue were detected by ELISA. The results showed that:1) Compared with the sham operation group, the rats in the model group had a relatively poor general survival condition, and a significant swelling and hyperemia of the pancreatic tissue under the microscope. The protein expression of HMGB1 and RAGE, the phosphorylation of NF-κBp65 and IκBα, and the contents of TNF-α, IL-1β and IL-6 in pancreatic tissue were significantly increased (P<0.05); 2) Compared with the model group, the general living conditions of the rats in the treatment groups were improved in different degrees, interstitial edema and necrosis were significantly improved under the microscope. The protein and gene expression of HMGB1 and RAGE, the phosphorylation of NF-κB p65 and IκBα, and the contents of TNF-α, IL-1 β and IL-6 in pancreatic tissue were significantly decreased, especially in the large dose group of Dahuang Mudan decoction (P<0.05). This study indicate that Dahuang Mudan decoction can significantly improve acute pancreatic lesions in rats, and its mechanism is that it can inhibit the activation of HMGB1 /RAGE/ NF-κB pathway and block the inflammatory cascade.

Key words: Dahuang Mudan decoction, acute pancreatitis, HMGB1/RAGE/NF-κB signaling pathway

中图分类号:

宋冰, 辜吉秀, 汪永锋, 张延英, 余四九. 大黄牡丹汤调控HMGB1/RAGE/NF-κB信号通路干预急性胰腺炎模型大鼠的机制[J]. 畜牧兽医学报, 2021, 52(11): 3260-3269.

SONG Bing, GU Jixiu, WANG Yongfeng, ZHANG Yanying, YU Sijiu. Mechanism of Dahuang Mudan Decoction Regulating HMGB1/RAGE/NF-κB Signaling Pathway in Rats with Acute Pancreatitis[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3260-3269.

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