中药方剂葛根芩连汤缓解LPS诱导小鼠回肠炎症作用研究

Study on the Effects of the Chinese Herbal Formula Ge Gen Qin Lian Decoction in Alleviating LPS-Induced Ileal Inflammation in Mice

本文拟研究葛根芩连汤(GQD)对脂多糖(LPS)刺激诱导的小鼠回肠炎症的抑制作用与对肠道菌群的调节作用。雄性C57BL/6小鼠经口给予治疗剂量的GQD和抗生素(ABX) 7 d,第8天腹腔注射10 mg/kg的LPS。使用HE染色和AB-PAS染色进行回肠组织学评价;RT-qPCR检测回肠组织炎症因子(IL-1 β、IL-6、TNF-α、IL-10、IL-17和IL-22)、LPS受体(CD14、TLR4和MyD88) mRNA水平;ELISA测定超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;对盲肠内容物与粪便进行细菌革兰氏染色计数。GQD显著降低了小鼠回肠组织中由LPS诱导的炎症因子的mRNA水平(IL-1 β、IL-6、TNF-α、IL-17、IL-22、IL-10),LPS受体(CD14、MyD88、TLR4)的表达、SOD活性、MDA含量、修复了上皮细胞之间的紧密连接。GQD一定程度上改变了肠道菌群的结构。GQD预处理可以通过改变肠道菌群和肠屏障在一定程度上减轻LPS导致的肠损伤。This paper investigates the inhibitory effects of Ge Gen Qin Lian Decoction (GQD) on LPS-induced intestinal inflammation and dysbiosis. Male C57BL/6 mice were given therapeutic doses of GQD and antibiotics (ABX) orally for 7 days, followed by LPS challenge (10 mg/kg) intraperitoneally on day 8. Histological evaluation was performed using HE and AB-PAS staining;RT-qPCR was performed to detect inflammatory factors (IL-1β, IL-6, TNF-α, IL-10, IL-17 and IL-22), LPS receptors (CD14, TLR4 and MyD88) at mRNA levels;Superoxide Dismutase (SOD) activity and Malonic dialdehyde (MDA) content were determined using ELISA;Cecum and fecal bacterial Gram staining and 16S rDNA-Seq were used to analyze the changes of intestinal flora. GQD significantly reduced LPS-induced inflammatory factors (IL-1β, IL-6, TNF-α, IL-17, IL-22, and IL-10), expression of LPS receptors (CD14, TLR4) and MyD88. GQD decreased SOD activity and MDA content, and repaired tight junctions between epithelial cells in mouse ileal tissues. GQD somewhat altered the structure. Pretreatment with GQD can change the intestinal flora and the intestinal barrier, and reduce the intestinal injury caused by LPS.