Medical School of Chinese People's Liberation Army, The General Hospital of People's Liberation Army, Beijing, 100853, China.
Beijing Institute of Radiation Medicine, Beijing, 100850, China.
Chin J Integr Med. 2021 Mar;27(3):198-205. doi: 10.1007/s11655-020-3261-x. Epub 2020 Apr 1.
To observe the effect of Modified Xijiao Dihuang Decoction (, MXDD) on rats with radiation enteritis, and explore its action mechanism.
Thirty female Sprague Dawley rats were divided into the control, model, dexamethasone (DXM), golden bifid (GB) and MXDD groups using random number table, 6 rats in each group. Except the control group, the other rats were developed into radiation enteritis model by exposing to a single Co-γ ray at a dose of 11 Gy. The rats in the DXM, GB and MXDD groups were treated with DXM (1.425 mg/kg), GB (0.8 g/kg) and MXDD (36.0 g/kg) for 3 days, respectively. Body weight and diarrhea condition of rats were evaluated daily. On day 3, the feces of rats were collected for intestinal flora detection and the small intestinal tissues were also collected. Bacterial species annotation, alpha and beta diversities as well as composition of intestinal flora were detected and compared. The protein and mRNA expressions of interleukin 17 (IL-17), retinoid-related orphan nuclear receptor gamma t (ROR-γt) and forkhead/ winged helix transcription factor p3 (FoxP3) were determined by Western blot and polymerase chain reaction, respectively. The abundance and diversity of intestinal flora as well as the proportion at the phylum and genus levels were assayed by 16S rRNA metagenome sequencing. Correlation between intestinal flora and Th17/Treg was analyzed by heatmap method.
On day 1 to 3 after radiation, compared with the control group, the body weight in model group was decreased (P<0.05 or P<0.01). Compared with the model group, MXDD could alleviate weight loss and diarrhea caused by irradiation. At the phylum level, MXDD cause a significant increase in Firmicutes, and a decrease in Proteobacteria (P<0.05 or P<0.01). At the genus level, MXDD reduced the proportion of Escherichia Shigella (P<0.01). In addition, IL-17 and FoxP3 mRNA and protein expression levels were down-regulated and ROR-γt was up-regulated by MXDD treatment (P<0.05). Besides, Firmicutes and Lactobacillus were positively correlated with FoxP3 (r=0.73, 0.79, respectively; P<0.01), negatively correlated with IL-17 (r=0.66, 0.64, respectively; P<0.01 or P<0.05) and ROR-γt (r0.73, 0.81, respectively; P<0.01). Proteobacteria and Escherichia Shigella both had positive correlation with IL-17 (r 0.77, 0.57, respectively; P<0.01 or P<0.05 ) and ROR-γt (r=0.94, 0.79, respectively; P<0.01) and negative correlation with FoxP3 (r0.74, 0.65; P<0.01).
MXDD could improve the survival status of irradiated rats by regulating the richness, diversity and composition of intestinal flora, and restoring the balance of Th17/Treg.
观察加味犀角地黄汤对放射性肠炎模型大鼠的作用,并探讨其作用机制。
30 只雌性 SD 大鼠采用随机数字表法分为对照组、模型组、地塞米松组(DXM 组)、金双歧组(GB 组)和加味犀角地黄汤组(MXDD 组),每组 6 只。除对照组外,其余大鼠单次照射 Co-γ射线,剂量为 11 Gy,建立放射性肠炎模型。DXM 组、GB 组和 MXDD 组大鼠分别给予 DXM(1.425 mg/kg)、GB(0.8 g/kg)和 MXDD(36.0 g/kg)灌胃 3 天。每日评估大鼠体质量和腹泻情况。第 3 天收集大鼠粪便进行肠道菌群检测,取小肠组织,检测细菌种属注释、α和β多样性以及肠道菌群组成。采用 Western blot 和聚合酶链式反应检测白细胞介素 17(IL-17)、维甲酸相关孤儿核受体γ t(ROR-γt)和叉头/翅膀状螺旋转录因子 p3(FoxP3)的蛋白和 mRNA 表达。采用 16S rRNA 宏基因组测序检测肠道菌群的丰度和多样性以及门和属水平的比例。采用热图方法分析肠道菌群与 Th17/Treg 的相关性。
照射后 1~3 天,与对照组相比,模型组大鼠体质量降低(P<0.05 或 P<0.01)。与模型组相比,MXDD 可缓解照射引起的大鼠体质量减轻和腹泻。在门水平上,MXDD 可显著增加厚壁菌门,减少变形菌门(P<0.05 或 P<0.01)。在属水平上,MXDD 降低了大肠埃希菌/志贺菌属的比例(P<0.01)。此外,MXDD 下调了 IL-17 和 FoxP3 mRNA 和蛋白表达水平,上调了 ROR-γt 表达水平(P<0.05)。此外,Firmicutes 和乳杆菌与 FoxP3 呈正相关(r=0.73,0.79;P<0.01),与 IL-17(r=0.66,0.64;P<0.01 或 P<0.05)和 ROR-γt(r=0.73,0.81;P<0.01)呈负相关。变形菌门和大肠埃希菌/志贺菌属与 IL-17(r=0.77,0.57;P<0.01 或 P<0.05)和 ROR-γt(r=0.94,0.79;P<0.01)呈正相关,与 FoxP3 呈负相关(r=0.74,0.65;P<0.01)。
加味犀角地黄汤通过调节肠道菌群的丰度、多样性和组成,恢复 Th17/Treg 平衡,可改善照射大鼠的生存状态。