Lu Zhuo, Xiong Wanna, Xiao Simeng, Lin Yilong, Yu Kai, Yue Guihua, Liu Qiaoming, Li Fang, Liang Jianqin
1Guangxi University of Chinese Medicine, Nanning, 530001 China.
2Department of Pharmacy, Guangxi Medical College, Nanning, 530023 China.
Chin Med. 2020 May 8;15:45. doi: 10.1186/s13020-020-00327-9. eCollection 2020.
Ulcerative colitis (UC) is an intestinal disease which was characterized by intestinal inflammation, mucosal injury and fibrosis. In this paper, the effect of Huanglian Jiedu Decoction (HJD), a well-known traditional Chinese medicine with significant anti-inflammatory effect, on dextran sulphate sodium (DSS)-induced UC in mice and inhibition of JAK2/STAT3 pathway were investigated.
BALB/c mice were randomly divided into 6 groups: HJD group (high, medium and low dose), USAN group, UC group, and control group. UC in mice were induced through free access to 3% DSS solution. After being treated with HJD for 8 days, all animals were sacrifice. Pathological examination of colonic specimen was performed by H&E staining. Cytokines (TNF-α, IL-6, and IL-1β) in colon were assayed by ELISA and immunofluorescence, MPO in colon and ATT in serum were detected by ELISA. Moreover, mice in HJD group and UC group were treated with AG490 to inhibit the expression of JAK2 protein, then the expression of JAK2 and STAT3 protein in colon was determined by western blotting and immunofluorescence staining. Furthermore, KI67 in colon was examined by immunohistochemistry, and apoptosis was detected by TUNEL staining, and collagen deposition was assayed by Masson staining after JAK2/STAT3 pathway in UC mice was inhibited by HJD.
After mice being treated with HJD, the symptoms (weight loss and haematochezia) of UC were alleviated, and the contents of inflammatory cytokines (TNF-α, IL-6 and IL-1β) and MPO in colon were significantly decreased. The expression of JAK2 and STAT3 protein was reduced after administration with HJD. After JAK2/STAT3 pathway being inhibited with HJD, the cell apoptosis, collagen deposition and immunoreactivity of macrophage in colon were significantly reduced, but the expression of Ki67 was markedly enhanced in both UC group and HJD group compare with control group.
HJD treatment can alleviate intestinal mucosal damage and has the protective effect on UC by downregulating JAK2 and STAT3 expression to reduce inflammation via JAK2/STAT3 pathway.
溃疡性结肠炎（UC）是一种以肠道炎症、黏膜损伤和纤维化为特征的肠道疾病。本文研究了具有显著抗炎作用的著名中药黄连解毒汤（HJD）对葡聚糖硫酸钠（DSS）诱导的小鼠UC的影响以及对JAK2/STAT3通路的抑制作用。
将BALB/c小鼠随机分为6组：HJD组（高、中、低剂量）、美沙拉嗪组、UC组和对照组。通过自由饮用3% DSS溶液诱导小鼠发生UC。用HJD治疗8天后，处死所有动物。结肠标本经苏木精-伊红（H&E）染色进行病理检查。采用酶联免疫吸附测定（ELISA）和免疫荧光法检测结肠中的细胞因子（肿瘤坏死因子-α、白细胞介素-6和白细胞介素-1β），用ELISA检测结肠中的髓过氧化物酶（MPO）和血清中的丙氨酸氨基转移酶（ALT）。此外，HJD组和UC组小鼠用AG490处理以抑制JAK2蛋白的表达，然后通过蛋白质免疫印迹法和免疫荧光染色法检测结肠中JAK2和STAT3蛋白的表达。此外，HJD抑制UC小鼠的JAK2/STAT3通路后，通过免疫组织化学检测结肠中的增殖细胞核抗原（KI67），通过脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）染色检测细胞凋亡，并用Masson染色法检测胶原沉积。
用HJD治疗小鼠后，UC的症状（体重减轻和便血）得到缓解，结肠中炎性细胞因子（肿瘤坏死因子-α、白细胞介素-6和白细胞介素-1β）和MPO的含量显著降低。给予HJD后，JAK2和STAT3蛋白的表达降低。HJD抑制JAK2/STAT3通路后，结肠中的细胞凋亡、胶原沉积和巨噬细胞免疫反应性显著降低，但与对照组相比，UC组和HJD组中KI67的表达均明显增强。
HJD治疗可减轻肠道黏膜损伤，通过下调JAK2和STAT3表达，经JAK2/STAT3通路减轻炎症，对UC具有保护作用。
