Wu Na, Wan Zhi-Ping, Han Ling, Liu Hui-Ying, Li Hua-Shan
Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine Nanchang 330006,China.
Nanchang Hongdu Hospital of Traditional Chinese Medicine Nanchang 330000,China.
Zhongguo Zhong Yao Za Zhi. 2021 Mar;46(5):1191-1196. doi: 10.19540/j.cnki.cjcmm.20201117.402.
To explore the effect of Huangqin Decoction on ulcerative colitis(UC) pyroptosis, and to explain the mechanism of pyroptosis based on NOD-like receptor thermoprotein domain 3(NLRP3)/cysteine proteinase 1(caspase-1) pathway. The animal model of UC induced with 3% dextran sodium sulfate(DSS) was established. The experimental animals were divided into control group, model group, low-dose(4.55 g·kg(-1)), medium-dose(9.1 g·kg(-1)) and high-dose(18.2 g·kg(-1)) Huangqin Decoction groups and salazosulfapyridine group(0.45 g·kg(-1)). While modeling, intragastric administration was given for 7 consecutive days. On the 8 th day, the mice were euthanized, the colon length was collected, and the histopathological changes were observed by HE staining. The content of interleukin-18(IL-18) was observed by ELISA. The content of lactatedehydrogenase(LDH)was determined by microplate method. TUNEL assay kit was used to detect the cell death. The immunohistochemical staining was used to detect the expressions of NLRP3 and apoptosis-associated speck-like protein containing a CARD(ASC). Western blot was used to detect the expressions of interleukin-1β(IL-1β), caspase-1 and gasdermin D(GSDMD).The experimental study showed that compared with normal group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were significantly increased(P<0.05). Compared with model control group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were decreased, and these results were more significant in high-dose groups(P<0.05). The results of HE staining showed that Huangqin Decoction could improve the pathological changes of colon. Huangqin Decoction could inhibit UC cell pyroptosis, and the mechanism may be closely related to NLRP3/caspase-1 signaling pathway.
探讨黄芩汤对溃疡性结肠炎（UC）细胞焦亡的影响，并基于核苷酸结合寡聚化结构域样受体热蛋白结构域3（NLRP3）/半胱氨酸蛋白酶1（caspase-1）通路阐释细胞焦亡的机制。建立3%葡聚糖硫酸钠（DSS）诱导的UC动物模型。将实验动物分为对照组、模型组、低剂量（4.55 g·kg⁻¹）、中剂量（9.1 g·kg⁻¹）和高剂量（18.2 g·kg⁻¹）黄芩汤组以及柳氮磺吡啶组（0.45 g·kg⁻¹）。造模同时连续7天灌胃给药。第8天，处死小鼠，采集结肠长度，通过苏木精-伊红（HE）染色观察组织病理学变化。采用酶联免疫吸附测定（ELISA）法检测白细胞介素-18（IL-18）含量。采用微孔板法测定乳酸脱氢酶（LDH）含量。使用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）检测试剂盒检测细胞死亡情况。采用免疫组织化学染色法检测NLRP3和含半胱天冬酶激活和招募结构域的凋亡相关斑点样蛋白（ASC）的表达。采用蛋白质免疫印迹法检测白细胞介素-1β（IL-1β）、caspase-1和gasdermin D（GSDMD）的表达。实验研究表明，与正常组相比，LDH含量、TUNEL阳性染色、炎症因子（IL-18、IL-1β）以及与细胞焦亡相关的蛋白均显著升高（P＜0.05）。与模型对照组相比，LDH含量、TUNEL阳性染色、炎症因子（IL-18、IL-1β）以及与细胞焦亡相关的蛋白均降低，且在高剂量组中这些结果更显著（P＜0.05）。HE染色结果显示，黄芩汤可改善结肠的病理变化。黄芩汤可抑制UC细胞焦亡，其机制可能与NLRP3/caspase-1信号通路密切相关。
