Liu Meng-Ru, Li Hui, Wei Lan-Fu, Liu Xiao-Tong, An Zhen-Tao, Gu Li-Mei, Tian Yao-Zhou
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine Nanjing 210028, China Jiangsu Province Academy of Traditional Chinese Medicine Nanjing 210028, China.
Zhongguo Zhong Yao Za Zhi. 2023 Jan;48(1):226-233. doi: 10.19540/j.cnki.cjcmm.20221018.502.
The aim of this study was to explore the effects of Huangqin Tang(HQT) on the NLRP3/Caspase-1 signaling pathway in mice with DSS-induced ulcerative colitis(UC). C57BL/6J mice were randomly divided into a blank group, a model group(DSS group), and low-, medium-and high-dose HQT groups(HQT-L, HQT-M, and HQT-H), and western medicine mesalazine group(western medicine group). The UC model was induced in mice. Subsequently, the mice in the HQT-L, HQT-M, HQT-H groups, and the western medicine group were given low-, medium-, high-dose HQT, and mesalazine suspension by gavage, respectively, while those in the blank and DSS groups were given an equal volume of distilled water by gavage. After 10 days of administration, the body weight, DAI scores, and colonic histopathological score of mice in each group were determined. The levels of IL-6, IL-10, IL-1β, and TNF-α in serum were determined by ELISA. The mRNA expression of NLRP3 and Caspase-1 in colon tissues was determined by RT-qPCR. The protein expression of NLRP3 and Caspase-1 in colon tissues was detected by immunohistochemistry. The results showed that compared with the blank group, the DSS group showed decreased body weight of mice and increased DAI scores and intestinal histopathological score. Compared with the DSS group, the HQT groups and the western medicine group showed improved DAI scores, especially in the HQT-M, HQT-H, and the western medicine groups(P<0.05). The intestinal histopathological scores of the HQT groups and the western medicine group significantly decreased, especially in the HQT-M, HQT-H, and the western medicine groups(P<0.05). In addition, compared with the blank group, the DSS group showed elevated expression of NLRP3 and Caspase-1 in colon tissues, increased serum levels of IL-6, IL-1β, and TNF-α, and decreased IL-10 level. Compared with the DSS group, the HQT groups and the western medicine group displayed decreased expression of NLRP3 and Caspase-1 in colon tissues, reduced serum levels of IL-6, IL-1β, and TNF-α, and increased IL-10 level. The improvement was the most significant in the HQT-H group and the western medicine group(P<0.01). In conclusion, HQT may reduce the expression of NLRP3 and Caspase-1 in colon tissues, reduce the se-rum levels of IL-6, IL-1β, and TNF-α, and increase the expression of IL-10 by regulating the classic pyroptosis pathway of NLRP3/Caspase-1, thereby improving the symptoms of intestinal injury and inflammatory infiltration of intestinal mucosa in DSS mice to achieve its therapeutic effect.
本研究旨在探讨黄芩汤（HQT）对葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎（UC）小鼠NLRP3/半胱天冬酶-1信号通路的影响。将C57BL/6J小鼠随机分为空白组、模型组（DSS组）、低、中、高剂量HQT组（HQT-L、HQT-M、HQT-H）和西药美沙拉嗪组（西药组）。诱导小鼠建立UC模型。随后，分别对HQT-L、HQT-M、HQT-H组和西药组的小鼠灌胃给予低、中、高剂量HQT和美沙拉嗪混悬液，而空白组和DSS组的小鼠灌胃给予等体积的蒸馏水。给药10天后，测定各组小鼠的体重、疾病活动指数（DAI）评分和结肠组织病理学评分。采用酶联免疫吸附测定（ELISA）法测定血清中白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的水平。采用逆转录定量聚合酶链反应（RT-qPCR）法测定结肠组织中NLRP3和半胱天冬酶-1的mRNA表达。采用免疫组织化学法检测结肠组织中NLRP3和半胱天冬酶-1的蛋白表达。结果显示，与空白组相比，DSS组小鼠体重下降，DAI评分和肠道组织病理学评分升高。与DSS组相比，HQT组和西药组的DAI评分有所改善，尤其是HQT-M、HQT-H组和西药组（P<0.05）。HQT组和西药组的肠道组织病理学评分显著降低，尤其是HQT-M、HQT-H组和西药组（P<0.05）。此外，与空白组相比，DSS组结肠组织中NLRP3和半胱天冬酶-1的表达升高，血清中IL-6、IL-1β和TNF-α水平升高，IL-10水平降低。与DSS组相比，HQT组和西药组结肠组织中NLRP3和半胱天冬酶-1的表达降低，血清中IL-6、IL-1β和TNF-α水平降低，IL-10水平升高。HQT-H组和西药组的改善最为显著（P<0.01）。综上所述，HQT可能通过调节NLRP3/半胱天冬酶-1经典的细胞焦亡途径，降低结肠组织中NLRP3和半胱天冬酶-1的表达，降低血清中IL-6、IL-1β和TNF-α水平，增加IL-10的表达，从而改善DSS小鼠的肠道损伤症状和肠黏膜炎症浸润，达到治疗效果。
