Huang Jianbo, Jiang Tao, Kang Jingyi, Xu Jiayi, Dengzhang Yiting, Zhao Zhengqi, Yang Chuqi, Wu Mengting, Xu Xiaoping, Zhang Guangji, Lou Zhaohuan
School of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, China.
The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
Front Pharmacol. 2022 Jul 6;13:933070. doi: 10.3389/fphar.2022.933070. eCollection 2022.
To demonstrate the effectiveness of Huangqin decoction (Huangqin Tang in Chinese, HQT) combined with Radix (Tengligen in Chinese, TLG) under the guidance of "dampness-heat theory" in preventing and treating colorectal cancer (CRC) with dampness-heat accumulation and to preliminarily reveal its mechanism. : The mice model of CRC was established by intraperitoneal injection of AOM combined with consumption of 2.5% DSS solution, and celecoxib, HQT, TLG, and their combination (HQT + TLG) were administered at the same time. The physical signs and death of the mice were observed daily. At the end of the experiment, the colorectal tissue was dissected, and the tumor was observed and counted. HE staining and Masson's staining were used to observe the histopathological changes of colon. Expression levels of TNF-α, IL-6, and IL-10 in colorectal tissue were detected by ELISA, and the expression of TNF-α was observed by immunofluorescence. TUNEL assay was used to observe the apoptosis of tumor tissues, and immunohistochemistry was used to observe the expression of Ki-67 and occludin. The mRNA expression levels of claudin-1, occludin, ZO-1, and IL-6 and IL-17 were detected by RT-PCR, and occludin, ZO-1, NF-κB, and STAT3 protein levels were detected by Western blot. The composition of intestinal flora was analyzed by 16S rRNA. HQT + TLG could significantly reduce the mortality of model mice and improve the intestinal mucosal inflammatory cell infiltration and high-grade intraepithelial neoplasia in model mice. All administration groups show a great reduction in the levels of IL-6 and TNF-α in the colorectal tissues of model mice, and increase in the level of IL-10, the total number of CD3 T cells, the proportion of CD3CD4 T cells, and the ratio of CD4/CD8. HQT and HQT + TLG could significantly change the composition of intestinal flora and increase the abundance of Firmicutes and Patescibacteria. HQT and TLG combination has a good effect on inhibiting AOM-DSS-induced CRC. This function may be related to improving the composition of the intestinal flora, regulating the proportion of T-cell subsets in colorectal lymphoid tissue to improve inflammatory response, and downregulating the expression of claudin-1, inhibiting the activation of IL-6/STAT3 signaling pathway to improving abnormal hyperplasia.
为探讨在“湿热理论”指导下，黄芩汤（HQT）联合藤梨根（TLG）防治湿热蕴结型结直肠癌（CRC）的有效性，并初步揭示其作用机制。通过腹腔注射氧化偶氮甲烷（AOM）联合饮用2.5%葡聚糖硫酸钠（DSS）溶液建立CRC小鼠模型，同时给予塞来昔布、HQT、TLG及其联合用药（HQT+TLG）。每日观察小鼠的体征及死亡情况。实验结束后，解剖结直肠组织，观察并计数肿瘤。采用苏木精-伊红（HE）染色和Masson染色观察结肠组织病理学变化。采用酶联免疫吸附测定（ELISA）法检测结直肠组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-10（IL-10）的表达水平，采用免疫荧光法观察TNF-α的表达。采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）法观察肿瘤组织凋亡情况，采用免疫组织化学法观察Ki-67和闭合蛋白的表达。采用逆转录-聚合酶链反应（RT-PCR）检测闭合蛋白-1、闭合蛋白、紧密连接蛋白-1（ZO-1）、IL-6和IL-17的mRNA表达水平，采用蛋白质免疫印迹法检测闭合蛋白、ZO-1、核因子-κB（NF-κB）和信号转导与转录激活因子3（STAT3）蛋白水平。采用16S核糖体RNA（rRNA）分析肠道菌群组成。HQT+TLG可显著降低模型小鼠死亡率，改善模型小鼠肠道黏膜炎症细胞浸润及高级别上皮内瘤变。各给药组均使模型小鼠结直肠组织中IL-6和TNF-α水平大幅降低，IL-10水平、CD3 T细胞总数、CD3CD4 T细胞比例及CD4/CD8比值升高。HQT及HQT+TLG可显著改变肠道菌群组成，增加厚壁菌门和帕氏菌门丰度。HQT与TLG联合应用对抑制AOM-DSS诱导的CRC具有良好作用。该作用可能与改善肠道菌群组成、调节结直肠淋巴组织中T细胞亚群比例以改善炎症反应、下调闭合蛋白-1表达、抑制IL-6/STAT3信号通路激活以改善异常增生有关。
