Min Si,1– 4,* Yan Wang,3,* Hao Wu,4 Yuzhen Huang,1,2 Mengqing Ma,1,2 Kunhan Su,4 Dan Qiao,3 Weiwei Tao,3 Wanli Liu1,2 

1Department of Gastroenterology, Nanjing First Hospital, Nanjing University of Chinese Medicine, Nanjing, 210014, People’s Republic of China; 2Department of Gastroenterology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, People’s Republic of China; 3Nanjing University of Chinese Medicine, Nanjing, 210023, People’s Republic of China; 4Department of Gastroenterology, Nanjing Integrated Traditional Chinese and Western Medicine Hospital, Nanjing University of Chinese Medicine, Nanjing, 210014, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Wanli Liu, Department of Gastroenterology, Nanjing First Hospital, Nanjing University of Chinese Medicine, Nanjing, 210006, People’s Republic of China, Email njzxjh001@njucm.edu.cn Weiwei Tao, Nanjing University of Chinese Medicine, Nanjing, 210023, People’s Republic of China, Email taoww@njucm.edu.cn

Background: Jianpi Lishi Jiedu Decoction (JLJD) are used in China to prevent colorectal adenoma recurrence, but their molecular mechanism is unclear.
Objective: Evaluate JLJD’s preventive and therapeutic effects on colorectal adenomas and elucidate its molecular mechanisms.
Methods: JLJD components were identified via HPLC. The ApcMin/+ mouse model assessed therapeutic efficacy. Effects on colorectal tissue proliferation/apoptosis were analyzed. Flow cytometry evaluated Th17 cells; ELISA quantified inflammatory cytokines (IL-17, IL-6, IL-1β, IL-18). Jak2/Stat3/RORγt pathway proteins were detected by Western blot (WB) and immunohistochemistry (IHC). A Jak2 activator (RO8191) validated functional targets. Key components (chlorogenic acid, atractylenolide I) were tested in vitro for: (1) non-toxic concentrations (MTT), (2) IL-17A levels, (3) Th17 differentiation, (4) p-Stat3/RORγt expression (WB).
Results: JLJD significantly reduced adenoma number and progression in ApcMin/+ mice. Anti-Ki67 IHC showed suppressed proliferation; TUNEL assay confirmed induced apoptosis. ELISA indicated JLJD significantly decreased pro-inflammatory cytokine levels. WB/IHC demonstrated JLJD inhibited Th17 cell differentiation by downregulating Jak2/Stat3/RORγt pathway proteins in colon tissues and mesenteric lymph nodes. RO8191 abrogated JLJD’s anti-adenoma effects, reversed Th17 suppression, and nullified pathway inhibition. In vitro, chlorogenic acid and atractylenolide I significantly reduced IL-17A, reversed Th17 expansion, and decreased p-Stat3/RORγt expression.
Conclusion: JLJD inhibits the differentiation of Th17 cells by suppressing the Jak2/Stat3/RORγ pathway, thereby exerting an inhibitory effect on colorectal adenomas.

Keywords: Jianpi Lishi Jiedu decoction, colorectal adenomas, Th17 cell differentiation, Jak2/Stat3/RORγt pathway, ApcMin/+ mouse model