Context . Anti-mitochondrial antibody M2+ (AMAM2+) primary bile cholangitis ( PBC) is difficult to diagnose, and early diagnosis is the key to ensure effective treatment and the safety of patients. Objective . The study intended to investigate the role of T helper type 17 (Th17) cells and their transcription factors in the early diagnosis of AMA-M2+ PBC to provide an effective guarantee of the ability to predict the prognosis of patients in the future. Design . The research team designed a prospective controlled study. Setting . The study took place at the Affiliated Hospital of Hebei University in Baoding, Hebei, China. Participants . Participants were 30 patients with AMAM2+ PBC at the hospital between November 2020 and August 2021 and 30 healthy controls who concurrently underwent physical examinations. Outcome Measures . The study measured liver function (LF) and secretion of Th17 and its transcription factorsforkhead box P3 (Foxp3) and RAR-related orphan receptor gamma (ROR.t)-and inflammatory factorsinterleukin-17 IL-17 and IL-22-in participants' peripheral blood. The study also evaluated Th17 and its transcription factors in AMA-M2+ PBC, determined the expression of phosphorylated proteins using Western blotting, and analyzed the relationship between Th17 and LF. Results . The Th17 in the intervention group's peripheral blood was significantly higher than that of the control group (P <.05), and the sensitivity and specificity of the AMA-M2+ PBC were 63.33% and 96.67%, respectively. The expression of Foxp3 and p-Foxp3 proteins for the intervention was significantly lower (P <.001), while ROR.t and P-ROR. T were significantly higher (P <.001). The levels of interleukin-17 (IL-17) and IL-22 for the intervention group were significantly higher than those for the control group. The Pearson correlation coefficient showed that alanine aminotransferase (ALT), alkaline phosphatase (ALP), and.-glutamyl transpeptidase (GGT) were positively correlated with Th17 cells, ROR.t, IL-17, and IL-22 and negatively correlated with Foxp3. Conclusions . Th17 plays an important role in the early diagnosis of AMA-M2+ PBC, and Th17 and its transcription factors are highly effective for the early diagnosis of AMA-M2+ PBC, which is expected to be a breakthrough in the future diagnosis of the disease. (Altern Ther Health Med. 2023;29(2):168-173)