FIGURE 6.

Berberine exhibited anti‐tumor effect in orthotopic pancreatic cancer implantation model. (A) Oral administration of berberine had minimal effect on the body weight of C57BL/6N mice. (B) Compared to the control (CTL, n = 16) group, berberine (5 mg/kg, n = 16 and 10 mg/kg, n = 16) can prolong survival in pancreatic cancer model (P = 0.0294). (C) Representative images of the dissected pancreas and liver at the end of berberine treatment showing that berberine intervention can regress orthotopic growth of implanted pancreatic cancer and suppress liver metastasis. The graph represents the orthotopic tumor size of the controlled group mice (CTL, n = 6), 5 mg/kg berberine group (n = 9) and 10 mg/kg berberine group (n = 7) by the end of the 4‐week intervention (**P < 0.01). (D) Compared to the control group, berberine treatment (5 mg/kg, n = 9 and10 mg/kg, n = 7) suppresses serum LDH level (*P < 0.05). (E) a shift of serum LDH isoenzyme to LDH‐5 was found and berberine treatment group exhibit lower serum LDH‐5 than that of the control (*P < 0.05). (F) Compared to the control (CTL, n = 6) group, berberine (5 mg/kg, n = 6, and 10 mg/kg, n = 6) treatment suppresses serum L‐lactate level (*P < 0.05, **P < 0.01) in murine in orthotopic pancreatic cancer implantation model. (G) Compared to the control (CTL) group, berberine (5 mg/kg and 10 mg/kg) treatment suppresses intratumoral L‐lactate level (n = 6 each group, **P < 0.01) in murine in orthotopic pancreatic cancer implantation model. (H) Representative histological and immunohistochemistry (IHC) photomicrographs of the tumor in individual groups. Paraffin‐embedded tumor tissues were stained with hematoxylin and eosin (H&E), anti‐Ki67 antibody and anti‐LDHA antibody to determine the pathological type, proliferation, and LDHA expression of the tumor cells upon different treatments. The graph represents the percentage of Ki‐67 and LDHA‐positive cells of pancreatic and liver lesions in control group (CTL, n = 6), 5 mg/kg berberine group (n = 6) and 10 mg/kg berberine group (n = 6) (**P < 0.01, ***P < 0.001). (I) Representative histological and immunofluorescent (IF) photomicrographs of the pancreatic orthotopic lesions. Paraffin‐embedded tumor tissues were stained with DAPI (Blue), anti‐CD3 (Red), anti‐CD4 (Green), and anti‐CD8 (Green) antibody in individual groups upon different treatments. (J) The schematic representative of regulatory mechanism underlying LDHA‐mediated inhibition of pancreatic cancer by berberine. CTL: control group; LDH: lactate dehydrogenase; HE: hematoxylin and eosin staining; LDHA: lactate dehydrogenase A