ABSTRACT Gentiopicroside (GPS) is a bioactive iridoid glycoside isolated from Gentianaceae plants. In recent years, GPS has received increasing attention due to its multiple pharmacological activities. This review encapsulates the botanical origin, pharmacological activity, toxicity, and underlying mechanisms of GPS in the treatment of various diseases. The results have proved that GPS possesses a wide spectrum of pharmacological properties, including hepatoprotective, anti‐arthritic, analgesic, antidiabetic, anticancer, and neuropsychopharmacological activities. Specifically, GPS can alleviate liver injury caused by alcohol, d ‐galactosamine, or lipopolysaccharide by reducing liver function impairment, improving hepatic lipid disorders, and inhibiting oxidative stress and inflammation. GPS also has good antitumor activity by inducing apoptosis and inhibiting the proliferation of cancer cells. Additionally, GPS can improve diabetes by maintaining glucose homeostasis, reducing lipid disorders, improving insulin resistance, inhibiting lipogenesis, and alleviating oxidative stress and inflammation. As for the toxicity of GPS, studies have shown that GPS has low toxicity. Collectively, given its promising pharmacological activity, GPS holds potential as a drug candidate for the treatment of liver disease, arthritis, diabetes, and cancer. In the future, more research is still needed to uncover its long‐term toxicity, oral bioavailability, and molecular mechanisms for treating these diseases.