Hyperglycemia is a condition in which the blood glucose levels exceed normal limits. Ethanol extract of Jamaican cherry leaves (Muntingia calabura L.) contains active components that have the potential to lower blood sugar levels and heal pancreatic cell damage in rats. Gut microbiota imbalance can cause immune system abnormalities, illnesses, and metabolic disorders, including diabetes. The aim of the present study was to determine whether an ethanol extract of Jamaican cherry leaves can repair pancreatic cell damage, as well as influence the total coliforms and lactic acid bacteria in hyperglycemic mice. In this study, 25 mice were randomly placed into five groups, which were then provided with food and drinking water. Alloxan in a dose of 160 mg/kg bw was administered to the positive control group and three treatment groups. Each of the three groups was then treated with the ethanol extract of Jamaican cherry leaves at doses of 300, 400, or 500 mg/kg bw for 14 days. The groups that received the extract were able to repair the pancreatic damage considerably by increasing the number of normal pancreatic cells. This was supported by histological observations. The total abundance of lactic acid bacteria in hyperglycemic mice was 1.08 ± 45 x 10¹⁰ CFU/g, which was substantially lower than that in mice treated with the extract. The overall coliform abundance in hyperglycemic mice was 4.37 ± 41 x 10¹⁰ CFU/g, which was higher than that in mice supplemented with the extract. Therefore, the administration of Jamaican cherry leaf ethanol extract on a regular basis can increase the abundance of cecum microbiota in hyperglycemic mice. Furthermore, this extract can be used as a natural alternative treatment for recovering pancreatic cell damage.