Pharmacological Effects of CuiRu Formula on Lactation and Immune Function in a Rat Model of L-DOPA-Induced Postpartum Hypogalactia

Postpartum hypogalactia impairs lactation and may compromise offspring immunity. CRF, derived from the traditional Tonggan Shengru Tang, was developed to address this condition. However, its pharmacological mechanisms remain unclear. Network pharmacology was used to screen candidate constituents and predict potential targets of CRF. CRF was chemically characterized by untargeted LC-MS/MS, targeted flavonoid analysis, and monosaccharide composition analysis. An L-DOPA-induced rat model of postpartum hypogalactia was established. The pharmacological effects of CRF were evaluated by measuring milk yield, pup weight gain, serum hormones, including prolactin (PRL), estradiol (E2), and progesterone (P), and serum immunoglobulins (IgA, IgG, sIgA). Mammary gland histopathology (H&E) and the mRNA expression of lactation-related genes (PRLR, CSN2, LALBA) and PI3K/AKT-related genes were analyzed. Additionally, 16S rRNA sequencing was used to assess changes in gut microbiota composition following CRF treatment. Network pharmacology suggested flavonoids as potentially important constituents of CRF and implicated the PI3K/AKT pathway. Several predicted flavonoids were also detected in CRF by LC-MS/MS. In vivo, CRF (notably mid-dose) significantly increased milk yield, showed a trend toward increased pup weight gain, alleviated mammary gland atrophy, and increased serum PRL levels. CRF elevated serum immunoglobulin levels in dams and pups while upregulating the mRNA expression of PRLR, CSN2, LALBA, PI3K, and AKT in mammary tissues. Additionally, 16S rRNA sequencing revealed that CRF partially reversed the changes in gut microbiota richness and community structure observed after L-DOPA treatment. CRF improved lactation performance and increased immunoglobulin levels in postpartum hypogalactic rats and their offspring, accompanied by increased serum PRL levels, upregulated mRNA expression of PRLR and PI3K/AKT-related genes, and partial reversal of L-DOPA-associated changes in gut microbiota composition.