To investigate whether electroacupuncture at "Ciliao" (BL32) can improve the mitochondrial homeostasis imbalance of periurethral tissues in mice by promoting the homing of endogenous bone marrow mesenchymal stem cell (BMMSC) to the periurethral region of mice, and ultimately improve stress urinary incontinence (SUI), with the aim of providing a reference for the treatment of pelvic floor dysfunction disorders.

In experiment 1, 25 C57BL/6 female mice were randomly divided into the sham operation, model, electroacupuncture, acupuncture, and electrical stimulation groups, with five mice per group. Based on the largest improvement in the indexes of the electroacupuncture group in experiment 1, experiment 2 was conducted. The other 15 C57BL/6 female mice were randomly divided into the model, electroacupuncture, and electroacupuncture + ADM3100 groups, with five mice per group. In the sham operation group, the catheter was only placed in the vagina without balloon injection expansion, whereas vaginal dilation was used in the other groups to construct the SUI model. Experiment 1: the sham operation and the model groups received no intervention; the "Ciliao" (BL32) was stimulated in the electroacupuncture group; the acupuncture group was only acupunctured at "Ciliao" (BL32); and the electrode sheet was attached to the "Ciliao" (BL32) in the electrical stimulation group. Experiment 2: the model group received no intervention; the electroacupuncture+ADM3100 group received ADM3100 solution injected peritoneally 24 h before the first intervention, followed by electroacupuncture at "Ciliao" (BL32); and the electroacupuncture group received an equal volume of phosphate-buffered saline injected peritoneally 24 h before the first intervention, followed by electroacupuncture at "Ciliao" (BL32). Each intervention lasted 20 min, once daily for 7 consecutive days. After the intervention, the modified simulated human urethral bladder intubation method was used to record the bladder leakage point pressure (BLPP) and abdominal leakage point pressure (ALPP). Masson staining was used to detect collagen deposition in periurethral tissue. The content of mitochondrial adenosine triphosphate (ATP) in periurethral tissue was detected using a luciferase luminescence assay. Immunofluorescence staining was used to detect the homing of BMMSC in periurethral tissue. JC-1 staining method was used to detect mitochondrial membrane potential in periurethral tissue. The expression of reactive oxygen species (ROS) in periurethral tissue was detected using the fluorescent probe DCFH-DA method. Real-time PCR was used to detect the mRNA expression of stromal cell-derived factor-1α(SDF-1α) in periurethral tissue.

Experiment 1: compared with the sham operation group, the BLPP and ALPP of the mice in the model group decreased (P < 0.05); the blue staining of collagen fibers in the periurethral tissue was lighter, the distribution was disordered, and the deposition was reduced; the collagen volume fraction, mitochondrial membrane potential and mitochondrial ATP content in the periurethral tissue decreased, whereas the ROS expression increased (P < 0.05). Compared with the model group, BLPP and ALPP in the electroacupuncture, acupuncture, and electrical stimulation groups were increased (P < 0.05); the blue staining of collagen fibers in the periurethral tissue deepened, the distribution disorder was reduced, and the deposition increased; and the collagen volume fraction of the periurethral tissue, BMMSC ratio, mitochondrial membrane potential, mitochondrial ATP content, and the mRNA expression of SDF-1α increased, the ROS expression decreased (P < 0.05). Compared with the electroacupuncture group, BLPP and ALPP decreased in the acupuncture and electrical stimulation groups (P < 0.05); and the collagen volume fraction, BMMSC ratio, mitochondrial membrane potential, mitochondrial ATP content, and SDF-1α mRNA expression in periurethral tissue decreased, whereas ROS expression increased (P < 0.05). Experiment 2: compared with the electroacupuncture group, the BLPP and ALPP of mice in the electroacupuncture+ADM3100 group decreased (P < 0.05); and the collagen volume fraction of periurethral tissue, BMMSC ratio, mitochondrial membrane potential, mitochondrial ATP content, and SDF-1α mRNA expression decreased, whereas the ROS expression increased (P < 0.05).

Electroacupuncture at "Ciliao" (BL32) may improve mitochondrial homeostasis imbalance in periurethral tissues by promoting endogenous BMMSC homing in SUI mice, thereby improving SUI.

To observe changes in the molecular expression of the NLR Family Pyrin Domain Containing Protein 3 (NLRP3) pathway in depressed rats after treatment with Xiaoyaosan, and identify the regulatory mechanism of this compound.

Male Sprague–Dawley rats were randomly divided into five groups with 12 rats in each group, including the control group, model group, Fluoxetine group, Xiaoyaosan group, and MCC950 group. A depression model was generated by chronic immobilization stress (induced by 3 h of restraint immobilization every day), and the drugs were administered at the same time in each group for 21 days. The effects of Xiaoyaosan on behavioral changes of depressed rats were observed through macroscopic characterization, body mass, open field experiments, and a sucrose preference test. The mRNA and protein expression of the NLRP3 signaling pathway was examined by fluorescence real-time quantitative PCR and Western blot assays.

The Xiaoyaosan group, Fluoxetine group, and MCC950 group rats showed improved depressive behavior and an increased weight of sucrose water consumption. The protein and mRNA expression levels of NLRP3, Caspase-1, and IL-1β were also decreased in the Fluoxetine, Xiaoyaosan, and MCC950 groups.

NLRP3, Caspase-1, and IL-1β protein and mRNA expression levels were increased in the cortex of depressed rats, while Xiaoyaosan protected cortical tissue in these rats by decreasing NLRP3, Caspase-1, and IL-1β protein and mRNA expression.

To investigate the effect of Xiaoyao San on the brain-gut axis in rats exposed to chronic immobilization stress (CIS).

Rats were divided into control, model, and treatment groups. The rats belonging to the model and treatment groups were subjected to CIS for 21 consecutive days, during which they were administered Xiaoyao San decoction [3.854 g/(kg·d)] or vehicle by gavage, and their body weight gain, food intake and water intake were monitored. The rats were subsequently subjected to the open field test (OFT) and d-xylose absorption test, and the expression levels of neuropeptides secreted by the hypothalamus and stomach were determined by enzyme-linked immunosorbent assay (ELISA), radioimmune analysis, or real-time fluorescence quantitative polymerase chain reaction. Gastric mucosal morphology was also assessed.

The model rats exhibited complex brain-gut axis abnormalities following exposure to CIS, abnormalities signified by decreases in food intake, reductions in digestive absorption, decreases in body weight, decreases in the total distances traveled and increases in the time in the central zone during the OFT, gastric mucosal lesion development and decreases in gastrointestinal hormone secretion. These changes were reversed after treatment with Xiaoyao San, which also regulated the secretion of both peripheral (serum and stomach) and central (hypothalamus) brain-gut peptides. Specifically, the levels of neuropeptide Y (NPY) and neuropeptide Y receptor Y5, which are secreted by the hypothalamus and promote digestive function, were increased in the Xiaoyao San-treated group compared with the model group. Furthermore, the levels of pro-opiomelanocortin (POMC) and its receptor, melanocortin-4 receptor (MC4R), which are secreted by the hypothalamus and inhibit digestive function, were significantly decreased in the treatment group compared with the model group. However, the levels of ghrelin (GHRL), gastrin (GAS) and motilin (MTL), which are secreted by the stomach, were significantly increased in the serum and stomach of the treatment group compared with the serum and stomach of the model group following Xiaoyao San treatment (P < .05 vs. the model group).

Xiaoyao San attenuates CIS-induced gastrointestinal dysregulation by regulating the peptides secreted by both the hypothalamus and the gastrointestinal tract (GIT), suggesting that its effects are associated with the brain-gut axis.