OBJECTIVES: To investigate the molecular mechanism by which He's Yangchao Decoction (HSYC) improves ovarian function in a mouse model of premature ovarian insufficiency (POI). METHODS: Forty ICR mice were used to establish a POI model via intraperitoneal injection of cyclophosphamide and were randomly assigned to four groups: model control group, low-dose HSYC group, high-dose HSYC group, and estradiol group (positive control). Additionally, 10 age-matched ICR mice were selected as the blank control group. After intragastric intervention, the ovarian index, serum follicle-stimulating hormone (FSH) levels, and ovarian tissue expression of the FSH receptor (FSHR) were measured. A POI cell model was established by treating the human granulosa tumor cell line (KGN) with 4-hydroxycyclophosphamide. The cells were divided into four groups: solvent control group, HSYC group, inhibitor control group, and inhibitor+HSYC group, which were respectively treated with TH5487 (an OGG1 inhibitor) and HSYC-containing serum. The expressions of OGG1, mitochondrial DNA (mtDNA) oxidative damage markers, and pyroptosis-related proteins were detected by molecular docking, Western blotting, and immunofluorescence. RESULTS: Compared with the blank control group, the model control group showed a decreased ovarian index (P<0.05) and increased serum FSH levels (P<0.01). The ovarian index was higher in both the low- and high-dose HSYC groups compared with the model control group (both P<0.05). FSHR expression in ovarian tissue was lower in the model control group than that in the blank control group, but was higher in the high-dose HSYC group compared with the model control group (P<0.05 or P<0.01). Molecular docking confirmed strong binding affinity between the active components of HSYC and OGG1 (binding energy: －6.3 to －8.3 kcal/mol). Western blotting analysis revealed that OGG1 protein expression in the ovaries of the model control group was significantly reduced compared with the blank control group, while it was increased in the low-dose HSYC group and the estradiol group (P<0.05 or P<0.01). Immunofluorescence results demonstrated that the expression levels of mito-chondrial transcription factor A (TFAM) and peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) were decreased in the model control group compared with the blank control group (P<0.01), whereas their expressions were significantly elevated in the high-dose HSYC group and the estradiol group (all P<0.01). Cell experiments showed that TH5487 intervention increased the expression of 8-oxoguanine (8-OxoG) (P<0.01), while HSYC-containing serum intervention reduced 8-OxoG expression and increased TFAM expression (P<0.01). The expression of pyroptosis-related proteins (GSDMD, N-GSDMD, caspase-1, IL-1β) increased after TH5487 intervention (P<0.05), whereas HSYC-containing serum suppressed their expression (all P<0.05). CONCLUSIONS HSYC improves POI by upregulating OGG1 expression, mitigating mtDNA oxidative damage, and inhibiting granulosa cell pyroptosis.

This article was aimed to study the dynamics of biocontrol endobacteria in roots, stems and leaves of Panax ginseng and soils adjacent roots. Gradient screening and root irrigation methods were used to develop Rif-resistance bacteria and inoculation. Dilution plate method was used to count the number of them in roots, stems and leaves of P. ginseng and soils adjacent roots. The results showed that bacteria which can endure 120 μg·mL-1 Rif was developed, and no change was found on their antagonistic activity against A lternaria panax Whetz and Phytophthora cactorum. In roots and stems of P. ginseng, ge15 and ge25 showed the highest number in the initial.Then, populations of ge15 and ge25 in ginseng roots and stems decreased to a stable status gradually. In ginseng leaves, populations of ge15 and ge25 increased to the highest 3 days after inoculation, then, populations of them were decreased and kept at certain concentration. In ginseng rhizosphere soil, populations of ge15 showed reciprocating change, and which of ge25 showed increase initial, and then decrease to a stable status. It was concluded that endobacteria can keep a certain population in ginseng roots, stems, and leaves. Biocontrol agents based on them will be helpful for ginseng diseases controlling in the field.

By nutrient solution culture method, effects of benzoic acid, diisobutyl phthalate, diisobutyl succinate, palmitic acid, 2,2-bis(4-hydroxyphenyl) propane on root exudates of Panax ginseng were studied. GC-MS results in-dicated that, 15 to 23 components were detected in treatments including control, which were organic acids, alcohol, amino acid esters, phenols, amines, condensed nucleus, saccharine, etc. Through compared with control we found that, phenol and phenolic acids in root exudates increased in most of stress treatments. So we deduced that, the re-laxation of autotoxins from roots to outside environment was stimulated by autotoxins released into soil, and which ac-celerated the autotoxicity to ginseng seedlings.