| Objective: To research the improvement of chronic low-grade inflammation and follicular development in obese polycystic ovary syndrome(PCOS)model rats by kidney-nourishing and phlegm-removing method(KNPR),and to explore the possible mechanism through NLRP3 inflammasome signaling pathway.Methods: Control group rats were randomly selected from all female SD rats aged 7-8weeks.Treated the remaining rats with Letrozole combined high-fat and cholesterol to induce the obesity PCOS rat models.After modeling,then,they were divided into model group,Chinese medicine high dose group(HDG),Chinese medicine middle dose group(MDG),Chinese medicine low dose group(LDG)and Metformin group.The doses of LDG,MDG,HDG were given Chinese medicine of 16.65 g /kg,41.63 g /kg and 83.25 g /kg every day,while the dose of Metformin group was given metformin of300 mg /kg every day,and the gavage volume was 5ml /kg.Hematoxylin and eosin(HE)staining of ovarian tissue,testosterone(T)and free fatty acids(FFAs)levels of serum were detected by enzyme linked immunosorbent(ELISA).Protein relative expressions of NLRP3,apoptosis associated speck-like protein-2 containing a CARD(ASC2),cysteine-requiring aspartate protease-1(Caspase-1)were detected by Western-blotting.Statistical software SPSS 25.0 was used to analyze all the data.The test level wasα=0.05.Results:1.Effects of KNPR on the body mass growth rate and estrous cycle in each group:compared with blank group,the body mass growth rate of rats in model group increased,and the difference was statistically significant(P<0.05);Compared with model group,metformin group and HDG,MDG,LDG decreased,and the difference was statistically significant(P<0.05).compared with the blank group,the rats in the model group lost the regular estrous cycle and were mostly in the Diestrus phase or proestrous phase;After the intervention of TCM compound of tonifying kidney and reducing phlegm,the rats in HDG,MDG and metformin groups recovered regular estrous cycle.2.Effects of KNPR on T,FFAS in serum of rats in each group: compared with blank group,the levels of T,FFAS in serum of model group were significantly increased(P<0.05);Compared with model group,the serum levels of T,FFAS in metformin group and HDG were significantly decreased(P<0.05).3.The effect of KNPR on the pathological sections of the ovaries of rats in each group:obvious luteum and dominant follicles were observed in the blank group;In the model group,there was proliferation of interstitial cells and fibroblasts in the ovarian tissues,with multiple small follicles and no dominant follicle growth.There were obvious luteal bodies or growing follicles in metformin group and HDG,MDG.In LDG,there was no dominant follicle growth in the ovary,but proliferation of fibrous tissues and interstitial cells in the ovary.4.The effect of KNPR on the expression of NLRP3,Caspase-1 and ASC2 in ovarian tissue of rats in each group: compared with blank group,protein expressions of Caspase-1,NLRP3 and ASC2 in model group were significantly increased(P<0.05 or P<0.01);Compared with model group,the protein expressions of Caspase-1,NLRP3 and ASC2 in ovarian tissues of metformin group and HDG and MDG were significantly decreased(P<0.05 or P<0.01).Conclusion:1.The obesity PCOS animal model was successfully induced by quantitative gavage of letrozole combined with high-fat emulsion.KNPR can reduce the weight of obese PCOS rats and improve the obesity status of obesity PCOS rats.The method can also restore the estrous cycle of model rats.2.KNPR can improve reproductive endocrine disorders and lipid metabolism disorders in obesity PCOS model rats,and promote follicle maturation and luteal formation.3.KNPR can improve chronic low-grade inflammation and promote follicular development in obese PCOS model rats by inhibiting the NLRP3 inflammatory corpus-signaling pathway in ovarian tissue. |
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