| Objective: To observe the effect of electroacupuncture on NLRP3/caspase-1/GSDMD pathway in the uterus of primary dysmenorrhea(PDM)model rats,and to explore the mechanism of electroacupuncture treatment of PDM.Methods: Forty healthy non-pregnant female SD rats were randomly divided into four groups: control group,model group,electroacupuncture group and ibuprofen group,with 10 rats in each group.(1)Modeling: After 7 days of adaptive feeding,rats in model group,electroacupuncture group and ibuprofen group were injected subcutaneously with estradiol benzoate for 10 days,and intraperitoneally injected with oxytocin on the 11 th day to establish PDM model rats;Rats in the control group were injected with 0.9%sodium chloride(Na Cl)solution at corresponding sites.(2)Intervention: The rats in the electroacupuncture group were selected at Guanyuan and Sanyinjiao points and treated with electroacupuncture after estradiol benzoate was injected in the first 10 days during modeling;The rats in ibuprofen group were given ibuprofen(concentration: 1.25mg/m L)by gavage.Control group and model group were only given the same way and time to grasp and bind.(3)Index detection: Within 30 minutes after the 11 th day of oxytocin injection,the behavior of the rats was observed,and the twisting response of the rats was taken as the symbol of the success of the PDM model.The twisting incubation period was observed,and the twisting frequency,twisting score and twisting incubation period were statistically analyzed.After the behavioral observation,all rats were anesthetized with pentobarbital sodium.Blood was collected from the abdominal aorta of the rats,and serum PGF2 α and PGE2 contents were detected by ELISA after centrifugation.The uterus of the rats was removed,and the left upper segment was placed in 4% paraformaldehyde.After embedding and sectionalization,the uterus was stained by HE method.The morphology of the uterus was observed under the optical microscope,and the pathological score of the uterus was performed according to the findings of the light microscope.The other uterus was placed in liquid nitrogen,and the relative protein expression levels of NLRP3,caspase-1,ASC,GSDMD,GSDMD-N,IL-1 β and IL-18 in the uterine tissues of rats were detected by WB.Results: 1.Behavior: Compared with the control group,the twisting times(P<0.01)and twisting scores(P<0.01)of rats in the model group were significantly increased,indicating successful modeling.Compared with model group,the number of twisting(P<0.05)and score of twisting(P<0.05)were significantly decreased in electroacupuncture group,while the latency of twisting(P<0.01)was significantly prolonged.Compared with model group,the writhing times(P<0.05)and writhing score(P<0.05)of rats in ibuprofen group were significantly decreased,while the writhing latency period(P<0.01)was significantly prolonged.Compared with ibuprofen group,there was no statistical difference in the above indexes in electroacupuncture group(P>0.05),indicating that the effect of electroacupuncture on PDM was similar to that of ibuprofen group.2.HE staining: HE staining was performed on all rat uteruses,and morphology was observed using an optical microscope.The endometrial epithelial cells in the PDM group were massively vacuolated and dead,many spiral arterioles were congested,and neutrophil infiltration was observed.In the electroacupuncture group,there was a small amount of vacuolar degeneration and cell death in the uterine epithelium,a few spiral arterioles were congested,and a small amount of neutrophil infiltration.In the ibuprofen group,the endometrial epithelial cells showed a small amount of vacuolar degeneration and cell death,and no obvious congestion was observed in the basal layer,and occasionally neutrophil infiltration was observed.Compared with the control group,pathological injury score of uterus in model group was significantly increased(P<0.01).Compared with model group,pathological injury scores of rats in electroacupuncture group and ibuprofen group were significantly decreased(P<0.05,P<0.05).The uterine tissue structure of PDM rats was damaged,and electroacupuncture could reduce the uterine tissue damage.Compared with ibuprofen group,there was no statistical difference in uterine pathological injury score in electroacupuncture group(P>0.05),indicating that the effect of electroacupuncture on reducing uterine tissue injury in rats was comparable to ibuprofen.3.ELISA test: Compared with the control group,the content of PGF2 αand the value of PGF2 α/PGE2 in serum of rats in model group was significantly increased(P<0.01),while the content of PGE2 was significantly decreased(P<0.01).Compared with model group,the content of PGF2 α(P<0.01)and the value of PGF2 α/PGE2 in serum of rats in electroacupuncture group and ibuprofen group were significantly decreased(P<0.01),while the content of PGE2 was significantly increased(P<0.01).PG content can be adjusted by electroacupuncture to treat PDM.Compared with ibuprofen group,there was no statistical difference in the above indexes in electroacupuncture group(P>0.05),indicating that the effect of electroacupuncture on PDM was similar to that of ibuprofen group.4.Western blotting: Compared to the control group,The relative expression levels of NLRP3(P<0.01),caspase-1(P<0.01),ASC(P<0.01),GSDMD(P<0.01),GSDMD-N(P<0.01),IL-1 β(P<0.01)and IL-18(P<0.01)in uterine tissue of model group were significant To rise;Compared to the model group,Relative levels of NLRP3(P<0.01),caspase-1(P<0.01),ASC(P<0.05),GSDMD(P<0.05),GSDMD-N(P<0.05),IL-1 β(P<0.05)and IL-18(P<0.05)in uterine tissues of rats in electroacupuncture and ibuprofen groups.The results indicated that electroacupuncture could inhibit NLRP3/caspase-1/GSDMD pathway,and inhibit the release of caustic death and inflammatory factors in the uterus of PDM rats.Compared with ibuprofen group,there was no statistical difference in the above indexes in electroacupuncture group(P>0.05),indicating that the effect of electroacupuncture on PDM was similar to that of ibuprofen group.Conclusion: 1.The effect of electroacupuncture on PDM was significant.2.The mechanism of electroacupuncture in the treatment of PDM may be realized by inhibiting NLRP3/caspase-1/GSDMD pathway and inhibiting pyroptosis,so as to reduce the inflammation of uterus and tissue damage. |
PDF Full Text Request