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Objective To observe the therapeutic effects and mechanisms of Guanyuan Mingmen Sequential Acupuncture on rats with premature ovarian insufficiency(POI)model.Methods Female SD rats were divided into the blank group,the model group,the protein kinase A(PKA)inhibitor(H89)+acupuncture group,and the acupuncture group,with 12 rats in each group.Except for the blank group,the POI model was prepared by gavage with Tripterygium Glycosides Tablets in the other three groups of rats.After the model was successfully established,the blank group and the model group were bundled once a day;in the acupuncture group,Guanyuan(RN4)point was taken during the intermotility period,and in the pre-motility period,Mingmen(DU4)point was taken;in the H89+acupuncture group,the intervention was performed in accordance with the acupuncture protocol of the acupuncture group,and H89 was injected intraperitoneally for 30 minutes prior to each acupuncture session.Continuous intervention was performed for 20 days.Samples were taken from each group of rats in the first estrus period and in proestrus period after intervention.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of follicle stimulating hormone(FSH)and estradiol(E2)during the estrous phase,Western Blot was used to measure the protein expressions of follicle stimulating hormone receptor(FSHR)and aromatase P450(P450arom)during the estrous phase,and the activity of granulocytes during the estrous phase and the proestrus phase were measured using the cell-counting kit 8(CCK-8)method.The immunohistochemistry method was used to detect the protein expression of pre-motility proliferating cell nuclear antigen(PCNA).Results(1)Compared with the blank group,the serum FSH level of the model group and H89+acupuncture group was significantly increased(P＜0.01),and the E2 level was significantly decreased(P＜0.001);there was no difference between the FSH level of the H89+acupuncture group and that of the model group(P＞0.05),and the E2 level of the H89+acupuncture group was lower than that of the model group(P＜0.05);the FSH level of the acupuncture group was lower than that of the model group and that of the H89+acupuncture group(P＜0.05),had no difference with the blank group(P＞0.05),E2 level was significantly higher than the model group and H89+ acupuncture group(P＜0.01),still being lower than the blank group(P＜0.05).(2)The protein expressions of FSHR and P450arom in the model group and H89 + acupuncture group was lower than those in the blank group;the protein expression of FSHR in the H89 + acupuncture group was not different from that in the model group(P＞0.05),while the protein expression level of P450arom was lower than that in the model group(P＜0.05);the protein expression levels of FSHR and P450arom in the acupuncture group were higher than those in the model group and H89 + acupuncture group,but still lower than those in the blank group(P＜0.05).(3)Both GCs activity and average optical density value of PCNA in the model group and H89+acupuncture group were lower than the blank group(P＜0.05);both GCs activity and average optical density value of PCNA in the H89+acupuncture group were lower than the model group(P＜0.05);the activity of GCs and average optical density value of PCNA of the acupuncture group were significantly higher than that of the model group and H89+acupuncture group(P＜0.05 or P＜0.01).Conclusion Guanyuan Mingmen Sequential Acupuncture can regulate sex hormone levels,increase GCs activity and promote GCs cell proliferation by up-regulating protein expressions of follicle stimulating hormone receptor(FSHR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)pathway FSHR,P450arom,thus improving POI.
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Objective To sequence follicle stimulating hormone receptro (FSHR) promoter of the ovarian granulocyte and initially research the molecular mechanism of the poor ovarian response. Methods To study the relationship between FSHR promoter mutation of ovarian granulocyte and ovarian respone. The 263 bp DNA fragments before FSHR 5'initiation site in 70 cases of patients with poor ovarian respone and 88 cases of patients with ovarian normal respone who were in the cycle of IVF-ET were sequenced, Results There were 63 cases which occurred 29th site G → A point mutation in 158 women and the mutation rate was 40. 0%. Mutation rate [ 60. 0% ( 42/70 ) ] of 29th site G → A in group of poor ovarian respone was significantly higher(χ2 = 21. 450,P < 0. 01 ) than normal response group [ 23.9% ( 21/88 ) ]. There was no obviously variability ( t = 0. 457, P 0. 05 ) of basic FSH values between two groups [ G/G group was (7.2 ± 2. 3) U/L, G/A & A/A group was (7. 1±2. 0) U/L];there was obviously variability (t = 35. 81 ,P < 0. 05 ) in the number of follicles sinus between two groups ( G/G group was 14. 2±1.3, G/A & A/A group was 4. 5±0. 8 ) ;there was obviously variability ( t = 40. 35, P < 0. 05 ) in the number of ovum pick-up between two groups ( G/G group was 14. 0±1.2, G/A & A/A group was 4. 5±1.1 ) ;there was obviously variability (t =25. 80,P <0.05) of FE2-peak value between two groups [G/G group was (2 865±557) pmol/L, G/A & A/A group was (880±211 ) pmol/L] ;there was obviously variability (t =40. 22 ,P <0. 05) in the number of mature eggs ( G/G group was 13.6±1.2, G/A&A/Agroupwas4.3±0. 9).Conclusion The 29th site of FSHR promoter significantly affect the activity of FSHR promoter. Mutation of G→A can weaken promoter activity, so that ovarian granulocyte poor respone to FSH.
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Objective To observe the localization and distribution of follicle stimulating hormone receptor(FSHR) in the rat submandibular gland and stomach. Methods Immunohistochemical SABC method was used in the experiment. Results The serous glandular cells,granular convoluted epithelial cells and all other duct epithelial cells in the submandibular gland and the parietal cells of gastric gland showed FSHR positive immunoreactivity.The positive substance was distributed in the cytoplasm with negative nuclei.Conclusion The function of rat submandibular gland and gastric gland might be regulated by follicle stimulating hormone(FSH) through FSHR.