Ameliorative effect of Yishen Tiaojing pill suspension on ovarian function in rats with POI and its mechanism
Objective To observe the improvement effect of intragastric administration of Yishen Tiaojing pill sus-pension on ovarian function in rats with premature ovarian insufficiency(POI)and to explore its possible mechanism.Methods Seventy rats were randomly divided into seven groups,with 10 rats in each.In the high-dose,medium-dose,and low-dose Yishen Tiaojing pill groups,the estradiol valerate group,and the model group,we prepared the POI models by intraperitoneal injection of VCD solution,and rats in the adjuvant group were intraperitoneally injected with the same vol-ume of sesame oil for 15 consecutive days,while rats in the normal group received no treatment.After successful modeling,rats in the high-dose,medium-dose,and low-dose Yishen Tiaojing pill groups were given the suspension of Yishen Tiaojing pill at doses of 3.6,1.8,and 0.9 g/(kg d)by gavage,rats in the estradiol valerate group were given a suspension of the es-tradiol valerate at a dose of 0.09 mg/(kg d)by gavage,and rats in the model group were given 0.1 mL/kg of 0.9%sodium chloride solution by gavage.ELISA was used to detect serum FSH,LH,AMH,E2,IL-10,and IL-17 levels;HE staining was used to observe the morphology of ovarian tissues;immunohistochemistry was used to detect TGF-β1,TGF-βRⅠ protein in ovarian tissues,and Western blotting was used to detect Smad2,Smad3,and Smad7 protein in the ovarian tissues.Re-sults Compared with the normal group,the ovarian tissue structure of rats was disorganized,the number of follicles at all levels were reduced,the atretic follicles increased,the serum levels of FSH,LH and IL-17 increased,the levels of AMH,E2 and IL-10 decreased,the protein expression levels of TGF-β1,TGF-βRⅠ,Smad2 and Smad3 in the ovarian tissues de-creased,and the protein expression of Smad7 increased in the model group(all P<0.05).After the drug intervention,com-pared with the model group,the number of follicles at all stages in the ovaries of rats increased,the atretic follicles de-creased,the serum levels of FSH,LH and IL-17 decreased,the levels of AMH,E2 and IL-10 increased,the protein expres-sion levels of TGF-β1,TGF-βRⅠ,Smad2 and Smad3 in the ovarian tissues increased,and the protein expression of Smad7 decreased in the treatment groups(the serum levels of AMH and E2 in the medium-and low-dose Yishen Tiaojing pill groups,as well as the protein expression levels of Smad2 and Smad3 in the low-dose Yishen Tiaojing pill group,showed no statistically significant difference,and significant differences were found in the remaining indexes,all P<0.05).In the dif-ferent dose groups of Yishen Tiaojing pill,the levels of serum FSH,LH,and IL-17 in the high-dose group were lower than those in the medium-dose and low-dose groups,while the levels of AMH and IL-10 were higher than those in the medium and low-dose groups(all P<0.05),and there was no statistically significant difference in other detection indicators(all P>0.05).There were no significant differences in various detection indicators between the high-dose Yishen Tiaojing pill group and the estradiol valerate group(all P>0.05).Conclusions Yishen Tiaojing pill can regulate the levels of serum sex hormones and inflammatory factors,thereby improving ovarian function in POI rats.Its mechanism of action may be re-lated to the regulation of the TGF-β/Smads signaling pathway.
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