Ovarian protective effect of catalpol on premature ovarian insufficiency rats by regulating Hedgehog pathway
Objective To investigate the ovarian protective effect of catalpol on premature ovarian insufficiency(POI)rats by regulating the Hedgehog pathway.Methods Rats were randomly grouped into control group,model group,catalpol low and high dose(30 and 60mg·kg-1)group,and catalpol(60mg·kg-1)+GANT-61(Hedgehog pathway inhibitor,40 mg·kg-1)group.Except for the control group,a 50 mg·kg-1 suspension of Tripterygium glycosides was administered orally once a day for 14 consecutive days to prepare a POI model.After the model was successful,it was administered via ip once a day for four consecutive weeks.The wet weight and ovarian index of rat ovaries were calculated.ELISA was applied to measure serum hormones estradiol(E2),luteinizing hormone(LH),and follicle stimulating hormone(FSH).The reagent kits were applied to detect reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD),and peroxidase(CAT)in ovarian tissue.HE staining was applied to observe ovarian tissue structure and perform follicle counting.TUNEL was applied to detect apoptosis in ovarian tissue cells.Western blottinng was applied to detect the expression of Cleaved caspase-3,Bax,Bcl-2,Shh,and Glil proteins in ovarian tissue.Results Compared with the control group,the ovarian tissue morphology of rats in the model group was obviously damaged,the ovarian index,serum E2 level,SOD and CAT levels in ovarian tissue,number of growing follicles,and Bcl-2,Shh,and Glil protein expression levels were obviously reduced(P＜0.05),the serum FSH and LH levels,ROS and MDA levels in ovarian tissue,number of atresia follicles,cell apoptosis rate,and Cleaved caspase-3 and Bax protein expression levels were obviously increased(P＜0.05).Compared with the model group,the morphological damage of ovarian tissue of rats in the catalpol groups was obviously reduced,the ovarian index,serum E2 level,SOD and CAT levels in ovarian tissue,number of growing follicles,and Bcl-2,Shh,and Glil protein expression levels were obviously increased(P＜0.05),the serum FSH and LH levels,ROS and MDA levels in ovarian tissue,number of atresia follicles,cell apoptosis rate,and Cleaved caspase-3 and Bax protein expression levels were obviously reduced(P＜0.05).GANT-61 was able to alleviate the protective effect of catalpol on the ovaries of POI rats(P＜0.05).Conclusion Catalpol can reduce oxidative stress and cell apoptosis in ovarian tissue of POI rats,regulate serum hormone level,and thereby improve ovarian function,which is possibly achieved by activating the Hedgehog pathway.
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