Study On The Intervention Effect Of Iodine-rich Chinese Medicine On Graves' Goiter Based On TSHR And Related Signaling Pathway

Purpose: In this experiment,iodine-rich Chinese herbal was used for intervention treatment,to verify its effect on the proliferation and apoptosis of FRTL-5 cells induced by thyroid-stimulating antibody(TRAb),and to provide a new theoretical basis for the prevention and treatment of GD goiter.Material and method:Experimental animals 18 Wistar rats(6-8 weeks old)were randomly divided into three groups: normal group,iodine-rich Chinese herbal group,and Na I group,and drug-containing serum was prepared respectively.The normal group was gavaged with distilled water,and the latter two groups were gavaged with iodine-rich traditional Chinese medicine and Na I respectively.Consecutive gavage for 2 weeks,once a day,blood was collected from the abdominal aorta 1 h after the last gavage,animal serum was separated,sterilized by 0.22 um filtration,inactivated at 56 °C for 30 min,and stored at-20 °C for later use.FRTL-5 cells were cultured in vitro and randomly divided into 4 groups,normal group,model group,iodine-rich Chinese herbal group,and Nal group.Except the normal group,the other groups were added with 1 μg /ml M22 to stimulate FRTL-5 cells for 24 hours,while the normal group and the model group were added with normal serum,and the other drug groups were added with the corresponding drug-containing serum for intervention.Observe the growth of cells after dosing.The proliferation and apoptosis of thyroid cells were detected by flow cytometry and MTT,and protein expression PKA C-?,p-PKA C(Thr197),CREB,p-CREB(Ser133),ERK,and p-ERK(Thr202/Tyr204)were detected by Western blot.Results:1.Cell proliferation detected by MTT assay: Both the iodine-rich Chinese herbal group and the Na I group had inhibitory effects on the proliferation of FRTL-5 cells.Compared with the normal group,the cell proliferation in the model group was significantly increased(P<0.01);Compared with the model group,the proliferation of cells in the iodine-rich Chinese herbal group and Na I group was significantly decreased(P<0.01);There was no significant difference in cell proliferation between the iodine-rich Chinese herbal compound group and the Na I group(P>0.05).2.Apoptosis results detected by flow cytometry: Both iodine-rich Chinese herbal and Na I can induce apoptosis of FRTL-5 cells.Compared with the normal group,the apoptosis rate of the model group decreased(P<0.01);Compared with the model group,the apoptosis rate of iodine-rich Chinese herbal group and Na I group was significantly increased(P<0.01);There was no significant difference in the apoptosis rate between the iodine-rich Chinese herbal compound group and the Na I group(P>0.05).3.Expression of p-PKA C and PKA C-α in FRLT-5 cells: Compared with the normal group,the expression of p-PKA C protein in the model group was significantly increased(P<0.01);Compared with the model group,the expression of p-PKA C protein in the iodine-rich Chinese herbal group decreased(P<0.01);Compared with the Na I group,the expression of p-PKA C protein in the iodine-rich Chinese herbal group decreased(P<0.01);There was no significant difference in the protein expression of PKA C-α among the groups(P>0.05).4.Expression of p-CREB and CREB in FRLT-5 cells:Compared with the normal group,the expression of p-CREB protein in the model group was significantly increased(P<0.01);Compared with the model group,the expression of p-CREB protein in the iodine-rich Chinese herbal group decreased(P<0.01);The expression of p-CREB protein decreased in Na I group(P<0.01);Compared with the Na I group,there was no significant difference in the expression of p-CREB protein in the iodine-rich Chinese herbal group(P>0.05).There was no significant difference in the protein expression of CREB among the groups(P>0.05).5.Expression of p-ERK and ERK proteins in FRLT-5 cells: Compared with the normal group,the expression of p-ERK protein in the model group was significantly increased(P<0.01);Compared with the model group,the expression of p-ERK protein in the iodine-rich Chinese herbal group decreased(P<0.01);The expression of p-ERK protein decreased in Na I group(P<0.01);Compared with the Na I group,there was no significant difference in the expression of p-ERK protein in the iodine-rich Chinese herbal group(P>0.05).There was no significant difference in the protein expression of ERK among the groups(P>0.05).Conclusion:1.Iodine-rich Chinese herbal containing serum can inhibit FRTL-5 cell excessive proliferation and induce apoptosis of FRTL-5 cells.2.The iodine-rich Chinese herbal can effectively downregulate the protein expression of PKA,CREB and ERK,Iodine-rich Chinese herbal affects thyroid levels by inhibiting the expression of related proteins.