| Part 1 A Case Report of Suspicious Malignant Thyroid Nodule and Literature Review Background:Thyroid cancer has become the fastest growing malignant tumor.Thyroid nodules are the first thyroid sign of thyroid cancer.About 7%-15% of nodules are malignant and can be identified by thyroid color ultrasonography or thyroid fine needle puncture cytology.Currently,there is no effective non-invasive treatment for suspicious malignant thyroid nodules.Surgery or medical follow-up is of more options.This paper reports the data of a patient with suspicious malignant thyroid nodule admitted to the General Hospital of the Central Theater Command,and reviews the relevant literature at home and abroad,in order to explore the effective non-surgical treatment for this disease.Objective:To observe the clinical outcome of suspected malignant thyroid nodule treated with traditional Chinese medicine.Methods:The clinical data of a patient with suspicious malignant thyroid nodule admitted to the General Hospital of the Central Theater Command was retrospectively analyzed: an elderly female patient was presented to the doctor with progressive enlargement of thyroid nodules.The thyroid color Doppler ultrasound suggested the possibility of malignant thyroid disease.The patient refused the operation and was given oral traditional Chinese medicine berberine.After a follow-up visit after 6 months,the changes of thyroid function and thyroid nodule before and after berberine treatment were compared,and the relevant literature was analyzed and discussed.Results:There was no significant change in thyroid function,and the thyroid nodules did not enlarge.The re-examination of thyroid color Doppler ultrasound showed that the TI-RADS classification was changed from 4b to 4a.Conclusion:Berberine may have the effect of inhibiting thyroid nodules.Part 2 Berberine,Induces Apoptosis in Human Papillary Thyroid Cancer Cells through Inhibition of Nrf2 Signalling Pathway Objective:To investigate the effect of Berberine(BBR)on apoptosis of human Papillary thyroid cancer(PTC)K1 cells by inhibiting Nuclear factor erythroid 2-related factor 2(NRF2)related signaling pathway and to explore its mechanism.Methods:K1 cells were classified into 10 groups based on different treatment factors:(1)0umol/L berberine group(NC group),(2)40umol/L berberine group(BBR group),(3)Nrf2 overexpression group(Nrf2-OE group),(4)Nrf2 overexpression +40umol/L berberine group(Nrf2-OE+BBR group),(5)Nrf2 small interference group(Nrf2-si RNA group),(6)Nrf2 small interference+40umol/L berberine group(Nrf2-si RNA+BBR group),(7)PI3K excitement Drug group(IGF-1 group),(8)PI3K agonist+40umol/L berberine group(IGF-1+BBR group),(9)PI3K inhibitor group(LY294002 group),(10)PI3K inhibitor+40umol/L berberine group(LY294002+BBR group).After K1 cells were treated according to the above groups,relevant indexes were detected by the following methods: Cell apoptosis was detected by Annexin V-FITC/PI double staining combined with flow cytometry.The protein expression levels of Bcl-2,Caspase-3,PI3 K,p-PI3 K,Akt,p-Akt,nuclear Nrf2,plasma Nrf2,HO-1 and NQO1 were detected by Western blot.The intracellular reactive oxygen species(ROS)content was detected by flow cytometry,SOD activity was detected by chemiluminescence assay,GSH-Px content was detected by fluorescence assay.And the expression of Nrf2 m RNA was detected by RT-PCR,the nuclear translocation of Nrf2 was observed by fluorescence microscopy.Results:Compared with the NC group,the early apoptosis rate,late apoptosis rate,the expression level of pro-apoptotic protein Caspase-3 and the content of ROS of K1 cells in the BBR group were significantly increased,and the expression level of anti-apoptotic protein Bcl-2 and the content of SOD and GSH-Px were markedly diminished(all p<0.05).It had the same effect with Nrf2-si RNA group and Nrf2-si RNA+BBR group(all p<0.05),but had the opposite effect with Nrf2-OE group(all p<0.05).Compared with the NC group,there was significant improvement in the early cell apoptosis rate and ROS level in the Nrf2-OE+BBR group,the GSH-Px and Bcl-2 levels were markedly diminished(all p<0.05),and the late cell apoptosis rate,SOD and Caspase-3 levels were not perceptibly altered(all p>0.05).Compared with the NC group,the levels of HO-1 and NQO1 protein in the BBR group were markedly diminished(all p<0.05),and there was no statistical difference between the BBR group and the Nrf2-si RNA group(all p>0.05).Compared with the NC group,the protein expression levels of PI3 K and Akt in the BBR group did not perceptibly altered(all p>0.05),and the levels of p-PI3 K and p-Akt were markedly diminished(all p<0.05).Compared with the NC group,the nuclear Nrf2 protein level and Nrf2 nucleoplasmic protein ratio in the BBR group were markedly diminished,plasma Nrf2 protein level was significantly elevated,Nrf2 m RNA expression and Nrf2 nuclear translocation were significantly inhibited(all p<0.05).It had the same effect with LY294002 group and LY294002 + BBR group(all p<0.05),but had the opposite effect with IGF-1 group(all p<0.05).Compared with the NC group,the nuclear Nrf2 protein level and Nrf2 nuclear/plasma protein ratio in the IGF-1+BBR group were improved significantly(all p<0.05),and there were no significant changes in the plasma Nrf2 protein level,Nrf2 m RNA expression and Nrf2 nuclear translocation(all p>0.05).Conclusion:Berberine can inhibit the phosphorylated level of PI3K/Akt,down-regulate the protein expression of Nrf2 and nuclear translocation,inhibit the expression of Nrf2 downstream proteins HO-1 and NQO1,and induce apoptosis of K1 cells possibly by up-regulating the level of ROS,the expression of Caspase-3,down-regulating the level of GSH-Px and SOD and the expression of Bcl2. |
