| Objective The model rats with hypothyroidism and yang deficiency model replicated by intragastric administration of 0.1% PTU solution were used as the research objects.The correlation between the pathogenesis of kidney injury and the protective mechanism of Yougui Wan and the TGF-? / Smad signaling pathway provides experimental basis for the prevention and treatment of hypothyroidism kidney injury by traditional Chinese medicine,inherits and discovers the application value of ancient prescriptions.Method 1.Theoretical research: By consulting relevant domestic and foreign literature,the name of TCM disease,etiology and pathogenesis,the relationship with the five internal organs,the traditional Chinese medicine treatment methods,and the theoretical basis of Yougui Wan acting on hypothyroidism,as well as the theoretical basis of Western medicine Epidemiological research,pathogenesis,diagnosis and western medical treatment methods are systematically studied.2.Experimental research: Thirty-two SD male rats at 6 weeks of age were randomly divided into 4 groups,8 in each group,namely normal group,model group,Youjiale group and Youguiwan group.In the latter three groups,0.1% propylthiouracil(PTU)solution(1 mg/100 g)was administered by intragastric administration to replicate the hypothyroidism rat model.Enzyme-linked immunosorbent assay(ELISA)was used to detect thyroid function free triiodothyronine(FT3),free thyroxine(FT4),thyrotropin(TSH)and yang deficiency index cyclic adenosine phosphate(c AMP),cyclic Guanosine phosphate(c GMP),to determine whether the model of hypothyroidism is successful.After the model was successful,the model group,Youjiale group,and Yougui wan group continued to administer PTU at the same dose,and the dosage frequency was changed to once every two days.On this basis,the Youjiale group was given Youjiale solution(At a dose of 9 ?g/kg/d),the Yougui wan group was given Yougui wan solution(At a dose of 2.43 g/kg/d),and the normal group was given 1 ml/100 g of drinking water,which was given by gavage every day for 6 weeks.After 6 weeks of treatment,the blood was taken from the heart,thyroid function FT3,FT4,TSH were detected by ELISA,serum creatinine(SCr)and urea nitrogen(BUN)were detected by biochemical method;kidney tissue was taken for HE staining for morphological observation,ELISA method Detect the expression of transforming growth factor-?1(TGF-?1),Smad3,collagen I(Collagen I)protein in kidney tissue,and detect the TGF-?1,Smad3,Collagen in kidney tissue by reverse transcription-polymerase chain reaction(RT-PCR)method I m RNA expression.Collect data and use statistical software SPSS20.0 for statistical analysis.Results 1.Thyroid function test: After 4 weeks of modeling,FT3 and FT4 of the model group were significantly lower than that of the normal group,and TSH was significantly higher than that of the normal group.The difference was statistically significant(P <0.01).After 6 weeks of drug treatment,compared with the model group,Youjiale group and Yougui pill group can increase FT3 and FT4 levels and lower TSH level;compared with Youjiale group,Yougui pill group has higher FT3 and FT4 levels than Ujia The Le group was obvious,and the difference was statistically significant(P <0.01).The lowered TSH of Youguiwan group was not as obvious as that of Youjiale group,and the difference was statistically significant(P <0.01).2.Detection of Yang deficiency index: After 4 weeks of modeling,c AMP expression in the model group was lower than that in the normal group(P<0.05),c GMP expression was significantly higher than that in the normal group(P<0.05),the ratio of c AMP/c GMP was significantly lower than that in the normal group,suggesting The rat model of hypothyroidism and yang deficiency model was successfully replicated.3.HE staining: HE staining slices were observed under light microscope.The glomeruli,tubules and tubule interstitial morphology of the normal group were clear,the structure was complete,and no abnormalities were found.In the model group,renal interstitial edema was accompanied by a large amount of inflammatory cell infiltration,the glomerular volume increased,and a large number of renal tubular epithelial cells were necrotic and shed.Youjiale group and Youguiwan group had slight renal interstitial edema,infiltration of a small amount of inflammatory cells,and the shedding of renal tubular epithelial cells was significantly improved.4.Renal function test: Compared with the normal group,the creatinine and urea nitrogen levels in the model group were significantly increased;compared with the model group,Youjiale and Yougui pills significantly decreased the creatinine and urea nitrogen levels;compared with the Youjiale group,Yougui pills There was no statistically significant difference in serum creatinine concentration(P>0.05),while Yougui Pill had lower urea nitrogen level than Ugala,but no statistical difference(P>0.05).5.TGF-?/Smad signaling pathway protein detection: After 6 weeks of treatment,the expression of TGF-?1,Samd3,and Collagen I protein of hypothyroidism rats increased significantly,and Yougui wan can significantly inhibit the expression of TGF-?1,Samd3,and Collagen I protein in kidney tissue of hypothyroidism rats.6.TGF-?/Smad signaling pathway gene detection: After 6 weeks of treatment,the expression of TGF-?1,Samd3,and Collagen I m RNA of hypothyroidism rats increased significantly,and Yougui wan can significantly inhibit the expression of TGF-?1,Samd3,and Collagen I m RNA in kidney tissue of hypothyroidism rats.Conclusion 1.Using Yougui wan,a warm kidney yang prescription,could nourish the essence and blood,which is effective in the treatment of hypothyroidism,both from the improvement of western medicine thyroid function indicators or the overall regulation of traditional Chinese medicine.Therefore,warming the kidneys and aphrodisiac,nourishing essence and blood are important rules of hypothyroidism treatment.2.After being administrated with 0.1% PTU,the thyroid function of rats could be decreased,and a series of hypothyroidism symptoms appeared.The rat model of hypothyroidism was successfully replicated.3.Increased levels of BUN and Scr of hypothyroidism rats suggest that hypothyroidism can affect renal function.4.The expressions of TGF-?1,Smad3,Collagen I and genes of hypothyroidism rats were increased,suggesting that the pathogenesis of hypothyroidism renal damage may be achieved through the TGF-?/Smad signaling pathway.5.The traditional Chinese medicine compound Yougui wan can effectively inhibit the high expression of TGF-?1,Smad3,and Collagen I proteins and genes in the TGF-?/Smad signaling pathway.It is speculated that the mechanism of Yougui Wan's improvement of hypothyroidism and kidney damage may be related to its regulation of TGF-?/Smad signaling pathway. |
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