Effects Of 1,25-dihydroxy Vitamin D₃ On Treg Cells In Rats With Renal Interstitial Fibrosis

Objective: To explore the effects of 1,25-dihydroxy vitamin D₃?1,25?OH?₂D₃?on Treg cells in rats with renal interstitial fibrosis.Methods:?1?The 54 healthy male wistar rats with weight?200 + 10?g was choosed.They were randomly divided into control group?group A?,unilateral ureteral obstruction grop?UUO group,group B?and 1,25?OH?₂D₃ treatment group?group C?and the number of each group was 18.Group B and group C were treated with unilateral ureteral ligation.The group A was treated with ureteral free,but not ligated.Group C was endowed 1,25?OH?₂D₃ with 3 ng / 100 g/d.Group A and group B were endowed peanut oil for 0.5 ml.They were lavaged in the daily morning.During the experiment,each group had a normal diet.Group A,group B and group C respectively in postoperative 1 day,3 days,7 days were randomly killed.The number of Each group was 6.The blood specimens and surgical side kidney tissues was acquired.?2?The level changes blood serum creatinine,blood calcium,and phosphorus were detected.The changes of kidney tissue pathological was observed by HE and Masson staining.Human forkhead box P3 m RNA level in the rat kidney tissue was detected by real-time fluorescent quantitative polymerase chain reaction?Real-time PCR,RT-PCR?.The levers changes of the alpha-smooth muscle acyin and human forkhead box P3 in the rat kidney tissue were detected by immunohistochemical.Results:?1?B group?group UUO?compared with A group?sham operation group?at the same time point,the level of creatinine increased significantly,and the difference was statistically significant?P < 0.05?.C group?1,25?OH?₂D₃ treatment group?compared with B group at the same time point,the level of creatinine was significantly reduced,and the difference was statistically significant?P < 0.05?.The statistical results showed that at the same time point,A group,B group and C group rats had no significant difference between the serum calcium,serum phosphorus level?P>0.05?.?2?Kidney tissue HE and Masson staining: The results of HE showed that under light microscope,there were no obvious changes in the renal tubules of the A group?sham operation group?at different time points,and there was no infiltration of inflammatory cells such as monocytes.B group?group UUO?for first days and third days can be seen in renal tubular epithelial cells with mild dilatation,occasional degeneration,edema,a few epithelial cells necrosis,renal interstitial width increased and visible inflammatory cells such as monocyte infiltration amount.In the seventh day,the renal tubular expansion was more obvious,and the epithelial cells were vacuolar degeneration.There were a large number of mononuclear cells infiltration in the renal interstitium.The interstitial width was increased and the interstitial structure was destroyed.Compared with the B group at the same time point,the pathological changes and the degree of fibrosis were reduced in C group.Masson staining and semi quantitative analysis showed that at the same time B groupcompared with A group,the relative area increased significantly,and the degree of fibrosis,the difference was statistically significant?P < 0.05?.C group compared with B group,the relative area decreased,and the degree of fibrosis is reduced,the difference was statistically significant?P < 0.05?.?3?The relative expression of FOXP3 m RNA in rat kidney: B group compared with A group at the same time point,the level of FOXP3 m RNA in renal tissue was significantly decreased,and the difference was statistically significant?P < 0.05?.Compared with B group at the same time point,the level of FOXP3 m RNA in renal tissue of C group increased significantly,and the difference was statistically significant?P < 0.05?.?4?B group compared with A group at the same time point,expression of-SMA in renal tissue was significantly increased,and the expression of FOXP3 was decreased.The difference was statistically significant?P < 0.05?.With prolonged obstruction,the expression of B in renal tissue of FOXP3 group decreased significantly,and alpha the expression of ?-SMA was significantly increased.C group compared with B group at the same time point,?-SMA expression was decreased in renal tissue,and FOXP3 expression was significantly increased.The difference was statistically significant?P < 0.05?.Conclusion:?1?Renal interstitial fibrosis may be accompanied with the decrease of Treg cells.?2?By an immunoregulation,1,25?OH?₂D₃ can slow down the progression of renal interstitial fibrosis through raising Treg cells.