Enhanced Expression of ABCG2 by saRNA increases Uric Acid Excretion of Renal and Intestinal Cells in Mice
OBJECTIVE To screen and study the effects of small activating RNA(saRNA)enhancing the expression of ABCG2 gene on renal and intestinal cell excretion of uric acid(UA)to explore a new type of therapeutic mechanism drugs to promote UA excretion.METHODS saRNAs targeting mice and human ABCG2 genes were designed and screened by real-time quantitative reverse transcription PCR(RT-qPCR)and Western blot in mice TCMK-1 and human HK-2 cells.The effect of the selected saRNA on the ex-cretion of UA was examined in TCMK-1 and HK-2 cells.The mice saRNA was injected into the tail vein of hyperuricemia(HUA)model mice.The levels of serum uric acid(SUA),urinary uric acid(UUA),intestinal uric acid,blood urea nitrogen(BUN)and ser-um creatinine(SCr),and the activity of liver xanthine oxidase(XOD)in the mice were detected to verify the ability of selected saRNA to promote UA excretion.The pathological changes of kidney and small intestine were analyzed through H&E.The expression of ABCG2 in kidney and small intestine was analyzed by RT-qPCR,Western blot and immunohistochemistry.RESULTS Four saRNAs targeting ABCG2 were selected,2 for mice and 2 for human,named saRNA-1/2 and hsaRNA-1/2 respectively,which increased the expression of ABCG2 in TCMK-1 and HK-2 cells(P＜0.05),and promoted the extracellular excretion of UA(P＜0.05).In HUA model mice,saR-NA-1/2 increased the levels of UUA and intestinal UA(P＜0.05),reduced the levels of SUA,BUN,and SCr(P＜0.05),decreased the damage of HUA to kidney and small intestine,and enhanced the expression of ABCG2 in kidney and small intestine,but did not show significant effect on the activity of liver XOD(P＞0.05).CONCLUSION The selected saRNAs could enhance the expression of ABCG2 and promote the excretion of UA in cells and mice,reducing SUA level.
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