Study On Traditional Chinese Medicine Screening For Sjogren’s Syndrome Based On Aquaporin-1 And Aquaporin-5 Promoter Targets

Sjogren’s syndrome, is an autoimmune chronic disease which violate salivary glands, lacrimal glands and other exocrine. The clinical manifestations of Sjogren’s syndrome are salivary gland dysfuncion, saliva secretion reduction and thus cause mouth drying, affected the wearing and the retention of dentures. The core issue of Sjogren’s syndrome was glandular secretion disorders. Aquaporin (Aquaporin, AQP) is closely related to glands. Studies have shown that Sjogren’s syndromes’expression of AQP significantly reduced, which leaded water transport decompensation. Therefore, restored expression and regulation of AQP 1 and AQP5, the amount of saliva secretion would be increased. There would be provide a new ideas for the treatment of Sjogren’s syndrome. Therefore, the main purpose of this study was to make AQP1 and AQP5 promoters as a target to screen effective Chinese medicines on treating Sjogren’s Syndrome.AQP1 and AQP5 was specific aquaporins in salivary gland cells. Our group successfully constructed both of the recombinant plasmid promoter sequences. Including the sequence of AQP 1 promoter (from-1717 to+151) of and AQP5 promoter (from-852 to+111). Then, we set drug screen model on two targets for screening salivary secretion-promoting drug about more than many kinds natural medicines.First, we examined from 200 kinds of medicines which traditional Chinese medicine ingredients can significantly activated AQP1 and AQP5 promoter biological activity on the transcriptional level. The results showed that American ginseng and ginseng can significantly promote AQP1 and AQP5 promoter activity.On the base, we examined the effects of ginseng and American ginseng components on the expression of AQP1 protein from the expression levels. AQP1 was a 28kD glycosylated water channel protein, which glycosylated proteins generally between 40-60kD. The produce of prototype AQP1 proteins were depended on its accumulation in the cells. The results showed that American ginseng can promote the expression of prototype AQP1 protein, meanwhile the proterins had dose-dependent.Next, we would furtherly test the impact of the screened drugs on the secretion of water. Cl-was a main anions in cells, which would change with the concentration of water and cause an instantaneous change.The fluorescence quenched after EYFP-H148Q-V163S binded with Cl-. So we applied EYFP-H148Q-V163S fluorescence to detect the water secretory function in the cells.Found that ginseng and American ginseng ingredients can significantly promote the secretion of water, there was a significant difference compared with the control group. After adding water channel protein specific blocker HgCl2, water secretion was inhibited obviously.The study found ginseng structure has the characteristics of steroid hormones. Steroid hormones had a effect by binding to intracellular receptors. Nextly, we examined which receptors by binding to play a catalytic role in the expression of AQP1 about the ingredients of ginseng and American ginseng. By overexpression of ERα, ERβ and AR, we found that afert over expressed ERα, American ginseng can significantly promote the expression of AQP1. When ERα gene silenced, American ginseng was suppressed the expression of AQP1 protein. We detected the AQP5 by the same way. When overexpressed ERβ, American ginseng can significantly activate AQP5 promoter biological activity. When ERβ gene silenced, American ginseng inhibited the AQP5 promoter transcriptional activity.On the basis, we screened which components of American ginseng and ginseng activated AQP1 and AQP5 promoter that played a major role.We found out that ginseng glycol, glycerol, and panaxatriolcan ginseng significantly promote AQP1 and AQP5 promoter activity.These experimental results proved that American ginseng and ginseng ingredients could increase salivary secretion of key genes-AQP1 and AQP5 to regulate the secretion of saliva, increased salivation. Provided a novel molecular targeting and new ideas for the treatment of Sjogren’s syndrome.The study may guide the clinical application of American ginseng and ginseng treatment Sjogren’s syndrome. There would have be a important theoretical significant and practical value.