Methods: Present study is an extension of a randomized placebo controlled double-blinded clinical trial of T2DM (n = 56/group) given two capsules/d of cellulose/olive oil (placebo), or NS-3 for 24 weeks. Pure mRNAs and miRNAs of plasma of pre-dose versus post-dose of NS-3 treated samples were analyzed by next generation sequencing (NGS). Data was uploaded into “Ingenuity Pathways Analyses”. 

Results: A total of 4000 genes are considered significant, based on > 2-fold gene expression changes. Out of which 1373 genes are significantly differentially expressed in pre-dose vs post-dose (p < 0.02) samples. Out of those, 20 are up-regulated and 27 are down-regulated of NS-3 treated RNAs of T2DM. Gene expression of up-regulated miR-29b-3p modulates (GLUT4, insulin resistance), miR-624-5p (nephropathy biomarker), miR-361-5p (chronic inflammation), miR-130a-3p (glucose metabolism, insulin secretion), miR-3912-3p (lipid metabolism), and miR-11401 (cellular transcription). The miR-374c-5p (insulin resistance), miR-4326 (HbA1c level)), miR-874-3p (β -cell function) are down-regulated of NS-3 treated people with T2DM. Whereas messengerR-ML-1621513 (oxidative/stress), mR-CTD-2349P217 (insulinmediated glucose-uptake), are up- regulated, and mR-CTC-246B1810 (β-cell/biology) are down-regulated in T2DM after NS-3 treatment. Venn diagrams have established genetic regulatory network images and canonical signaling pathways for mRNA, miRNA, and paired mRNA-miRNA of gene expression profiles of pre-dose vs. post-dose of NS-3 treatment group. 

Conclusions: The NS-3 treatment of people with T2DM indicates up- or down-regulation of several new miRNAs (miR-29b-3p, miR-624- 5p, miR-361-5p, miR-130a-3p, miR-3912-3p, miR-374c-5p, miR-4326 [HbA1c], miR-1247-3p, miR-874-5p) which may be used to identify onset T2DM. Overexpression of mRNA-AL1621513 indicates oxidative stress in people with T2DM, resulting in complications of diabetes (neuropathy, retinopathy, and stroke).