Exosomal Circular RNA hsa_circ_0046060 of Umbilical Cord Mesenchymal Stromal Cell Ameliorates Glucose Metabolism and Insulin Resistance in Gestational Diabetes Mellitus via the miR-338-3p/G6PC2 Axis

Figure 6

Exosomal hsa_circ_0046060 of hUMSC regulates glucose homeostasis and induces insulin resistance through hsa-miR-338-3p in vitro and in vivo. (a) mmu-miR-338-3p expression level (, vs. Control; ^  ^  ^  ^ , vs. GDM; ^####, vs. GDM + G-mUMSC-EXOs; ^&&&&, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-mUMSC-EXOs) and (b) G6PC2 mRNA expression level (, , vs. Control; ^ , GDM vs. GDM + si-circ-G-hUMSC-EXOs; ^##, ^###, vs. GDM + G-mUMSC-EXOs; ^&&&&, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-hUMSC-EXOs) in GDM mice administered with PBS, C-mUMSC-EXOs, G-mUMSC-EXOs and si-circ-G-mUMSC-EXOs as evaluated by RT-qPCR. U6 was used as an internal control. (c, d) Protein expression levels of G6PC2 in GDM mice administrated with PBS, C-mUMSC-EXOs, G-mUMSC-EXOs and si-circ-G-mUMSC-EXOs as determined by western blot. GAPDH was used as an internal control (, , , vs. Control; ^  ^  ^ , ^  ^  ^  ^ , vs. GDM; ^####, vs. GDM + G-mUMSC-EXOs; ^&, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-mUMSC-EXOs). Expression levels of (e) hsa-miR-338-3p (, vs. si-circ-G-hUMSC-EXOs; ^  ^  ^  ^ , vs. si-circ-G-hUMSC-EXOs + inhibitor NC), (f) G6PC2 (, vs. si-circ-G-hUMSC-EXOs; ^  ^  ^  ^ , vs. si-circ-G-hUMSC-EXOs + inhibitor NC) and (g) hsa_circ_0046060 (, vs. si-circ-G-hUMSC-EXOs; ^  ^ , vs. si-circ-G-hUMSC-EXOs + inhibitor NC) following treatment with inhibitor NC and hsa-miR-338-3p inhibitor of si-circ-G-hUMSC-EXOs in L-02 cells as determined by RT-qPCR assay. Glucose concentration: (h) in si-circ-G-hUMSC-EXOs group alone and with inhibitor NC and hsa-miR-338-3p inhibitor (, vs. si-circ-G-hUMSC-EXOs; ^{^^^^}, vs. si-circ-G-hUMSC-EXOs + inhibitor NC), and (i) PA-induced insulin resistance cell model alone and with inhibitor NC and hsa-miR-338-3p inhibitor of L-02 cells (PI : PA + insulin, , vs. PI + si-circ-G-hUMSC-EXOs; ^  ^  ^, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC) as determined using glucose content assay kit. (j) Glycogen levels in insulin resistance model of L-02 cells as determined using Glycogen Content Assay Kit (PI : PA + insulin, , vs. PI + si-circ-G-hUMSC-EXOs; ^  ^ , vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC). (k) Protein expression levels of (l) G6PC2 (PI : PA + insulin, , vs. PI + si-circ-G-hUMSC-EXOs; ^  ^ , vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC), (m) p-IRS-1 and IRS-1 (PI : PA + insulin, , vs. PI + si-circ-G-hUMSC-EXOs; ^  ^, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC) in PA-induced insulin resistance cell model alone and with inhibitor NC and hsa-miR-338-3p inhibitor of in L-02 cells as determined by western blot analysis. (n) Morphological changes of liver from GDM mice as evaluated by HE staining. Scale bar = 100 μm.

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