1.Effect of berberine combined with bone marrow mesenchymal stem cells on energy metabolism of HUVECs damaged by high glucose
Ling LI ; Jiajia TAN ; Huiping LIU ; Rong YU ; Sha CHEN ; Puzhen ZHANG ; Qian WU
Chinese Pharmacological Bulletin 2016;32(11):1585-1591
Aim To investigate the effects of berberine combined with bone marrow mesenchymal stem cells ( BM-MSCs) on the energy metabolism of human um-bilical vein endothelial cells ( HUVECs ) under the condition of high glucose. Methods ①The state of cell reproduction and cell proliferating activity were de-termined by MTT assay. ②The cell cycle was detected by flow cytometry ( FCM ) . ③DNA damage of cells was measured by comet tail assay. ④The contents of ATP, ADP and AMP were determined by high perform-ance liquid chromatography ( HPLC ) and the level of energy charge ( EC) was calculated. ⑤The expression of CCR and COX mRNA was detected by reverse tran-scription-polymerase chain reaction ( RT-PCR) . ⑥The expressions of COX and CCR were detected by Western blot. Results ①The proliferating activity of HUVECs declined apparently and the proliferation decreased af-ter high glucose intervention. Meanwhile, the quantity of cells during S +G2 dropped dramatically and there was certain degree of damage to DNA. The berberine and BM-MSCs respectively improved the proliferating activity and the proliferation in different degrees, in-creased the quantity of cells during S+G2 and promo-ted the repair of DNA ( P <0 . 01 ) , and so did the combination of the two, with a better effect than each of them alone. ②After high glucose intervention and the damage caused, the content of both ATP and ADP of HUVECs was reduced, and EC level also declined significantly, while the content of AMP increased. The berberine and BM-MSCs respectively up-regulated the content of ATP and ADP ( P<0. 01 ) , and so did the combination of the two, with a better effect than each of them alone. ③After high glucose intervention and the damage caused, the expression of COX, CCR mR-NA and protein decreased obviously. Yet, all of the three gained a dramatic increase when the berberine, BM-MSCs or the combination of the two were added ( P<0. 01 or P <0. 05 ) , among which the combination worked more effectively. Conclusions The berber-ine, BM-MSCs and the joint use of the two could im-prove the energy metabolism of HUVECs, which had been damaged by high glucose, probably because the berberine and BM-MSCs could up-regulate the expres-sion of COX, CCR mRNA and protein, which leads to the hydrolyzation of glucose oxide and thus the im-provement of blood environment and the enhancement of glucose's supply and intake of HUVECs. Then, here comes the final result: the improvement of the energy metabolism of damaged vascular endothelial cells by high glucose.