Objective To investigate the effects of Danggui Sini Decoction containing serum on mitochondrial division and Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing kinase(ROCK) pathway of dorsal root neurons of SD rats cultured with high glucose.
Methods Six SD rats were given intragastric administration of normal saline, and another six given Danggui Sini Decoction, the blood was taken from the carotid artery to prepare normal serum and drugs containing serum. Bilateral dorsal root neurons of normal SD rats were cultured in vitro and randomly divided into control group, high glucose group, and Danggui Sini Decoction group. The cells in high glucose group and Danggui Sini Decoction group were treated with high glucose, cells in control group were cultured normally, and cells in the Danggui Sini Decoction group were cultured with 10% drug-containing serum. After treatment with normal serum and containing serum, the cell viability of each group was detected with CCK-8 experiment; the levels of cellular reactive oxygen species (ROS), release lactate dehydrogenase (LDH), interleukin-6 (IL-6), and interleukin-18 (IL-18) levels in each group were determined with kits; the expressions of mitochondrial division proteins dynamin-related protein 1(Drp1) and fission protein 1(Fis1), fusion proteins mitofusin 1(MFN1) and mitofusin 2(MFN2) and RhoA/ROCK pathway proteins such as RhoA and ROCK in each group were detected with western blotting.
Results Compared with the control group, the levels of ROS, the levels of LDH, IL-6 and IL-18, and the expression of Drp1, Fis1, RhoA and ROCK proteins in the dorsal root neurons of rats in the high glucose group were significantly increased (P < 0.05), while the expressions of cell viability, mitochondrial membrane potential, MFN1 and MFN2 protein in cells were significantly reduced (P < 0.05); compared with the high glucose group, the levels of ROS, the levels of LDH, IL-6 and IL-18, and the expression of Drp1, Fis1, RhoA and ROCK proteins in the dorsal root neurons of rats in the Danggui Sini Decoction group were significantly reduced (P < 0.05), while the expressions of cell viability, mitochondrial membrane potential, MFN1 and MFN2 protein in cells were significantly increased (P < 0.05).
Conclusion Danggui Sini Decoction containing serum can down-regulate the expression of RhoA/ROCK pathway proteins, inhibit the expressions of ROS and inflammatory factors induced by high glucose, reduce inflammation reaction, weaken mitochondrial division, and alleviate the damage of dorsal root neurons of SD rats.
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