Effect of sanguinarine on the bone metabolism and inflammatory injury in diabetic osteoporosis rats through regulating the RhoA/ROCK signaling pathway

ZHOU Fei; YI Song; YUE Jianbiao; ZHANG Shangshang

doi:10.13431/j.cnki.immunol.j.20260012

Abstract：Objective To investigate the effects of sanguinarine on bone metabolism and inflammatory injury in diabetic osteoporosis rats by regulating the Ras homologous gene family member A(RhoA)/Rho-associated kinase(ROCK)signaling pathway.Methods A total of 60 healthy male 8-week-old SD rats in this study were divided into normal group(without any treatment),model group(intraperitoneal injection of 60 mg/kg 0.2％Streptozotocin+reduced bone mineral density),low-dose sanguinarine group(L-sanguinarine group),high-dose sanguinarine group(H-sanguinarine group),and H-sanguinarine+inhibitor group,with 12 rats per group.In L-sanguinarine group,H-sanguinarine group and H-sanguinarine+inhibitor group,the rats with successful modeling were given 2.50 mg/(kg·d)sanguinarine by gavage+intraperitoneal injection of the same volume of 0.9％sodium chloride injection,6.25 mg/(kg·d)sanguinarine by gavage+intragastric injection of the same volume of 0.9％sodium chloride injection,and 6.25 mg/(kg·d)sanguinarine by gavage+intraperitoneal injection of 40 mg/(kg·d)Rhosin.After the rats in each group were treated for 8 weeks,a dual-energy X-ray absorptiometry scanning system was used to measure bone density,and serum Ca,osteocalcin(OC),osteoprotegerin(OPG),and alkaline phosphatase(ALP)levels were detected.Biomechanical testing was performed to detect the maximum load of the femur of rats in each group,and HE staining was used to detect the histopathological changes of pancreatic tissues and femoral tissue morphology in rats in each group.ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6 in each group,and Western blot was used to detect the expression of RhoA,ROCK1,and ROCK2 proteins in femoral tissue of rats in each group.Results Compared with the normal group,the model group showed decreased bone density and maximum load on the femur,destruction of pancreatic and femoral tissues,decreased levels of the serum Ca,OC,OPG,reduced expression of RhoA,ROCK1,and ROCK2 proteins,and increased levels of the serum ALP,IL-1β,TNF-α,and IL-6(P＜0.05).Compared with the model group,the L-sanguinarine group and H-sanguinarine group showed increased bone density and maximum load on the femur in rats,reduced pancreatic and femoral tissue damage,increased levels of the serum Ca,OC,OPG,expression of RhoA,ROCK1,and ROCK2 proteins in femoral bone tissue,and decreased levels of the serum ALP,IL-1β,TNF-α,and IL-6,and the changes in the above indicators in the H-sanguinarine group were more significant(P＜0.05).Compared with the H-sanguinarine group,the H-sanguinarine+inhibitor group reversed the effects of sanguinarine on bone metabolism and inflammatory injury in rats(P≤0.05).Conclusion Sanguinarine improves bone metabolism in diabetic osteoporosis rats and reduces its inflammatory injury by activating the RhoA/ROCK signaling pathway.