“Baihui”(DU20)-penetrating“Qubin”(GB7)acupuncture on blood–brain barrier integrity in rat intracerebral hemorrhage models via the RhoA/ROCKⅡ/MLC 2 signaling pathway

Background:Blocking the Rho A/ROCKⅡ/MLC 2(Ras homolog gene family member A/Rho kinaseⅡ/myosin light chain 2)signaling pathway can initiate neuroprotective mechanisms against neurological diseases such as stroke,cerebral ischemia,and subarachnoid hemorrhage.Nevertheless,it is not clear whether and how disrupting the Rho A/ROCKⅡ/MLC 2 signaling pathway changes the pathogenic processes of the blood–brain barrier(BBB)after intracerebral hemorrhage(ICH).The present investigation included the injection of rat caudal vein blood into the basal ganglia area to replicate the pathophysiological conditions caused by ICH.Methods:Scalp acupuncture(SA)therapy was performed on rats with ICH at the acupuncture point“Baihui”-penetrating“Qubin,”and the ROCK selective inhibitor fasudil was used as a positive control to evaluate the inhibitory effect of acupuncture on the Rho A/ROCKⅡ/MLC 2 signaling pathway.Post-assessments included neurological deficits,brain edema,Evans blue extravasation,Western blot,quantitative polymerase chain reaction,and transmission electron microscope imaging.Results:We found that ROCKⅡacts as a promoter of the Rho A/ROCKⅡ/MLC 2 signaling pathway,and its expression increased at 6 h after ICH,peaked at 3 days,and then decreased at 7 days after ICH,but was still higher than the preintervention level.According to some experimental results,although 3 days is the peak,7 days is the best time point for acupuncture treatment.Starting from 6 h after ICH,the neurovascular structure and endothelial cell morphology around the hematoma began to change.Based on the changes in the promoter ROCKⅡ,a 7-day time point was selected as the breakthrough point for treating ICH model rats in the main experiment.The results of this experiment showed that both SA at“Baihui”-penetrating“Qubin”and treatment with fasudil could improve the expression of endothelial-related proteins by inhibiting the Rho A/ROCKⅡ/MLC 2 signaling pathway and reduce neurological dysfunction,brain edema,and BBB permeability in rats.Conclusion:This study found that these experimental data indicated that SA at“Baihui”-penetrating“Qubin”could preserve BBB integrity and neurological function recovery after ICH by inhibiting Rho A/ROCKⅡ/MLC 2 signaling pathway activation and by regulating endothelial cell–related proteins.

Catharmus tinctorius volatile oil promote the migration of mesenchymal stem cells via ROCK2/Myosin light chain signaling

MSC transplantation has been explored as a new clinical approach to stem cell-based therapies for bone diseases in regenerative medicine due to their osteogenic capability. However, only a small population of implanted MSC could successfully reach the injured areas. Therefore, enhancing MSC migration could be a beneficial strategy to improve the therapeutic potential of cell transplantation. Catharmus tinctorius volatile oil(CTVO) was found to facilitate MSC migration. Further exploration of the underlying molecular mechanism participating in the pro-migratory ability may provide a novel strategy to improve MSC transplantation efficacy. This study indicated that CTVO promotes MSC migration through enhancing ROCK2 mRNA and protein expressions. MSC migration induced by CTVO was blunted by ROCK2 inhibitor, which also decreased myosin light chain(MLC) phosphorylation.Meanwhile, the si RNA for ROCK2 inhibited the effect of CTVO on MSC migration ability and attenuated MLC phosphorylation,suggesting that CTVO may promote BMSC migration via the ROCK2/MLC signaling. Taken together, this study indicates that C.tinctorius volatile oil could enhance MSC migration via ROCK2/MLC signaling in vitro. C. tinctorius volatile oil-targeted therapy could be a beneficial strategy to improve the therapeutic potential of cell transplantation for bone diseases in regenerative medicine.