Protective Effects And Mechanisms Of The Formula Of Herba Erigerontis And Radix Paeoniae Rubra On Cerebral Ischemia-reperfusion Injury

[ABSTRACT] Objective To determine the optimal formula of Herba Erigerontis and Radix Paeoniae Rubra and then to study the protective effects and possible mechanism on cerebral ischemia-reperfusion injury treated with the optimal. Methods ①The PC 12 cells induced by oxygen-glucose deprivation was estabished. To research the protection on PC 12 cells induced by OGD treated with different formula of Herba Erigerontis and Radix Paeoniae Rubra. The cell viability and the LDH content were studied to find the optimal formula. ②The PC 12 cells induced by oxygen-glucose deprivation was used. The general morphology, cell viability, SOD activity, LDH, MDA and ROS content, mitochondrial membrane potential were studied to determine the protective effects and mechanisms of the formula in vitro. ③The middle cerebral artery occlusion in rats was established. The neurological deficits score, infarct volume percentage and the level of MDA, SOD, GSH-PX, TNOS, iNOS, NO in the serum were studied. Pathological changes of neurons in hippocampal CA1 region was observed. The expressions of Bcl-2, Bax and Caspase-3 were observed through immunohistochemistry in hippocampal CA1 region. The protective effects and mechanisms of the formula was studied in vivo. Results ① Experiment including that different formula have certain protective effect and the 2:3 is the optimal formula to the PC12 cell induced by OGD. ② In vitro, compared with the model group, the activity of SOD, the cell viability and mitochondrial membrane potential were significantly increased, the content of LDH, ROS and MDA were obviously decreased in the formula group, and the PC 12 cell morphology was significantly improve. ③ In vivo, compared with the model group, the formula significantly improve neurological deficits score in brain injury rats. In addition, the infarct volume percentage was effectively reduce, the activity of SOD and GSH-PX were obviously increased, the activity of TNOS and iNOS were significantly inhibited, the level of MDA and NO were observably reduced in the formula group. Immunohistochemistry shows that the formula can enhance the expression of Bcl-2 and weaken that of Bax and Caspase-3 in hippocampal CA1 region suffered ischemia-reperfusion injury. As a whole, the formula is better than the Herba Erigerontis or Radix Paeoniae Rubra. Conclusion The formula of 2:3 take certain protective effect on cerebral ischemia-reperfusion injury and it is the optimal formula. The possible mechanism may partly owe to the anti-free radical effect and it also may be related to regulating the expression of apoptosis -related proteins.