ABSTRACT
OBJECTIVE: Using the method of lactate dehydrogenase (LDH) assay, to observe the activities of rat cerebral microvascular endothelial cells (CMECs) intervened by Tongluo Jiunao Injection (TLJNI), a traditional Chinese compound drug removing toxin to dredge brain collaterals, and then further study the effects of different kinds of conditioned mediums (CMECs-CM) of cerebral microvascular endothelial cells on ischemia and ischemia/reperfusion cerebral cortex cells, and to probe into the drug pharmacological mechanisms of CMECs in modulating the neurons. METHODS: Three kinds of CMECs (normal, ischemic and ischemic/reperfusional) were all treated by TLJNI previously, and then the three pairs of CMECs-CM without serum were collected respectively for LDH assay. Rat cerebral cortex neurons were also primarily cultured and then divided into similar three groups (normal, ischemic and ischemic/reperfusional). The neuron responses caused by CMECs-CM at different concentrations were observed by using LDH transudation rate assay. RESULTS: The LDH release values of ischemic and ischemic/reperfusional CMECs with TLJNI treatment were obviously reduced (P<0.01) compared with the same kinds of CMECs untreated. For ischemic neurons, both conditioned medium of ischemic CMECs (Is-CM) and conditioned medium of ischemic CMECs with drug treatment (IsT-CM) in high concentration of 100% increased the LDH transudation rate (P<0.01), while in low concentration of 10%, IsT-CM reduced the transudation rate (P<0.05). For ischemia/reperfusion neurons, all kinds of CMECs-CM reduced the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, 10% or 50% showed relatively stronger effects, and both conditioned medium of normal CMECs (N-CM) group and conditioned medium of ischemic/reperfusional CMECs (Rp-CM) group had statistical significance (P<0.05 or P<0.01). For normal neurons, all kinds of CMECs-CM increased the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, only conditioned medium of normal CMECs (N-CM) had statistical significance (P<0.05 or P<0.01). CONCLUSION: The study shows that TLJNI is capable of preventing the damage of CMECs from both ischemia and ischemia/reperfusion states. Chinese drug can restrain the brain ischemia and ischemia/reperfusion damage by the media that CMECs modulate the neurons, demonstrating the pharmacological mechanisms of TLJNI. This work also indicates that there exist some active substances against ischemia/reperfusion injury secreted from CMECs-CM with TLJNI treatment.
ABSTRACT
Objective: To observe the protective effects of Gastrodine on the cultivated rat brain microvessel endothelial cells damage by mimic cerebral ischemia.Methods: The endothelial cells activity,survival rate,the change in NO content,and the effects of Gastrodine were observed in the cultivated rat brain microvessel endothelial cells(BMEC) damaged by mimic cerebral ischemia.Results: The activity and survival rate of BMEC in the ischemia groups are obviously lower than that in the normal groups;compared with the normal groups,the activity,survival rate and NO content of BMEC in the Gastrodin groups have the increasing tendency;comparing to the ischemia groups,the activity of BMEC in the Gastrodin groups obviously increasing(P
ABSTRACT
Objective: To study the influence of conditioned medium of rat brain microvascular endothelial cells on the activity of cortical neurons and the protective effect of Tong Luo Jiu Nao Injection.Methods: We collected the conditioned media from 4 different cultured endothelial cell groups,which were normal endothelial cells,the normal ones and treated with Tong Luo Jiu Nao Injection,the injured ones damaged by simulated cerebral ischemia,as well as the injured ones and treated with Tong Luo Jiu Nao Injection,respectively(N-CM,NT-CM,I-CM,IT-CM).Then,the conditioned medium was added into the cultures of the normal neurons and the damage ones which are injured by simulated cerebral ischemia as well,respectively.The effect of each type of conditioned medium on the activities of neurons was determined through the measurement of MTT and the transduction rate of LDH.Results:(1) N-CM has no obvious effect on the normal neurons,but does show some protective effect on the injured ones by increasing its activity significantly;(2) I-CM could decrease the activity of the normal neurons as well as aggravate the damage on the injured ones,while this injury effect can be reversed remarkably by IT-CM.Conclusion: The paracrine secretion of the brain microvascular endothelial cells might be one of the vital mechanisms in cerebral ischemic injury,indicating that the brain microvascular endothelial cells could be the therapeutic targets of Chinese medicine,which are not able to permeate through the Blood-Brain Barrier.