The Neuroprotective Effects Of Injection Stauntoniae (IS) And The Extracts Of Stauntonia Chinesis DC On The Oxidative Damaged Neurons Caused By H₂O₂

Objective To study the neuroprotective effects of Injection Stauntoniae (IS) and the extracts of Stauntonia chinesis DC (AI₁, AI₂, AI₃) on the oxidative damaged cortical neurons and spinals neurons caused by H₂O₂ and then analysis the differences of these two neurons. To determine the antioxidant activity of IS and AI₁, AI₂ and AI₃. To analysis the content of flavonoid glycoside and triterpenoid saponin in AI₁, AI₂ and AI₃. This study is aimed at to be the underlying reason of antioxidant effects of Stauntonia chinesis DC. Methods: The MTT assay, SOD activity and the leakage of LDH were used to observed the effects of IS on unmarred cortical neurons and spinals neurons. The oxidative damage model used by H₂O₂ was established to make sure the experimental damaged concentration. The measurements of the leakage of LDH, SOD activity and the MTT assay were used to observe the neuroprotective effects of IS on oxidative damaged cortical neurons and spinals neurons. Besides, the changes of Ca²⁺ concentration in cytoplasm between damaged group and protected group was detected by LSCM. Theα-deoxyribose oxidation assay was used to elucidate the·OH clearance of IS in vitro. The contents of flavonoid glycoside and triterpenoid saponin in AI₁, AI₂ and AI₃ were respectively measured by standard curve method. The ?OH clearance of AI₁, AI₂ and AI₃ were observed byα-deoxyribose oxidation assay and the differences of antioxidant activity were compared among AI₁, AI₂ and AI₃. The MTT assay was used to observe the neuroprotective effects of AI₁, AI₂ and AI₃ on oxidative damaged cortical neurons and spinals neurons. Results: IS had no conspicuous effects on the growing of cortical neurons while it could promote the growing of spinal neurons evidently with the concentration of 100-200 mg/L. The cell viability of cortical neurons and spinals neurons were about 80% and 70% respectively after 3 days exposed to 100μmol/L H₂O₂. There were no evident changes of unmarred cortical neurons on the cell viability, SOD activity in intracellular and the leakage of LDH in extracellular fluid after 3days affected by IS. But IS could increase the cell viability and SOD activity of oxidative damaged cortical neurons, and decrease the leakage of LDH. To the unmarred spinal neurons, IS had no evident effect on the SOD activity, but it could decrease the leakage of LDH in extracellular fluid. IS also could increase the cell viability and SOD activity of oxidative damaged spinal neurons, and decrease the leakage of LDH in extracellular fluid. In addition, IS could inhibit the Ca²⁺ concentration in cytoplasm of oxidative damaged spinal neurons in case Ca²⁺ overloading. All of IS and Stauntonia chinesis DC extracts had high ?OH clearance activity with the does dependent in a certain extent. The antioxidant activity of AI₃ was better than AI₁ and AI₂. The content of flavonoid glycoside in AI₁, AI₂ and AI₃ were 9%, 11% and 27% respectively; and the content of triterpenoid saponin in AI₁, AI₂ and AI₃ were 26.9%, 41.7% and 52.3%. All of AI₁, AI₂ and AI₃ had neuroprotective effects on the oxidative damaged neurons, and the effects on cortical neurons were better than that on spinal neurons, the cell viability of cortical neurons was up to 112.7%. Conclusion: IS could promote the growing of spinal neurons, while it had no evident effects to cortical neurons. IS could protect neurons from oxidative injury via accommodating the activity of a series of antioxidases and inhibiting the overloading of Ca²⁺ concentration in cytoplasm. Besides, IS and AI₁, AI₂ and AI₃ had high·OH clearance activity, so they could protect neurons by scavenging free radicals directly. With the separation and purification of extracts, the content of glycosides in AI₃ was up to 79.3%, so it was deduced that the neuroprotective effects of AI₃ on damaged cortical neurons was relative to the content of glycosides. It also needs further additional study to observe the component which plays a major role, because the flavonoid glycoside and triterpenoid saponin weren't separated completely.