Study On The Effect Of Qi Zhi Ke Bi Tong Capsuleon On High Glucose-induced RSC96 Schwann Cell Injury And Its Mechanism

Objective:To explore the effect and mechanism of Qi Zhi Ke Bi Tong capsuleon on diabetic neuropathy model induced by high glucose in RSC96 cells.Methods:The appropriate dose of RSC96 cell damage was determined by using multiple glucose concentrations to determine the viability of RSC96 cells by MTT hair assay.The cell apoptosis was detected by flow cytometry(FCM),the level of reactive oxygen species(Ros)in the cells was detected by DCF-DA fluorescence probe,and the intracellular Ca+concentration was measured by Fluo-4.Enzyme-linked immunosorbent assay(Elisa)was used to detect the intracellular cleaved-caspase-9,cleaved-caspase-3,Bax,Bcl-2,NGF and other key factors.And determine its influence and action mechanism.Results: Glucose of 50 mM could decrease the viability of RSC96 cells to a certain extent.Compared with the blank group,the viability of reconstituted cells decreased by11.4±1.52%.With the increase of dose,the viability of RSC96 cells decreased further.Middle dose(50?g / mL,100?g / mL)could effectively inhibit the decrease of RSC96 cell viability induced by high glucose,which was restored to 82.2±3% and 88.3±4.1% of the control group,respectively.When RSC96 cells were induced by high glucose,the late apoptosis increased to21.1% and the early apoptosis was detected by flow cytometry.It rose to 10.5 percent.However,this phenomenon was improved after adding Qizhi to quench thirst arthralgia,in which 50?g / mL could effectively reduce the early apoptosis and late apoptosis of RSC96 cells induced by high glucose,which were 3.77% and 14.5%,respectively.Qi Zhi Ke Bi Tong capsuleon could decrease the increase of cleaved-caspase3 and cleaved-caspase9 in RSC96 cells induced by high glucose in a concentration-dependent manner.At the concentration of 100 ?g / mL,the level of Qi Zhi Ke Bi Tong capsuleon decreased to117.5±3.6% and 121.±5.1% of that of blank group,respectively.Stilbene Both doses(50?g,100?g / mL)could significantly reduce the increase of Ca+ in RSC96 cells induced by high glucose,the level of which decreased to 135.45±2.87% and 115.71±3.19%,respectively.Qi Zhi Ke Bi Tong capsuleon could decrease the increase of Bax induced by high glucose in aconcentration-dependent manner.At the same time,we detected that the negative regulatory protein of Bax,Bcl-2,had no obvious effect on Bcl-2,but only showed significant difference at the concentration of 100 ?g / mL.The level of Bcl-2 was 81.1% of that of the blank group.2±2.5%.Qi Zhi Ke Bi Tong capsuleon could decrease the increased ROS in RSC96 cells induced by high glucose to 130.2±8.3% and 109.5±3.2%,respectively.When the high concentration of Qi Zhi Ke Bi Tong capsuleon quenched thirst was restored to a certain degree of RSC96 cell viability,its NGF secretion gradually returned to normal,which was about 82.4±3.6% of that of the blank group.Conclusion: Qi Zhi Ke Bi Tong capsuleon can inhibit the activation of Bax and the release of Ca in RSC96 cells by decreasing the increase of intracellular ROS induced by high glucose,thus preventing the activation of caspase-3 and caspase-9,and thus inhibiting the apoptosis of RSC96 cells.Finally,the expression of NGF in RSC96 was restored,and the function of the cell was maintained.