Study On The Mechanism Of Berberine’s Effects In Alleviating Ferroptosis Of Nerve Cells Induced By Palmitic Acid

Objective: To study the mechanism of berberine’s effects in alleviating ferroptosis of nerve cells induced by palmitic acid.Methods: The model of type 2 diabetes was built in vitro by using HT22 cells,with Ctrl group,PA treatment group and BBR therapy group set up.The CCK-8 was used to detect the proliferation rate of cells.The lactate dehydrogenase detection kit was used to detect the extent of cell damage.The fluorescence confocal microscope was used to observe the morphological change of mitochondria.The mitochondrial function was observed by detecting the content of ATP,the mitochondrial membrane potential and the content of ROS in different groups.The expression of proteins related to ferroptosis was detected by Western Blot.Results: According to experimental results,the proliferation rate of HT22 cells in PA treatment group was significantly lower than that in Ctrl group(P<0.0001),but increased to a certain extent after BBR treatment(P<0.001).Based on the test result of the LDH kit,the LDH content of cell supernatant in PA group was significantly higher than that in Ctrl group(P<0.01),indicating that the cells were seriously damaged.The observation result from the fluorescence confocal microscope showed that mitochondria in Ctrl group were in slender,elongated and threadlike forms,while those in PA treatment group were in fragmented and dot-like forms.In BBR therapy group,the forms of fragmented mitochondria gradually recovered with the increase of BBR concentration.In accordance with the mitochondrial function test result,compared with Ctrl group,the content of ATP and the mitochondrial membrane potential in PA treatment group decreased significantly,and the content of ROS increased(P<0.01).After BBR treatment,the content of ATP and the mitochondrial membrane potential increased significantly,and the content of ROS decreased(P<0.01),indicating that BBR can improve the function of damaged mitochondria.WB test results showed that compared with Ctrl group,the expression of proteins related to ferroptosis(ACSL4 and HO-1)in PA treatment group went down significantly(P<0.001).After BBR treatment,the expression of ACSL4 went up(P<0.01),indicating that BBR may reduce the ferroptosis of cells by alleviating the over-expression of ACSL4.At the same time,it was found that BBR could dephosphorylate AMPK caused by PA(P<0.0001).Conclusion: BBR can restrain the over-expression of ACSL4 protein induced by PA,reduce the content of ROS in cells,regulate the mitochondrial function,and alleviate the ferroptosis of HT22 cells caused by PA.This mechanism may be related to the regulation of AMPK signaling pathway.