Lipoic Acid For High Hcy, High Glucose Induced Mice Podocytes Apoptosis Of ER Stress

Objective:To cultivate mice podocyte with high glucose、high homocysteine and thioctic acid,to observe the apoptosis rate and the change of endoplasmic reticulum stress (ERS) marker:GRP78and Caspase-12.Methods:Already differentiated mice podocyte were divided into Hcy group and glucose group.The Hcy group has3subgroups:normal control group、high Hcy group、high Hcy+thioctic acid group; the Glucose group has5subgroups:normal concentration glucose group、hypertonic control group、high concentration glucose group、high concentration glucose+thioctic acid group。 The reagents concentration as follows:high Hcy:200umol/l、thioctic acid:500umol/l、normal concentration glucose:5.6mmol/l、mannitol:25mmol/l、high concentration glucose:25mmol/l、Indexs and detection method:after48h, to detect apoptosis rate by flow cytometer, to detect the GRP78and Caspase-12mRNA expression by RT-PCR technology, to detect the GRP78and Caspase-12protein expression by Western blot technology。Results:(1) The normal control group apoptosis rate was very low in the Hcy group, compared with the control group, the high Hcy group apoptosis rate was obviously higher (P<0.05); compared with the high Hcy group, the thioctic acid+high Hcy apoptosis rate reduced obviously (P<0.05)。 At the same time, the glucose group have similar results。the normal concentration glucose group and the hypertonic group apoptosis rate was very low, the apoptosis rate between the normal concentration glucose group and the hypertonic group was not statistically significant, the apoptosis rate of the high glucose group was significantly higher than the control group (P<0.05), the thioctic acid+high glucose group apoptosis rate was lower than the high glucose group (P<0.05).(2) Compared with the control group, GRP78and Caspase-12mRNA expression of the high Hey group increased significantly (P<0.05), GRP78and Caspase-12mRNA expression in the thioctic acid+Hey group were significantly lower than the high Hey group (P<0.05); Compared with the normal glucose concentration group, GRP78and Caspase-12mRNA expression of high glucose group increased significantly in the glucose group (P<0.05); GRP78and Caspase-12mRNA expression of thioctic acid+glucose group were significantly lower than the high glucose group (P<0.05)。(3) compared with the control group, GRP78and Caspase-12protein expression of the high Hey group increased significantly (P<0.05); GRP78and Caspase-12protein expression of thioctic acid+high Hcy group was significantly lower than high Hey group (P<0.05); The results were also demonstrated in glucose group:GRP78and Caspase-12protein expression of the high glucose group was higher than the normal glucose concentration group (P<0.05); the GRP78and Caspase-12protein expression of thioctic acid+glucose decreased significantly than the high glucose group (P<0.05).Conclusion:(1) HHcy, high glucose can increase apoptosis of podocyte by the endoplasmic reticulum stress reaction.(1) Lipoic acid can reduce the endoplasmic reticulum stress reaction to protect the podocyte, but can not protect fully.