1.The effects of let-7c-1 on the learning and memory of epileptic rats induced by PTZ
Yayun LIAO ; Xixia LIU ; Yiqing HUANG ; Yuhan LIAO ; Yuan WU
Chinese Journal of Nervous and Mental Diseases 2017;43(4):239-244
Objective To explore the effect of let-7c-1 on the learning and memory of PTZ-induced epileptic rats and its relevant mechanism.Methods A model of temporal lobe epilepsy (TLE) was induced via PTZ kindling in SD male rats.The epileptic rats were divided into epilepsy group,agomir-control group,let-7c-1 agomir group (12 rats for each).Twelve rats were served as a negative control group.The behavior and the expression levesl of let-7c-1,Bcl-2 protein and Caspase3 were evaluated at 28 days following PTZ.Results Compared to the negative group,the escape latency of epilepsy group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P<0.05).However,those parameters were not significantly different between the epilepsy group and the agmoir-control group (P>0.05).Compared to the agmoir-control group,the escape latency of let-7c-1 agomir group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P< 0.05).The expression levels of let-7c-1 and let-7c-1 were 1.35±0.32 in agmoir-control group and 62.53±21.01 in agomir group (F=50.97,P<0.05).The expression levels of let-7c-1 were higher in let-7c-1 agomir group than in other groups (P<0.05).Compared to the negative group,the expressions of Bcl-2 protein in other groups were decreased (P<0.05) and the Caspase3 protein were increased (P<0.05).Compared to the agomir-control group,the expression of Bcl-2 protein was significantly decreased and the expression of Caspase3 protein was significantly increased in let-7c-1 agomir group (P<0.05).Conclusions The present study shows that let-7c-1 may impair the learning and memory of PTZ-induced epileptic rats through decreasing the Bcl-2 protein and increasing Caspase3 protein in the hippocampus.
2.Inhibitory Effects of Protopine on the Proliferation of Human Hepatic Stellate Cells HSC-LX 2 and Its Mechanism Study
Xinyu WU ; Jing LI ; Jinjuan ZHANG ; Shanggao LIAO ; Qing MEI ; Yayun WU ; Xiaolan XI
China Pharmacy 2019;30(23):3210-3215
OBJECTIVE: To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS: MTT assay was used to detect the effects of 25, 50, 100, 200, 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group (1640 medium containing 5% fetal bovine serum), protopine low-concentration, medium-concentration and high-concentration groups (100, 200, 400 μmol/L). After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA, Collagen Ⅰ, Collagen Ⅲ, MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA, Collagen Ⅰ, Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS: The inhibitory rates of 25, 50, 100, 200, 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0, 6.9%, 18.7%, 34.2%, 48.9%, 53.9%, respectively. Compared with control group, mRNA expression of Collagen Ⅰ, TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration, medium-concentration and high-concentration groups, while protein expression of MMP- 2 was increased significantly, with statistical significance (P<0.05 or P<0.01). Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups, mRNA expression of α-SMA and Collagen Ⅲ, protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly, with statistical significance (P<0.05 or P<0.01). CONCLUSIONS: Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation, and reduce the expression of a-SMA, Collagen Ⅰ, Collagen Ⅲ and TIMP-1, and increase the expression of MMP-2.