Cytotoxicity and underlying mechanism of evodiamine in HepG2 cells / 北京大学学报(医学版)

OBJECTIVE@#To investigate evodiamine (EVO)-induced hepatotoxicity and the underlying mechanism.@*METHODS@#HepG2 cells were treated with EVO (0.04-25 μmol/L) for different time intervals, and the cell survival rate was examined by cell counting kit-8 (CCK-8) method. After HepG2 cells were treated with EVO (0.2, 1 and 5 μmol/L) for 48 h, the alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) activities and total bilirubin (TBIL) content of supernatant were detected. A multifunctional microplate reader was used to detect the intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in HepG2 cells to evaluate the level of cell lipid peroxidation damage. The interactions between EVO and apoptosis, autophagy or ferroptosis-associated proteins were simulated by molecular docking. The HepG2 cells were stained by mitochondrial membrane potential (MMP) fluorescent probe (JC-10) and annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI), and MMP and apoptosis in HepG2 cells were detected by flow cytometry. The protein expression levels of caspase-9, caspase-3, bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2) were detected by Western blot.@*RESULTS@#The cell survival rate was significantly reduced after the HepG2 cells were exposed to EVO (0.04-25 μmol/L) in a time- and dose-dependent manner. The half maximal inhibitory concentration (IC50) of the HepG2 cells treated with EVO for 24, 48 and 72 h were 85.3, 6.6 and 4.7 μmol/L, respectively. After exposure to EVO (0.2, 1 and 5 μmol/L) for 48 h, the ALT, AST, LDH, ALP activities and TBIL content in the HepG2 cell culture supernatant, and the MDA content in the cells were increased, and SOD enzyme activity was decreased. Molecular docking results showed that EVO interacted with apoptosis-associated proteins (caspase-9 and caspase-3) better. JC-10 and Annexin V-FITC/PI staining assays demonstrated that EVO could decrease MMP and promote apoptosis in the HepG2 cells. Western blot results indicated that the protein expressions of cleaved caspase-9 and cleaved caspase-3 were upregulated in the HepG2 cell treated with EVO for 48 h. In contrast, the protein expressions of pro-caspase-3, BSEP and MRP2 were downregulated.@*CONCLUSION@#These results suggested that 0.2, 1 and 5 μmol/L EVO had the potential hepatotoxicity, and the possible mechanism involved lipid peroxidation damage, cell apoptosis, and cholestasis.

Effects of Zishen Yutai Pills on endocrine in DOR rats / 中成药

AIM To study the effects of Zishen Yutai Pills (Codonopsis Radix,Dipsaci Radix,Morindae officinalis Radix,etc.) on endocrine in diminished ovarian reserve (DOR) rats.METHODS Female SD rats with normal oestrous cycle were induced by intragastric administration of 55 mg/(kg · d) tripterygium glycosides for fourteen days to establish DOR model,and then divided into Zishen Yutai Pills high-,low-dose groups,Bujiale group,model control group and negative control group.After 35 days' administration,serum estradiol (E2),follicle-stimulating hormone (FSH),luteinizing hormone (LH) and progesterone (P) levels were detected by ELISA.And ovarian tissue estrogen receptor (ER),follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) expressions were detected by immunohistochemical method.RESULTS Compared with the negative control group,there were significant changes in levels of E2,FSH,LH,P,and expressions of ER,FSHR,LHR in the model control group (P ＜ 0.05),but there were no statistical significances in body weight,ovary index,uterus index and serum FSH/LH level between the two groups (P ＞ 0.05).Compared with the model control group,E2 level was increased in the Zishen Yutai Pills high-,low-dose groups and Bujiale group,FSH,LH levels were decreased,and they also promoted ER expression and restrained FSHR,LHR excessive expressions,which were statistically significant (P ＜ 0.05).P level was significangly increased in different dose groups (P ＜ 0.05),FSH/LH in the high-dose group was obviously decreased (P ＜ 0.05),and body weight,ovary index,uterus index in various intervening groups showed no statistical significances (P ＞ 0.05).CONCLUSION Zishen Yutai Pills can improve ovarian reserve function in DOR rats by affecting the expression of ovarian tissue hormone receptor and regulating endocrine function.

Analyzing the Chinese medicine pathogenesis of stroke / 中国中西医结合杂志

Both ischemic and hemorrhage stroke pertain to the category of wind stroke in Chinese medicine (CM). Up to date, it is deemed that the etiology and pathogenesis of wind stroke are wind, fire, sputum, qi, stasis, and deficiency. Among them, it is regarded that wind and fire are the key factors triggering wind stroke. By analyzing the time order and causality, it is found that wind stroke is prior to the onset of wind and fire, wind and fire are the secondary outcomes of wind stroke. By parallel comparing stroke with thromboembolism and hemorrhagic diseases in other Zang-organs, it can be comprehended that the reason why wind symptoms appear in stroke is due to its physiological feature of brain itself. Based on Neijing, the pathogenesis of wind stroke is proposed as follows. Tunnels of viscera (vessels) get lesions. The old pathogenic factors of sputum and stasis or the stasis formed by bleeding inside viscera consume qi, and blood of viscera and damage the spirits hidden in them. The damage of Gan-spirit causes symptoms of stroke, such as hemiplegia, deviation of eyes and mouth, and so on. Wind and fire symptoms are caused by the injury of Gan blood and yin, and/or the stagnation of fire in pericardium (the pathway organ) due to obstruction by old pathogenic factors and stasis (formed by bleeding).