1.Study on Preinducing the Adult Animal Hepatocytes for Boosting the Metabolic Activity of Bilirubin
Nanqing CHEN ; Jiamin SUN ; Tonghe WU
Journal of Medical Research 2006;0(10):-
Objective To probe whether the preinduced adult animal hepatocytes (AHH) by phenobarbital sodium (PBS) in vivo could boost the metabolic activity of bilirubin in vitro for seeking the optimal biomaterial for the extracorporeal bioartificial liver support system (EBLSS). Methods Twelve adult male rats with 34g mean weight were randomly divided into preinduce group and control group, the rats of the preinduce group were intraperitoneal injected daily with 45mg/kg PBS for 5 days, while the control group with normal sodium. 0.5g wet weight liver was taken from every rat after 24 hours of last administer, the suspensions were made by mixing six wet weight livers in two groups and put in the cultivate board of 96 apertures, 30ul per aperture, then 50?l jaundice serum and 20?l culture fluid were added into every aperture, which were statically cultured in the incubaton of humidity 95% at 37℃, 5% CO2 for 3 hours, and then revolved, the metabolic activity of bilirubins were determined by Beckmann auto-analyzer. Results The total bilirubins and indirect bilirubins fell 60%, 71.42% respectively in the preinduced group, and 34.78%, 54.87% in the control group (P0.05, t=1.571). There were no obvious changes in direct bilirubins in two groups. Conclusions The preinduced AHH by PBS in vivo can boost the metabolic activity of bilirubin in vitro, it may be the optimal biomaterial for EBLSS.
2.The Analysis of Mouse Blood Serum and Colture of Primary Hepatocytes Induced by Phenobarbital Sodiu
Tonghe WU ; Nanqing CHEN ; Yongqing HUANG ; Aijing CHEN ; Yanfen SHI
Journal of Medical Research 2006;0(10):-
Objective To investigate the biological characteritics of adult animal hepatocytes induced by phenobarbital sodium(PBS) in vivo,and to study the potential value of biological artificial liver of effective hepatocytes.Methods 12 adult male mice,are yandomly divided in to preinducing group and controll group.The preinducing group are intraperitoneally injected PBS per day,45mg/kg for 7 times in total;the controll ware injected NS.after that,we detected the blood serum TP,BUN,CHOL,HDLC.And the same amount of isolative hepatocytes was developed after being developed 48h;MTT was used to investigate the proliferation of hepatocytes after being developed 24h;chromosome was investigated to observe the cell division;and the survival deadline and morphology was also investigated.Results The TP had remarkable difference between the two groups(t=2.678,P
3.Effect of Zishen Huoxue Prescription on Regulation of Mitophagy and Neurogenesis in Hippocampal CA1 Region of Vascular Dementia Rats
Ziting ZHAO ; Dahua WU ; Xiuli ZHANG ; Le XIE ; Tonghe LIU ; Shanshan ZENG ; Jiayi SHI ; Ting YAO ; Yao XIE ; Yinchen LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(20):45-52
ObjectiveTo investigate the effect of Zishen Huoxue prescription on promoting neurogenesis in hippocampal CA1 region of vascular dementia (VD) rats by regulating mitophagy. MethodThe 2-VO method was used to establish the VD rat model and 60 SD rats were randomly divided into sham operation group, model group, donepezil hydrochloride group, and Zishen Huoxue prescription low-dose(8.9 g·kg-1), medium-dose(17.8 g·kg-1) and high-dose(35.6 g·kg-1) groups. Morris water maze test was performed to detect the escape latency and the number of crossing platform in each group. The expression of phosphatase and tensin homology-induced kinase 1 (PINK1) and Parkinson protein (Parkin) mRNA in hippocampal CA1 region was detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR). Western blot was used to determine the expression of mitochondrial autophagy signaling pathway-related proteins Parkin, prohibitin 2 (PHB2), mitofusin 2 (Mfn2) and dynamin-related protein 1 (Drp1) in hippocampal CA1 region. The neurogenesis in hippocampal CA1 region was tested by Brdu method. ResultCompared with the conditions in the sham operation group, the learning and spatial memory abilities of the model group were decreased (P<0.05), with damaged mitochondrial structure and autolysosome formation in the hippocampal CA1 region. The expressions of Parkin, Pink1 mRNA and Parkin, PHB2, and Drp1 proteins were up-regulated (P<0.05), while the expression of Mfn2 protein and the neuronal regeneration in hippocampal CA1 region were reduced (P<0.05, P<0.05). Compared with the conditions in the model group, Zishen Huoxue prescription enhanced the learning and spatial memory abilities of VD rats (P<0.05), increased the number of autophagosomes in hippocampal CA1 region and improved the mitochondrial structure. The expression of Parkin, Pink1 mRNA and Parkin, PHB2, and Drp1 proteins in hippocampal CA1 region was up-regulated (P<0.05, P<0.01)while the expression of Mfn2 protein was down-regulated(P<0.05, P<0.01). The number of new neurons in hippocampal CA1 region was also increased(P<0.05, P<0.01). ConclusionThe promoting effect of Zishen Huoxue prescription on the neurogenesis in hippocampal CA1 region of VD rats was related to the mitophagy mediated by Pink1/Parkin signaling pathway.