ABSTRACT
Objective To study the function and mechanism of receptor-interacting protein kinase 3 (RIPK3)-mediated necroptosis in liver ischemia-reperfusion injury (IRI) in mice.Methods Sixty mice were randomly divided into four groups using Stata statistical software:Wild-type (WT)-sham group,WT-IRI group,HKO (HKO:RIPK3 liver-specific knockout)-sham group and HKO-IRI group.Sham operation was used as a control in which only the hepatic portal blood vessels were freed after laparotomy,and blood flow was not blocked.In the WT-IRI group and the HKO-IRI group,the hepatic portal vein was freed,and the blood supply of left hepatic lobe and the mid-hepatic lobe wer blocked for 90 min,then the blood vessels were opened for 6 h.Blood and liver tissue samples of each group of mice were taken to detect liver function.Inflammatory infiltration and liver injury were detected by immunohistochemistry and hematoxylin and eosin (HE) staining,and autophagyassociated protein LC3-Ⅱ and P62 were detected by Western blotting.The primary hepatocytes of WT mice and HKO mice were extracted and divided into control group and hypoxia-reoxygenation group (HIR group).After attachment of primary hepatocytes,the HIR group was given hypoxia for 6 h and reoxygenated for 4 h.The supernatant was taken for detecting ALT and AST,and the cell extract protein was used to detect LC3-Ⅱ and P62.Results As compared with the control groups,the liver functions of the IRI groups were significantly impaired,and as compared with the WT-IRI group,the liver damage was significantly aggravated in the HKI-IRI group (P ＜ 0.05),and the LC3-Ⅱ protein content was significantly decreased and the P62 protein content was increased.Similarly,after hepatocytes were were given hypoxia and reoxygenated,HKO-derived hepatocytes were more severely damaged than WT-derived hepatocytes.Conclusions Blocking RIPK3-mediated necroptosis of hepatocytes could induce autophagy inhibition,which aggravates hepatic ischemia-reperfusion injury.
ABSTRACT
OBJECTIVE:To establish HPLC fingerprints of Potentilla discolor,and to conduct authenticity identification.METHODS:HPLC method was adopted.The determination was performed on InertSustain C18 column with mobile phase consisted of 0.1％ formic acid solution-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min,detection wavelength of 360 nm,colunn temperature of 30 ℃,sample size of 10 μL.Using rutin as reference,HPLC chromatograms of 19 batches of P.discolor and 2 batches of P.chinesis were determined.TCM Fingerprint Similarity Evaluation System (2004) was used for similarity evaluation of 21 batches of samples,and common peak identification of 19 batches of P discolor SPSS 21.0 statisticl software was used for main component analysis and cluster analysis.RESULTS:There were 18 common peaks in HPLC fingerprints of 19 batches of P.discolor,the similarity was higher than 0.9.-HPLC chromatogram was in good agreement with control fingerprint.The similarity of 2 batches of P chinesis was lower than 0.7.The 21 batches of medicinal materials could be grouped into 2 categories,2 batches of P chinesis could be grouped into a category,19 batches of P.discolor could be grouped into a category.P discolor could be grouped into 4 categories.Rutin and quercitrin were main ingredients in 19 batches of P discolor.CONCLUSIONS:Established fingerprint can provide reference for authenticity identification and quality evaluation of P.discolor.
ABSTRACT
Purpose To investigate the anti-inflammatory effect of Radix Rehmanniae Praeparata(RRP)on the footpad inflammation induced by complete Freund's adjuvant(CFA)in rats.Methods CFA 100 μL were injected subcutaneously to the Wistar rats at the pad of right hindfoots.19 days later,the rats were daily siven RRP water extract(0.625,1.250,2.500 g/kg·bw)or dexamethasone(0.5mg/ks·bw)intragastrically.The changes of body weight and foot volume were measured.The indexes of organ and blood were determined at the 29th day,the foot pad was removed,and routine paraffin section was performed.Results The model rats kept foot swelling and lymphocyte infiltrating,and the platelet number decreased.The other indexes were statistically insignificant when compared to the controls.RRP did not display any anti-inflammatory effect on the swollon foots,but thoracic gland and spleen indexes were rescued,and platelet number and creatinine content were increased by RRP administration in a dose-dependent manner.The anti-inflammation of dexamethasone was conspicuous,but the side effects were also significant.Conclusion RRP may be plays an adjunctive action in herbal recipes to treat rheumatoid arthritis.