Establishment and genotype identification of hepatic stellate cell-specific Grk2 gene knockout mouse model / 中国药理学通报

Aim To establish a stable hepatic stellate cell ( HSC ) -specific G protein-coupled receptor kinase 2 ( GRK2 ) knockout mice and provide the important animal model for further studying the biological function of GRK2 in HSC. Methods The loxP-labeled Grk2 gene mouse (Grk2

Comparison of two methods for establishing non-alcoholic fatty liver disease model in mice / 中国药理学通报

Aim To compare the effects of two different methods of establishing non-alcoholic fatty liver disease ( NAFLD) model, and to explore a more efficient method for establishing NAFLD model that conforms to the characteristics of human disease.Methods C57BL/6J mice were randomly divided into NI) group, WD group, WD combined with CC14 group.'Hie NI) group was fed a normal maintenance diet, and WD group was fed WD.On the basis of WD feed, mice in WD + CC14 group were intraperitoneally injected with CC14 oil solution.'Hie mice were sacrificed on 6, 11 and 16 weeks after modeling.HE staining and oil red 0 staining were performed to observe the pathological changes of liver.The serum levels of ALT, AST, TG and T- CHO were detected by automatic biochemical analyzer, and the levels of IL-lp, IL-6 and TNF-a in liver homogenate were detected by ELISA.The protein expression of FAS and a-SMA was detected by Western blot.Results As the development of model, pathological results showed that NAFLD model was success-fully established by these two methods.At the same time point of modeling, compared with WD group, the liver pathology of WD + CC14 group was more serious, liver steatosis appeared since 6th week.The serum ALT, AST levels and the contents of TG and T-CHO significantly increased.Meanwhile, the levels of inflammatory cytokines obviously increased in the liver, the expression of fibrosis-associated protein a-SMA increased, and the model could progress to the stage of NASH on 16th week.The course of NAFLD in the WD group progressed slowly, and steatosis appeared on 1 1 th week, and it was further aggravated till 16th week.The pro-tein level of FAS was significantly higher than that in WD + CC14 group, and no obvious inflammatory cell infiltration was observed.Conclusions WD feed combined with CC14 to establish NAFLD model takes shorter time and exerts better effect than feeding WD a- lone.It can progress to NASH on 16th week, which can be used as an ideal method to establish NAFLD model.

Breeding and genotype identification of Arrb2 gene knockout mice / 中国药理学通报

Aim To explore the optimal way of breeding and genotype identification of Arrb2 knockout mice, and to find a simple and quick polymerase chain reaction ( PCR) method for the genotyping of Arrb2 knockout mice. Methods Breeding homozygote genotype of Arrb2 gene knockout mice were copula-ted with wild-type C57BL/6J mice, and then the heterozygous mating were used for mating. The growth and development of off-spring were observed. The genomic DNA was extracted from the tail of two-week-old mice. PCR was employed to amplify the Arrb2 gene fragment, and electrophoresis was used to present the gene type. Results The breeding and reproducing were successful and three genotype offspring, including wild-type,heterozygous and homozygous knockout mice were obtained. Agarose gel electrophoresis results showed the size of PCR prod-ucts was about 186 bp and 224 bp, which was consistent with the expected target gene fragment, and identified Arrb2 gene knockout mice of different genotypes successfully. Western blot analysis demonstrated the lack ofβ-arrestin2 protein in the major organs from Arrb2 -/ - mice compared with Arrb2 +/ + and Arrb2 +/ - mice. Conclusions It is feasible to obtain the homo-zygous Arrb2 knockout mice by inbreeding heterozygotes. It is simple, rapid and reliable to identify mouse genetype by PCR.

Effects and mechanisms of shaoqiduogan on mice with chemical liver injury / 中国中药杂志

<p><b>OBJECTIVE</b>To study the therapeutic effects and mechanisms of SQDG on carbon tetrachloride-induced chemical liver injury in mice as well as its possible mechanisms. At the same time the pharmacodynamics of SQDG was compared with TGP or ASTs of effective dose.</p><p><b>METHOD</b>The model of carbon tetrachloride-induced chemical liver injury in mice was prepared. The levels of ALT, AST, MDA content, SOD and GSH-Px activities in liver homogenate were assayed by spectrophotometry; Meanwhile, hepatic pathological examination was observed.</p><p><b>RESULT</b>Protective effect of SQDG on carbon tetrachloride-induced chemical liver injury: SQDG was able to significantly decrease serum transaminase levels of chemical liver injury's mice induced by carbon tetrachloride, decreased MDA content and improved the reduced SOD and GSH-px levels in liver homogenate. Furthermore, SQDG also attenuate the area and extent of necrosis and reduce the infiltration of inflammatory cell. Compared with TGP or ASTs of effective dose, SQDG has a better effect on carbon tetrachloride-induced chemical liver injury in mice.</p><p><b>CONCLUSION</b>SQDG can protect mice injured by carbon tetrachloride-induced chemical.</p>

Therapeutic effects of recombinant human interleukin 1 receptor antagonist on type II collagen-induced arthritis rats / 中国药理学通报

Aim: To study the therapeutic effects of recombinant human interleukin 1 receptor antagonist (rhIL-1ra) on type II collagen-induced arthritis (CIA) rats. Methods: SD rats were divided randomly into six groups including normal, model, rhIL-1ra (7.5,30,120 mg·kg-1) and anakinra(120 mg· kg-1) groups. Collagen II emulsion was used to induce CIA model in rats. The body weight was observed once a week. Paw swelling of CIA rats was measured with volume meter. C II induced delayed-type hypersensitivity (DTH) was measured. Meanwhile, the level of anti-C II IgG antibody in serum was assayed by ELISA. Results: The onset of paw swelling was on dlO after injection of emulsion. The level of serum anti-C II IgG antibody was increased significantly in CIA. Pathological changes in joints of CIA rats showed hyperplastic synovium of CIA, inflammatory cells infiltration, pannus, destruction of cartilage and bone. rhIL-1ra(7.5,30,120 mg·kg-1·d-1 x 7 d) and anakinra (120 mg·kg-1·d-1 x 7 d) subcutaneous injection (sc) inhibited inflammatory swelling. rhIL-1ra(30, 120 mg·kg-1·d-1 x 7 d) significantly suppressed the DTH reaction induced with C II in CIA rats. Moreover, rhIL-1ra reduced the level of anti-C II IgG antibody in serum. Pathological examination showed rhIL-1ra(120 mg·kg-1·d-1 x 7 d) significantly improved subchondral inflammation, synovium hyperplasia, pannus and cartilage damage. Conclusion: rhIL-1ra has therapeutic effects on CIA rats.