To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.

The purpose of this study was to investigate the effect of medium-chain fatty acids(MC-FAs)caprylic acid(C8∶0)on lipid metabolism of calf hepatocytes.Primary calf hepatocytes were extracted and cultured,and 1.2 mmol/L nonesterified fatty acids(NEFAs)were added to the hep-atocytes to construct a model of hepatic lipid deposition in primary calf hepatocytes,Five process-ing groups have been set up:Control group(Ctrl),NEFA added group(NEFA),C8∶0 1.2 mmol/L treatment group(C8∶0 1.2),NEFA+C8∶0 0.2 mmol/L treatment group(NEFA+C8∶00.2),C8∶0 0.2 mmol/L treatment group(C8∶0 0.2).Stimulate calf liver cells for 12 hours,and the levels of triglyceride(TG),lipid oxidation(MDA),hydrogen peroxide(H2O2)and total SOD activity were detected by biochemical kit,and FAS,a protein related to lipid synthesis,was detec-ted by Western blot.The results showed that compared with the control group,the concentrations of TG,MDA and H2O2 in NEFA group increased significantly(P＜0.01),and the activity of SOD decreased significantly(P＜0.05).The protein expression levels of FAS,ACC1,DGAT2 and SREBP-1C were significantly up-regulated(P＜0.01),while the expression level of CPT1A was significantly down-regulated(P＜0.01).Compared with the NEFA group,the protein expression levels of SREBP-1C and DGAT2 in the NEFA+C8∶0(concentration 0.2 mmol/L)group de-creased significantly(P＜0.05),and the protein expression level of fatty acid β-oxidation related molecule CPT1A was slightly higher than that in the NEFA group,but there was no statistical sig-nificance(P＞0.05),and the MDA level in hepatocytes decreased significantly(P＜0.05).In a word,the results of this study show that C8∶0 has antioxidant effect,which can effectively reduce the liver injury caused by oxidative stress,regulate the expression of liver fat gene,and then pro-tect liver injury.

To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.

The purpose of this study was to investigate the effect of medium-chain fatty acids(MC-FAs)caprylic acid(C8∶0)on lipid metabolism of calf hepatocytes.Primary calf hepatocytes were extracted and cultured,and 1.2 mmol/L nonesterified fatty acids(NEFAs)were added to the hep-atocytes to construct a model of hepatic lipid deposition in primary calf hepatocytes,Five process-ing groups have been set up:Control group(Ctrl),NEFA added group(NEFA),C8∶0 1.2 mmol/L treatment group(C8∶0 1.2),NEFA+C8∶0 0.2 mmol/L treatment group(NEFA+C8∶00.2),C8∶0 0.2 mmol/L treatment group(C8∶0 0.2).Stimulate calf liver cells for 12 hours,and the levels of triglyceride(TG),lipid oxidation(MDA),hydrogen peroxide(H2O2)and total SOD activity were detected by biochemical kit,and FAS,a protein related to lipid synthesis,was detec-ted by Western blot.The results showed that compared with the control group,the concentrations of TG,MDA and H2O2 in NEFA group increased significantly(P＜0.01),and the activity of SOD decreased significantly(P＜0.05).The protein expression levels of FAS,ACC1,DGAT2 and SREBP-1C were significantly up-regulated(P＜0.01),while the expression level of CPT1A was significantly down-regulated(P＜0.01).Compared with the NEFA group,the protein expression levels of SREBP-1C and DGAT2 in the NEFA+C8∶0(concentration 0.2 mmol/L)group de-creased significantly(P＜0.05),and the protein expression level of fatty acid β-oxidation related molecule CPT1A was slightly higher than that in the NEFA group,but there was no statistical sig-nificance(P＞0.05),and the MDA level in hepatocytes decreased significantly(P＜0.05).In a word,the results of this study show that C8∶0 has antioxidant effect,which can effectively reduce the liver injury caused by oxidative stress,regulate the expression of liver fat gene,and then pro-tect liver injury.

Objective To observe the clinical efficacy of electroacupuncture plus oral administration ofZeling Guanjie Xiaozhong Heji in treating rheumatoid arthritis (RA) due to liver-kidney yin deficiency.Method Totally 126 patients with active RA due to liver-kidney yin deficiency were randomized into a treatment group and a control group, 63 cases in each group. The control group was prescribed with orally taking Methotrexate tablets and Leflunomide tablets, while the treatment group was intervened by electroacupuncture plus oral administration ofZeling Guanjie Xiaozhong Heji in addition to the medications given to the control group. The Disease Activity Score 28 (DAS28) was evaluated before and after intervention, and the therapeutic efficacies were compared based on the criteria of the American College of Rheumatoid (ACR) and syndrome of traditional Chinese medicine (TCM).Result The ACR total effective rate was 87.3% in the treatment group versus 65.1% in the control group, and the difference was statistically significant (P<0.01). The total effective rate based on TCM syndrome was 87.3% in the treatment group versus 73.0% in the control group, and the difference was statistically significant (P<0.05). There was a significant difference in comparing the DAS28 score between the two groups after intervention (P<0.01).Conclusion Electroacupuncture plus oral administration of Chinese medication and western medication is an effective approach in treating RA due to liver-kidney yin deficiency, and it can significantly enhance the therapeutic efficacy based on ACR20 and TCM syndrome.

OBJECTIVE: To study the influence of PD173074 on proliferation and apoptosis of nasopharyngeal carcinoma. METHOD: With immunoblotting and RT-PCR, FGFR1 expression was detected in CNE, PONE1 and C666-1 cell lines. With MTT assay,the time-effect and dose-effect correlation between PD173074 and inhibition of CNE proliferation was evaluated. After PD173074 stimulation, the phosphorylation level of FGFR1 and AKT was detected with immunoblotting assay. Furthermore, influence of PD173074 on the activation of Caspase3 and Caspase9 was detected to study the underlying mechanism of why PD173074 could inhibit CNE proliferation. RESULT: FGFR1 has the highest expression in CNE cell line. Under incubation of 10 nmol/L PD173074 stimulation for 36 hours to 72 hours, the phosphorylation of FGFR1 and AKT was impaired significantly, which further reduced the proliferation of CNE. Moreover, PD173074 can activate the intrinsic apoptotic pathway by stimulating Caspase9,which activated Caspase3 and induced the apoptosis. CONCLUSION PD173074 could inhibit proliferation of nasopharyngeal carcinoma cell through reducing the phosphorylation of FGFR1 and AKT. Additionally, PD173074 can induce CNE apoptosis by activating intrinsic apoptotic pathway via cleaving Caspase9 and Caspase3.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; Pyrimidines ; pharmacology