Objective To establish a new method for detection of β-glucuronidase (β-G) mRNA in human liver and kidney tissues. Methods β-G mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR) in 10 cases of normal liver tissues, 10 cases of normal kidney tissues and 8 cases of hepatocellular carcinoma tissues. Results The expand products of β-G mRNA were expressed in liver and kidney tissues with similar size of 422 bp. The expression contents of β-G mRNA in liver and kidney normal tissues were different (1.71±0.32 vs 1.83±0.22) but without statistical significance (Pgt;0.05). However, the expression content of β-G mRNA in hepatocellular carcinoma tissues was 3.88±0.86， which was significantly higher (P＜0.01) than that in normal liver tissues. Conclusion β-G mRNA determination is feasible by checking β-G gene alignment in gene bank and designing draw matter in the tissue of liver and kidney. It may be very significant to explore the change of β-G mRNA in various tissues in studying of molecular mechanism.
To study the relationship between the activity of endogenous β-glucuronidase (β-G) in hepatic tissue and the formation of bilirubin stone. We assessed the β-G activity in hepatic tissue of 44 cases of bilirubin stone, 8 cases of cholesterol stone and 25 cases of liver injury by using immunohistochemistry. Results: The cell percentage of β-G positive reaction in bilirubin stone group (49.2%±4.6%) was significantly higher than those in cholesterol stone group (32.5%±3.8%) and liver injury group (27.8%±4.2%), Ｐ＜0.05. There was no relationship between cell percentage of β-G positive reaction and age, patient history, size of stone. Conclusions: The activity of endogenous β-G is closely related to the formation of bilirubin stone. The difference of activity between individuals is possibly the intrinsic factor which may influence the formation of bilirubin stone.
ObjectiveTo investigate the effect of curcumin on the expression regulation of endogenousβ-glucoronidase (β-GD) induced by lipopolysaccharide (LPS).Methods① Human normal intrahepatic biliary epithelial cell line (HiBEpiC) cells in the logarithmic growth phase were divided into blank control group (0 h group) and 7 different stimulation time groups. The cell density was adjusted to 1×104/mL, and the cells were stimulated with 100 mg/mL LPS for 1, 3, 6, 18, and 24 hours respectively, including another two groups where the cells were cultured with LPS-free medium for 18 and 24 hours after LPS stimulation for 24 h. ② HiBEpiC cells in the logarithmic growth phase were divided into blank control group, LPS+low, medium, and high concentration curcumin group. The cell density was adjusted to 1×104/mL. In the blank control group, cells were not stimulated with any reagent; in the LPS group, cells were stimulated with 100 mg/mL LPS, in the other three groups, the cells were stimulated with 100 mg/mL LPS and simultaneously 20, 40, and 80 μmol/L curcumin, respectively, for 24 hours. The expressions of c-myc and endogenous β-GD were detected by Western blot method.Results① The expressions of endogenous β-GD and c-myc in HiBEpiC cells gradually increased with the prolongation of treatment time by LPS, and the expression levels of β-GD and c-myc at each time point group were significantly different from those in the 0 h group (P<0.05). ② There were significant difference between any two groups of the blank control group, LPS group, LPS+low concentration of curcumin group, LPS+medium concentration of curcumin group, and LPS+high concentration of curcumin group (P<0.05).ConclusionCurcumin is able to inhibit the increased expression of endogenous β-GD induced by LPS, possibly via inhibiting expression of c-myc.