Study About The Relationships Between The Formation Of Cholesterol Calculus And The ABCB4 Gene In Hepatocyte

Objective To explore the relationship between the expressionof ABCB4 gene in liver cell and the formation of cholesterol calculus in gallbladder, To provide a new clew of the formation of cholesterol calculus in molecular level. To provide the new theory foundation about how to prevent the formation of cholesterol calculus, further. Methods Fourteen hepatic tissue specimens were taken from patients with cholesterolcalculus. The patient's average age were forty-nine years old, of which nine were male, five were female. As comparison, ten were taken from patients without cholesterol calculus. These average age are forty-five years old, of which five were male, five were female. Calculus of bile duct were excluded. The group of cholesterol calculus were confirmed by operation and the cholesterol calculus were accredited if its cholesterol content was bigger than 70% by the analysis of infrared spectrometry. As control, the group without cholesterol calculus were verified by abdomen ultrasound inspection and operation. The two group already refused the cardiovascular system disease, hyperlipemia, diabetes, thyroid gland disease, pregnancy, malignant tumor, hepatorenal disease and so on. The experimentation comprised two components: The first, the RNA of liver tissue from all the samples was extracted and ultraviolet spectrophotometry was used to measure the content and purity of it. The mRNA expressions of ABCB4 gene were determined by reverse transcription-polymerase chain reaction (RT-PCR) analysis,respectively. The second, the expressions intensity of Pgp3 in hepatictissue frozen section were determined by immunity histochemistry staining method. Result The electrophoresis result showed: in the experimentation, all production comprised the straps ofβ-actin gene and ABCB4 gene. The expressions of straps in patients with cholesterol calculus were duller than those in the liver of patients without calculus. Computed the optical density value of production in each swimming lane by Quantity One Software, to get the relative ratio of ABCB4 gene andβ-actin gene. At last, calculated therelative content of ABCB4 gene's mRNA. The experimental data was needed to correct three times under the same level process condition and taked the final mean value as the statistics data. RT-PCR showed that the mRNA expressions of ABCB4 gene in liver were significantly lower in patients withcholesterol calculus (0.9075±0.0606) than those in the liver of patients withoutcalculus (1.1538±0.0420) (t=-11.062 P<0.05). The cholangiole cells of liver tissue were accredited the masculine cells if they were dyed yellowish brownunder the high power objective. Each slice selected five fields of vision randomly, in each field observed one hundred cells, the masculine cell≥10% wasaccredited the masculine result, otherwise it was negative. the immunity histochemistry staining method showed that the expressions of Pgp3 in liver were significantly lower in patients with cholesterol calculus (14.29%) than those in the liver of patients without calculus (80%) (X2=9.932 P<0.05). Conclusion The mRNA expressions of ABCB4 gene were decreased in the liver of patients with cholesterol calculus. The expressions of Pgp3 in liver were decreased with cholesterol calculus. The decreased expression of Pgp3, which encoded by ABCB4 gene, may correlate with the formation of cholesterol calculus.