The Expression of Transthyretin and Its Nucleating Role in Tissues of Human Gallbladder with Cholesterol Stones_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 QIANChang-lin ¹ , LIUHua ¹ ,  SHENZhi-yong ¹ , JIFu ¹ , CAIDuan ²

1. Department of General Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao tong University, Shanghai 201112, China;
2. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China;

Corresponding?author：

SHENZhi-yong, Email: doctorszy1970@163.com

Keywords：

Transthyretin; Cholesterol gallstones; Nucleation role

DOI：

10.7507/1007-9424.20140170

Video：

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Objective To explore the expressions of transthyretin(TTR) mRNA and its protein in tissues of human gallbladder with cholesterol gallstones, and to explore its role in the formation of cholesterol gallstones. Methods Gallbladder were got from cases of cholesterol gallstones(cholesterol gallstones group, n=25) and cases underwent liver transplantation with normal gallbladder(normal control group, n=9) respectively, who were treated in Ren Ji Hospital and Huashan Hospital. Real time PCR(RT-PCR) and Western blot method were used to determine the expressions of TTR mRNA and its protein respectively. In addition, 2 kinds of artificial model bile system were established to test nucleation time(NT) and nucleation activity, which added TTR and albumin(ALB). Results The expression levels of TTR mRNA and protein in cholesterol gallstones group were 1.51±0.78 and 3.95±0.09 respectively, which were both higher than those of normal control group(P<0.05). The NT were(14.5±1.3)d and(18.0±0.8)d in TTR group and ALB group in small model bile system(P<0.01), which was similar with comprehensive model bile system[(13.5±0.6)d vs. (18.5±1.3)d]. The nucleation activity were 0.81 and 0.73 in small model bile system and comprehensive model bile system respectively. Conclusions Expression of TTR up-regulates in human gallbladder tissues of patients with cholesterol gallstones, and TTR plays role of nucleation in model bile system, which is related to the formation of cholesterol gallstones.

Citation： QIANChang-lin, LIUHua, SHENZhi-yong, JIFu, CAIDuan. The Expression of Transthyretin and Its Nucleating Role in Tissues of Human Gallbladder with Cholesterol Stones. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2014, 21(6): 716-720. doi: 10.7507/1007-9424.20140170 Copy

1.	?Schreiber G.The evolutionary and integrative roles of transthyretin in thyroid hormone homeostasis[J].J Endocrinol, 2002, 175(1):61-73.
2.	Hou X, Mechler A, Martin LL, et al.Cholesterol and anionic phospholipids increase the binding of amyloidogenic transthyretin to lipid membranes[J].Biochim Biophys Acta, 2008, 1778(1):198-205.
3.	Dincer A, Onal S, Timur S, et al.Differentially displayed proteins as a tool for the development of type 2 diabetes[J].Ann Clin Biochem, 2009, 46(Pt 4):306-310.
4.	韋麗婭,翟永貞,馮國和.血清前白蛋白對肝功能評估的研究進展[J].世界華人消化雜志, 2013, 21(15):1387-1393.
5.	Sousa MM, Saraiva MJ.Internalization of transthyretin.Evidence of a novel yet unidentified receptor-associated protein (RAP)-sensitive receptor[J].J Biol Chem, 2001, 276(17):14420-14425.
6.	Noguchi H, Kaname T, Sekimoto T, et al.Naso-maxillary deformity due to frontonasal expression of human transthyretin gene in transgenic mice[J].Genes Cells, 2002, 7(10):1087-1098.
7.	Busch N, Matiuck N, Sahlin S, et al.Inhibition and promotion of cholesterol crystallization by protein fractions from normal human gallbladder bile[J].J Lipid Res, 1991, 32(4):695-702.
8.	Zhang D, Xiang J, Wang L, et al.Comparative proteomic analysis of gallbladder bile proteins related to cholesterol gallstones[J].PLoS One, 2013, 8(1):e54489.
9.	張殿彩,丁永斌,夏建國,等.視黃醇結合蛋白的促膽固醇成核效應[J].中華消化外科雜志, 2012, 11(5):476-479.
10.	錢昌林,季福,劉驊,等.人去漿膜層膽囊組織蛋白質表達譜的建立與分析[J].肝膽胰外科雜志, 2014, 26(1):33-37, 54.
11.	Holan KR, Holzbach RT, Hermann RE, et al.Nucleation time:a key factor in the pathogenesis of cholesterol gallstone disease[J].Gastroenterology, 1979, 77(4 Pt 1):611-617.
12.	Holzbach RT, Kibe A, Thiel E, et al.Biliary proteins.Unique inhibitors of cholesterol crystal nucleation in human gallbladder bile[J].J Clin Invest, 1984, 73(1):35-45.
13.	Kibe A, Holzbach RT, Larusso NF, et al.Inhibition of cholesterol crystal formation by apolipoproteins in supersaturated model bile[J].Science, 1984, 225(4661):514-516.
14.	朱雷明,蔡端,張延齡.模擬膽汁體系中游離膽紅素的熱力學研究[J].外科理論與實踐, 1999, 4(1):13-17.
15.	Bradford MM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding[J].Anal Biochem, 1976, 72:248-254.
16.	Planté-Bordeneuve V, Kerschen P.Transthyretin familial amyloid polyneuropathy[J].Handb Clin Neurol, 2013, 115:643-658.
17.	劉燁,郗光霞.轉甲狀腺素蛋白與2型糖尿病[J].中國藥物與臨床, 2011, 11(6):691-692.
18.	Refai E, Dekki N, Yang SN, et al.Transthyretin constitutes a functional component in pancreatic beta-cell stimulus-secretion coupling[J].Proc Natl Acad Sci U S A, 2005, 102(47):17020-17025.
19.	Sundsten T, Ostenson CG, Bergsten P.Serum protein patterns in newly diagnosed type 2 diabetes mellitus-influence of diabetic environment and family history of diabetes[J].Diabetes Metab Res Rev, 2008, 24(2):148-154.
20.	Zanlungo S, Rigotti A, Miquel JF.Abnormalities of lipid metabolism, gallstone disease and gallbladder function[J].Clin Lipidol, 2011, 6(3):315-325.
21.	Tsai CH, Wu JS, Chang YF, et al.The number of metabolic abnormalities associated with the risk of gallstones in a non-diabetic population[J].PLoS One, 2014, 9(3):e90310.
22.	李立,唐繼紅,孫峰,等.膽囊結石患者血脂及蛋白測定的意義[J].中國普外基礎與臨床雜志, 2000, 7(4):255-256.
23.	Han T, Zhang D, Fu Z, et al.Retinol-binding protein 4 as a risk factor for cholesterol gallstone formation[J].Mol Cell Biochem, 2013, 377(1-2):219-227.
24.	張殿彩,項建斌,蔡端,等.膽汁泡相與微膠粒相的蛋白質表達譜對比觀察[J].中華醫學雜志, 2008, 88(46):3298-3301.
25.	Mody N, Graham TE, Tsuji Y, et al.Decreased clearance of serum retinol-binding protein and elevated levels of transthyretin in insulin-resistant ob/ob mice[J].Am J Physiol Endocrinol Metab, 2008, 294(4):E785-E793.
26.	Choi SA, Yun JW, Park HS, et al.Hypoglycemic dipeptide cyclo (His-Pro) significantly altered plasma proteome in streptozocininduced diabetic rats and genetically-diabetic (ob/ob) mice[J].Mol Biol Rep, 2013, 40(2):1753-1765.
27.	Pullakhandam R, Palika R, Ghosh S, et al.Contrasting effects of type 2 and type 1 diabetes on plasma RBP4 levels:the significance of transthyretin[J].IUBMB Life, 2012, 64(12):975-982.

1. ?Schreiber G.The evolutionary and integrative roles of transthyretin in thyroid hormone homeostasis[J].J Endocrinol, 2002, 175(1):61-73.
2. Hou X, Mechler A, Martin LL, et al.Cholesterol and anionic phospholipids increase the binding of amyloidogenic transthyretin to lipid membranes[J].Biochim Biophys Acta, 2008, 1778(1):198-205.
3. Dincer A, Onal S, Timur S, et al.Differentially displayed proteins as a tool for the development of type 2 diabetes[J].Ann Clin Biochem, 2009, 46(Pt 4):306-310.
4. 韋麗婭,翟永貞,馮國和.血清前白蛋白對肝功能評估的研究進展[J].世界華人消化雜志, 2013, 21(15):1387-1393.
5. Sousa MM, Saraiva MJ.Internalization of transthyretin.Evidence of a novel yet unidentified receptor-associated protein (RAP)-sensitive receptor[J].J Biol Chem, 2001, 276(17):14420-14425.
6. Noguchi H, Kaname T, Sekimoto T, et al.Naso-maxillary deformity due to frontonasal expression of human transthyretin gene in transgenic mice[J].Genes Cells, 2002, 7(10):1087-1098.
7. Busch N, Matiuck N, Sahlin S, et al.Inhibition and promotion of cholesterol crystallization by protein fractions from normal human gallbladder bile[J].J Lipid Res, 1991, 32(4):695-702.
8. Zhang D, Xiang J, Wang L, et al.Comparative proteomic analysis of gallbladder bile proteins related to cholesterol gallstones[J].PLoS One, 2013, 8(1):e54489.
9. 張殿彩,丁永斌,夏建國,等.視黃醇結合蛋白的促膽固醇成核效應[J].中華消化外科雜志, 2012, 11(5):476-479.
10. 錢昌林,季福,劉驊,等.人去漿膜層膽囊組織蛋白質表達譜的建立與分析[J].肝膽胰外科雜志, 2014, 26(1):33-37, 54.
11. Holan KR, Holzbach RT, Hermann RE, et al.Nucleation time:a key factor in the pathogenesis of cholesterol gallstone disease[J].Gastroenterology, 1979, 77(4 Pt 1):611-617.
12. Holzbach RT, Kibe A, Thiel E, et al.Biliary proteins.Unique inhibitors of cholesterol crystal nucleation in human gallbladder bile[J].J Clin Invest, 1984, 73(1):35-45.
13. Kibe A, Holzbach RT, Larusso NF, et al.Inhibition of cholesterol crystal formation by apolipoproteins in supersaturated model bile[J].Science, 1984, 225(4661):514-516.
14. 朱雷明,蔡端,張延齡.模擬膽汁體系中游離膽紅素的熱力學研究[J].外科理論與實踐, 1999, 4(1):13-17.
15. Bradford MM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding[J].Anal Biochem, 1976, 72:248-254.
16. Planté-Bordeneuve V, Kerschen P.Transthyretin familial amyloid polyneuropathy[J].Handb Clin Neurol, 2013, 115:643-658.
17. 劉燁,郗光霞.轉甲狀腺素蛋白與2型糖尿病[J].中國藥物與臨床, 2011, 11(6):691-692.
18. Refai E, Dekki N, Yang SN, et al.Transthyretin constitutes a functional component in pancreatic beta-cell stimulus-secretion coupling[J].Proc Natl Acad Sci U S A, 2005, 102(47):17020-17025.
19. Sundsten T, Ostenson CG, Bergsten P.Serum protein patterns in newly diagnosed type 2 diabetes mellitus-influence of diabetic environment and family history of diabetes[J].Diabetes Metab Res Rev, 2008, 24(2):148-154.
20. Zanlungo S, Rigotti A, Miquel JF.Abnormalities of lipid metabolism, gallstone disease and gallbladder function[J].Clin Lipidol, 2011, 6(3):315-325.
21. Tsai CH, Wu JS, Chang YF, et al.The number of metabolic abnormalities associated with the risk of gallstones in a non-diabetic population[J].PLoS One, 2014, 9(3):e90310.
22. 李立,唐繼紅,孫峰,等.膽囊結石患者血脂及蛋白測定的意義[J].中國普外基礎與臨床雜志, 2000, 7(4):255-256.
23. Han T, Zhang D, Fu Z, et al.Retinol-binding protein 4 as a risk factor for cholesterol gallstone formation[J].Mol Cell Biochem, 2013, 377(1-2):219-227.
24. 張殿彩,項建斌,蔡端,等.膽汁泡相與微膠粒相的蛋白質表達譜對比觀察[J].中華醫學雜志, 2008, 88(46):3298-3301.
25. Mody N, Graham TE, Tsuji Y, et al.Decreased clearance of serum retinol-binding protein and elevated levels of transthyretin in insulin-resistant ob/ob mice[J].Am J Physiol Endocrinol Metab, 2008, 294(4):E785-E793.
26. Choi SA, Yun JW, Park HS, et al.Hypoglycemic dipeptide cyclo (His-Pro) significantly altered plasma proteome in streptozocininduced diabetic rats and genetically-diabetic (ob/ob) mice[J].Mol Biol Rep, 2013, 40(2):1753-1765.
27. Pullakhandam R, Palika R, Ghosh S, et al.Contrasting effects of type 2 and type 1 diabetes on plasma RBP4 levels:the significance of transthyretin[J].IUBMB Life, 2012, 64(12):975-982.

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The Expression of Transthyretin and Its Nucleating Role in Tissues of Human Gallbladder with Cholesterol Stones

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