2001 Fiscal Year Final Research Report Summary
THE ADHERENCE OF CALCIUM OXALATE CRYSTAL ON UROTHELIUM AND CYTOKINE PRODUCTION
| Project/Area Number |
12671554
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Keio University |
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Principal Investigator |
NAKAJIMA Fumio Keio University School of Med. Urology, Lecturer, 医学部, 専任講師 (10159064)
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| Co-Investigator(Kenkyū-buntansha) |
SATOH Akinori Keio University School of Med. Urology, Staff, 医学部, 助手 (00296675)
MURAI Masaru Keio University School of Med. Urology, Professor, 医学部, 教授 (90101956)
NAKASHIMA Jun Keio University School of Med. Urology, Lecturer, 医学部, 専任講師 (10167546)
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| Project Period (FY) |
2000 – 2001
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| Keywords | calcium oxalate / urothelium / adherence / cytokine / allopurinol |
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| Research Abstract |
It is known that oxalate and calcium oxalate monohydrate (COM) crystals induce urothelium injury. Although xanthine oxidase inhibitor (allopurinol, oxypurinol) can protect the cell injury which is induced by oxalate and calcium crystals, whether xanthine oxidative inhibitor can inhibit crystal adhesion onto urothelium remains unclear. Therefore we investigate whether xanthine oxidative inhibitor can inhibit adhesion of COM on urotheium cells. On the basic experiment, the adhesion of COM seed onto urothelium cell lines were found to be more in the growth state than the static state of the cell lines. The cell lines used were MDCK, LLCPK_1 and HK-2. These cells were cultured to confluent on round glass cover slips in a 12-well culture plate in MEM with 10% FCS at 37 ℃ and 5% C0_2. These cells were incubated at the presence or absence of 0.2 or 2 mmol. of allopurinol or oxypurinol for 4 hours before attachment with COM crystal. One mg/ml of ^<14>C COM crystal suspension were added and incubated for 10 minutes. Non-adherent crystals were washed in PBS and adherent radioactive crystals were measured by liquid scintillation counter. The cytotoxicity induced by COM crystal was measured by LDH release assays. Allopurinol and oxypurinol significantly reduce the COM crystal adhesion in dose dependent manner. Two mmol. of allopurinol and oxypurinol showed approximately 60% of adhesion inhibition. Allopurinol and oxypurinol also prevent the release of LDH from COM injured cell lines. However, only 10 minutes incubation with COM seed showed less than 2% of cytotoxicity. Therefore the inhibition of cell cytotoxicity caused by COM is not the major factor in anti-adherence effect of allopurinol. Xanthine oxidative inhibitor can prevent the COM crystal adhesion on urothelium. Therefore xanthine oxidative inhibitor may be expected as an agent for prevention of urolithiasis. Regarding the cytokine production by urothelium cell lines, we can not show any evidence.
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Research Products
(5 results)
