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The Effect of Hexavalent Chromium on the Expression of ABP、Tf and INH mRNA and Protein in Rat Sertoli Cells

Author: LiuHaiDong
Tutor: LiShiEn
School: Zhengzhou University
Course: Occupational and Environmental Health
Keywords: Cr(Ⅵ) Sertoli Cell ABP Tf INH
CLC: R114
Type: Master's thesis
Year: 2011
Downloads: 22
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Abstract

Chromium (Cr) is one of the most important metalloid element existing in environment. It has been always closely related to our activities both in natural environment and industrial production. Cr and its compound, especially hexavalent chromium [Cr(Ⅵ)], are also used in many industrial processes. Through the advancement of the society, Cr and its compound have imposed increasingly grave harm on social and ecological environment. Experiment show that Cr and its compound are immune toxicity, reproductive toxicity, neurotoxicity, carcinogenicity, nephrotoxicity, of which reproductive toxicity has been paid more and more attention and known.There are many factors contributing to the male infertility, among which pollution factor of environment has attracted more and more attention.Testis is one of the most important gonad of male reproductive system, and is one of the primary target organs of most out chemical agents. Sertoli cell (SC) as the only somatic in tubuli contorti of testis plays an important role in male reproductive system. SC can secrete ABP, Tf and INH which are essential for male reproductive system and are already widely used as measures of SC function.In this paper, Rat SC in primary culture with potassium dichromate at different concentration was carried out to have knowledge of the effect of Cr(Ⅵ) on protein and mRNA expression of ABP, Tf and INH in SC. The results provide basic data for effects of potassium dichromate on the reproductive system of male, and valuable theoretical basis for further research on the mechanism of male reproductive toxicity induced by Cr(VI).Methods:1. Establishing rat SC in primary culture models.2. Determination of Cr(Ⅵ) on the activity of SC in MTT Colorimetry. And use activity measurement to discover the concentration of Cr(Ⅵ).3. The mRNA levels of ABP/Tf/INH were determined by Real-time RT-PCR.4. Western blot analysis was used to detect expression of ABP/Tf/INH protein. 5. statistical analysis. SPSS 12.0 software was used for statistical analysis. The results were measured by mean+SD. Multiple groups means were compared with single factor analysis of variance, and the comparison among groups was performed with Bonfferoni method. a=0.05 was the level of test.Results:1. As a demonstration, the rat SC survival rate was above 95%.24 hours later SC began to adhere to the wall and 48 hours later SC was growing well. The purity of SCs was above 90%.2. MTT assay indicated that Cr(Ⅵ) SC had obvious effect on SC survival rate above 10-6mol/L, and there was significant difference comparing with control group. The results show that 10-8mol/L、10-7mol/L and 10-6mol/L were appropriate for further study.3. PCR products were verified and didn’t have non specific fluorescent staining. ABP、Tf and INH could continuously expressed. Compared with control group, the ABPmRNA in group 10-7mol/L and 10-6mol/L were significantly increased (P<0.05), the TfmRNA in group 10-6mol/L was significantly increased (P<0.05), INHmRNA in group 10-7mol/L and 10-6mol/L were significantly increased (P<0.05).4. Compared with control group, the expression of ABP protein in group 10-6mol/L was significantly increased (P<0.05), the expression of Tf protein in group 10-7 mol/L and 10-6 mol/L were significantly increased (P<0.05), the expression of Tf protein in group 10-6mol/L was significantly increased (P<0.05).Conclusion:1. Cytotoxic effect was hardly ever observed when the concentration is from 10-8 mol/L to 10-6 mol/L. The toxic effect on cells appeared while PTF concentration was 10-5mol/L2. Cr(Ⅵ) may affect the ability of secretory functions of rat sertoli cell.

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