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肿节风防护腮腺放射性损伤的作用机理研究

The Research for Mechanism of Sarcandra Glabra Anti-Radiation Damage to the Parotid Gland

【作者】 马姗姗

【导师】 王仁生;

【作者基本信息】 广西医科大学 , 肿瘤放射治疗学, 2011, 硕士

【摘要】 目的:观察肿节风对腮腺放射性损伤的防护作用,研究其可能的防护机制以及腮腺放射性损伤的作用机理。方法:(1)采用实验用巴马小型猪作为实验对象,建立肿节风干预下的腮腺放射性损伤实验动物模型:将45头小型猪随机分入空白对照组(空白组)、单纯照射组(单照组)以及肿节风+照射组(药照组),每组15头;照射方式采用γ射线双侧单后野垂直照射腮腺组织,DT 15Gy/1次;给药方式为照射前一周开始按照0.3g/Kg体重的药量经口腔给予肿节风配方颗粒溶液,直到观察结束。建立起肿节风干预下的腮腺放射性损伤实验动物模型,同时观察小型猪的行为体征以及体重变化,取照射后10天、40天、90天3个时间点,随机摘取每组5头小型猪的腮腺进行研究。(2)用RT-PCR法及Western blotting法分别检测照射后10天、40天、90天腮腺中Bcl-2、Bax、P53和Caspase3 mRNA和蛋白表达的变化:比较各组相同时段不同基因mRNA及蛋白的表达差异,组内不同时段同一基因mRNA及蛋白表达的变化,研究腮腺放射损伤后细胞的凋亡和增殖,明确腮腺放射性损伤作用机理的同时探寻肿节风抗放射性损伤的防护机制。结果:(1)成功建立起肿节风干预下的腮腺放射性损伤实验动物模型;药照组的小型猪对放射性损伤的耐受较单照组好,恢复较单照组快。(2) RT-PCR与Western blotting的结果呈现出相同变化趋势:与空白组相比,单照组和药照组中Bax、P53及Caspase3的mRNA和蛋白表达水平均有所提高,三者的组间差异具有统计学意义(P<0.05),其中单照组高于药照组; Bcl-2的表达,空白组高于药照组,药照组高于单照组,差异具有统计学意义(P<0.05)。各组内,空白组中各mRNA和蛋白的表达无明显变化;单照组中Bax、P53及Caspase3的表达水平呈逐渐增高趋势,Bcl-2则为逐渐降低趋势;药照组中,Bax、P53及Caspase3的表达水平呈先高后低的改变,Bcl-2则为先低后高的变化趋势。结论:(1)单次15Gy双侧单后野垂直照射腮腺组织可成功建立起腮腺放射性损伤的实验动物模型;(2)射线导致的腺细胞凋亡增加可能是腮腺放射性损伤的机制之一;(3)肿节风能够较好的防护放射线引起的腮腺损伤,其可能的防护机制为抑制腺细胞的过度凋亡、促进细胞的修复增殖。

【Abstract】 Objective: To observe the effect of anti-radiation damage of Sarcandra glabra to the parotid gland and figure the mechanism of it, and discover the mechanism of damage of the salivary gland.Methods: (1) Model parotid radioactive damage: 45 pigs randomly divided into the placebo, radiotherapy group (R group) and Sarcandra glabra+radiotherapy group (SR group), 15 pigs per group. Illuminating parotid gland issues with theγray DT 15Gy single dose and oral administration with 0.3g/Kg till the end of experiment. Observing and recording the changes of pig’s weight and behaviors of daily living, and researching the parotid of pig’s which being irradiated after 10days 40days and 90days.(2) Using RT-PCR and Western blotting method to detect Bcl-2,Bax, P53 and Caspase3 mRNA and protein expression of parotid gland of pig’s which being irradiated after 10 days 40 days and 90 days.Results: (1) Established the model of parotid radioactive damage intervened by Sarcandra glabra successfully. And the recovery of pigs in SR group were better than R group.(2) The result of RT-PCR and Western blotting showed the same trend: The expression of mRNA and protein of Bax, P53 and Caspase3 in radiotherapy group (R group) and Sarcandra glabra+radiotherapy group (SR group) were higher than placebo, and the R group was higher than SR group (P<0.05); But the expression of Bcl-2 in placebo was higher than SR group, and SR group was higher than R group (P<0.05). The expression of each factor was stable. But it became higher increasingly in R group, and lower after firstly high in SR group. The trend of Bcl-2 took the opposite way in each group.Conclusions:(1) It can model parotid radioactive damage successfully with illuminating parotid gland issues with theγray DT 15Gy single dose;(2) The increased apoptosis might be one of the mechanisms of radioactive damage result from ray. (3)Sarcandra glabra can protect the function of parotid gland very well and the mechanism may be that restrained the excessive apoptosis and facilitated the repair of normal cell.

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