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为了解低蛋白条件对燃煤型氟中毒雌鼠卵巢颗粒细胞的影响,选取120只断乳2周的60~80 g清洁级SD雌性大鼠,随机分为对照组、低蛋白组、染氟组、低蛋白染氟组,每组30只。对照组和低蛋白组进食75%非病区玉米饲料(含氟量为5.2 mg/kg),染氟组和低蛋白染氟组进食75%燃煤型氟中毒病区玉米饲料(含氟量为96 mg/kg);对照组及染氟组饲料中大豆蛋白质含量为18 g/kg,低蛋白组和低蛋白染氟组为9 g/kg。每组分别于60、120、180 d动态观察雌鼠氟斑牙变化情况,测定尿氟、骨氟水平,采用透射电镜观察卵巢颗粒细胞超微结构变化,以末端转移酶标记技术(TUNEL法)检测卵巢颗粒细胞凋亡情况。结果显示,成功建立燃煤型氟中毒雌鼠模型,低蛋白染氟组较染氟组电镜下病理损伤更明显。与对照组比较,低蛋白组120、180 d卵巢颗粒细胞凋亡率升高,染氟组和低蛋白染氟组60、120、180 d卵巢颗粒细胞凋亡率升高,差异均有统计学意义(P<0.05);低蛋白染氟组60、120、180 d卵巢颗粒细胞凋亡率高于低蛋白组,60、120 d卵巢颗粒细胞凋亡率高于染氟组,差异均有统计学意义(P<0.05)。染氟组180 d时卵巢颗粒细胞凋亡率高于60 d,差异有统计学意义(P<0.05)。析因分析结果显示,饲养60、120、180 d时氟对雌鼠卵巢颗粒细胞凋亡率有影响(F值分别为1 180.051,1 187.294,999.577,P<0.05);饲养60、120 d时低蛋白对雌鼠卵巢颗粒细胞凋亡率有影响(F值分别为17.613,12.706,P<0.05),而饲养180 d时无影响(F=4.017,P>0.05),氟与低蛋白对雌鼠卵巢颗粒细胞凋亡率的影响无交互效应(F值分别为3.479,0.088,0.941,P>0.05)。提示低蛋白条件下燃煤型氟中毒雌鼠卵巢颗粒细胞病理损伤有所加重,但低蛋白与氟染毒对凋亡率的影响无明显相关性。
In order to understand the effect of low protein on ovarian granulosa cells in coal-induced fluorosis in female rats, 120 female SD rats of 60 ~ 80 g clean grade were weaned for 2 weeks, and were randomly divided into control group, low protein group, Group, low-protein fluoride group, 30 rats in each group. The control group and low protein group fed 75% non-ward corn feed (the content of fluoride was 5.2 mg / kg), while the group fed with fluoride and low protein drank fluoride fed corn containing 75% coal-fired fluorosis area 96 mg / kg). The contents of soy protein in the control group and the group fed with fluoride were 18 g / kg and 9 g / kg for the low-protein and low-protein groups. The changes of dental fluorosis were observed dynamically at 60, 120 and 180 days in each group. Urinary fluoride and bone fluoride levels were measured. The ultrastructure of ovarian granulosa cells was observed by transmission electron microscopy. TUNEL method Detection of ovarian granulosa cell apoptosis. The results showed that the successful establishment of a model of coal-induced fluorosis in female rats, fluoride-stained with low-protein group than the staining of the pathological damage under the microscope more obvious. Compared with the control group, the apoptotic rates of ovarian granulosa cells in low protein group increased on the 120th and 180th days, and the apoptotic rate of ovarian granulosa cells increased on the 60th, 120th and 180th days in the fluorochrome group and the low protein-coated fluoride group, with statistical differences (P <0.05). The apoptotic rates of ovarian granulosa cells in low-protein-exposed fluoride group were higher than those in low-protein group at 60,120,180 d, and higher than those in fluoride group at 60 and 120 d Significance (P <0.05). The apoptosis rate of ovarian granulosa cells in the fluoridated group was higher than that of the 60 d after 180 d (P <0.05). Analysis of factorial analysis showed that fluorine had effects on the apoptosis rate of ovarian granulosa cells in the females at 60,120,180 d (F = 180.051,1 187.294,999.577, P <0.05, respectively), and at 60 and 120 d Low protein had effects on the rate of apoptotic ovarian granulosa cells (F = 17.613,12.706, P <0.05, respectively), but had no effect on 180 d (F = 4.017, P> 0.05) There were no interaction effects on apoptosis rate of ovary granulosa cells (F = 3.479,0.088,0.941, P> 0.05). It is suggested that the pathological damage of ovarian granulosa cells in the coal-exposed fluorosis poisoning rats was aggravated under the condition of low protein, but there was no significant correlation between the low protein and fluorine exposure on the apoptotic rate.