中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2015年
11期
790-794
,共5页
申庆丰%夏英鹏%徐天同%田融%李辉南
申慶豐%夏英鵬%徐天同%田融%李輝南
신경봉%하영붕%서천동%전융%리휘남
氟中毒,牙%脊髓%细胞凋亡%p53%脱氟
氟中毒,牙%脊髓%細胞凋亡%p53%脫氟
불중독,아%척수%세포조망%p53%탈불
Fluorosis,tooth%Spinal cord%Apoptosis%p53%Defluorination
目的 探讨慢性氟中毒大鼠脊髓神经细胞凋亡及p53变化,并观察短期脱氟(脱氟12 w)后对脊髓神经细胞凋亡及p53蛋白表达的影响.方法 120只Wistar大鼠按体质量采用随机数字表法分为4组:高氟组,饮含氟离子为200 mg/L的蒸馏水12 w;高氟对照组,饮蒸馏水12 w;脱氟组,先饮含氟离子为200 mg/L的蒸馏水12 w,再改为饮用蒸馏水12 w;脱氟对照组,饮蒸馏水24w,每组30只.分别于第4、8、12周后测定4组大鼠尿氟含量,于第16、20、24周后测定脱氟组和脱氟对照组尿氟含量.第12周后,将高氟组和高氟对照组大鼠处死,脱氟组改饮蒸馏水.第24周后将脱氟组及脱氟对照组大鼠处死,所有大鼠均取脊髓,流式细胞仪观察脊髓神经细胞凋亡情况;免疫组织化学染色观察p53的表达;蛋白质印迹法(Western blot)检测p53蛋白的含量.结果 流式细胞仪检测细胞凋亡率显示,高氟组[(3.36±0.71)％]大鼠脊髓神经细胞凋亡率与高氟对照组[(0.78±0.65)％]比较明显增加(t=14.680,P＜0.01);脱氟组[(3.47±0.56)％]大鼠脊髓细胞凋亡率明显高于脱氟对照组[(0.83±0.64)％,t=17.003,P＜0.01],但与高氟组比较差异无统计学意义(P＞0.05).免疫组化结果及Western blot结果显示,高氟组大鼠脊髓组织中p53蛋白表达与高氟对照组比较表达明显增多,差异有统计学意义(422.69±12.35比177.82±14.16;253.37±10.42比87.14±7.39,t=77.212、72.988,P均＜0.01);脱氟组大鼠脊髓组织中p53蛋白表达与脱氟对照组比较表达明显增多(418.75±11.84比163.47±8.57;248.29±10.23比98.74±11.52,t=95.663、53.167,P均＜0.01),而与高氟组比较差异无统计学意义(418.75±11.84比422.69±12.35;248.29±10.23比253.37±10.42,t=1.261、1.906,P均＞0.05).结论 慢性氟中毒大鼠的脊髓神经细胞存在明显凋亡,p53表达增多可能是造成损害的原因之一,脱氟后短时间内脊髓神经细胞凋亡的数量未见减少,p53的持续高表达可能是脊髓神经细胞凋亡未见降低的原因之一.
目的 探討慢性氟中毒大鼠脊髓神經細胞凋亡及p53變化,併觀察短期脫氟(脫氟12 w)後對脊髓神經細胞凋亡及p53蛋白錶達的影響.方法 120隻Wistar大鼠按體質量採用隨機數字錶法分為4組:高氟組,飲含氟離子為200 mg/L的蒸餾水12 w;高氟對照組,飲蒸餾水12 w;脫氟組,先飲含氟離子為200 mg/L的蒸餾水12 w,再改為飲用蒸餾水12 w;脫氟對照組,飲蒸餾水24w,每組30隻.分彆于第4、8、12週後測定4組大鼠尿氟含量,于第16、20、24週後測定脫氟組和脫氟對照組尿氟含量.第12週後,將高氟組和高氟對照組大鼠處死,脫氟組改飲蒸餾水.第24週後將脫氟組及脫氟對照組大鼠處死,所有大鼠均取脊髓,流式細胞儀觀察脊髓神經細胞凋亡情況;免疫組織化學染色觀察p53的錶達;蛋白質印跡法(Western blot)檢測p53蛋白的含量.結果 流式細胞儀檢測細胞凋亡率顯示,高氟組[(3.36±0.71)％]大鼠脊髓神經細胞凋亡率與高氟對照組[(0.78±0.65)％]比較明顯增加(t=14.680,P＜0.01);脫氟組[(3.47±0.56)％]大鼠脊髓細胞凋亡率明顯高于脫氟對照組[(0.83±0.64)％,t=17.003,P＜0.01],但與高氟組比較差異無統計學意義(P＞0.05).免疫組化結果及Western blot結果顯示,高氟組大鼠脊髓組織中p53蛋白錶達與高氟對照組比較錶達明顯增多,差異有統計學意義(422.69±12.35比177.82±14.16;253.37±10.42比87.14±7.39,t=77.212、72.988,P均＜0.01);脫氟組大鼠脊髓組織中p53蛋白錶達與脫氟對照組比較錶達明顯增多(418.75±11.84比163.47±8.57;248.29±10.23比98.74±11.52,t=95.663、53.167,P均＜0.01),而與高氟組比較差異無統計學意義(418.75±11.84比422.69±12.35;248.29±10.23比253.37±10.42,t=1.261、1.906,P均＞0.05).結論 慢性氟中毒大鼠的脊髓神經細胞存在明顯凋亡,p53錶達增多可能是造成損害的原因之一,脫氟後短時間內脊髓神經細胞凋亡的數量未見減少,p53的持續高錶達可能是脊髓神經細胞凋亡未見降低的原因之一.
목적 탐토만성불중독대서척수신경세포조망급p53변화,병관찰단기탈불(탈불12 w)후대척수신경세포조망급p53단백표체적영향.방법 120지Wistar대서안체질량채용수궤수자표법분위4조:고불조,음함불리자위200 mg/L적증류수12 w;고불대조조,음증류수12 w;탈불조,선음함불리자위200 mg/L적증류수12 w,재개위음용증류수12 w;탈불대조조,음증류수24w,매조30지.분별우제4、8、12주후측정4조대서뇨불함량,우제16、20、24주후측정탈불조화탈불대조조뇨불함량.제12주후,장고불조화고불대조조대서처사,탈불조개음증류수.제24주후장탈불조급탈불대조조대서처사,소유대서균취척수,류식세포의관찰척수신경세포조망정황;면역조직화학염색관찰p53적표체;단백질인적법(Western blot)검측p53단백적함량.결과 류식세포의검측세포조망솔현시,고불조[(3.36±0.71)％]대서척수신경세포조망솔여고불대조조[(0.78±0.65)％]비교명현증가(t=14.680,P＜0.01);탈불조[(3.47±0.56)％]대서척수세포조망솔명현고우탈불대조조[(0.83±0.64)％,t=17.003,P＜0.01],단여고불조비교차이무통계학의의(P＞0.05).면역조화결과급Western blot결과현시,고불조대서척수조직중p53단백표체여고불대조조비교표체명현증다,차이유통계학의의(422.69±12.35비177.82±14.16;253.37±10.42비87.14±7.39,t=77.212、72.988,P균＜0.01);탈불조대서척수조직중p53단백표체여탈불대조조비교표체명현증다(418.75±11.84비163.47±8.57;248.29±10.23비98.74±11.52,t=95.663、53.167,P균＜0.01),이여고불조비교차이무통계학의의(418.75±11.84비422.69±12.35;248.29±10.23비253.37±10.42,t=1.261、1.906,P균＞0.05).결론 만성불중독대서적척수신경세포존재명현조망,p53표체증다가능시조성손해적원인지일,탈불후단시간내척수신경세포조망적수량미견감소,p53적지속고표체가능시척수신경세포조망미견강저적원인지일.
Objective To study the spinal neurocyte apoptosis and the changes of p53 in chronic fluorosis rats,and the improvement after drinking no fluoride water.Methods One hundred twenty Wistar rats were divided into 4 groups by random number table method according to body mass,30 rats in one group fed with high concentration NaF water (200 mg/L) to make fluorosis model and classified as high fluoride group;other 30 rats were fed with distilled water as control group;another 30 rats were fed with high concentration NaF water (200 mg/L) for 12 weeks,then fed with distilled water for 12 weeks and classified as defluorination group;the rest 30 rats were classified as defluorination control group.The content of fluoride in urine was tested after the 4th,8th,and 12th weeks.Then the content of fluoride in urine of defluorination group and defluorination control group was tested.The high fluoride group rats and control group rats were killed after 12th week.Defluorination group rats and defluorination control group rats were killed after 24th week.Their spinal cord was collected.The expression of p53 protein in spinal cord was detected by immunohistochemistry and Westem blotting.Apoptosis of the neurocyte was quantitatively analyzed by flow cytometry (FCM).Results By FCM,apoptosis of neurocyte was increased in both high fluorosis group rats and defluorination group rats compared with those in control group rats [(3.36 ± 0.71)％ vs.(0.78 ± 0.65)％;(3.47 ± 0.56)％ vs.(0.83 ± 0.64)％,t =14.680,17.003,all P ＜ 0.01)],but no difference was found between these two groups [(3.47 ± 0.56)％ vs.(3.36 ± 0.71)％,P ＞ 0.05)].Immunohistochemistry and Western blotting revealed that p53 expression in spinal cord of high fluorosis group rats was increased compared with those in control group rats (422.69 ± 12.35 vs.177.82 ± 14.16;253.37 ± 10.42 vs.87.14 ± 7.39,t =77.212,72.988,all P ＜ 0.01).And p53 expression in spinal cord of defluorination group rats was increased compared with those in control group rats (418.75 ± 11.84 vs.163.47 ± 8.57;248.29 ± 10.23 vs.98.74 ± 11.52,t =95.663,53.167,all P＜ 0.01).But the differences were not statistically significant (418.75 ± 11.84 vs.422.69 ± 12.35;248.29 ± 10.23 vs.253.37 ± 10.42,t =1.261,1.906,all P ＞ 0.05).Conclusions There is apoptosis of neurocytes in the spinal cord of chronic fluorosis rats;overexpression of p53 probably plays an important role in the mechanism of damage induced by excessive fluorine.Apoptosis can not be recovered after defluorination for a short time,and persistent overexpression of p53 may be one of the reasons that apoptosis of neurocytes in the spinal cord can not decrease.
