CAJ | 학술논문

目的研究丹蓝仙硼疗氟胶囊对饮水型氟中毒大鼠骨组织中骨保护蛋白配体(OPGL)和巨噬细胞集落刺激因子(M-CSF)mRNA及其蛋白质表达的影响.方法以72只SD大鼠为研究对象,按体重均衡随机分为6组(组内雌雄各半):氟中毒组、高剂量药物干预组、中剂量药物干预组、低剂量药物干预组、对照组、硼砂治疗组.除对照组外,各实验组均饮用50 mg/L含氟水,复制氟中毒动物模型.各药物组应用中药丹蓝仙硼疗氟胶囊,高、中、低剂量药物干预组按大鼠每千克体质量每天分别给药0.8、0.4和0.2 g,阳性药物对照(硼砂)剂量为大鼠每千克体质量每天0.8 g.氟电极法检测尿氟、骨氟含量,HE切片观察骨骼病变,并测定骨小梁厚度、骨小梁密度及骨皮质厚度.应用逆转录聚合酶链式反应(RT-PCR)方法和免疫组织化学(EnVision法)染色,观察骨组织中OPGL和M-CSFmRNA及其蛋白表达的变化.结果 (1)氟中毒大鼠10/12发生氟斑牙;低剂量药物干预大鼠2/12发生氟斑牙.中、高剂量药物干预组,正常对照组及硼砂治疗组均无氟斑牙发生.氟中毒组骨氟尿氟明显高于对照组,高中剂量干预组及硼砂治疗组显著降低.(2)与对照组比较,氟中毒大鼠骨小梁厚度、骨皮质厚度及骨小梁密度均明显减少,而各药物组大鼠骨小梁厚度、骨皮质厚度及骨小梁密度均与对照组无明显差异.(3)与对照组比较,氟中毒大鼠骨组织OPGL、M-CSF mRNA表达均增高,差异有统计学意义(P＜0.05).与氟中毒组比较,高、中剂量药物干预组及硼砂治疗组OPGL mRNA表达降低,差异有统计学意义(P＜0.05);高、中、低剂量药物干预组及硼砂治疗组M-CSFmRNA表达降低,差异有统计学意义(P＜0.05).(4)与对照组比较,氟中毒大鼠OPGL蛋白表达增高,M-CSF蛋白表达降低;与氟中毒大鼠比较,高、中、低剂量药物干预组及硼砂治疗组OPGL蛋白表达降低,M-CSF蛋白表达有所升高,差异均有统计学意义(P＜0.05).结论氟可能通过上调OPGL和M-CSFmRNA及其蛋白的共同表达而影响骨代谢;丹蓝仙硼疗氟胶囊能通过下调OPGL mRNA和M-CSF mRNA的表达影响骨重建. 目的研究丹藍仙硼療氟膠囊對飲水型氟中毒大鼠骨組織中骨保護蛋白配體(OPGL)和巨噬細胞集落刺激因子(M-CSF)mRNA及其蛋白質錶達的影響.方法以72隻SD大鼠為研究對象,按體重均衡隨機分為6組(組內雌雄各半):氟中毒組、高劑量藥物榦預組、中劑量藥物榦預組、低劑量藥物榦預組、對照組、硼砂治療組.除對照組外,各實驗組均飲用50 mg/L含氟水,複製氟中毒動物模型.各藥物組應用中藥丹藍仙硼療氟膠囊,高、中、低劑量藥物榦預組按大鼠每韆剋體質量每天分彆給藥0.8、0.4和0.2 g,暘性藥物對照(硼砂)劑量為大鼠每韆剋體質量每天0.8 g.氟電極法檢測尿氟、骨氟含量,HE切片觀察骨骼病變,併測定骨小樑厚度、骨小樑密度及骨皮質厚度.應用逆轉錄聚閤酶鏈式反應(RT-PCR)方法和免疫組織化學(EnVision法)染色,觀察骨組織中OPGL和M-CSFmRNA及其蛋白錶達的變化.結果 (1)氟中毒大鼠10/12髮生氟斑牙;低劑量藥物榦預大鼠2/12髮生氟斑牙.中、高劑量藥物榦預組,正常對照組及硼砂治療組均無氟斑牙髮生.氟中毒組骨氟尿氟明顯高于對照組,高中劑量榦預組及硼砂治療組顯著降低.(2)與對照組比較,氟中毒大鼠骨小樑厚度、骨皮質厚度及骨小樑密度均明顯減少,而各藥物組大鼠骨小樑厚度、骨皮質厚度及骨小樑密度均與對照組無明顯差異.(3)與對照組比較,氟中毒大鼠骨組織OPGL、M-CSF mRNA錶達均增高,差異有統計學意義(P＜0.05).與氟中毒組比較,高、中劑量藥物榦預組及硼砂治療組OPGL mRNA錶達降低,差異有統計學意義(P＜0.05);高、中、低劑量藥物榦預組及硼砂治療組M-CSFmRNA錶達降低,差異有統計學意義(P＜0.05).(4)與對照組比較,氟中毒大鼠OPGL蛋白錶達增高,M-CSF蛋白錶達降低;與氟中毒大鼠比較,高、中、低劑量藥物榦預組及硼砂治療組OPGL蛋白錶達降低,M-CSF蛋白錶達有所升高,差異均有統計學意義(P＜0.05).結論氟可能通過上調OPGL和M-CSFmRNA及其蛋白的共同錶達而影響骨代謝;丹藍仙硼療氟膠囊能通過下調OPGL mRNA和M-CSF mRNA的錶達影響骨重建.
목적 연구단람선붕료불효낭대음수형불중독대서골조직중골보호단백배체(OPGL)화거서세포집락자격인자(M-CSF)mRNA급기단백질표체적영향.방법 이72지SD대서위연구대상,안체중균형수궤분위6조(조내자웅각반):불중독조、고제량약물간예조、중제량약물간예조、저제량약물간예조、대조조、붕사치료조.제대조조외,각실험조균음용50 mg/L함불수,복제불중독동물모형.각약물조응용중약단람선붕료불효낭,고、중、저제량약물간예조안대서매천극체질량매천분별급약0.8、0.4화0.2 g,양성약물대조(붕사)제량위대서매천극체질량매천0.8 g.불전겁법검측뇨불、골불함량,HE절편관찰골격병변,병측정골소량후도、골소량밀도급골피질후도.응용역전록취합매련식반응(RT-PCR)방법화면역조직화학(EnVision법)염색,관찰골조직중OPGL화M-CSFmRNA급기단백표체적변화.결과 (1)불중독대서10/12발생불반아;저제량약물간예대서2/12발생불반아.중、고제량약물간예조,정상대조조급붕사치료조균무불반아발생.불중독조골불뇨불명현고우대조조,고중제량간예조급붕사치료조현저강저.(2)여대조조비교,불중독대서골소량후도、골피질후도급골소량밀도균명현감소,이각약물조대서골소량후도、골피질후도급골소량밀도균여대조조무명현차이.(3)여대조조비교,불중독대서골조직OPGL、M-CSF mRNA표체균증고,차이유통계학의의(P＜0.05).여불중독조비교,고、중제량약물간예조급붕사치료조OPGL mRNA표체강저,차이유통계학의의(P＜0.05);고、중、저제량약물간예조급붕사치료조M-CSFmRNA표체강저,차이유통계학의의(P＜0.05).(4)여대조조비교,불중독대서OPGL단백표체증고,M-CSF단백표체강저;여불중독대서비교,고、중、저제량약물간예조급붕사치료조OPGL단백표체강저,M-CSF단백표체유소승고,차이균유통계학의의(P＜0.05).결론 불가능통과상조OPGL화M-CSFmRNA급기단백적공동표체이영향골대사;단람선붕료불효낭능통과하조OPGL mRNA화M-CSF mRNA적표체영향골중건.
Objective To investigate mRNA and protein expressions of OPGL and M-CSF mRNA in bones of rats with experimental fluorosis induced by intake of fluoride in the drinking water, and to study the antagonizing effects of Danlan Xianpeng Liaofu capsules on the gene expression. Methods Totally 72 SD rats were randomly assorted into 6 groups including the control group, the fluoride group, the high-dosage (0.8 g/kg·d), mid-dosage (0.4 g/kg·d) and low dosage(0.2 g/kg·d) medication groups and the borax group (borax, 0.8 g/kg·d). The distribution of female and male rats in each group was divided up on a fifty-fifty basis. Except the control group, a NaF containing water (NaF 50 mg/L in concentration) was supplied as the drinking water for all the experimental rats in order to establish experimental fluorosis. The thickness and density of trabecula and the thickness of bone cortex were measured by light microscopy. The fluoride content in urine and bone were analyzed by using fluoride ion selective electrode method. Expressions of OPGL and M-CSF mRNA and protein were studied using RT-PCR and immuno histochemistry, respectively. Results (1) 10/12 of the experimental fluorosis rats developed dental fluorosis, and 2/12 of dental fluorosis rats occurred in the low-dosage medication group. Fluoride content in urine and bone of the fluorosis rats increased (P＜0.05). (2) Compared with that of the control rats, the bone trabecular depth,cortical thickness and trabecular density in experimental fluorosis rats were remarkably reduced. (3) Compared with that of the control group, mRNA expression of both OPGL and M- CSF was increased in the fluoride group rats. The difference was statistically significant ( P＜0.05).(4) Compared with that of the fluoride group animals, the expression intensity of OPGL mRNA decreased in animals of the control group, the high, mid- and low- dosage medication groups and the borax group. Among them, except the low-dosage group, the difference between all the other groups and the fluoride group was statistically significant, respectively (P＜0.05).There was also a decrease of M-CSF mRNA in all the 3 medication groups and the borax group animals in comparing with that of the fluoride group and the difference was also statistically significant(P＜0.05 ), respectively. ( 5) Compared with that of the control group. There were an increase of OPGL and a decrease of M-CSF protein expression; and in addition, there were a decrease of OPGL and an increase of M-CSF protein expression in all 3 medication groups and the borax groupi in comparing with that of the fluoride group anima ( P ＜ 0. 05 ). Conclusions Excessive fluoride induces an accelerated bone turnover and may promote the absorption activity of osteoclasts by increasing the expression of OPGL and M-CSF.Danlan Xianpeng Liaofu capsule may be capable of regulating bone remodeling through a down-regulation on OPGL and M-CSF expression.