中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
28期
5539-5543
,共5页
耿丽芬%刘坤申%马华%焦保权%李振彬
耿麗芬%劉坤申%馬華%焦保權%李振彬
경려분%류곤신%마화%초보권%리진빈
关节炎大鼠%类风湿关节炎%雌激素受体%补肾活血方
關節炎大鼠%類風濕關節炎%雌激素受體%補腎活血方
관절염대서%류풍습관절염%자격소수체%보신활혈방
背景:药理学研究显示,补肾活血方中的补肾中药如熟地、葛根等含有黄酮类物质,具有植物雌激素的作用特点,其中异黄酮具有雌激素受体调控剂的作用.目的:观察自拟补肾活血方对胶原诱导性关节炎大鼠血清雌二醇水平及垂体雌激素受体αmRNA表达的影响.设计、时间及地点:随机分组设计、对照动物实验,于2006-05/2008-05在白求恩国际和平医院中心实验室完成.材料:健康Wistar雌性大鼠60只,随机分为2组:假手术组(n=10)和手术组(n=50).手术组在卵巢切除术后2周复制胶原诱导性关节炎模型,选取关节炎指数≥5的大鼠再随机分为4组,每组10只:模型组、补肾活血方、甲氨蝶呤组和补肾活血方+甲氨蝶呤组.补肾活血方所用中药骨碎补、红花、熟地、仙灵脾、丹参、葛根等购自石家庄乐仁堂药店.常规煎煮,离心去杂,浓缩为1.3 kg/L的水煎液,密封分装,4℃冷藏备用.方法:假手术组和模型组灌胃给予生理盐水,2 mL/d;补肾活血组:补肾活血方煎液2 mL/d灌胃;甲氨蝶呤组按2.6 mg/kg剂量灌胃1次/周,体积为2 mL,其余时间生理盐水灌胃2 mL/d;补肾活血方+甲氨蝶呤组按甲氨蝶呤2.6 mg/kg剂量灌胃1次/周,体积为2 mL,另外补肾活血方煎液2 mL/d;各组均持续6周.主要观察指标:腹主动脉采血用磁分离酶联免疫法检测血清雌二醇水平,同时取大鼠垂体采用反转录一聚合酶链反应检测大鼠垂体雌激素受体αmRNA的表达.结果:与模型组比较,甲氨蝶呤组雌激素受体α mRNA表达增加,但差异无显著性意义;补肾活血方+甲氨蝶呤组表达明显增加(P<0.01);补肾活血方表达量高于甲氨蝶呤组,低于补肾活血方+甲氨蝶呤组(P<0.05);补肾活血方、补肾活血方+甲氨蝶呤组与假手术组差异无显著性意义.与假手术组比较,模型组雌二醇水平明显降低.与模型组比较,3个治疗组雌二醇水平均有不同程度的提高,其中甲氨蝶呤组偏低,与模型组差异无显著性意义(P>0.05),补肾活血方+甲氨蝶呤组提高最明显,接近正常水平(P<0.01).雌二醇与垂体雌激素受体αmRNA呈显著正相关(r=0.483,P<0.05).结论:自拟补肾活血方具有拟雌激素样作用,可提高雌二醇水平,上调垂体雌激素受体α mRNA表达.
揹景:藥理學研究顯示,補腎活血方中的補腎中藥如熟地、葛根等含有黃酮類物質,具有植物雌激素的作用特點,其中異黃酮具有雌激素受體調控劑的作用.目的:觀察自擬補腎活血方對膠原誘導性關節炎大鼠血清雌二醇水平及垂體雌激素受體αmRNA錶達的影響.設計、時間及地點:隨機分組設計、對照動物實驗,于2006-05/2008-05在白求恩國際和平醫院中心實驗室完成.材料:健康Wistar雌性大鼠60隻,隨機分為2組:假手術組(n=10)和手術組(n=50).手術組在卵巢切除術後2週複製膠原誘導性關節炎模型,選取關節炎指數≥5的大鼠再隨機分為4組,每組10隻:模型組、補腎活血方、甲氨蝶呤組和補腎活血方+甲氨蝶呤組.補腎活血方所用中藥骨碎補、紅花、熟地、仙靈脾、丹參、葛根等購自石傢莊樂仁堂藥店.常規煎煮,離心去雜,濃縮為1.3 kg/L的水煎液,密封分裝,4℃冷藏備用.方法:假手術組和模型組灌胃給予生理鹽水,2 mL/d;補腎活血組:補腎活血方煎液2 mL/d灌胃;甲氨蝶呤組按2.6 mg/kg劑量灌胃1次/週,體積為2 mL,其餘時間生理鹽水灌胃2 mL/d;補腎活血方+甲氨蝶呤組按甲氨蝶呤2.6 mg/kg劑量灌胃1次/週,體積為2 mL,另外補腎活血方煎液2 mL/d;各組均持續6週.主要觀察指標:腹主動脈採血用磁分離酶聯免疫法檢測血清雌二醇水平,同時取大鼠垂體採用反轉錄一聚閤酶鏈反應檢測大鼠垂體雌激素受體αmRNA的錶達.結果:與模型組比較,甲氨蝶呤組雌激素受體α mRNA錶達增加,但差異無顯著性意義;補腎活血方+甲氨蝶呤組錶達明顯增加(P<0.01);補腎活血方錶達量高于甲氨蝶呤組,低于補腎活血方+甲氨蝶呤組(P<0.05);補腎活血方、補腎活血方+甲氨蝶呤組與假手術組差異無顯著性意義.與假手術組比較,模型組雌二醇水平明顯降低.與模型組比較,3箇治療組雌二醇水平均有不同程度的提高,其中甲氨蝶呤組偏低,與模型組差異無顯著性意義(P>0.05),補腎活血方+甲氨蝶呤組提高最明顯,接近正常水平(P<0.01).雌二醇與垂體雌激素受體αmRNA呈顯著正相關(r=0.483,P<0.05).結論:自擬補腎活血方具有擬雌激素樣作用,可提高雌二醇水平,上調垂體雌激素受體α mRNA錶達.
배경:약이학연구현시,보신활혈방중적보신중약여숙지、갈근등함유황동류물질,구유식물자격소적작용특점,기중이황동구유자격소수체조공제적작용.목적:관찰자의보신활혈방대효원유도성관절염대서혈청자이순수평급수체자격소수체αmRNA표체적영향.설계、시간급지점:수궤분조설계、대조동물실험,우2006-05/2008-05재백구은국제화평의원중심실험실완성.재료:건강Wistar자성대서60지,수궤분위2조:가수술조(n=10)화수술조(n=50).수술조재란소절제술후2주복제효원유도성관절염모형,선취관절염지수≥5적대서재수궤분위4조,매조10지:모형조、보신활혈방、갑안접령조화보신활혈방+갑안접령조.보신활혈방소용중약골쇄보、홍화、숙지、선령비、단삼、갈근등구자석가장악인당약점.상규전자,리심거잡,농축위1.3 kg/L적수전액,밀봉분장,4℃랭장비용.방법:가수술조화모형조관위급여생리염수,2 mL/d;보신활혈조:보신활혈방전액2 mL/d관위;갑안접령조안2.6 mg/kg제량관위1차/주,체적위2 mL,기여시간생리염수관위2 mL/d;보신활혈방+갑안접령조안갑안접령2.6 mg/kg제량관위1차/주,체적위2 mL,령외보신활혈방전액2 mL/d;각조균지속6주.주요관찰지표:복주동맥채혈용자분리매련면역법검측혈청자이순수평,동시취대서수체채용반전록일취합매련반응검측대서수체자격소수체αmRNA적표체.결과:여모형조비교,갑안접령조자격소수체α mRNA표체증가,단차이무현저성의의;보신활혈방+갑안접령조표체명현증가(P<0.01);보신활혈방표체량고우갑안접령조,저우보신활혈방+갑안접령조(P<0.05);보신활혈방、보신활혈방+갑안접령조여가수술조차이무현저성의의.여가수술조비교,모형조자이순수평명현강저.여모형조비교,3개치료조자이순수평균유불동정도적제고,기중갑안접령조편저,여모형조차이무현저성의의(P>0.05),보신활혈방+갑안접령조제고최명현,접근정상수평(P<0.01).자이순여수체자격소수체αmRNA정현저정상관(r=0.483,P<0.05).결론:자의보신활혈방구유의자격소양작용,가제고자이순수평,상조수체자격소수체α mRNA표체.
BACKGROUND: Pharmacological researches demonstrate that Chinese crude drugs of invigorating the kidney contain flavonoids such as prepared rhizome of rehmannia and kudzuvine root, they possess phytoestrogenic effect, among them, isoflavone acts as a regulator of estrogen receptor.OBJECTIVE: To approach the effects of self-planned Bushen Huoxue Fang (BSHXF) on the serum estradiol and estrogen receptor α mRNA expression in pituitary gland in rats with collagen-induced arthritis. DESIGN, TIME AND SETTING: Randomized grouping controlled animal experiment was performed in the central laboratory of Bethune Intemational Peace Hospital of Chinese PLA between May 2006 and May 2008. MATERIALS: Sixty female and healthy Wistar rats were randomly divided into sham operated group (n=10) and operation group (n=50). Operation group was reproduced collagen-induced arthritis model at 2 weeks following ovariectomy. The rats of arthritic index ≥ 5 were assigned into model group, BSHXF group, methotrexate (MTX) group, and MTX + BSHXF group, each group included 10 rats. The Chinese crude drugs were bought from LERENTANG drugstore in Shijiazhuang, including rhizoma drynariae, safflower, prepared rhizome of rehmannia, Herba Epimedii, danshen root, kudzuvine root and so on. After routine decoction, centrifugalization, rogue and condensing 1.3 kg/L decoction, the solution was sealed up, divided pack and reserved at 4℃.METHODS: Sham operated group and model group were intragastrically administered with normal saline 2 mL per day; BSHXF group with BSHXF decoction 2 mL per day; MTX group with MTX 2.6 mg/kg (2 mL) once a week, and normal saline 2 mL per day at other time points; MTX + BSHXF group with MTX 2.6 mg/kg (2 mL) once a week and BSHXF juice 2 mL per day. Each group was given 6 weeks.MAIN OUTCOME MEASURES: After blooding by abdominalis aorta, magnetic-split enzyme linked immunosorbent assay was applied to detect the level of serum estradiol. Reverse transcription-polymerase chain reaction detected the expression of estrogen receptor a mRNA in rat pituitary gland.RESULTS: Compared with model group, the expression of estrogen receptor a mRNA in pituitary gland increased in the MTX group, but there was not statistically significant. BSHXF + MTX group showed a significantly increased expression (P < 0.01). The expression of estrogen receptor α mRNA in BSHXF group was higher than MTX group, but lower than BSHXF + MTX group (P < 0.05). No significant difference existed between BSHXF group, BSHXF + MTX group and sham operation group. Compared with sham operation group, there were statistically significant decrease of estradiol in the model group. Compared with model group, the estradiol levels were shown to increase in three treatment groups. Among them, MTX group had slightly slower level with no significant difference (P > 0.05), while BSHXF + MTX group had the most obvious increase, which was close to normal level (P < 0.01). There was a significantly positive correlation between estradiol and the expression of estrogen receptor o mRNA (r=0.483, P < 0.05).CONCLUSION: Serf-planned BSHXF possessing oestrogenic hormonelike role can elevate the level of serum estradiol and upregulate the expression of estrogen receptor a mRNA in pituitary gland.
