本文選題：巴戟天 + 微波輻射　； 參考：《福建醫(yī)科大學(xué)》2014年博士論文

【摘要】：【目的】通過微波輻射構(gòu)建雄性SD大鼠生殖系統(tǒng)損傷模型,觀察巴戟天水提液經(jīng)乙酸乙酯萃取后水層和酯層修復(fù)微波輻射致生殖系統(tǒng)損傷的功效,探討巴戟天活性成分對下丘腦-垂體-睪丸軸(Hypothalamic-Pituitary-Testis axis,HPTA)的可能作用機(jī)制,為巴戟天有效成分的進(jìn)一步篩選和對生殖系統(tǒng)損傷的修復(fù)作用及機(jī)制提供依據(jù)。第一部分【方法】12只健康成年雄性SD大鼠隨機(jī)分為2組,每小組6只。A組為空白對照組,B組為輻射組。B組應(yīng)用頻率為900MHz、輻射密度為218μW/cm2的微波信號發(fā)生器,輻射24h/d,持續(xù)10天,以獲得微波輻射致生殖系統(tǒng)損傷模型。觀察大鼠生長發(fā)育、性行為學(xué)差異,比較睪丸、附睪指數(shù)和組織形態(tài)學(xué)的改變,觀察下丘腦Gn RH熒光表達(dá),以評估模型是否成功建立�！窘Y(jié)果】1.體重:微波輻射10天后,輻射組大鼠體重增加(19.83±12.32)g,體重較之空白對照組(80.17±29.01)g差別有顯著性意義(P0.05)。2.性行為學(xué):與空白對照組相比,輻射組大鼠撲捉潛伏期(CIP)顯著延長(P0.05),撲捉次數(shù)(CT)顯著減少(P0.01)。3.與空白對照組相比,輻射組大鼠睪丸和附睪指數(shù)均顯著降低(P0.05)。睪丸和附睪HE染色可見輻射組睪丸生精小管管徑減小,排列疏松,未見精原細(xì)胞、各級生精細(xì)胞排列松散、脫落、壞死,附睪管內(nèi)充滿脫落的細(xì)胞及細(xì)胞碎片,未見精子。4.免疫熒光組織化學(xué)染色法:Gn RH主要表達(dá)在位于第三腦室底兩側(cè)的弓狀核;較之空白對照組,Gn RH熒光強(qiáng)度在輻射模型組表達(dá)有所減弱�！窘Y(jié)論】以頻率為900MHz、輻射密度為218μW/cm2的低功率微波持續(xù)輻射10天可成功建立雄性SD大鼠生殖損傷模型。第二部分【方法】66只健康成年雄性SD大鼠隨機(jī)分為11組(其中D、E組各有4個(gè)不同劑量組10g/kg、20 g/kg、30 g/kg、40 g/kg),每小組6只。A組為空白對照組,B組為輻射未給藥組,C組為輻射+生理鹽水組,D組為萃取水層梯度濃度液治療組,E組為萃取酯層梯度濃度液治療組。制備巴戟天水提液,經(jīng)乙酸乙酯萃取后得到萃取水層和萃取酯層的梯度濃度液(每ml各含2、4、6和8g生藥量)。以電感耦合等離子體質(zhì)譜法(Inductively coupled plasma massspectrometry,ICP-MS)測定和對比與HPTA軸修復(fù)作用相關(guān)的微量元素鋅(Zinc,Zn)、錳(Manganese,Mn)、硒(Selenium,Se)、鐵(Iron,Fe)、銅(Copper,Cu)在巴戟天水提液各萃取層的分布情況。輻射后C、D、E組分別給予等量生理鹽水或相應(yīng)濃度的巴戟天提取物萃取液持續(xù)灌胃12天。觀察大鼠性行為學(xué)差異,同時(shí)在下丘腦、垂體水平檢測Gn RH表達(dá),血清LH、FSH、Gn RH和T變化;比較睪丸、附睪指數(shù)以及組織形態(tài)學(xué)的改變�！窘Y(jié)果】1.微量元素分布:ICP-MS測定結(jié)果顯示微量元素鋅、錳、鐵在巴戟天水提液和巴戟天水提液乙酸乙酯萃取水層中含量一致,在巴戟天水提液乙酸乙酯萃取酯層中含量極少(P0.05)。銅含量在以上三份樣品中差別無顯著性意義。硒含量差別亦無顯著性意義。2.性行為學(xué)巴戟天治療后,D3、D4組撲捉潛伏期/D4組撲捉次數(shù)與B組、C組差別有統(tǒng)計(jì)學(xué)意義,余治療組性行為指標(biāo)與B組、C組差別均無統(tǒng)計(jì)學(xué)意義。3.性腺水平:(1)B~E組睪丸指數(shù)均低于A組,差別有統(tǒng)計(jì)學(xué)意義;B組附睪指數(shù)低于A組,差別有統(tǒng)計(jì)學(xué)意義;D4組睪丸、附睪指數(shù)與B組、C組差別有統(tǒng)計(jì)學(xué)意義,余治療組與B組、C組差別均無統(tǒng)計(jì)學(xué)意義。(2)血清睪酮水平比較:A組與B組差別無統(tǒng)計(jì)學(xué)意義。D3、D4組睪酮水平與B組、C組差別有統(tǒng)計(jì)學(xué)意義,余治療組與B組、C組差別均無統(tǒng)計(jì)學(xué)意義。(3)睪丸和附睪組織形態(tài)學(xué)改變:輻射組形態(tài)學(xué)改變見第一部分。D組中,隨著給藥劑量從20~40g/kg逐漸增加,大鼠睪丸生精小管上皮各級生精細(xì)胞及生精小管內(nèi)精子數(shù)量逐漸增多;B、C、E組形態(tài)學(xué)無明顯差別。輻射組大鼠附睪管管腔空虛、未見精子,可見少量脫落的生精細(xì)胞。D3、D4組大鼠附睪管內(nèi)可見密集的精子,但仍少于空白對照組數(shù)的精子量。4.血清FSH、LH含量差異:各組間FSH差別均無統(tǒng)計(jì)學(xué)意義。A組與B組LH差別無統(tǒng)計(jì)學(xué)意義。D3、D4組LH水平與B組、C組差別有統(tǒng)計(jì)學(xué)意義,余治療組與B組、C組差別均無統(tǒng)計(jì)學(xué)意義。5.Gn RH表達(dá)量對比:(1)經(jīng)ELISA法測定各組血清Gn RH含量,顯示B組大鼠血清Gn RH含量低于A組;D3、D4組Gn RH水平與B組、C組差別有統(tǒng)計(jì)學(xué)意義,余治療組與B組、C組差別均無統(tǒng)計(jì)學(xué)意義。(2)免疫熒光組織化學(xué)染色法標(biāo)記可見Gn RH主要表達(dá)在位于第三腦室底兩側(cè)的弓狀核;B組Gn RH水平較A組下降,但差別無統(tǒng)計(jì)學(xué)意義。D3、D4組Gn RH與B組差別有統(tǒng)計(jì)學(xué)意義,余治療組與B組差別均無統(tǒng)計(jì)學(xué)意義。(3)Real-time PCR結(jié)果顯示。D4組大鼠下丘腦Gn RH-m RNA表達(dá)較之B、C組降低(P0.05)。余治療組Gn RH-m RNA表達(dá)與B、C組差距無統(tǒng)計(jì)學(xué)意義�！窘Y(jié)論】1.巴戟天水提液及巴戟天水提液乙酸乙酯萃取水層中富含微量元素鋅、錳、鐵,可能與生殖功能保護(hù)有關(guān)。2.巴戟天水提液乙酸乙酯萃取水層可改善大鼠性行為學(xué),其機(jī)制可能與調(diào)節(jié)HPTA下丘腦、垂體以及性腺各個(gè)水平上相關(guān)激素分泌有關(guān),其中最佳治療劑量為40g/kg。3.巴戟天水提液乙酸乙酯萃取酯層對微波輻射誘導(dǎo)的大鼠生精功能障礙無明顯修復(fù)作用。
[Abstract]:[Objective] to construct the model of radiation injury of reproductive system in male SD rats by microwave induced by extract of Morindae Officinalis, observe efficacy of injury of reproductive system after extracted by ethyl acetate layer and the ester layer repair microwave radiation of Morinda active components on the hypothalamic pituitary testicular axis (Hypothalamic-Pituitary-Testis, axis, HPTA) the possible mechanism, to provide the basis for further screening the constituents of Morinda officinalis and on the reproductive system damage repair effects and mechanism of the first part. [Methods] 12 healthy adult male SD rats were randomly divided into 2 groups, 6 in each group group.A group B group.B group applied radiation frequency is 900MHz, the density of radiation microwave signal generator, 218 W/cm2 radiation 24h/d, lasting 10 days, in order to obtain the microwave radiation induced reproductive system injury model. To observe the growth of rat, differences in behavior, compared with testis, epididymis The morphological index and tissue changes were observed in the hypothalamus Gn RH expression, to assess whether the model was successfully established. [results] 1. weight: 10 days after the microwave radiation, the radiation group weight increase (19.83 + 12.32) g, body weight compared with the blank control group (80.17 + 29.01) had significant difference (P0.05 g.2.) of behavior: compared with the control group, radiation group rats Puzhuo latency (CIP) significantly prolonged (P0.05), capture times (CT) was significantly reduced (P0.01).3. compared with the blank control group, radiation group of rat testis and epididymis index were significantly lower (P0.05) in testis and epididymis. HE staining decreased, visible radiation group seminiferous tubule diameter were arranged loosely, spermatogonial stem cells, spermatogenic cells arranged loose, fall off, necrosis, cell and epididymis tube filled with falling debris, no sperm.4. immunofluorescence staining method: Gn RH was mainly expressed in the in The end of the third ventricle on both sides of the arcuate nucleus; compared with the control group, the Gn fluorescence intensity of RH was reduced in the model group. [Conclusion] the frequency of 900MHz, low power microwave radiation density was 218 W/cm2 irradiation for 10 days can be successfully established model of reproductive damage in male SD rats. The second part [method] 66 healthy adult male SD rats were randomly divided into 11 groups (D, E group each have 4 different doses of 10g/kg, 20 g/kg, 30 g/kg, 40 g/kg), each group 6.A group was control group, B group was not radiation dose group, C group and normal radiation saline group, D group to extract water concentration gradient liquid treatment group, E group for the extraction of the ester layer concentration gradient liquid treatment group. Preparation of Morindae Officinalis Extract, the concentration gradient of liquid extraction and water extraction ester layer was extracted by ethyl acetate (each containing 2,4,6 per ml and 8g crude drug) by inductively coupled. Plasma mass spectrometry (Inducti Vely coupled plasma massspectrometry, ICP-MS) determination of trace elements zinc and compared with HPTA axis repair related (Zinc, Zn), Mn (Manganese, Mn), Se (Selenium, Se), iron (Iron, Fe), copper (Copper, Cu). The distribution of each layer in the liquid extract from Morindae officinalis radiation. After C, D, E group were given normal saline or the corresponding concentrations of Morinda officinalis extract extract continuous gavage for 12 days. Observe the behavior of rats and the difference in the hypothalamus, pituitary Gn RH expression, detect the level of serum LH, FSH, Gn, RH and T change; comparison of testis, epididymis index and the change of morphology. [results] 1. trace element distribution: ICP-MS measurement results show that the trace elements of zinc, manganese, and iron in Morindae Officinalis Extract from Morindae Officinalis Extract ethyl acetate extract water content in Morindae Officinalis Extract, ethyl acetate ester layer with low content (P0.05). The content of copper in the above 涓変喚鏍峰搧涓樊鍒棤鏄捐憲鎬ф剰涔，

本文編號：1756097