口服高氧液抗小鼠運(yùn)動(dòng)性疲勞的作用及機(jī)制研究

1、口服高氧液抗小鼠運(yùn)動(dòng)性疲勞的作用及機(jī)制研究     口服高氧液抗小鼠運(yùn)動(dòng)性疲勞的作用及機(jī)制研究【摘要】 目的：研究高氧液(HO)對(duì)小鼠的抗疲勞作用及其機(jī)制。方法：將昆明小鼠120只平均分為對(duì)照組（C組）和高氧液組（HO組），再根據(jù)劑量分為三組，每10克體重分別給予0.15ml、0.25ml和0.5ml的平衡鹽或高氧液。灌胃后即刻和游泳30min后檢測(cè)肌糖原含量、血清乳酸含量、三磷酸腺苷酶活力、丙二醛含量和總抗氧化能力并且觀察一般情況。結(jié)果：高氧液組小鼠較對(duì)照組泳動(dòng)活躍，下沉率低。高氧液組血清乳酸含量（8.9110.42±1.41 mmol/L）

2、明顯少于對(duì)照組（11.9112.01±1.34 mmol/L, P<0.05），高氧液組肌糖原貯備（1.572.07±0.15 mg/g）明顯多于對(duì)照組（1.03±0.14 mg/g，P<0.05)，高氧液組三磷酸腺苷酶活力（1.101.49±0.12molPi/mgprot/hour）比對(duì)照組（0.791.08±0.13molPi/mgprot/hour）增高明顯(P<0.05)，高氧液組丙二醛含量（17.7418.93±1.26 nmol/ mgprot）明顯少于對(duì)照組(20.5020.55±1.24n

3、mol/ mgprot, P<0.05)，高氧液組總抗氧化能力（175.87202.87±22.00nkat/ mgprot）比對(duì)照組（134.53137.36±16.00nkat/ mgprot）增高明顯(P<0.05)。結(jié)論：HO對(duì)小鼠有顯著的抗疲勞作用,其機(jī)制可能與HO增加氧供及抗脂質(zhì)過(guò)氧化有關(guān)?！娟P(guān)鍵詞】 高氧液；疲勞；乳酸；肌糖原;三磷酸腺苷酶活力；丙二醛；總抗氧化能力A experimental study of hyperoxia solution taken orally on fatigue and its mechanism in mice

4、induced by exerciseLIU Chun-Ran1, ZHANG Guo-Liang1, XU L i-Xian11Department of Anesthesiology,College of Stomatology, Fourth Military Medical University , Xian 710032 , China【Abstract】AIM：To investigate the antifatigue effect of hyperoxia solution (HO) and its mechanism. METHODS：The mice were divide

5、d randomly into two groups: control group(C group) , hyperoxia solution group(HO group).Then each group was divided into three smaller groups by dose, which were infused with balanced salt solution or HO at 0.15ml、0.25ml and 0.5ml per 10g.After the mice were infused intragastricly, the contents of m

6、uscle glucogen, lactic acid  and malondialdehyde，the general ability of antioxidation and the energy of adenosine triphosphate enzyme were determined instantly and after swimming 30 minute while they were observed about common state. RESULTS：Hyperoxia solution group swam more actively and sank

7、fewer than control group. The contents of lactic acid of hyperoxia solution group(8.9110.42±1.41mmol/L) were smaller significantly than those of control group (11.9112.01±1.34mmol/L,P<0.05);the content of muscle glucogen of hyperoxia solution group(1.572.07±0.15mg/g) was bigger dis

8、tinctly than those of control group (1.03±0.14mg/g,P<0.05) ; the activity of adenosine triphosphate enzyme of hyperoxia solution group(1.101.49±0.12molPi/mgprot/hour) increased more than those of control group(0.791.08±0.13molPi/mgprot/hour,P<0.05);the content of malondialdehyde

9、 of hyperoxia solution group(17.7418.93±1.26 nmol/ mgprot) were smaller significantly  than those of control group(20.5020.55±1.24nmol/mgprot,P<0.05);the general ability of antioxidation of hyperoxia solution group(175.87202.87±22.00nkat/mgprot )increased more than those of co

10、ntrol group (134.53137.36±16.00nkat/mgprot,P<0.05). CONCLUSION:HO can increase moving capability of mice significantly, and has antifatique effect. The mechanism may be that HO can increase the supply of oxygen and has anti-lipid peroxidation effect.【Keywords】hyperoxia solution ; fatigue ; l

11、actic acid ; muscle glucogen ; the activity of adenosine triphosphate enzyme ; malondialdehyde ; the general ability of antioxidation0   引言疲勞是機(jī)體在一定環(huán)境條件下，由于長(zhǎng)時(shí)間或過(guò)于繁重、緊張的勞動(dòng)（包括腦力和體力勞動(dòng)）而引起的工作效率暫時(shí)明顯降低的一種生理病理現(xiàn)象。疲勞的發(fā)生機(jī)制目前尚不十分明確,可能主要與機(jī)體短期內(nèi)因大運(yùn)動(dòng)量消耗過(guò)多、缺氧、乳酸堆積引起人體內(nèi)環(huán)境紊亂有關(guān)，同時(shí)伴有低血糖、肌糖原和肝糖原耗竭1,2。既往對(duì)于抗疲勞作用主

12、要著重于補(bǔ)充營(yíng)養(yǎng)物質(zhì)、降低消耗和給予適量興奮方面的研究，尚無(wú)從液體給氧途徑探索抗疲勞效果。本文運(yùn)用液體攜氧方法，以血清乳酸含量、肌糖原含量、三磷酸腺苷酶活力、丙二醛含量、總抗氧化能力為指標(biāo)，探索高氧液體口服對(duì)小鼠的抗疲勞作用和耐受力，從而為增強(qiáng)機(jī)體運(yùn)動(dòng)機(jī)能，延緩疲勞提供新的途徑。1   材料和方法1.1 材料 健康昆明小鼠120只（第四軍醫(yī)大學(xué)實(shí)驗(yàn)動(dòng)物中心提供）,雄性,質(zhì)量1822g。以平衡鹽液為基液,用高氧液治療儀(西安高氧醫(yī)療設(shè)備有限公司生產(chǎn)，專(zhuān)利號(hào)：922412936)進(jìn)行光化學(xué)溶氧技術(shù),以氧流量為3 L/min,溶氧15min制成HO。乳酸含量，肌糖原含量，三磷酸腺

13、苷酶活力，丙二醛含量，總抗氧化能力試劑盒為南京建成生物工程研究所產(chǎn)品，其它試劑均為分析純。80-2離心沉淀器(上海手術(shù)器械廠),低溫離心機(jī)(Biofuge Heraeus ,德國(guó)產(chǎn)),752C紫外可見(jiàn)分光光度計(jì),電熱恒溫水浴鍋(上海躍進(jìn)醫(yī)療器械廠) 。1.2方法  選擇健康昆明小鼠籠養(yǎng)，每籠5只，自由進(jìn)食和飲水。各組動(dòng)物在實(shí)驗(yàn)前均籠養(yǎng)2d。將昆明小鼠隨機(jī)均分為對(duì)照組（C組）和高氧液組（HO組），再根據(jù)劑量分為三組，每10克體重分別給予0.15ml、0.25ml和0.5ml的平衡鹽或高氧液。如此共分為12組,每組10只。灌胃后即刻和游泳30min后檢測(cè)血清乳酸含量、肌糖原含量、三磷酸腺

14、苷酶活力、丙二醛含量和總抗氧化能力。1.2.1負(fù)重游泳實(shí)驗(yàn)：給藥后小鼠尾根部負(fù)荷5體重的鉛皮即刻置游泳箱中游泳，泳箱60cm×40cm×50cm，水溫25左右。在游泳時(shí)觀察小鼠的一般情況：泳動(dòng)靈敏性、?？繒r(shí)間、次數(shù)、下沉率。    1.2.2 給藥后即刻和游泳30min后摘眼球取血測(cè)定血清乳酸含量；取腦組織測(cè)定丙二醛含量、總抗氧化能力；取右后肢股四頭肌測(cè)定肌糖原含量、三磷酸腺苷酶活力，按照試劑盒說(shuō)明操作。統(tǒng)計(jì)學(xué)處理  計(jì)量數(shù)據(jù)以c±s表示, 用SPSS10.0統(tǒng)計(jì)軟件,采用單因素方差分析和LSD-t檢驗(yàn)比較小鼠運(yùn)動(dòng)

15、前后血清乳酸含量、肌糖原含量、三磷酸腺苷酶活力、丙二醛含量和總抗氧化能力的差別，P<0.05認(rèn)為有顯著差異。2  結(jié)果2.1 一般情況 高氧液組小鼠較對(duì)照組泳動(dòng)較靈敏，泳動(dòng)有力，對(duì)外界反應(yīng)性較好，30min下沉率：對(duì)照組為93.33，而高氧液組為56.67，明顯低于C組（P<0.05）。2.2 HO對(duì)肌糖原和血清乳酸含量含量的影響  結(jié)果顯示運(yùn)動(dòng)后小鼠的肌糖原各組均減少，但是服用高氧液組減少的量顯著小于對(duì)照組(P<0.05)，而血清乳酸含量各組均增加，但是服用高氧液組顯著少于對(duì)照組 (P<0.05，見(jiàn) Tab 1)。表1  口服高氧液對(duì)運(yùn)動(dòng)

16、小鼠肌糖原含量和血清乳酸含量的影響（n=10,c±s）Tab 1 Effects of hyperoxia solution taken orally on the contents of lactic acid and muscle glucogen of the mice induced by exercise（n=10,c±s）group             muscle glucogen     

17、;          lactic acid(mg/g )                 (mmol/L)                    &

18、#160;  0min       30 min          0 min           30 minC1           2.68±0.29    1.03±0.12

19、0;       7.51±0.91      12.01±1.30        C2           2.66±0.29     1.03±0.14        7.13

20、±0.90      11.96±1.34C3           2.64±0.26     1.03±0.13        6.98±0.96       11.91±1.05   &

21、#160; HO1           2.63±0.22    1.61±0.14*           7.31±0.92       10.42±1.33*    HO2     &#

22、160;     2.71±0.31     2.07±0.14*        7.61±0.78        8.91±1.41*     HO3            2.61

23、77;0.22     1.57±0.15*        7.24±0.72       10.31±1.19*P<0.05 vs C. C：control group；HO：hyperoxia solution group；1:0.015ml/g;2：0.025ml/g ;3：0.05ml/g .2.3  HO對(duì)Na+，K+-ATP酶活力和Ca2+-ATP酶活力的影響 

24、; 結(jié)果顯示運(yùn)動(dòng)后小鼠的Na+，K+-ATP酶活力和Ca2+-ATP酶活力各組均增加，但是服用高氧液組均較對(duì)照組增加顯著(P <0.05 ,見(jiàn)Tab 2)。表2 口服高氧液對(duì)運(yùn)動(dòng)小鼠Na+，K+-ATP酶活力和Ca2+-ATP酶活力的影響（n=10,c±s）Tab 2 Effects of hyperoxia solution taken orally on the activity of Adenosine Triphosphate enzyme of the mice induced by exercise（n=10,c±s） group &#

25、160;           Na+，K+-ATP                      Ca2+-ATP               （molP

26、i/ mgprot /hour）         （molPi/mgprot /hour）                  0 min              30 min   

27、0;    0 min             30 minC1        0.70±0.11         1.05±0.13      0.50±0.11    

28、0;   0.79±0.10     C2        0.70±0.11         1.02±0.10      0.50±0.12        0.80±0.10C3   &#

29、160;    0.70±0.10         1.08±0.13      0.50±0.11        0.81±0.10          HO1      0.69±

30、;0.10         1.21±0.10*      0.50±0.11        1.10±0.12*     HO2       0.70±0.12        

31、1.49±0.12*      0.51±0.11        1.32±0.11*      HO3      0.70±0.12         1.22±0.10*       0.

32、51±0.10           1.12±0.12*P<0.05 vs C.C，HO，1，2 and 3 same as Tab 1.2.4 HO對(duì)丙二醛含量和總抗氧化能力的影響   結(jié)果顯示運(yùn)動(dòng)后小鼠的丙二醛含量各組均增加，但是服用高氧液組均顯著少于對(duì)照組 (P <0.05)。而總抗氧化能力各組均減少，但是服用高氧液組減少的量顯著小于對(duì)照組(P <0.05 Tab 3)。表3 口服高氧液對(duì)運(yùn)動(dòng)小鼠丙二醛含量和總抗氧化能力的影響（n=

33、10,c±s）Tab 3 Effects of hyperoxia solution taken orally on the contents of malondialdehyde and the general ability of antioxidation of the mice induced by exercise（n=10,c±s）group     malondialdehyde            the g

34、eneral ability of antioxidation                （nmol/ mgprot）                      （nkat/ mgprot）     

35、60;          0 min          30 min                 0 min          30 minC1  

36、;       16.16±1.06    20.50±1.15           111.39±21.17    134.53±16.00C2        16.17±0.96    20.51±1.12 

37、;          109.39±21.67     135.03±14.00C3        16.21±1.09    20.55±1.24           108.89±17.00  &

38、#160;  137.36±15.67  HO1         16.21±1.00    18.91±1.26*           110.89±20.67     176.04±16.17* HO2      16.17±0.99     17.74±1.0