【摘要】：目的: 自然衰老SD大鼠經(jīng)常作為腎陽(yáng)虛證動(dòng)物模型應(yīng)用于補(bǔ)腎陽(yáng)藥物的研究開(kāi)發(fā)以及腎陽(yáng)虛與衰老關(guān)系的研究中,但在應(yīng)用時(shí)存在著月齡的差異,所使用的大鼠月齡主要集中在24月齡到30月齡之間,究竟哪個(gè)月齡段的大鼠,相對(duì)來(lái)說(shuō)更適宜作為腎陽(yáng)虛證動(dòng)物模型,文獻(xiàn)上沒(méi)有明確的指出。本課題使用篩選的腎陽(yáng)虛證動(dòng)物模型評(píng)價(jià)標(biāo)準(zhǔn)及溫補(bǔ)腎陽(yáng)的中藥對(duì)不同月齡自然衰老SD大鼠進(jìn)行初步的驗(yàn)證和治療,并根據(jù)不同月齡的自然衰老SD大鼠的檢測(cè)結(jié)果以及反證療效的差異,篩選出較適宜性別和月齡的自然衰老SD大鼠的腎陽(yáng)虛模型,為中醫(yī)證候動(dòng)物模型規(guī)范化研究及抗衰老的藥物研究提供支持和幫助。 方法:本實(shí)驗(yàn)分為三部分進(jìn)行 實(shí)驗(yàn)一:自然衰老SD大鼠腎陽(yáng)虛的觀察 選取雄性的青年大鼠作為對(duì)照組,雄性的自然衰老大鼠作為模型組通過(guò)觀察兩組動(dòng)物的癥狀、體征以及血清皮質(zhì)醇(COR)、三碘甲狀腺原氨酸(T_3)、甲狀腺素(L_4)、睪酮(T)/雌二醇(E_2)、臟器指數(shù)、病理改變等指標(biāo)來(lái)評(píng)價(jià)自然衰老雄性大鼠是否具有腎陽(yáng)虛證的實(shí)質(zhì)。 實(shí)驗(yàn)二:連續(xù)月齡的自然衰老SD大鼠腎陽(yáng)虛損程度的比較 取連續(xù)三個(gè)月齡的衰老早期的大鼠進(jìn)行實(shí)驗(yàn),實(shí)驗(yàn)結(jié)束后取血、取材對(duì)比它們之間的血清皮質(zhì)醇(COR)、三碘甲狀腺原氨酸(L_3)、甲狀腺素(L_4)、睪酮(T)/雌二醇(E_4)、臟器指數(shù),通過(guò)對(duì)比研究尋找出差異的最小間隔期為下一次試驗(yàn)提供依據(jù)。 實(shí)驗(yàn)三:腎陽(yáng)虛證自然衰老SD大鼠月齡的篩選 按照實(shí)驗(yàn)二的結(jié)果作為實(shí)驗(yàn)周期將實(shí)驗(yàn)分成三個(gè)不同的月齡段進(jìn)行,首先取一定數(shù)量的24月齡大鼠隨機(jī)分為兩組,一組為空白組,另一組為藥物治療反證組,空白組每天灌胃兩毫升無(wú)菌水,藥物組灌胃兩毫升右歸丸水煎劑,期間觀察兩組的癥狀體征,實(shí)驗(yàn)結(jié)束后采集血清血漿—60℃冰箱保存,第二次取26月齡的自然衰老SD大鼠、最后取28月齡的自然衰老SD大鼠進(jìn)行相同過(guò)程的實(shí)驗(yàn)。 結(jié)果: 1.實(shí)驗(yàn)一:自然衰老SD雄性大鼠具有腎陽(yáng)虛的癥狀體征,檢測(cè)結(jié)果基本符合所篩選腎陽(yáng)虛的指標(biāo)。 2.實(shí)驗(yàn)二:用所篩選的血清學(xué)指標(biāo)檢測(cè)連續(xù)月齡的自然衰老SD雄性大鼠時(shí),出現(xiàn)差異的時(shí)間至少間隔一個(gè)月。 3.實(shí)驗(yàn)三:26到28月齡段為最佳自然衰老腎陽(yáng)虛模型。 結(jié)論:
[Abstract]:Objective: natural aging SD rats are often used as an animal model of kidney-yang deficiency syndrome in the research and development of kidney-yang drugs and the relationship between kidney-yang deficiency and aging. The age of the rats is mainly between 24 months and 30 months old. It is not clearly pointed out in the literature that which segment of the rats is more suitable for the animal model of kidney yang deficiency syndrome. The objective of this study was to use the selected animal model of kidney-yang deficiency syndrome and the Chinese medicine to warm up the kidney-yang to verify and treat the natural aging SD rats of different months of age. According to the test results of natural aging SD rats of different ages and the difference of curative effect, the kidney yang deficiency model of natural aging SD rats with suitable gender and month age was selected. To provide support and help for the standardization of animal models of TCM syndromes and the research of anti-aging drugs. Methods: this experiment was divided into three parts. One was the observation of kidney-yang deficiency in natural aging SD rats. Young male rats were selected as the control group. Male natural aging rats were used as model group to observe the symptoms, signs and serum cortisol (COR), triiodothyronine (T _ S _ 3) and thyroxine (L _ 4) in the two groups. Testosterone (T) / estradiol (E _ 2), viscera index and pathological changes were used to evaluate the essence of kidney yang deficiency in natural aging male rats. Experiment 2: comparison of the degree of deficiency of Kidney Yang in naturally aged SD Rats aged 3 months. Blood samples were taken from the rats at the early stage of aging, and the blood samples were taken after the end of the experiment. Serum cortisol (COR), triiodothyronine (LS3), thyroxine (L4), testosterone (T) / estradiol (E4), organ index, A comparative study was conducted to find the minimum interval for the next experiment. Experiment 3: the screening of natural aging SD rats with kidney-yang deficiency syndrome divided the experiment into three different age segments according to the results of experiment 2. At first, a certain number of 24-month-old rats were randomly divided into two groups: one group was blank group, the other group was drug therapy reverse syndrome group. The blank group was given two ml of aseptic water daily, and the drug group was given two ml right Gui Pill decoction. The symptoms and signs of the two groups were observed during the experiment. After the experiment, the serum plasma was collected and stored in the refrigerator at -60 鈩，

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