艾灸对D-半乳糖致衰老大鼠松果体氧化损伤影响的实验研究

【作者】 梁欣；

【导师】 罗永芬；

【作者基本信息】 成都中医药大学 ， 针灸推拿学， 2008， 博士

【摘要】 目的为进一步深入探讨艾灸延缓衰老机理,以D-半乳糖致衰老大鼠为研究对象,以中医衰老理论为指导,以现代衰老松果体学说为切入点,通过观察艾灸对D-半乳糖致衰老大鼠松果体中SODmRNA表达、HSP70表达、LF含量、松果体细胞数的影响,探讨了艾灸对D-半乳糖致衰老大鼠松果体氧化损伤的干预作用,从分子转录及表达水平深入揭示艾灸延缓衰老机理,为艾灸延缓衰老应用于临床提供新的实验依据。方法实验大鼠随机分为空白组、模型组、模型+艾灸组,空白组每日颈部皮下注射生理盐水1次,模型组每日颈部皮下注射D-半乳糖溶液1次,模型+艾灸组在每日颈部皮下注射D-半乳糖溶液后,选用双侧脾俞、肾俞进行艾灸治疗,连续治疗6周,6周结束时,采集松果体,经固定、包埋、组织切片处理后,采用原位杂交技术、多克隆抗体免疫组化技术及组织化学染色技术,分别测定各组大鼠松果体SODmRNA表达、HSP70表达、LF含量、松果体细胞数,应用病理图文图像分析系统及细胞医学图像分析系统对结果进行图像采集及分析处理。结果1与空白组相比,模型组大鼠松果体中SODmRNA及HSP70表达显著降低(P＜0.001,P＜0.01)、LF明显含量增加(P＜0.001)、松果体细胞数显著减少(P＜0.001)。2与模型组相比,模灸组大鼠松果体中SODmRNA及HSP70表达显著增加(P＜0.01,P＜0.05)、LF含量显著减少(P＜0.001)、松果体细胞数显著增多(P＜0.01)。结论1艾灸可以在分子转录、表达水平上影响机体衰老过程中松果体的氧化损伤,其可能的作用机理是:艾灸通过减缓LF在松果体中沉积,上调松果体中SODmRNA及HSP70的表达而干预松果体的氧化损伤,进而延缓松果体衰退进程。2艾灸通过对松果体氧化损伤的干预作用,减缓机体衰老过程中松果体细胞减少的速度,在一定程度上维持松果体的正常形态及功能,从而延缓松果体衰老。这可能是艾灸延缓衰老的又一作用机理。更多还原

【Abstract】 Purpose: To study the mechanism of moxibustion on slowing down the process of aging more profoundly, taking D-galators induced aged rats as observe goals, taking the TCM aging theory as instruction, taking the modern aging hypothesis of pineal gland as basis, by observing the effect of moxibustion on the expression of SODmRNA and HSP70, the amount of LF and the quantity of pinealocyte in pineal gland of D-galators induced aged rats, studying the interference role of moxibustion on oxidation derogation in pineal gland. Then explain the mechanism of moxibustion on slowing down the process of aging more profoundly on the level of molecule transcription and expression, providing new experimental basis for the clinical treatment of moxibustion on slowing down the process of aging.Methods: Dividing the exprimemtal rats into control group, model groups , model combined moxibustion group, randomly. Dealing control group rats with hypodermic injecting 0.9% normal seline once everyday; deal model group rats with hypodermic injecting D-galators once everyday; deal model combined moxibustion group with hypodermic injecting D-galators , then moxibustion two sides shen-shu point and pi-shu point once everyday, respectively, for six weeks. By the end of the sixth week, gathering pineal gland , treating pineal gland with fixing, embedding, sliceding sections, then testing the expression of SODmRNA and HSP70, the amount of LF and the quantity of pinealocyte in pineal gland of each group rat by us the method of in situ hybridization, polycloing antibody immunohistochemical and histochemical pigmentation. Analysing the results with pathological image analysis system and cell medicine image analysis system.Result: 1 Compared with control group rats, the expression of SODm RNA and HSP70 in pineal gland of model group rats declined significantly(P<0.001, P<0.01), the the amount of LF in pineal gland of model group rats increased significantly (P<0.001) , the quantity of pinealocyte in pineal gland of model group rats reduced significantly (P<0.001). 2 Compared with model group rats, the expression of SODm RNA and HSP70 in pineal gland of model combine moxibustion group rats rised significantly(P<0.01, P<0.05), the the amount of LF in pineal gland of model combine moxibusition group rats decreased significantly (P<0.001) , the quantity of pinealocyte in pineal gland of model group rats increased significantly (P<0.01) .Conclusion: 1 moxibusition has the role of slowing down the process of aging by interfere oxidation derogation in pineal gland on the level of molecule transcription and expression. The possible mechanisms include: moxibusition up-rugulate the expressions of SODmRNA and HSP70, delay the accumulation of LF in pineal gland. then interfering the oxidation derogation in pinea gland. postponing the decline of the pineal gland. 2 moxibusition has the role of delaying the quantity reducing of pinealocyte through interfere oxidation derogation in pineal gland, so as erert the action of slowing down the process of senescence of pineal gland on certain degree. This is one of the possible functional mechanisms of moxibusition delaying aging.更多还原