| ObjectiveAs the object of study in ovariectomized female rats of3months, to explore the effect of tonify the kidney, fortify the spleen and regulate the liver on OVX female rats of aging factors and ER. Supply the modern experimental basis for the theory Liu Wuansu:"heavenly tenth not to, but belong to middle Yin; heavenly tenth have been already, belong to Pole Yin; heavenly tenth has unique, belong to first yin", and enrich the prevention and treatment of menopausal syndrome of traditional Chinese medicine theory.MethodsFemale rats of80days, the weight between108～250g, adaptability feeding10days later, were blocked randomization into blank group and control group, sham group, the fortify spleen group of OVX, the regulate liver group of OVX, the tonify kidney group of OVX. The medicine groups of OVX prevent gavage medicine for30days. The drug concentration:fortify spleen group: guipi bolus,0.4g/ml; tonify Kidney:menopause nourishing kidney oral liquid,4.8g/ml; regulate liver:nourishing yin to sooth the liver capsule0.4g/ml. Others group shall be gavage in0.9%of normal saline, once a day. Then the administration(concluding the fortify spleen group, the regulate liver group and the tonify kidney group) and model groups were carried out the ovariectomy surgery, sham group only take out fat tissue around the ovaries.10days later, the dosage groups continue to gavage for2months. After the last delivery24hours using the0.35%chloral hydrate to excessive anesthesia those rats, adopted the blood sample through the abdominal aortic, then centrifugal those blood sample at3000r/min, in10min for serum. For detection in the serum MDA, H2O2, ROS (reactive oxygen species), SOD (Superoxide dismutase), CAT (Peroxidase), GPX (glutathione peroxidase) and ER in intestine and RT-QPCR detection of bowel aging gene P21, P16, cyclinDl and the expression of ER. According to take in the rat uterus, hypothalamus, hippocampus, pituitary, spleen and thymus weight index.Results1. In the model groups, The BMI of the organs was down, like the uterus、 hypothalamus、pituitary、adrenal gland (P<0.05). The BMI spleen was deceased with the aging. Whereas the BMI of thymus gland was increased.(1)In terms of uterus coefficient:compared with the blank and sham groups, the model group of uterus coefficient was significantly decreased(P<0.05). Whereas those administration the tonify the kidney, the uterus coefficient can be improved (P<0.05), The other groups was found no significance(P>0.05).(2)In terms of hippocampus BMI:There was no significance between the six groups(P<0.05).(3)In terms of hypothalamus BMI:compared with the blank and sham groups, the model group of hypothalamus BMI was significantly decreased(P<0.01). Compared with the model group, the fortify spleen and tonify the kidney can significantly increased the hypothalamus BMI (P<0.05).But the significance was not found in the regulating liver group.(4) In terms of pituitary BMI:Compared with the blank and sham groups, the pituitary BMI was significantly decreased in the model group (P<0.01). Compared with the model group, the fortify spleen、tonify the kidney and regulate the liver can significantly increased the pituitary BMI (P<0.05).However those administration herbs groups’pituitary BMI were significantly lower than the blank group.(5)In terms of adrenal gland BMI:Compared with the blank and sham groups, adrenal gland BMI, there was no significance tin he model group (P>0.05), whereas according the means, the adrenal gland BMI were decreased. Compared with the model group, the fortify spleen and regulate the liver can significantly increased the adrenal gland BMI (P<0.05).However there was no significance in the tonify the kidney group. Compared with the blank group, the fortify spleen can significantly increased the adrenal gland BML However the regulate the liver and tonify the kidney was not found significance(P>0.05). Compared with the tonify kidney, the way of fortify spleen can increased the adrenal gland BMI(P<0.05).(6) In terms of spleen BMI:Compared with the blank and sham groups, spleen BMI, there was no significance in the model group (P>0.05), the spleen BMI changed with the aging. Compared with the model group, the group of fortify the spleen、regulate the liver and tonify the kidney can significantly increased the spleen BMI (P<0.05).Compared with the blank group, the way of fortify spleen、regulate the liver and tonify the kidney can significantly increased the spleen BMI (P<0.05)(7)In terms of thymus gland BMI:Compared with the blank and sham groups, the thymus gland BMI of model group was significantly increased(P<0.05). Compared with the model group, the way of fortify spleen and regulate the liver can decreased the thymus gland BMI (P<0.05).Compared with the blank group, the fortify spleen, the regulate the liver and tonify the kidney can significantly increased the thymus gland BMI. However compared with the tonify kidney, the way of fortify the spleen and regulate the liver can decrease the thymus gland BMI(P<0.05).2. The changes of aging factors(H202、MDA and ROS)and anti-aging factor(SOD, CAT, GPX) in the OVX female rats.(1)aging factors①H2O2:Compared with the blank and sham groups, the content of serum H202in the model was increased(P<0.05). Compared with the model group, the way of fortify the spleen、regulate the liver and tonify the kidney can significantly decreased the content of serum H2O2(P<0.05). Compared with the blank group, the fortify spleen、regulate the liver and tonify the kidney was not found significance on the content of serum H2O2(P>0.05)②MDA:Compared with the blank and sham groups, the content of serum MDA in the model was significantly increased(P<0.01).Compared with the model group, through the way of regulate the liver can significantly decreased the content of serum MDA (P<0.05).Whereas the others groups cannot be found significance(P>0.05). Compared with the blank group, the fortify spleen、 regulate the liver and tonify the kidney was not found significance on the content of serum MDA (P>0.05)③ROS:Compared with the blank group, the content of serum ROS in the model was increased(P<0.05). Compared with the model group, through the way of regulate the liver and fortify the spleen can significantly decreased the content of serum ROS (P<0.05).Whereas the tonify kidney cannot be found significance(P>0.05). Compared with the blank group, the fortify spleen、 regulate the liver was not found significance on the content of serum ROS (P>0.05).(2)Aging factors①SOD:Compared with the blank and sham groups, the activity SOD in the model was not found significance (P>0.05). Whereas according the means the SOD activity was decreased. Compared with the model group, through the way of tonify the kidney and fortify the spleen can significantly increased the activity SOD (P<0.05).however through the way of regulate the liver was not found significance (P>0.05). Compared with the blank group, the fortify spleen and tonify the kidney was not found significance on the SOD activity (P>0.05)②CAT:There was not found significance between those six groups (P>0.05).③GPX:Compared with the blank and sham groups, the activity GPX in the model was significantly decreased(P<0.01). Compared with the model group, through the way of tonify the kidney and regulate the liver the activity GPX can been significantly increased (P<0.05), however through the way of fortify the spleen was not found significance (P>0.05). Compared with the blank group, the regulate liver and tonify the kidney was not found significance on the GPX activity (P>0.05). Compared with the spleen, the way of tonify kidney can increased the GPX activity(P<0.05).3. The neurotransmitter5-HT and DA(1)5-HT:Compared with the blank and sham groups, the content of5-HT of model group was significantly decreased(P<0.01). Compared with the model group, through the way of fortify the spleen can significantly increased the content of5-HT (P<0.05), however through the way of tonify the kidney and regulate the liver, there was not found significance (P>0.05). Compared with the blank group, the regulate liver and tonify the kidney was not found significance on the GPX activity (P>0.05).Compared with the way of tonify kidney and the regulate liver, the fortify the spleen can increased the GPX activity(P<0.05).(2)DA:There was no significance between the six groups (P>0.05).4. The senescence genes and the ER genes in the small intestine (1)The senescence genes CyclinDl,P21, P16①CyclinDl genes:Compared with the model group, through the way of regulate the liver, the expression of CyclinDl gene can be up-regulated(P<0.05), whereas the two others was not found significance(P>0.05).②P16genes:Compared with the model group, through the way of regulate the liver, tonify kidney and the fortify the spleen.there was no significance (P>0.05).③P21genes:Compared with the model group, through the way of tonify the kidney and fortify the spleen, the expression of P21genes down-regulated(P<0.05). However, through the way of regulate the liver, the expression of P21genes can be up-regulated (P<0.05). Compared with the regulate the liver, the way of tonify kidney and fortify spleen can down-regulate the expression of P21genes(P<0.05).(2)ER genes:Compared with the model, the three others was not found significance(P>0.05). Conclusion1. The effect of The way of fortify the spleen on the OVX rats:The way of fortify the spleen can improved the hypothalamus、pituitary、adrenal gland spleen BMI. Meantime controlled the thymus gland hypertrophy. So this way can strengthen the immunity. The effect on the adrenal gland and thymus gland was better than the tonify kidney and the regulate the liver groups. The way of fortify can also decreased the content of serum ROS and H2O2, down-regulate the P21genes, meantime increased the activity SOD, so this way can reinforce the ability of anti-aging. In addition,it can regulate the content neurotransmitter of serum5-HT.2.The effect of The way of tonify the kidney on the OVX rats:The way of tonify the kidney can improved the uterus、hypothalamus、pituitary and the spleen BMI. This way can also decreased the content of serum H2O2, down-regulate the P21genes, meantime increased the activity SOD、GPX.So it can reinforce the ability of anti-aging for the OVX rats.3. The effect of The way of regulate the liver on the OVX rats:The way of the liver can improved the pituitary、spleen and the adrenal gland BMI. This way can also decreased the content of serum MDA、ROS and H202, up-regulate the P21genes, meantime increased the activity GPX and up-regulated the expression of CyclinDl. So it can reinforce the ability of anti-aging for the OVX rats.Above of all, although three ways also improved the aging status of OVX rats, but the way of fortify the spleen was the best. The way of fortify the spleen can improve the aging status from5aspects, and more effect on4aspects. The way of tonify the kidney an improve the aging status from4aspects, and more effect on2aspects. The regulate the liver can improve the aging status from4aspects, and more effect on1aspects. So these experiment preliminary discussed the theory of Liu wansu "heavenly tenth has unique, belong to first yin", and supply the basic for the treatment of climacteric and its aging status. |
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