ABSTRACT
ObjectiveTo explore the therapeutic effect and mechanism of Guipitang on rats with myocardial ischemia. MethodFifty SD rats were divided into five groups： a control group， a model group， low and high-dose Guipitang （7.52， 15.04 g·kg-1） groups， and a trimetazidine group （0.002 g·kg-1）. By intragastric administration of vitamin D3 and feeding rats with high-fat forage and injecting isoproterenol， the rat model of myocardial ischemia was established. After drug treatment of 15 d， an electrocardiogram （ECG） was performed to analyze the degree of myocardial injury. A fully automatic biochemical analyzer was used to detect the changes in the serum levels of total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）. Hematoxylin-eosin （HE） staining and Masson staining were used to observe myocardial histopathological changes. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect cardiomyocyte apoptosis. Western blot was adopted to detect the protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， phospho-ERK1/2 （p-ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， phospho-p38 MAPK （p-p38 MAPK）， B-cell lymphoma-2 （Bcl-2）-associated X （Bax）， Bcl-2， and cleaved cysteine aspartate proteolytic enzyme （cleaved Caspase-3）. ResultCompared with the control group， the ECG S-T segment decreased in the model group. The serum levels of TC， TG， and LDL-C were increased significantly （P<0.05）. The arrangement of myocardial tissue was disordered， and the proportion of cardiomyocyte apoptosis increased. The protein levels of cleaved Caspase-3， Bax， and p-p38 MAPK in the heart were increased， and the Bcl-2 expression was decreased （P<0.05）. Compared with the model group， the S-T segment downward shift was restored in the low and high-dose Guipitang groups and trimetazidine group， and the levels of TC， TG， and LDL-C were decreased. The protein expression of cleaved Caspase-3 and Bax in the heart dropped， and p-p38 MAPK and p-ERK1/2 protein expressions increased significantly （P<0.05）. The degree of myocardial injury was alleviated， and the proportion of cardiomyocyte apoptosis decreased. Bcl-2 protein expression was increased significantly in the low-dose Guipitang group （P<0.05）. ERK1/2 and p38 MAPK proteins had no significant difference among different groups. ConclusionGuipitang could alleviate myocardial injury and inhibit cardiomyocyte apoptosis in rats by activating the expression of ERK1/2 and p38 MAPK.
ABSTRACT
BACKGROUND: According to correlation of "acu-points-meridians-viscera-target-organs", two administration ways of acu-point application and oral medication have been used to treat osteoporosis, both ways can effect via "kidney-meridian tropism"; however whether there is a specificity of target-administration?OBJECTIVE: To investigate the effect of different administration ways of kidney-strengthening remedy on "yang abating and yin hoarding" of osteocytes in experimental osteoporosis by observation the correlation between "receptor and meridian-tropism".DESIGN: A randomized and controlled trial of different administration ways of kidney-strengthening remedy, taking rats with osteoporosis as subject.SETTING: Departments of Prescription Science and Biochemistry of Traditional Chinese Medicine (TCM) Collage, Hebei Medical University, and Qiaodong District Hospital of Shijiazhuang City.MATERIALS: The experiment was completed from June 2004 to May 2005 at the Laboratory of Integrative Medicine Basis of Hebei Medical University. Totally 120 SD rats, aged 3 months and of clean grade, were at random divided into 6 groups: normal control, pathological model, kidney strengthening, bladder meridian application, kidney meridian application and non-special application groups, with either sex being 10 in each group.The kidney strengthening remedy mainly consisted of fortune's Dry Naria Flavoniods and Icarine, each gram of fluid extract contained 9.543 g of the crude drug. The application recipe for anti-osteoporosis consisted of Radix Rehmanniae Praeparata, Cortex Moutan Radicis, Herba Epimedii, etc.METHODS: Except for those in normal group, the rats in the other groups were all given injection of dexamethasone to set up the osteoporosis models. The rats in normal control and pathological model groups were not given medicine, the rats in kidney strengthening group were by gavage given kidney strengthening remedy at 0.8 g/100 g wet. The rats in bladder meridian group were treated by anti-osteoporosis application at the points of Shenshu (BL 23) and Feiyang (BL 58) of the bladder meridian; the rats in kidney meridian group were treated by anti-osteoporosis application at the points of Taixi (KI 3 23) and Dazhong (KI 4) of the kidney meridian;and the rats in non-special application group were treated by anti-osteoporosis application at the areas of non-point-meridian. After 13 weeks administration, the rats were put to death, the concentrations of serum estradiol, testosterone, calcitonin and parathyroid hormone were detected with radio-immunoassay, and both the dynamic and static status parameters about bone morphosis were also measured.MAIN OUTCOME MEASURES:① Estradiol,testosterone,estradiol/testosterone, bone mineral density, calcitonin, parathyroid hormone, and calcitonin/parathyroid hormone of rats in each group after administration ② Both the dynamic and static status parameters about bone morphosis of rats in each group after administration.RESULTS: All rats involved in the trial entered the final result analysis.① The results of biochemical indexes of the rats in each group after administration: Compared with normal control group, the levels of estradiol,testosterone, estradiol/testosterone, bone mineral density, calcitonin and calcitonin/parathyroid hormone of the rats in pathological and non-special application groups after administration were remarkably decreased (P ＜ 0.01), but the level of parathyroid hormone was obviously increased (P ＜ 0.01); however for the levels of the above indexes of rats in kidney strengthening, bladder meridian application and kidney meridian application groups there were obvious changes (all P ＞ 0.05).② The results ofdynamic and static status parameters about bone morphosis of rats in each group after administration: Compared with normal group, in pathological and non-special application groups the volume percentage of bone trabecula, surface percentage of bone trabecula, formation surface percentage of bone trabecula, activity formation surface percentage of bone trabecula,mineralization deposition rate of bone trabecula, longitudinal bone growth rate, and velocity of bone trabeculation were all decreased (P ＜ 0.01), but the absorbance surface percentage of bone trabecula remarkably increased (P ＜ 0.01); however for the levels of the above indexes of rats in kidney strengthening, bladder meridian application and kidney meridian application groups there were obvious changes (all P ＞ 0.05).CONCLUSION: The anti-osteoporosis application can inhibit bone absorption and promote bone formation. It can be effectively controlled in quantity of drug entering the blood circulation, and with the same effect as that given orally. It can check the state of "bone destruction being more than bone formation" in osteoporosis, reverse the process of "yang abating and yin hoarding", making the bone absorption of osteoclast decrease, and the ability of the bone formation of osteoblast increase, enhancing bone mineral density, so that the "amplifying effect of point-meridian-passageway" comes into play.
ABSTRACT
BACKGROUND: In the research, the recipe for kidney tonification is prepared into "topical form" and "oral form". According to the interrelationship of "acupoints-meridans and collaterals-internal organs-target organs", osteoporosis is treated by the approaches of point compress and oral application respectively. According to the traditional theory of Chinese medicine, it is to infer that the herbs bring the adjustment into play by"being attributive to kidney meridian", but,whether does the specialty of herbs on "targeting medication" happenpractically or not? Hormones in "hypothalamus-pituitary- target gland" system is assayed to further analyze estrogen receptor and androgen receptor of target organs,such as kidney,bone, uterus, thyroid gland and testis so as to observe the relativity between "tropism and receptor".DESIGN: Complete random design with the objects of osteoporosis rats, and the control experimental study with kidney tonification recipe administrate with different approaches.OBJECTIVE: To probe into the effect of kidney tonification recipe on pituitary-thyroid axis in experimental osteoporosis.SETTING: College of Traditional Chinese Medicine of Hebei Medical University.MATERIALS: The experiment was performed in Chinese-Western integrated Basic Experiment Room of Hebei Medical University from January 2000 to December 2004. Sixty SD healthy female rats of 3-month old were employed,weighted(300 ± 20) g. The experimental animals were provided by Hebei Experimental Animal Center. After bred routinely for 1 week in Experimental Room, the rats were randomized into 6 groups,namely normal control(10rats), pathological model group(10 rats), infusing group(10 rats) in which kidney tonification recipe was applied based on 0. 8 g/100 g body mass,acupoint paste on bladder meridian group (10 rats), acupoint paste on kidney meridian group(10 rats) and paste on non-meridian group(10 rats). METHODS: Osteoporosis model was prepared with injection of dexamethasonc. Antagonism osteoporosis acupoint paste(AOAP)on acupoints and non-neridian places was administrated to compare with oral recipe for kidney tonification in the treatment of osteoporosis. It took the changes in serum estrogen, bone density, calcitonin/parathyroid hormone(CT/PTH), thyrotropic hormone(TSH), freeT3andfreeT4asthemeasuringindexes. Immunocyto-chemistry stain and morphological measurement of TSH cell,follicular adenoma of thyroid gland(C cell) and thyroid globulin(Tg) were applied in pituitary gland and thyroid gland to observe the therapeutic results.MAIN OUTCOME MEASURES: ① Adjustment of the recipe for kidney tonification on thyroid function. ② Adjustment of the recipe for kidney tonification on bone density.RESULTS: AOAP improved the levels of estrogen, CT/PTH, TSH, free T3and free T4, increased bone density. The counts of TSH cell, C cell and Tg positive reaction substance were increased and they were colored deeply.CONCLUSION: AOPA, by "multiple relevant convergence reaction", adjusts and affects systematically the endocrinal function of bone metabolism,improves the levels of estrogen and CT/PTH, reverses the declining tendency of pituitary-thyroid axis function and improves thyroid function so as to weaken osteocyte in the bony absorption, enhance osteoblast in osteogenesis and increase bone density.