BACKGROUND: Since chronic liver disorders are associated with bone loss commonly, it is very significant to probe into the prevention of such bone loss for the treatment of osteoporosis.OBJECTIVE: To study the mechanism of glycyrrhizic acid on prevention and treatment of bone loss induced by liver fibrosis in mice.DESIGN: Randomized and controlled study in which the experimental animals were taken as the objects.SETTING: Experimental animal center, central experimental room and pharmacological research room of a universityMATERIALS: The experiment was performed from January to September 2001. Forty common-grade PCR mice of either sex were employed, weighted varied from 20 to 22 g, provided by Experimental Animal Center of the Institution. According to mass-equivalence principle, 4 groups were randomized, named as control, model group, the treatment with colchicine group(CL group) and the treatment of glycyrrhizic acid group(GA group-) with 10 mice in each group.METHODS: Except the control group, in the model group, CL group and GA group, the subcutaneous injection with 400 g/L carbon tetrachloride prepared with peanut oil was given for 5 weeks to induce liver fibrosis in mice. Afterward, the treatment was applied with carbon-tetrachloride peanut-oil solution 10 mL/kg, colchicum autumnale solution 0. 1 g/kg and glycyrrhizic acid solution 0. 1 mg/kg successively. At the end of the experiment, the eyeball was extirpated for blood collection. The serum was separated to assay the various relevant biochemical indexes of liver injury, observe the changes in liver pathological tissues and measure bone calcium(Ca2+) content of femur and the contents of other bone trace elements as well as bone oxyproline hydroxyproline (Hyp).Effects of glycyrrhizic acid on the contents of bone trace elements in mice of every group.RESULTS: The levels of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in CL group and GA group were lower than those in the model group( P < 0. 01 ). The level of albumin and ratio between albumin and globulin in the model group were lower than those in CL group and GA group( P < 0.01, P < 0.05) . The content of bone calcium in CL group and GA group were lower than that in the control group ( P < 0. 05),but that was higher than the model group( P < 0.05) . The unit contents of copper(Cu2+ ), magnesium(Mg2+) and zinc(Zn2+) in the right femur of the model group were all higher than the control group ( P < 0.01 ), but the contents of those in GA group were not indicated significant differences compared with the control group ( P > 0.05 ). The liver pathological changes of mice in GA group were obviously milder compared with the model group ( P < 0.01 ) and it was shown with VG staining the severity of hyperplasia of liver collagenous fibers was remarkably milder compared with the model group( P < 0.01).CONCLUSION: The extractive solution of glycyrrhizic acid induces medical metabolic enzyme in the liver, enhances detoxification of liver, protects liver to maintain protein metabolic level and maintains the normal metabolism of bone to promote bone Ca2+ and balance between oxyproline hydroxyproline (Hyp) and trace elements of bone so as to prevent and treat bone loss.

AIM: To study the cerebra damaged by long-term administration of prednisone,and investigate the preventive effects of compound danqi (CD) in rats. METHODS: 40 SD rats were randomly divided into 5 groups with 8 rats each group. Group 1 was control (NS group),other groups were oral gavages prednisone ( 2.7 mg?kg -1 ?d -1 ) first,and then plus vehicle (GC group),or plus CD at dose of 2.5 , 5.0 or 10.0 g?kg -1 ?d -1 ,respectively,once a day for 12 weeks. At the experimental endpoint,animals were drawn blood from right ventricle under anesthesia. The left half cerebrum was milled in 10% homogenate to test the content of mono-amine oxidase (MAO),superoxide dismutase (SOD) and acetylcholinesterase (AchE). The right half cerebrum was for histological observation. RESULTS: The concentration of SOD and AchE decreased,yet MAO increased significantly in GC group. Moreover,histopathology showed that the structure of cerebrum cortex became thinner and hippocampi was in disorder. Dropsical and necrotic nerve cells were found. Yet CD could prevent the changes of nerve centre in prednisone tats. CONCLUTION: Cerebra damage occurs in four-month-old male Sprague-Dawley rats after prednisone-treated for 12 weeks. The treatment of CD in different dose can prevent the damage.

Aim To study the characteristics of osteoporosis induced by long-term administration of prednisone, and investigate the preventive effects of compound stanozolol (CS) on the adverse reactions in male rats. Methods Twenty-four four-month-old male Sprague-Dawley rats were randomly divided into 3 groups with 8 rats per group. Group 1 was control (NS group), other groups were oral gavages prednisone (2.7 mg?kg -1?d -1) first, and then plus vehicle (GC group) or plus CS (combination of Stanozolol 0.5 mg?kg -1?d -1 + Calciofon 0.5 g?kg -1?d -1 + Vitamin D 3 250 U?kg -1?d -1), once a day for 12 weeks. At the experimental endpoint, animals were drawn blood from right ventricle under anesthesia. The tibia, ulna and thighbone were collected to test for parameters related to bone.The undecalcified longitudinal proximal tibia metaphyseal sections were cut and stained with Masson-Goldner's Trichrome (5-?m thickness) for osteoclasts analysis, unstained sections (8-?m thickness) for the cancellous bone histomorphometric and fluorochrome labeling analysis. The measured parameters were used to calculate the percent trabecular area (Tb.Ar%), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), percent labeled perimeter (L.Pm%), Mineral apposition rate (MRA), bone formation rate (BFR/TV, BFR/BV, BFR/BS), unit area osteoclast number (Oc.N/BV) and Percent osteoclast number surface perimeter (OcP/BS). The desiccant left ulnar was dissolved in hydrochloric acid to test the content of Ca 2+ and hydroxyproline. Length and width of the thigh bone were tested to observe the instance of bone growing. Results Compared with NS group, bone weight, contents of bone hydroxyproline and Ca 2+decreased remarkably in GC group and the bone histomorphometric parameters also showed bone lose significantly.The effects of CS in prednisone rats showed that CS prevented bone loss effectively.Conclusions Bone loss occured in four-month-old male Sprague-Dawley rats after prednisone-treated for 12 weeks. CS could prevent bone loss effectively by inhibiting bone resorption and advancing bone formation.

AIM:To study the (rat's) osteoporosis induced by long-term administration of prednisone,and investigate the prophylactic effects of Salvia miltiorrhiza Beg in male rats. METHODS: Twenty-four SD rats were randomly divided into four groups such as control group,prednisone group and Danshen(Salvia miltiorrhiza Bge) group,with 8 rats per group.control group was treated with normal saline,other groups with prednisone at dose of(2.7) mg/kg first,and then prednisone group with normal saline group with Salvia miltiorrhiza Bge at dose of 5.0 g/kg,respectively,once a day for 12 weeks.At the experimental endpoint,bone histomorphometric analysis of thighbone were performed in undecalcified sections,the contet of Ca~(2+) and oxyproline hydroxyproline of ulnar bone were to be measured.length and width of the thighbone were to be tested. RESULTS: Bone histomorphometric parameters showed bone loss was significant to prednisone group(P

AIM To study the relationship between hepatic fibrosis and osteoporosis and to determine the preventive effects of Glycyrrhizin on bone loss in mice. METHOD Forty PCR Mice were randomly divided into 4 groups. Group A mice were controls; Group B mice were given sc injection of 40% CCl 4 10 ml?kg -1 once per 5 days as fibrosis model group; Group C were given orally colchicine of 0 01 mg?kg -1 plus CCl 4 sc, Group D were given Glycyrrhizin(GL) of 100 mg?kg -1 orally plus CCl 4 sc. The four groups were treated for 42 days. The liver injury indexes were measured and the mineral elements and Hydroxyproline of the femur were determined. RESULT Compared with group A, the serum enzymes of alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) were markedly increased and serum albumin (Alb) and A/G(Alb/(total protein Alb) were decreased significantly in group B whose liver slides also showed typical liver cirrhosis. The dried weight of femur in the cirrhosis mice was markedly reduced and the bone Calcium content and bone Hydroxyproline content were also significantly decreased in group B. Bone copper and bone magnesium were increased in group B. In group D, GL inhibited markedly the decrease of the serum enzymes and increased Calcium content and Hydroxyproline content of the bone compared with group B. The bone mass loss was prevented effectively by Glycyrrhizin. CONCLUSION The bone mass was lost in mice with chronic hepatic injury induced by CCl 4 and Glycyrrhizin can prevent bone loss which was accompanied by chronic hepatic injury in mice.

AIM: To study the time dose effects of liver damnification induced by CCl 4 in rats. METHODS: The SD rats were given sc injection 60% CCl 4 twice a week to induced liver damage. After the first week, one proportion animals were killed. The contents of serum alanine aminotransferase (ALT), aspartic acid aminotransferase (ALT), hyaluronic Acid (HA), total protein (TP) and albumin (Alb) were investigated, and the thymus, liver, spleen, kidney, and the adrend gland were weighed. On the other hand, the liver tissue was studied with histopathological to observe the degrees of inflammatory activity and fibrous hyperplasia. In the coming the third, fifth, seventh, ninth week, the different proportion animals were done as the above. RESULTS: In the first week, the level of serum ALT were significantly increased; in the third week, the hepatic cells were occurred fattiness denaturalization; in the fifth week, the histopathological showed the necrotic hepatic cells obviously; in the seventh week, the histopathological displayed that the liver turned into fibrotic; and in the ninth week, the liver changed into hepaticfibrosis induced by CCl 4. CONCLUSION: The injection (sc) of 60% CCl 4 will cause diverse hepatic injury with the dissimilar durative time in rats.

BACKGROUND: Long-term large dose application of glucocorticoid can cause osseous loss and even femoral head necrosis,which is one of the reasons of pharmaceutical damages. Researches on its intervention have practical significance.OBJECTIVE: To study the osteoporosis induced by long-term large dose administration of glucocorticoid, and investigate the preventive effects of compound danshen(CD) in male rats.DESIGN: A randomized and controlled study by employing experimental animals as subjectsSETTING: An Experimental Animal Center, a Central Laboratory and an Institute of Pharmacology of a Medical CollegePARTICIPANTS: The study was conducted in the Experimental Animal Center, the Central Laboratory and the Institute of Pharmacology of Guangdong Medical College between 2002 and 2003. Totally 40 SD rats were employed.INTERVENTION: SD rats were treated with prednisone(2.7 mg/kg per day) by oral gavages and CD including danshen huangqi , baishu and yinyanghuo at dose of 2.5 g/kg per day,5.0 g/kg per day or 10.0 g/kg per day respectively,once a day for 12 weeks. At the experimental endpoint,the impacts of long term large dose (beyond physiologic dose) application of glucocorticoid on bone metabolism and the preventive effects of CD were observed through the measurement of the static and dynamic indicators for bone growth in un-decalcified superior tibia,the detection of Ca2 + and hydroxyproline contents in ulna,and the length and width of thighbone.MAIN OUTCOME MEASURES: Principal consequences: the impacts of CD on quantitative static and dynamic parameters of osseous morphology in rats with prednisone-induced osteoporosis; Secondary consequences: the comparison of the impacts of CD on bone biochemical indictors and femoral physical indicators in rats with prednisone-induced osteoporosis.RESULTS: In glucocorticoid control group (GC group),bone mass significantly decreased(P<0. 01); as indicated by bone morphological indicators,the number of bone trabecula[(1.98±0.20) / mm]and the percentage of bone trabecular size [(8.83 ±0.98)%] significantly reduced; the ratio of osteogenesis rate at bone surface (8.91±3.97) /neogenesis bone trabacular size to total bone trabecular size(332. 8±142.5)/neogenesis bone trabecular size to bone size(29.6±13.2) significantly decreased; bone absorption perimeter significantly increased(P<0. 01); osseous content in ulna reduced[ (0. 155±0. 01) g]; and femoral length[ (32.64±0.51) mn]significantly shortened (P<0. 05) . But in CD group,CD had certain preventive effects on bone injury induced by prednisone while there was no significant difference among each subgroup with different dose.CONCLUSION: Long-term application of prednisone can significantly inhibit bone growth and induce bone loss. CD has favorablepreventive effects on bone loss through its promotion of osteogenesis and inhibition of osteoclast bone resorption.

AIM To determine the effects of different dose of cyclophosphamide (CP) on the contents of bone calcium and hydroxyproline and the weight of immune organ thymus. METHOD CP at dose of 1 5, 4 5 and 12 5mg?kg -1 were given to the male rats orally everyday for 15 days respectively. At the endpoint the right femoral was dried constantly and weighed (w), the calcium content (Ca.C) was assayed by the method of Atomic Absorption Spectrometry and the femoral hydroxyproline content (Hp.C) were assayed by Colorimetry. The leukocyte, thymus, liver and spleen were tested and weighed separately. RESULTS CP induced inhibiting effect on body weight, leukocyte and thymus in a dose dependent when compared with the vehicle control. Three doses of CP significantly decreased Ca.C and Ca.C/w of femoral ( P0 05). CONCLUSION CP stimulated bone calcium loss, induced shrinkage of thymus. The proper dose at 4 5mg?kg -1 of CP reduced both of Hp.C and Ca.C in bone leading to osteoporosis which is related to the decrease of bone mineral and bone matrix.

AIM: To observe the changes of different parts of bones in rats 90 days after ovariectomized (OVX). METHODS: Twenty 4.5 month old virgin female Sprague Dawley rats were randomly divided into 2 groups: sham group and OVX group. Rats in sham group were sham operated, while rats in OVX group were bilaterally OVX. Rats in two groups were treated with 5 ml?kg -1 ?d -1 ethanol for 90 days. Parameters of the proximal tibial metaphysis (PTM), tibial shaft (Tx) and the fifth lumbar vertebral body (LVB) were analyzed by bone histomorphometry method. RESULTS: %Tb.Ar and Tb.N were decreased by 80.5 % and 76.1 %, Tb.Sp and Oc.N were increased by 468.0 % and 356.6 %, and MAR and BFR/BV were increased by 43.9 % and 95.9 %, respectively, in PTM. The changes in LVB were not remarkable as those in PTM. %Tb.Ar and Tb.Th were decreased by 35.0 % and 31.0 %, while Oc.N and BFR/BV were increased by 106.9 % and 126.8 %, respectively. The cortical bone and marrow areas of tibial shaft did not change in Tx, but bone formation parameters (%P LPm, %E LPm) were increased. CONCLUSION: After OVX for 90 days, high bone turnover osteopenia model is duplicated successfully in rats. Different parts of the bones have different reaction to ovariectomization in rats.

AIM: To study the correlative changes of bone mineral contents and histomorphometry of lumbar vertebrae in ovariectomized rats by analysis of T-type correlation degree. METHODS: Forty 10.5-month-old virgin female Sprague-Dawley rats were randomly divided into five groups: (1) Basal: at 10.5 mon of age; (2) sham-1:sham-operated at 13 mon of age; (3) OVX-1: ovariectomized at 13 mon of age; (4) sham-2: sham-operated at 16 mon of age; (5) OVX-2:ovariectomized at 16 mon of age. After ovariectomy, all rats were treated orally with NS at 5 ml?kg -1?d -1. At the end-point of study, the undecalcified longitudinal fourth lumbar vertebra (LV4) sections were cut and stained with Goldner's Trichrome for bone histomorphometric analyses. The fifth lumbar vertebra (LV5) was dried with temperature and digested with acid for testing of bone mineral content. Then the effects of bone mineral contents on bone histomorphometry were assessed by analyzing the T-type correlation degree in the Grey system. RESULTS: All degrees of correlation between bone mineral contents and static histomorphometric parameters (trabecular bone volume) (BV/TV), trabecular thickness (Tb.Th) were positive, but for dynamic histomorphometric parameter (BFR/BV), the correlation degrees were negative. The effect of contents of calcium (Ca) and phosphorus (P) on histomorphometric parameters of lumbar vertebrae was much greater than that of the other bone mineral contents. CONCLUSIONS: Analysis of T-type correlation degree can evaluate the correlative changes of bone mineral contents and histomorphometry of lumbar vertebrae in ovariectomized objectively and fairly.