Genipin improves obesity through promoting bile secretion and changing bile acids composition in diet-induced obese rats.

OBJECTIVES: Bile acids (BAs), as signaling molecules to regulate metabolism, have received considerable attention. Genipin is an iridoid compound extracted from Fructus Gradeniae, which has been shown to relieve adiposity and metabolic syndrome. Here, we investigated the mechanism of genipin counteracting obesity and its relationship with BAs signals in diet-induced obese (DIO) rats. METHODS: The DIO rats were received intraperitoneal injections of genipin for 10 days. The body weight, visceral fat, lipid metabolism in the liver, thermogenic genes expressions in brown fat, BAs metabolism and signals, and key enzymes for BAs synthesis were determined. KEY FINDINGS: Genipin inhibited fat synthesis and promoted lipolysis in the liver, and upregulated thermogenic gene expressions in brown adipose tissue of DIO rats. Genipin increased bile flow rate and upregulated the expressions of aquaporin 8 and the transporters of BAs in liver. Furthermore, genipin changed BAs composition by promoting alternative pathways and inhibiting classical pathways for BAs synthesis and upregulated the expressions of bile acid receptors synchronously. CONCLUSIONS: These results suggest that genipin ameliorate obesity through BAs-mediated signaling pathways.

The morphology, biomechanics, and physiological function of the suboccipital myodural connections.

The myodural bridge (MDB) connects the suboccipital musculature to the spinal dura mater (SDM) as it passed through the posterior atlanto-occipital and the atlanto-axial interspaces. Although the actual function of the MDB is not understood at this time, it has recently been proposed that head movement may assist in powering the movement of cerebrospinal fluid (CSF) via muscular tension transmitted to the SDM via the MDB. But there is little information about it. The present study utilized dogs as the experimental model to explore the MDB's effects on the CSF pressure (CSFP) during stimulated contractions of the suboccipital muscles as well as during manipulated movements of the atlanto-occiptal and atlanto-axial joints. The morphology of MDB was investigated by gross anatomic dissection and by histological observation utilizing both light microscopy and scanning electron microscopy. Additionally biomechanical tensile strength tests were conducted. Functionally, the CSFP was analyzed during passive head movements and electrical stimulation of the suboccipital muscles, respectively. The MDB was observed passing through both the dorsal atlanto-occipital and the atlanto-axial interspaces of the canine and consisted of collagenous fibers. The tensile strength of the collagenous fibers passing through the dorsal atlanto-occipital and atlanto-axial interspaces were 0.16 ± 0.04 MPa and 0.82 ± 0.57 MPa, respectively. Passive head movement, including lateral flexion, rotation, as well as flexion-extension, all significantly increased CSFP. Furthermore, the CSFP was significantly raised from 12.41 ± 4.58 to 13.45 ± 5.16 mmHg when the obliques capitis inferior (OCI) muscles of the examined specimens were electrically stimulated. This stimulatory effect was completely eliminated by severing the myodural bridge attachments to the OCI muscle. Head movements appeared to be an important factor affecting CSF pressure, with the MDB of the suboccipital muscles playing a key role this process. The present study provides direct evidence to support the hypothesis that the MDB may be a previously unappreciated significant power source (pump) for CSF circulation.

[Effects of Genipin on the expression of uncoupling protein 1 in brown adipose and white adipose tissues in mice].

OBJECTIVE: To investigate the effects of genipin on promoting brown adipose tissue activation and white adipose tissue browning. METHODS: The male C57BL/6J mice were divided into three groups: normal control group, genipin group and cold-stimulus group.Genipin group were treated consecutively with genipin at a dose of 15 mg/kg once a day for 9 days, normal control group were treated with the saline.The mice with cold-stimulus were exposed to 4℃ environment for 5 days.Daily food amount and body weight were measured.Morphological changes were observed in the subscapular region, inguinal region and epididymis around the adipose tissue.The expression of uncoupling protein 1 (UCP1) was determined by real-time PCR and Western blot respectively. RESULTS: The wet weight of white fat in genipin-treated mice was decreased by 16% , and 28% in that of cold-stimulus mice, compared with the normal control group (P＜0.05).After treatments of genipin and cold-stimulus, the color of white adipose tissues was darker, and the size of lipid droplets in adipocytes was smaller, whereas the number was increased.Compared with the normal control group, UCP1 expression was increased obviously in fat tissues, including the subcutaneous and visceral white adipose tissues, and brown adipose tissue after treated with genipin and cold-stimulus (P＜0.05). CONCLUSION: Genipin promoted activation of brown adipose tissue and browning of white adipose tissue by upregulating UCP1 expression, which could contribute to the loss of body weight against obesity.

Genipin ameliorates diet-induced obesity via promoting lipid mobilization and browning of white adipose tissue in rats.

Genipin is the major active component of Gardeniae fructus and has been shown to ameliorate diabetes and insulin resistance in rat models. In this study, we first investigated the effect of genipin on obesity and the related lipid metabolism mechanisms in diet-induced obese rats. Our results showed that genipin reduced body weight, food intake, and visceral fat mass; ameliorated dyslipidemia, glucose intolerance, insulin intolerance, adipocyte hypertrophy, and hepatic steatosis; and reduced serum tumor necrosis factor-α level in diet-induced obese rats. Quantitative real-time reverse-transcription polymerase chain reaction results further illustrated that genipin promoted lipolysis and ß-oxidation of fatty acid by upregulating gene expressions of hormone-sensitive lipase and adipose triglyceride lipase in white adipose tissue (WAT) and peroxisome proliferator-activated receptor-α and carnitine palmitoyltransferase 1α in hepatic tissue. Moreover, genipin promoted browning of WAT by upregulating the mRNA and protein levels of uncoupling protein 1 and PRD1-BF1-RIZ1 homologous domain containing 16 in WAT. Additionally, genipin inhibited gene expressions of activin receptor-like kinase 7, tumor necrosis factor-α, and interlukin-6 in WAT. These results indicated that genipin had a potential therapeutic role in obesity, in which regulation of lipid mobilization and browning of WAT were involved.

Green tea polyphenols protect against okadaic acid-induced acute learning and memory impairments in rats.

OBJECTIVE: Green tea polyphenols (GTPs) are now being considered possible protective agents in neurodegenerative diseases such as Alzheimer's disease (AD). Previous studies suggested that GTPs could inhibit amyloid fibril formation and protect neurons from toxicity induced by ß-amyloid. However, whether GTPs can ameliorate learning and memory impairments and also reduce tau hyperphosphorylation induced by okadaic acid (OA) in rats remains unclear. The aim of this study was to determine if GTPs have neuroprotection against OA-induced neurotoxicity. METHODS: In this work, rats were pretreated with GTPs by intragastric administration for 4 wk. Then OA was microinjected into the right dorsal hippocampus. Morris water maze tests were used to test the ethologic changes in all groups, and tau protein hyperphosphorylation was detected both in vivo and in vitro. RESULTS: The ethologic test indicated that the staying time and swimming distance in the target quadrant were significantly decreased after OA treatment, whereas rats pretreated with GTPs stayed longer in the target quadrant. Methyl thiazolyl tetrazolium assay and lactate dehydrogenase leakage showed that GTPs greatly ameliorated primary hippocampal neurons damage induced by OA. Furthermore, reduced hyperphosphorylated tau protein was detected with GTPs pretreatment. CONCLUSION: Taken together, our results suggest that GTPs have neuroprotection against OA-induced neurotoxicity.

Effect of fucoxanthin alone and in combination with D-glucosamine hydrochloride on carrageenan/kaolin-induced experimental arthritis in rats.

The objective of the present study was to investigate the effect of the fucoxanthin (FUCO) alone and in combination with glucosamine hydrochloride (GAH) on carrageenan/kaolin-induced inflammatory arthritis model in rats and to explore its underlying mechanisms. Joint swelling, muscle weight ratio (%), histopathological examination and scoring, and proteoglycan degradation were examined. Pro-inflammatory interleukin (IL-1ß) and tumor necrosis (TNF-α) levels, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase(iNOS) protein expression and nitric oxide (NO) level in knee synovial tissue extract were analyzed using enzyme-linked immunosorbent assay, western blotting analysis, and Griess reagent assay, respectively. FUCO and FUCO + GAH not only may significantly reduce degrees of knee joint swelling and prevent against muscle atrophy, but also may significantly attenuate inflammation in synovial tissue, cartilage erosion, and proteoglycan loss. The efficacies of FUCO + GAH were stronger than that of GAH or FUCO. FUCO alone and FUCO + GAH can significantly inhibit upregulation of COX-2 and iNOS protein expressions, decrease of IL-1ß and TNF-α levels, and reduce NO production in knee synovial tissue extract. These results indicated that FUCO is an effective anti-arthritis agent through an antiinflammation mechanism. FUCO may enhance therapeutic effect of GAH on rat arthritis through mechanism of antiinflammation.

Genipin ameliorates age-related insulin resistance through inhibiting hepatic oxidative stress and mitochondrial dysfunction.

Insulin resistance (IR) increases with age and plays a key role in the pathogenesis of type 2 diabetes mellitus. Oxidative stress and mitochondrial dysfunction are supposed to be major factors leading to age-related IR. Genipin, an extract from Gardenia jasminoides Ellis fruit, has been reported to stimulate insulin secretion in pancreatic islet cells by regulating mitochondrial function. In this study, we first investigated the effects of genipin on insulin sensitivity and the potential mitochondrial mechanisms in the liver of aging rats. The rats were randomly assigned to receive intraperitoneal injections of either 25mg/kg genipin or vehicle once daily for 12days. The aging rats showed hyperinsulinemia and hyperlipidemia, and insulin resistance as examined by the decreased glucose decay constant rate during insulin tolerance test (kITT). The hepatic tissues showed steatosis and reduced glycogen content. Hepatic malondialdehyde level and mitochondrial reactive oxygen species (ROS) were higher, and levels of mitochondrial membrane potential (MMP) and ATP were lower as compared with the normal control rats. Administration of genipin ameliorated systemic and hepatic insulin resistance, alleviated hyperinsulinemia, hyperglyceridemia and hepatic steatosis, relieved hepatic oxidative stress and mitochondrial dysfunction in aging rats. Furthermore, genipin not only improved insulin sensitivity by promoting insulin-stimulated glucose consumption and glycogen synthesis, inhibited cellular ROS overproduction and alleviated the reduction of levels of MMP and ATP, but also reversed oxidative stress-associated JNK hyperactivation and reduced Akt phosphorylation in palmitate-treated L02 hepatocytes. In conclusion, genipin ameliorates age-related insulin resistance through inhibiting hepatic oxidative stress and mitochondrial dysfunction.

Monosodium iodoacetate induces apoptosis via the mitochondrial pathway involving ROS production and caspase activation in rat chondrocytes in vitro.

Monosodium iodoacetate (MIA) is an inhibitor of glyceraldehyde-3-phosphate dehydrogenase activity, and causes dose-dependent cartilage degradation resembling the pathological changes of human osteoarthritis (OA). In this study, we assessed the apoptosis induced by MIA and clarified the underlying mechanisms using the primary rat chondrocytes. The apoptosis of primary rat chondrocytes was analyzed by flow cytometry. The levels of mitochondrial membrane potential (ΔΨm) were evaluated using fluorescence spectrophotometer. The production of reactive oxygen species (ROS) was determined by fluorescence spectrophotometer. Apoptosis-related protein cytochrome c and procaspase-3 expressions were examined by Western blotting. We found that MIA treatment induces apoptosis in chondrocytes, as confirmed by increases in the percent of apoptotic cells, up-regulation of cytochrome c and caspase-3 protein levels. Treatment with MIA increases ROS production and decreases the levels of ΔΨm. The antioxidant, N-acetylcysteine (NAC), significantly prevented the production of ROS, the reduction of ΔΨm, the release of cytochrome c and the activation of caspase-3. Further, NAC completely protected the cells from MIA-induced apoptosis. Together these observations suggest that the mechanisms of MIA-induced apoptosis are primarily via ROS production and mitochondria-mediated caspase-3 activation in primary rat chondrocytes.

Effect of pyrroloquinoline quinone on neuropathic pain following chronic constriction injury of the sciatic nerve in rats.

Pyrroloquinoline quinone PQQ is a naturally occurring redox cofactor that acts as an essential nutrient, antioxidant, and redox modulator. PQQ has been demonstrated to oxidize the redox modulatory site of N-methyl-d-aspartic acid (NMDA) receptors. Such agents are known to be neuroprotective in experimental stroke models. However, there is not report about the therapeutic effect of PQQ on neuropathic pain. We tested the effects of oral administration of PQQ on neuropathic pain of rats with chronic constriction injury (CCI) of the sciatic nerve. The repeated oral administration of PQQ (20 and 40mg/kg, once a day for 4 weeks, from day 1 after the injury) attenuated both thermal and mechanical hyperalgesia, and also attenuated the muscle atrophy. The anti-hyperalgesic activity of PQQ was associated with a significant reduction of pro-inflammatory mediators such as tumor necrosis factor alpha (TNF-α) and lipid peroxide malondialdehyde (MDA) levels. In the present investigation, PQQ is shown to have analgesic effect which was found in the first time, probably through reducing the release of pro-inflammatory mediator and inhibiting oxidative stress.

[Establishment of the Chang liver cell line stably overexpressing human UCP2 gene and its effect on mitochondrial membrane potential and reactive oxygen species].

To establish the Chang liver cell line stably overexpressing human uncoupling protein 2 (UCP2) and observe the effect of UCP2 on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The Chang liver cell line was transfected with recombinant plasmid containing full-length human UCP2 cDNA (pcDNA3.1-hUCP2) or pcDNA3.1 empty vector. The stable cell line was established by antibiotic screening with Zeocin. UCP2 expression was detected by Western blotting and immunocytochemistry. The UCP2 overexpressing cells were pretreated with genipin at various doses (25, 50 and 100 munol/L). MMP and intracellular ROS were detected by fluorescence spectrophotometry. The total normalized protein content in UCP2 overexpressing cells was 1.6-fold higher than that in unmanipulated normal cells. The fluorescence intensities of Rhodamine123 and DCFH-DA in UCP2 overexpressing Chang liver cells (11.11+/-2.76 and 4.97+/-0.62, respectively) were significantly lower than those in unmanipulated normal cells (15.56+/-2.55, P less than 0.01 and 6.14+/-1.25, P less than 0.05, respectively) and in cells transfected with empty vector (16.11+/-2.93, P less than 0.01 and 6.23+/-1.13, P less than 0.05, respectively). Treatment of UCP2 overexpressing cells with 25, 50 and 100 munol/L genipin caused a dose-dependent increase in fluorescence intensities of Rhodamine123 (14.89+/-2.89, 17.89+/-2.93 and 24.00+/-2.55, respectively, all P less than 0.01) and DCFH-DA (9.16+/-0.78, 10.84+/-1.09 and 11.83+/-1.25, respectively, all P less than 0.01). The Chang liver cell line stably overexpressing UCP2 was established successfully. Using this cell system, UCP2 was found to play a role in mitochondrial function by regulating MMP and ROS.

Mechanisms of olive leaf extract-ameliorated rat arthritis caused by kaolin and carrageenan.

Olive leaf extract (OLE) has antioxidant and antiinflammatory actions. However, the role of OLE in mechanical inflammatory arthritis (osteoarthritis, OA) is unclear. This study investigated the effect of OLE on the development of kaolin and carrageenan-induced arthritis, a murine model of OA. Administration of OLE significantly ameliorated paw swelling, the paw Evans blue content and the histopathological scores. In the human monocyte cell line, THP-1, the OLE reduced the LPS-induced TNF-α production and was dose dependent. Croton oil-induced ear edema in mice also revealed that treatment with OLE suppressed ear edema, myeloperoxidase (MPO) production and was dose dependent. These results indicated that OLE is an effective antiarthritis agent through an antiinflammation mechanism. Also OLE may be beneficial for the treatment of OA in humans.

Olive leaf extract facilitates healing of experimental cartilaginous injuries in rabbits.

We investigated the restorative effect of orally administered olive leaf extract (OLE) on experimentally produced cartilaginous injuries in rabbits. In total, three holes in the left stifle joint, including one in the medial trochlear ridge and two in the trochlear sulcus (proximal and distal) of articular cartilage, were prepared surgically using a drill. For the control group only tap water alone was administered daily, and for the OLE group a water-based solution of OLE (500 mg/kg/day) was administered daily. The injured areas were observed macroscopically and histologically at 3 weeks after the operation. The results indicate that OLE facilitated healing of the three holes and increased the weight of the biceps femoris muscle. Histological examination revealed that in the OLE group, matured cartilage tissues and connective tissues were mixed with regenerated or maturing cartilage tissues with massive proliferation in the injured parts, around which the proliferation of undifferentiated blast cells and the tissue with cartilage substrates were observed. The histological score of the OLE group was significantly lower than that of the control group. The percentage of proliferating cell nuclear antigen-positive cartilage cells in the OLE group was higher than in the control group. Mean density of the restored area observed with Safranin O staining was higher in the OLE group than in the control group. Therefore, OLE is effective for enhancing the healing of cartilaginous injuries. OLE may also have a beneficial effect of slowing and reducing the pathogenesis of degenerative joint diseases in humans.

[Establishing the model of kappaB-decoy inhibiting the activity of NF-kappaB in PC12 cells].

OBJECTIVE: Observing the time course and establishing the model of kappaB-decoy oligodeoxynucleotides (rcB-decoy) inhibiting the activity of NF-kappaB in the PC12 cells. METHODS: PC12 cells cultivating in the 6 wells plate were divided into 3 groups, experimental group: adding kappaB-decoy complex (6 microg DNA/well), the control group: adding scrambled-decoy complex, the normal group: adding lipid-Lipofectamine 2000, transfer and cultivate 48 h, then lipopolysaccharide (LPS, 200 ng/ml) was added in the cells for 0.5-4 h. The immunocytochemistry and Western blot were used to measure the expression or the activity of NF-kappaB in PC12 cells. RESULTS: In PC12 cells, compared with normal group, the expression of NF-kappaB enhanced obviously with the time of the stimulation of LPS in scrambled-decoy treated control group (P < 0.01), in 2-4 h the level reached the peak; the expression of NF-kappaB showed the stable level with the time of the stimulation of LPS in kappaB-decoy treated experimental group, compared with the control group, the expression levels were obviously lower than the respective time point of control groups (P < 0.01). CONCLUSION: kappaB-decoy could reduce the expression of NF-kappaB in the normal PC12 cells and inhibit the activity of NF-cB in the pathologic PC12 cells.

Protective effects of estrogen against reperfusion arrhythmias following severe myocardial ischemia in rats.

BACKGROUND: Female sex hormones may have protective effects against arrhythmias, including reperfusion arrhythmias (RAs), but the mechanisms are still not completely known. METHODS AND RESULTS: Serial changes in rat hearts (rhythm, apoptosis and the its infuencing factors; cardiac vinculin mRNA expression and connexin43 (Cx43) dephosphorylation) were examined during periods of ischemia-reperfusion with and without estrogen treatment. After reperfusion, although the incidence of arrhythmias became higher in both the vehicle-group and estrogen-group, compared with the ischemia period, estrogen prevented reperfusion-induced upregulation of the incidence of arrhythmias, especially ventricular premature beats (VPB) and ventricular tachycardia (VT). The duration of VT and fibrillation, and the number of VPB and VT, were all significantly decreased in the estrogen-group. The expression of cardiac vinculin mRNA decreased significantly in the vehicle-group but not in the estrogen-group. Cx43 dephosphorylation and myocyte apoptosis increased in both groups, but the values for the estrogen-group were all markedly lower than those for the vehicle-group. A selective estrogen receptor (ER) beta agonist prevented reperfusion-induced upregulation of the incidence of both VPB and VT significantly; a selective ERalpha agonist had no significant influence. CONCLUSIONS: Estrogen can protect the heart against RAs, at least in part, mediated through gap junctions. Upregulation of ERbeta but not ERalpha mediated most of the estrogen-induced cardioprotection against RA.

Neural reflex hypotension induced by very small dose of hypertonic NaCl solution in rats.

Although the precise mechanisms of transient hypotension after intravenous infusion of hypertonic saline (HTS) are not yet clarified, a rapid infusion of HTS is widely used as the initial therapy of hypovolemia. We investigated the effect of the intravenous infusion of a small dose of 0.97-9.7% NaCl solutions in anesthetized rats. Intravenous infusion of HTS at a rate of 0.3 ml/kg/min for 1 min produced the transient hypotension lasting for several minutes. The depressor response to HTS was not abolished by bilateral cervical vagotomy. The HTS infusion into the femoral vein evoked the depressor response with a larger magnitude and a shorter latency than that into the aortic arch. While the arterial baroreceptor pressure was kept constant at the baseline level of systemic arterial pressure, HTS-induced hypotension was significantly augmented. The gain factor of the arterial baroreflex was reduced by intravenous HTS. Pretreatment with bretylium tosylate completely abolished the depressor response without affecting the baseline level of arterial pressure. These results suggest that the depressor response to the very small dose of intravenous HTS is the sympathosympathetic neural reflex with cardiopulmonary afferents and vasomotor efferents.

Cervical heterotopic kidney transplantation in rats using non-suturing and preserving-bag techniques.

BACKGROUND: This study describes a simple and stable cervical heterotopic kidney transplantation method in rats that uses artery sleeve anastomosis, vein cuff anastomosis and preserving-bag techniques. METHODS: The donor graft, consisting of kidney, renal vein (RV), renal artery (RA) and a ureterocystic flap, was removed en bloc and perfused in situ. The donor RA was end-to-end anastomosed to the recipient left common carotid artery (CCA) using a sleeve anastomosis, and the donor RV was connected to the recipient right external jugular vein (EJV) using a cuff technique. During the vascular anastomosis, the kidney graft was placed in a lactated Ringer's solution ice-water preserving bag. The donor bladder patch was exteriorized to form cervical cutaneous stoma. RESULTS: A total of 104 heterotopic renal transplantations were performed, which included pre-experimental (62 operations) and experimental stages (42 operations). The success rates of the two stages were 80.6% and 95.2%, respectively. The time for surgery was 40 +/- 6 min, the average time for donor surgery was 20 +/- 5 min, the preparation time for the graft was 8 +/- 2 min, the operative time for the recipient was 18 +/- 3 min that included the time for the arterial anastomosis (5 +/- 2 min) and venous anastomosis (2 +/- 1 min), the cold ischaemic time of the graft was 15 +/- 3 min and the warm ischaemic time of the graft was 2 +/- 1 min. CONCLUSION: We developed an easy and reliable model of cervical heterotopic kidney transplantation that may provide a useful tool for investigating kidney transplantation rejection and retransplantation pathology.

Protection of regenerating liver after partial hepatectomy from carbon tetrachloride hepatotoxicity in rats: roles of mitochondrial uncoupling protein 2 and ATP stores.

Uncoupling protein 2 (UCP(2)), an inner mitochondrial membrane protein, can limit the generation of reactive oxygen species (ROS) and protect cells from injuries mediated by oxidative stress. We investigated the effect of upregulation of UCP(2) in the regenerating liver 96 h after 68% partial hepatectomy (PH) on the self-protection of regenerating liver against carbon tetrachloride (CCl(4)) poisoning. Hepatotoxicity was induced in vivo by administering CCl(4) to rats that had undergone PH. After CCl(4) poisoning, the regenerating liver appeared to have less histological damage and lower serum alanine aminotransferase (ALT) levels. Lower malondialdehyde production and higher glutathione contents were also observed in the regenerating liver compared with the sham-operated liver after CCl(4) poisoning. UCP(2) expression was markedly elevated in the regenerating liver, and further increased after CCl(4) intoxication. Mitochondrial membrane potential and adenosine triphosphate stores maintained higher levels in the regenerating liver than in sham-operated liver after CCl(4) intoxication. The results showed that the regenerating liver exhibited a potent ability to resist CCl(4) intoxication, and the autoprotection of regenerating liver might result from reduction of ROS by UCP(2) and maintenance of higher ATP stores.

Effects of hydroxytyrosol-20 on carrageenan-induced acute inflammation and hyperalgesia in rats.

Hydroxytyrosol (HT) is a simple phenol compound extracted from olive leaves. The content of HT in the studied preparation was about 20%, and the preparation was called hydroxytyrosol-20 (HT-20). HT has antioxidant and antiinflammatory activities. There has been no report so far on the efficacy of HT-20 in carrageenan-induced acute inflammation and hyperalgesia in rats. Therefore, the aim of this study was to assess the inhibitory role of HT-20 on carrageenan-induced swelling and hyperalgesia of rat paw. Paw inflammation was assessed by the increase in paw volume and hyperalgesia. The rat paws were cut out under ether anesthesia at 270 min after administration of carrageenan. The tissue of the right paw was isolated separately from the individual rat. The levels of the tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta) and interleukin 10 (IL-10) mRNA in the tissue were estimated by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the paw pressure thresholds of rats orally administered HT-20 significantly increased at 210, 240 and 270 min after administration of carrageenan, compared with corresponding basal paw pressure thresholds; the degree of swelling of the right hind paw showed a statistically significant reduction, compared with rats in the carrageenan-treated control. In this model, HT-20 appears to decrease pro-inflammatory cytokines IL-1beta and TNF-alpha and not to increase the antiinflammatory cytokine mRNA expression of IL-10.

[Study on surgical techniques for cervical ectopic renal transplantation in rat].

OBJECTIVE: To establish a simple and stable cervical ectopic renal transplantation rat model that increase surgical successful rate. METHODS: A total of 208 male inbred Wistar rats (weighing 220-260 g) were randomly served as donors and recipients. The graft consisting of kidney, renal vein (RV) and renal artery (RA) was obtained, and perfused in situ. The donor RA was end-to-end anastomosed to the recipient left common carotid artery (CCA) by using of "sleeve" anastomosis, and the donor RV to the recipient right external jugular vein by using of "cuff" technique. The distal end of the ureter was brought out to form cervical cutaneous stomas. RESULTS: A total of 104 ectopic renal transplantations were performed in rats, including stages of the pre-experiment (62 operations) and experiment (42 operations). The success rates of the two stages were 80.6% and 95.2%, respectively. The causes of failure in the pre-experimental stage were anesthesia accidents, thrombosis of the arterial anastomosis, massive hemorrhage, air embolism and phlebemphraxis. In the experimental stage, 2 rats died due to late anastomotic hemorrhage and thrombosis. The remaining 40 transplanted kidney survived more than 6 months. The time for surgery was (40 +/- 6) minutes, the average time for donor surgery was (20 +/- 5) minutes, the preparation time for the graft was (8 +/- 2) minutes, the operative time for the recipient was (18 +/- 3) minutes, including the time for the arterial anastomosis (5 +/- 2) minutes and venous anastomosis (2 +/- 1) minutes, the cold ischemia time of graft was (15 +/- 3) minutes. CONCLUSION: The cervical ectopic renal transplantation technique has the advantages of easy-and fast-to-perform, shorter operation and cold ischemia time, higher successful rate.