| Objective: This study established animal models of pain, inflammatory and immune- abnormal respectively, and employed GSF's ethanol extract on models to observe the therapeutic effects of GSF's ethanol extract as well as the mechanisms. The purpose is to provide lab groundwork data and lay a foundation for the future research and development, clinical application of GSF's ethanol extract.Methods: 1. test of hot-plate and acetic acid induced mice peritonitis: NIH mice, develop them as the peritonitis models by celiac injection of acetic acid. The times of Painful writhing within ten minutes subsequently act as the observation index of painful reaction threshold after stimulation. Hot-pad test set the mice models of the reflecting pain. The time from touching hot-plate to licking aching hind paws is defined index of painful reaction threshold. The two tests above-mentioned divide mice into four groups at random: control group(administered with NS), aspirin group(administered with aspirin in hot-pad test, in peritonitis test with morphine),GSF Extracts high-dosage group(60g herb/kg), GSF Extracts low-dosage group(20g herb/kg).2. test of carrageen induced rats'hind paw swelling: Wistar rats, five groupsdivided at random: control group (administered with NS), indometain group(administered with indometain), GSF high-dosage group(42g herb/kg), GSF mid-dosage group(28g herb/kg), GSF low-dosage group(14g herb/kg), figure out the right paw volume on the first day and inject 0.1ml 1% carrageen solution in right hind paw volars lh after last administration, use paws swelling degree as estimating index, detect the PGE level in rats'hind paw of injection, denote by absorptivity in inflammatory tissue per gram.3. test of paws swell induced by egg white: Wistar rats, four groups divided at random :control group (administered with NS), indometain group(administered with indometain), GSF high-dosage group(42g herb/kg), GSF mid-dosage group(28g herb/kg), GSF low-dosage group(14g herb/kg), ig, figure out the right paw volume on the first day and inject 0.1ml 10% egg solution in right volars lh after last administration.Use paws swelling degree as estimating index.4. test of acute peritonitis induced by acetic acid: Kunming mice, five groups divided at random: control group (administered with NS),aspirin group(administered with aspirin), GSF high-dosage group(60g herb/kg), GSF mid-dosage group(40g herb/kg), GSF low-dosage group(20g herb/kg), ig, measure OD value of celiac effusion as anti-inflammation effect index.5. test of gasbag synovitis on mice's back induced by egg white: Kunming mice, five groups divided at random:control group (administered with NS), model group(administered with NS), aspirin Group(administered with aspirin), GSF high-dosage group(60g herb/kg), GSF mid-dosage group(40g herb/kg), GSF low-dosage group(20g herb/kg).Use the white blood cells count of inflammatory extravasate in gasbag on rats' back as anti-inflammation effect index.6. test of AA: Wistar rats, six groups divided at random: control group (administered with NS), model control group(administered with NS), Dexamethasonegroup(administered with dexamethasone), GSF high-dosage group(42g herb/kg), GSF mid-dosage group(28g herb/kg), GSF low-dosage group(14g herb/kg), figure out the right paw volume before and after test as well as the next hind paws, measure the right paws volume at different interval and administer again after secondary inflammation presence, calculate non-inflammatory paws swell degree, weigh the rats.When test finished weigh out the spleen and thymus gland, observe the pathological alteration of AA rats' ankle tissue structure under microsope, collect the blood samples three times respectively as before injection, after injection and without admistration drug, after the test finish. Centrifugal sera and measure the content of IL-1 P and TNF-a according to the reagent box requirement.7. test of paws swelling induced by carrageen in rats without adrenal gland: Wistar rats, remove adrenal glands, raise one week with NS, four groups divided at random: control group (administered with NS), model control group(administered with NS), Dexamethasone group(administered with), GSF high-dosage group(42g herb/kg), GSF low-dosage group(14g herb/kg), ig. figure out the right paw volume and estimate the anti-inflammation effect of drug.Choose other Wistar rats without adrenal gland removal and carrageen injection, grouping in same way as above-mentioned, ig, get sera from blood samples after last administration, detect the content of COR by radio-immune techniques according to the reagent box requirement.8. test of monocyte lick up:Kunming mice, four groups divided at random: control group (administered with NS), Tripterygium Glucosides group(administered with Tripterygium Glucosides), GSF high-dosage group (60g herb/kg), GSF low-dosage group of (20g herb/kg), ig. Detect the OD value of mice blood by spectrophotometer around 650nm. weigh out spleens and thymus glands, and determine the spleen and thymus gland index. Use K index to describe the immunoladjustment function of GSF extracts.9.test of hemolysin:NIH mice, four groups at random:control group(administered with NS), Tripterygium Glucosides group(administered with Tripterygium Glucosides), GSF high-dosage group(60g herb/kg), GSF low-dosage group(20g herb/kg), ig, celiac injection with 2% SRBC fluid 0.2ml on third day, execute mice after last administration on seventh day, weigh the thymus glands and spleens, calculate index of thymus gland and spleen, extract blood samples from eyeball removal and separate sera, detect colorimetric value around 540nm, compute HC50.Results: l.The result of hot-pad test revealed that times of wrest reaction distinctly decreased in GSF high-dosage or low-dosage group. It showed GSF had anti-inflammation effect(P<0.05).Meanwhile, the result demonstrated GSF appeared effects above dosage of 20 herb/kg 1.5h later. As a result, the pain threshold of mice was elevated (P<0.05).2. In test of paws swell induced by carrageen, the swelling degree of rats' paws in GSF mid-dosage group decreased distinctly 2h, 4h, 6h, 8h, 12h after carrageen injection(P<0.01> P<0.05^ P<0.01> P<0.0:L P<0.01). The swelling degree in model group spurred on the peak value in 4~6hrs after administration.3. In test of paws swell induced by egg white, the anti-inflammatory effect in GSF low-dosage group were remarkable 30ms and 90ms after injection. The swelling was restrained by GSF prominently(P<0.05 -. P<0.05). The mid-dosage group, however, presented the anti-inflammation effect 60ms and 90ms later (P<0.05, P<0.05) .4. In the capillary vessel penetration test, the OD value of celiac effusion in GSF mid-dosage group decreased distinctly (P<0.05) .5. The result of gasbag test showed that the WBC count of synovitis fluids in three GSF intervening groups reduced without exception (P<0.01) . There were nodistinct deference between three GSF groups and aspirin group.6. In test of AA , all three GSF intervening groups similarly alleviated the swelling degree of primary and secondary damage in AA rats (P<0.01, P<0.05) . No linear relation showed up between the drug's effect and dosage. The immune organs were influenced in various degree, such as spleen index decrease in GSF high-dosage group (P<0.05) , thymus index increase in GSF mid-dosage group(P<0.05).7. In test of paws swell induced by carrageen in rats without adrenal gland, the paws swelling degree reduced in GSF high-dosage and low-dosage groups at the interval of 4h~8h after administration (P<0.05 or P<0.01) . Moreover, GSF high-dosage and low-dosage groups had no influence on the content of steroid in sera.8. The result of paws swell induced by carrageen test discovered that PGE content of the inflammatory diffused fluid diminished in GSF low-dosage group(P<0.05) .9. In AA test, IL-lBsera content of blood sample in GSF high-dosage group was much lower than in model control group (P<0.05) .There were no distinct statistic discrepancy but tendency of reduction of TNF-a, IL-18 sera content between other therapeutic groups and model control group.lO.The result of macro-reticular resin test and hemolysin test revealed that GSF high-dosage group had lower paeonol. There was no statistic difference but reductive tendency of hemolysin between in GSF high, low dosage groups and model control group. The thymus gland index grew lower in both high and low dosage groups (P<0.05) . The spleen index tended to decline in GSF high-dosage group and Tripterygium Glucosides group. However, the spleen index increased statistically in GSF low-dosage group (P<0.05) .Conclusions :1.GSF could increase obviously the pain threshold in terms of chemicals and physical stimulation.2.GSF could fight against inflammation at different stages of acute inflammation, resulting from inhibiting extravasate WBC movement, depressing inflammatory capillary's penetrability and reducing swelling.3.GSF could amend the pathological process and symptoms of primary and secondary inflammation of AA rats. There were bidirectional modulating effect on the immune organs of model rats and GSF made the rats no weight loss. It revealed that GSF showed no apparent interference against rat's alimentary system.4.There were no linear relativity of anti-inflammation between GSF and HPPA, and probably GSF depressed the production of PGE> IL-16^ TNF-a and so on.5.GSF demonstrated bilateral functions in mice. One side, it restrained unspecial immune function. On the other hand, it possessed both enhancing and reducing special idio-immune function of humour.
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