ABSTRACT
BACKGROUND: Hibiscus Mutabilis L. Is a very effective Chinese traditional herb for non-specific inflammation in the clinic adopted by Professor Wei Zhixin, the eminent expert in Traumatology Department in China, lasting for near 60 years. In order to explain its pharmaceutical mechanism, the research of Hibiscus Mutabilis L. Was carried on in the model of ischemic reperfusion injury to explain its mechanism on anti-inflammation.OBJECTIVE: To observe the protection of the effective fraction of Hibiscus Mutabilis L. (MFR) on renal ischemic reperfusion injury in rats to probe into the mechanism of MFR on anti-inflammation.DESIGN: Randomized and controlled study on experimental animal.SETTING: A Municipal Institute of Traumatology and Orthopedic Department.MATERIALS: The experiment was performed in the Key Experiment Room (Institute of Traumatology and Orthopedic Department, Shanghai) of State Administration of Traditional Chinese Medicine, in which, 55 male Wistar rats were employed.METHODS: The rat model of renal ischemic reperfusion was adopted and MFR was applied for gastric perfusion to determine serum blood urea nitrogen(BUN) and serum creatinine(Scr) and the level of interleukin-1 (IL-1).MAIN OUTCOME MEASURES: ① Changes in serum BUN and Scr; ②Effect of MFR on level of serum IL-1.RESULTS: After MFR treatment, the renal function was improved remarkably after ischemic reperfusion in rats. In sham-operation group, serun BUN was (6.72 ± 1.30) mmol/L and serum Scr was (38.40 ± 6.23) μmol/L. In 24hours of reperfusion after one-hour ischemia, BUN was(60. 72±4.64)mmol/L in the control and(47.34 ± 8.32) mmol/L in the MFR treatment group, and the significant difference presented between two groups(t=2.562, P ＜ 0.05) .Serum Scr was(347.95±95) μmol/L and(518.20 ± 41.15) μmol/L in the treatment group and the control respectively, indicating significant difference ( t = 3.69, P ＜ 0.01 ) . Concerning to the effect on IL-1, in 1 hour of reperfusion after 1 hour renal ischemia,IL-1 was( 122.79 ±27.56) ng/L in the MER treatment group,(180. 28 ±33. 15) ng/L in the control group, indicating that IL-1 level in the treatment group was remarkably superior to that in the control group(t = 2.98, P ＜ 0.05). In 3 hours of reperfusion after 1 hour ischemia, level of IL-1 in the treatment group and control group was(15.58±8.59) ng/L and (34. 13±± 10. 02) ng/L respectively, indicating significant difference( t = 3.14, P＜ 0.05).CONCLUSION: MFR provides protection on renal ischemic reperfusion injury, which probably is related to its inhibition on IL-1 formation.
ABSTRACT
OBJECTIVE: To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. METHODS: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. RESULTS: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro alpha1 (III) in fibroblasts and some chondrocyte-like cells were dominant; and at the end of second week high expression of type-II procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type-I procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus. CONCLUSIONS: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional tec hnique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.