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Long-acting analgesia is a common clinical treatment method after surgery. The slow-release injection with long-acting analgesia has the advantages of less medication frequency and stable effect. In this study, the analgesic drug lappaconitine hydrobromide lyotropic liquid crystal injection was prepared, and its sustained release mechanism, drug release and pharmacodynamic characteristics were evaluated. The results of polarizing microscope and freeze-transmission electron microscope showed that the lyotropic liquid crystal injection of the liquid crystal precursor preparation of lappaconitine hydrobromide could be obtained by the combination of glycerol monooleate (GMO) and soybean lecithin (SPC) in different proportions. The results of dissolution study in vitro showed that the drug release rate of different forms of liquid crystal preparations was layered liquid crystal > cubic liquid crystal > hexagonal liquid crystal. The mathematical model fitting results of the release data showed that the external release of layered liquid crystal, cubic liquid crystal and hexagonal liquid crystal conforms to the Ritger-Peppas model, and the release mechanism was Fick diffusion. The results of pharmacodynamics study in vivo showed that the analgesic effect of lappaconitine hydrobromide lyotropic liquid crystal injection lasted for 3 days, and there was no abnormality in the incision and local tissue, showing good safety and tolerance. The study on drug release and elimination process of the in vivo gel repository showed that lappaconitine hydrobromide could be completely released from the lyotropic liquid crystal 3 days after administration, and the sustained-release materials could be gradually eliminated locally. All animal experiments were approved by the Experimental Animal Ethics Committee of the Shanghai Institute of Materia Medica, Chinese Academy of Sciences (No. 2021-08-GY-61) and the experiments were conducted in accordance with the relevant guiding principles and regulations. The lyotropic liquid crystal injection of lappaconitine hydrobromide prepared in this study presented a solution state at room temperature, and underwent phase transition in contact with the body fluid at the administration site, formed a drug depot and exerted a slow drug release effect. This preparation can reduce systemic toxicity, prolong the duration of analgesia, reduce the number of administrations, improve the compliance of postoperative patients, and provide a reference for the design of long-term sustained release analgesic preparations.
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Quantitative evaluation of analgesic efficacy improves understanding of the antinociceptive mechanisms of new analgesics and provides important guidance for their development. Lappaconitine (LA), a potent analgesic drug extracted from the root of natural Aconitum species, has been clinically used for years because of its effective analgesic and non-addictive properties. However, being limited to ethological experiments, previous studies have mainly investigated the analgesic effect of LA at the behavioral level, and the associated antinociceptive mechanisms are still unclear. In this study, electrocorticogram (ECoG) technology was used to investigate the analgesic effects of two homologous derivatives of LA, Lappaconitine hydrobromide (LAH) and Lappaconitine trifluoroacetate (LAF), on Sprague-Dawley rats subjected to nociceptive laser stimuli, and to further explore their antinociceptive mechanisms. We found that both LAH and LAF were effective in reducing pain, as manifested in the remarkable reduction of nocifensive behaviors and laser-evoked potentials (LEPs) amplitudes (N2 and P2 waves, and gamma-band oscillations), and significantly prolonged latencies of the LEP-N2/P2. These changes in LEPs reflect the similar antinociceptive mechanism of LAF and LAH, i.e., inhibition of the fast signaling pathways. In addition, there were no changes in the auditory-evoked potential (AEP-N1 component) before and after LAF or LAH treatment, suggesting that neither drug had a central anesthetic effect. Importantly, compared with LAH, LAF was superior in its effects on the magnitudes of gamma-band oscillations and the resting-state spectra, which may be associated with their differences in the octanol/water partition coefficient, degree of dissociation, toxicity, and glycine receptor regulation. Altogether, jointly applying nociceptive laser stimuli and ECoG recordings in rats, we provide solid neural evidence for the analgesic efficacy and antinociceptive mechanisms of derivatives of LA.
Asunto(s)
Animales , Ratas , Aconitina/farmacología , Analgésicos/farmacología , Preparaciones Farmacéuticas , Ratas Sprague-DawleyRESUMEN
Quantitative evaluation of analgesic efficacy improves understanding of the antinociceptive mechanisms of new analgesics and provides important guidance for their development. Lappaconitine (LA), a potent analgesic drug extracted from the root of natural Aconitum species, has been clinically used for years because of its effective analgesic and non-addictive properties. However, being limited to ethological experiments, previous studies have mainly investigated the analgesic effect of LA at the behavioral level, and the associated antinociceptive mechanisms are still unclear. In this study, electrocorticogram (ECoG) technology was used to investigate the analgesic effects of two homologous derivatives of LA, Lappaconitine hydrobromide (LAH) and Lappaconitine trifluoroacetate (LAF), on Sprague-Dawley rats subjected to nociceptive laser stimuli, and to further explore their antinociceptive mechanisms. We found that both LAH and LAF were effective in reducing pain, as manifested in the remarkable reduction of nocifensive behaviors and laser-evoked potentials (LEPs) amplitudes (N2 and P2 waves, and gamma-band oscillations), and significantly prolonged latencies of the LEP-N2/P2. These changes in LEPs reflect the similar antinociceptive mechanism of LAF and LAH, i.e., inhibition of the fast signaling pathways. In addition, there were no changes in the auditory-evoked potential (AEP-N1 component) before and after LAF or LAH treatment, suggesting that neither drug had a central anesthetic effect. Importantly, compared with LAH, LAF was superior in its effects on the magnitudes of gamma-band oscillations and the resting-state spectra, which may be associated with their differences in the octanol/water partition coefficient, degree of dissociation, toxicity, and glycine receptor regulation. Altogether, jointly applying nociceptive laser stimuli and ECoG recordings in rats, we provide solid neural evidence for the analgesic efficacy and antinociceptive mechanisms of derivatives of LA.
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OBJECTIVE: To prepare lappaconitine(LA)-loaded microemulsion with olive oil and study its morphology, particle size, drug loading capacity, drug release behavior and rheological characteristics. METHODS: Pseudo-ternary phase diagram method was used to screen and prepare LA-loaded microemulsion with olive oil. LA release properties in vitro were investigated by dynamic dialysis method. The rheological properties of LA-loaded microemulsion with olive oil were studied using MCR 301 rheometer. RESULTS: The optimal formulation of LA-loaded microemulsion with olive oil was as follows:olive oil was the oil phase, castor oil polyoxyethylene ether-40/span-80(4:1) was the surfactant, glycerin was the cosurfactant, and Km=3. The morphology of the microemulsion particles was round or oval, and the average particle size was(91±0.55) nm. The drug-loading rate of LA in the microemulsion was 1.85%.Drug release experiments in vitro showed that the microemulsion had a sustained release effect on LA, and the drug release behavior was more suitable to be described by Higuchi equation. Rheological experiments showed that the fluid of LA-loaded microemulsion with olive oil belong to pseudoplastic fluid of non-Newtonian fluid exhibiting thixotropic and shear-thinning fluid behavior as well as certain viscoelasticity. CONCLUSION: The LA-loaded microemulsion witholive oil are successfully prepared, and the microemulsion has ideal sustained release behavior and good rheological properties. The study provides a foundation for the developmenton preparation of LA-loaded microemulsion with olive oil.
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OBJECTIVE: To observe the effects of lappaconitine sulfate on proliferation, cycle and apoptosis of human cerical neoplasm Hela cells and its possible mechanism. METHODS: Cell Counting Kit-8 (CCK-8) was adopted to observe the inhibitory effects of lappaconitine sulfate on the proliferation of Hela cells and calculated its half maximal inhibitory concentration (IC50). The technology of flow cytometry, propidium iodide (PI) single stained method and Annexin-V-FITC/PI double stained method were employed to measure the cell cycle and apoptosis of Hela cells after lappaconitine sulfate treatment. RESULTS: Lappaconitine sulfate could inhibit the proliferation of human cerical neoplasm Hela cells in vitro, and the inhibition presented obvious dose-effect relationship (r=0.990 5, P<0.01). The IC50 of lappaconitine sulfate on Hela cells was 0.421 mg·mL-1. After dealt with lappaconitine sulfate, Hela cells indicated remarkable cell cycle arrest at G0/G1 phase, and the apoptotic ratio was gradually increased with the concentration of lappaconitine sulfate. CONCLUSION: Lappaconitine sulfate could inhibit the growth of human cerical neoplasm Hela cells, change the distribution of cell cycles and induce their apoptosis.
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Objective@#To explore the effects of lappaconitine (LA) on pain and inflammatory response of severely burned rats and the mechanism.@*Methods@#Forty SD rats were divided into healthy+ normal saline group, sham injury+ normal saline group, pure burn group, burn+ LA group, and healthy+ LA group according to the random number table (the same dividing method below), with 8 rats in each group. Rats in pure burn and burn+ LA groups were inflicted with about 32% total body surface area deep partial-thickness scald (hereinafter referred to as burn) on the back and right hind. Rats in sham injury+ normal saline group were sham injured. Rats in burn+ LA group were intraperitoneally injected with 1 g/L LA solution in the dosage of 4 mL/kg at 2.0 h before injury and post injury hour (PIH) 0 (immediately), 24.0, 48.0, and 72.0. Rats in healthy+ LA group were intraperitoneally injected with LA solution in the same dose at the same time points as above, and rats in healthy+ normal saline and sham injury+ normal saline groups were intraperitoneally injected with normal saline in the dose of 4 mL/kg at the same time points as above. At 1.5 h before injury and PIH 12.5, 24.5, 36.5, 48.5, and 72.5, the paw withdrawal mechanical threshold (PWMT) of injured rats was detected, and their pain behaviors were observed. The same observation and detection were conducted in rats without injury in the two groups at the same time points as above. Another 32 SD rats were divided into normal saline group, trinitrophenyl (TNP)-ATP group, minocyline group, pyridoxal-phosphate-6-azophenyl-2', 4'-disulfonic acid (PPADS) group, with 8 rats in each group, and all the rats were inflicted with the same burn injury as above. At PIH 48.0, rats in normal saline group were intrathecally injected with 10 μL normal saline; rats in TNP-ATP group were intrathecally injected with 10 μL TNP-ATP in the concentration of 30 nmol/μL; rats in minocyline group were intrathecally injected with 10 μL minocyline in the concentration of 5 g/L; rats in PPADS group were intrathecally injected with 10 μL PPADS in the concentration of 10 nmol/μL. The PWMT of rats was detected at 0.5 h before injection and 0.5 h after. At PIH 72.5, the tissue in the dorsal horn of spinal cord of rats in sham injury+ normal saline, pure burn, and burn+ LA groups was harvested to observe the co-expression of P2X4 receptor and OX42 receptor with immunofluorescent staining and to observe the expression of P2X4 receptor and count the positive cells with immunohistochemical staining. The venous blood was harvested for determination of serum content of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) with enzyme-linked immunosorbent assay. The same observation and determination were conducted in rats without injury in the two groups at the same time point as above. Data were processed with one-way analysis of variance, analysis of variance for repeated measurement, SNK test, paired t test, and Bonferroni correction.@*Results@#(1) There were no abnormal activity in rats of healthy+ normal saline, sham injury+ normal saline, healthy+ LA groups at all time points. Until PIH 72.5, rats in pure burn group were in poor mental state; red and swollen manifestation and blister were observed in burn wounds on the back and right hind; imbalance in gait, lick, bite, and scratch were observed occasionally. Fewer behaviors such as lick, bite, and limp were observed in rats in burn+ LA group than in pure burn group, and the red and swollen manifestation in wounds of rats in burn+ LA group dissipated faster than that in pure burn group. (2) At 1.5 h before injury, there were no significant differences in the PWMT values of rats in healthy+ normal saline, sham injury+ normal saline, pure burn, burn+ LA, and healthy+ LA groups (F=0.106, P>0.05). PWMT values of rats in pure burn group were significantly lower than those in the other 4 groups at all post injury time points (with P values below 0.05). PWMT values of rats in burn+ LA group were significantly lower than those in healthy+ normal saline, sham injury+ normal saline, and healthy+ LA groups at all post injury time points (with P values below 0.05). (3) At 0.5 h before injection, PWMT values of rats in normal saline, TNP-ATP, PPADS, and minocyline groups were close, respectively 15.3±0.8, 15.1±1.0, 15.3±0.9, and 15.6±1.1 (F=0.343, P>0.05). At 0.5 h after injection, PWMT values of rats in normal saline group and PPADS group were respectively 15.2±1.2 and 14.8±1.0, which were significantly lower than 20.8±1.4 and 26.3±1.0 in TNP-ATP group and minocyline group respectively (with P values below 0.05). PWMT values of rats in normal saline and PPADS groups were similar before and after injection (with t values respectively 0.073 and -0.772, P values above 0.05), while those of rats in TNP-ATP and minocyline groups were higher after injection than before injection (with t values respectively -10.180 and -20.813, P values below 0.01). (4) At PIH 72.5, co-expression of P2X4 receptor and OX42 receptor was observed in a few microglias of rats in healthy+ normal saline, sham injury+ normal saline, and healthy+ LA groups, while co-expression of P2X4 receptor and OX42 receptor was observed in a large number of microglias of rats in pure burn and burn+ LA groups. At PIH 72.5, more P2X4 receptor positive cells were observed in rats in pure burn group than in the other 4 groups (with P values below 0.05), and more P2X4 receptor positive cells were observed in rats in burn+ LA group than in healthy+ normal saline, sham injury+ normal saline, and healthy+ LA groups (with P values below 0.05). (5) At PIH 72.5, the serum content of TNF-α and IL-1β of rats in pure burn group was significantly higher than that in the other 4 groups (with P values below 0.001). The serum content of TNF-α and IL-1β of rats in burn+ LA group was significantly lower than that in healthy+ normal saline, sham injury+ normal saline, and healthy+ LA groups (with P values below 0.001).@*Conclusions@#LA has significant analgesic effects on severely burned rats, and it can ameliorate the excessive inflammational situation. The mechanism may be related to its inhibition of expression of P2X4 receptor in microglias in the dorsal horn of spinal cord and reduction in the release of inflammatory factors TNF-α and IL-1β.
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Objective To prepare lappaconitine hydrobromide push-pull osmotic pump controlled release tablets and screen for the optimal formulation. Methods The percent of cumulative release was used as the evaluation index for the drug release profile in vitro. The effects of PEO N750, PEO WSR303, and plasticizer amounts and coating weight gain on the releasing behavior were investigated through single-factor method. Based on single-factor study on the compositions, the optimal formulation for lappaconitine hydrobromide push-pull osmotic pump controlled release tablet was selected via orthogonal design. The in vitro release of the optimized formulation was also fitted to different models. Results The results of orthogonal design indicated that coating weight gain had a significant effect on the drug release in vitro —<0. 05). The optimal formula was as follows; lappaconitine hydrobromide 20 mg, PEO N750 160 mg, NaCl 30 mg in drug layer; PEO WSR303 75 mg, NaCl 20 mg in the push layer; and plasticizer PEG 4000 was 10% and weight gain was 5% in the coating composition The release rate of the tablets with optimized formulation was constant within 12 h, and the cumulative release could reach 95. 02 %. Conclusion The current method to prepare lappaconitine hydrobromide push-pull osmotic pump controlled release tablets is stable, and the in vitro drug release has an excellent zero-release profile within 12 h (r=0. 992 1), which meets the standard for controlled release.
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OBJECTIVE: To prepare lappaconitine (LA)-loaded chitosan/sodium β-glycerophosphate (CS/β-GP) thermosensitive hydrogels and investigate its phase transition mechanism of gel formation process and release properties in vitro. METHODS: The injectable CS/β-GP thermosensitive hydrogels were prepared with biodegradable CS as carrier material and β-GP as coagulation accelerator.The release behavior in vitro was studied by dynamic dialysis, and the phase transition mechanism of gel formation process was further investigated by rheologieal method. RESULTS: The optimized process condition was as follows;the concentration of β-GP and CS was 560 and 22 mg·mL-1, respectively, CS was dissolved by 0.1 mol·L-1 HOAc, and the valume ratio of CS to β-GP was 8.75:1.25 (V/V), the gelation time of CS/β-GP thermosensitive hydrogels with volume ratio of 8.75:1.25 (V/V) at 37℃ was 5 min 38 s.The in vitro release study showed that these injectable CS/β-GP thermosensitive hydrogels had sustained release effect for LA, and the release behavior could be well described by the Higuchi model and Korsmeyer-Peppas model.The mechanism of LA releasing from CS/β-GP thermosensitive hydrogels was attributed to drug dissolution and diffusion.Rheologieal studies showed that the CS/β-GP thermosensitive hydrogels belonged to thixotropic system and exhibited non-Newtonian and shear-thinning fluid behavior as well as "solid-like" gelatin behavior. CONCLUSION: LA-Loaded CS/β-GP injectable thermosensitive hydrogels with good elasticity and gel strength properties are prepared successfully, and they show sustained release effect of LA in vitro.
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OBJECTIVE: To prepare lappaconitine (LA)-loaded polylactic acid (PLA) nanoparticles (LA/PLA NPs) and investigate its release properties in vitro. METHODS: LA/PLA NPs were prepared by optimized emulsion-solvent evaporation method with biodegradable PLA as carrier material and polyvinyl alcohol (PVA) as emulsifier. The entrapment efficiency and drug loading rate of LA were used as the main evaluation indexes to optimize the preparation process by orthogonal design method. The mean particle size was measured by laser particle size analyzer; the morphology of LA/PLA NPs was observed by atomic force microscope; the in vitro release behavior was studied by dynamic dialysis. RESULTS: The optimized LA/PLA NPs were spherical. The mean particle size was (429±9.19) nm, the entrapment efficiency and drug loading rate were (86.34±2.15)% and (45.85±1.34)%, respectively. The in vitro release study showed that the LA/PLA NPs could provide a continuous release of LA for 15 d. CONCLUSION: LA/PLA NPs with high entrapment efficiency and drug loading rate are prepared successfully, and show sustained release effect for LA in vitro.
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Objective To explore the trend of variation and relationship of postoperative pain and serum complement C 3 and C4 in cancer patients undergoing rectum surgery .Methods 100 patients ,who were scheduled for rectum carcinoma surgery ,were se-lected to the study .Pain was assessed by a visual analog scale at 12 h before operation and 4 ,8 ,12 ,24 ,48 ,72 ,120 h after surgery . The blood samples were obtained at the same time .The contents of serum complement C3 and C4 were determined by immunoturbi-dimetry .Results The VAS values in 4 ,8 ,12 ,24 ,48 ,72 h post-operation were significantly higher than 12 h pre-operation(P<0 .01) ,and in 120 h post-operation returned to 12 h pre-operation level .Compared with 12 h pre-operation ,the contents of serum complement C3 and C4 in 4 ,8 ,12 ,24 ,48 h post-operation were significantly decreased (P<0 .01) .The contents of serum comple-ment C3 and C4 returned to 12 h pre-operation level in 72 h post-operation .The results of correlative study on VAS values and the contents of serum complement C3 showed a negative correlation(r= -0 .622 ,P<0 .01) .The results of correlative study on VAS values and the contents of serum complement C4 also showed a negative correlation(r= -0 .649 ,P<0 .01) .Conclusion Postoper-ative pain can induce complement activation ,reduce the levels of serum complement C3 and C4 ,and inhibit immunoreactions .
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OBJECTIVE:To study the immunoregularoty effect of Lappaconitine in S180 tumor-bearing mice.METHODS: Each mouse was inoculated hypodermically with S180 tumor cells and normal sodium suspension,24 hours later,all the mice were injected i.m.with Lappaconitine q.d for 10 consecutive days.The immune function of the S180 tumor-bearing mice was detected.RESULTS: In Lappaconitine-treated mice,serum IgG level was heightened significantly;the 2,4-DNFB-induced delayed type hypersensitivity was enhanced;the induction of transformation of lymphocytes into lymphoblast was achieved;the phagocytic function of reticuloendothelial system was enhanced significantly,and the immune function of red cells was promoted.CONCLUSION: Lappaconitine could inhibit the growth of S180 tumor cells and significantly enhance the cellular immune function of S180 tumor-bearing mice.
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0.05),but there were significant differences on the side-effects such as itch of skin,nausea and vomiting between two groups (P
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Lappaconitine hydrobromide can remove inflammation and swelling, lower temperature and relieve heat. It also can be used for local anesthesia. Clinically lappaconitine hydrobromide can be applied via i.v., i.m. or p.o. for analgesic treatment. Lappaconitine has better effect for moderate pain than for severe pain, which may be associated with the low dose. Future efforts should be made in increasing dosage, combining lappaconitine hydrobromide with other drugs and improving drug delivery technique.
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Objective To evaluate the effect and safety of point injection and em bedding therapy on postoperative pain of hemorrhoid patients. Me thods Totally 105 hemorrhoid patients were rand omized into a treatment group (54 cases) and a control group (51 cases). The tre atment group was treated by point injec tion and embedding therapy at Changqiang point (GV 1) under the local anesthesia . The control group was treated by Indometacin Suppository after operation. On the fir st, second, four th and seventh day of the treatment, the pain occurrence, pain severity, and com prehensive effect scores were recorded for the evaluation of therapeutic effect and safety. Results On all interview periods, the compr ehensive effect score, pain occurrence, pain severity, patient satisfaction, occurrence of untoward eff ect, complications, and analgesics combination in the treatment group were all superior to those in the control group (P
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Lappaconitine (LA) 0. 5 mg? kg-1had significant antagonistic action on the ar-rythmia induced by 3-acetylaconitine ( AA ) 0.07 mg?kg-1in rat. When LA and AA were used in combination (7:1) in mice, the analgesic activity of AA was not influenced marked-ly by LA but the LD50 of AA was increased by 50% ,therefore the therapeutic index of AA was increased and the margin of safety was widened.