| Pain after hemorrhoid surgery is the most common postoperative complication, which influenced Patients’quality of life. Severe postoperative pain may still occur and continues to concern patients who are often unwilling to undertake the procedure as a result. Anal sphincter hypertonicity has been identified as a major influencing factor in increased pain after surgery. The maintained internal anal sphincter spasm is considered to be a major factor in the genesis of pain, so relieving the internal anal sphincter spasm is considered to be the effective method to ease the pain. Now, multiple approaches, including pharmacological and surgical interventions, have been used to manage pain after haemorrhoidal surgery, such as sphincterotomy, botulinum toxin, glyceryl trinitrate, and Calcium channel blockers. sphincterotomy significantly improves the postoperative course after hemorrhoidectomy and that the procedure may lead to perianal infection or bleeding. As it is known to lead to permanent minor impairment of continence leading to incontinence of flatus, mucus, or occasionally stool. So many people have espoused the concept of chemical sphincterotomy with topical nitrates and calcium channel blockers. But the most common side effect of chemical sphincterotomy treatment is the development of headaches. This is the commonest reason for poor compliance of GTN in any form. Other side effects include headache, syncope, hypotension and perianal pruritus. So it will be the important research subject that how can reduce spasm of internal anal sphincter and side effects of treatment with minimal impair of internal anal sphincter. Haemorrhoidectomy coupled with spasmolysis treatment is a satisfactory treatment for hemorrhoid, and significantly reduces the incidence of postoperative complications, and improve the quality of life.The purpose of this study is to explore the effect of Guang Tong Xiao Foam Aerosol in relaxing spasm and relieving pain after hemorrhoidectomy, and to observe in vitro the effect of intestinal absorption solution of extracts of compound Guangtongxiao preparation on the tension of mice rectal ring. Meanwhile, a novel sensitive and selective HPLC-ESI-MS/MS method was developed and validated to simultaneously determinate and identify constituents in intestinal absorption solution. Then, a sensitive and selective LC-MS/MS method for the simultaneous determination of medicinal composition in rat intestinal absorption solution was firstly developed and validated to analyze intestinal absorption solution sample of the three analytes. Finally, a HPLC-MS method was established for the quantification of medicinal composition in rat intestinal absorption solution. In order to clarify the mechanism of intestinal absorption solution, the effect of THP and Pae on mice rectal ring contractility and the pharmacological mechanism were been studied. Part oneThe clinical research for the effect of Guang Tong Xiao Foam Aerosol in relaxing spasm and relieving pain after hemorrhoidectomyObjective To research the clinical efficacy of Guang Tong Xiao Foam Aerosol in relaxing spasm and relieving pain after hemorrhoidectomy.Methods:88patients with hemorrhoids of third and fourth degree were included in the study and randomized in three groups treatment group, control group and placebo group. After hemorrhoidectomy,88patients were randomly assigned to receive Guang Tong Xiao Foam Aerosol,0.2percent glyceryl trinitrate ointment or a placebo ointment. Anorectal manometry was performed preoperatively and5and14days after hemorrhoidectomy. Patients maintained a log to measure postoperative pain daily. Any postoperative morbidity noted during the follow-up period was recorded.Results:At the rest, the anal pain scores at the2nd-7rd days after operation in treatment group and control group were lower than that of the placebo group (P<0.05). During defecation, there was statistically significant among the three groups in the anal pain scores (P<0.05). Five days after hemorrhoidectomy, maximum resting pressure was significantly reduced compared with placebo values in both treatment group and control group (P<0.05). Analgesic consumption was highest in placebo group on2nd and3rd day (P<0.05). The incidence of edema in cut edge was lower in treatment group on2nd and3rd day (P<0.05). Adverse effects in the control group, such as headaches, was more than that of treatment group and placebo group (P<0.05). There was statistically significant among the three groups in the wound healing after three weeks (P<0.05).Conclusion:Guang Tong Xiao Foam Aerosol was more effective and safer in reducing anal rest pressure and the incidence of edema, relaxing the smooth muscle of the internal anal sphincter, promoting wound healing and relieving early postoperative pain at rest and during defecation.Part twoEffect of intestinal absorption solution of extracts of compound Guang tong xiao preparation on the tension of mice rectal ringObjective To observe in vitro effect of intestinal absorption solution of extracts of compound Guangtongxiao preparation on the tension of mice rectal ring.Methods Firstly, intestinal absorption solution of extracts of compound Guangtongxiao preparation was prepared by non-everted gut sac method in vitro at5different time points (30,45,75,105and120min). The rectal ring was quickly removed after rats were sacrificed by dislocation. Then rectal ring were pre-contracted by KC1or ACh, and the tension of rectal ring was observed by adding different volumes (200,400,800,1600and320μL) of the intestinal absorbed solution from extracts of compound Guangtongxiao preparation, experiments were performed in five replicates (n=5per group). Results The relaxation rate was enhanced with the increased volumes of intestinal absorption solution at120min, the results indicated that the relaxation was enhanced and a dose-effect relationship was found; Compared with control group of intestinal absorption solution without extracts of compound Guangtongxiao, when the volume of200,400,800,1600,3200μL was added, P<0.01. When the volume of3200μL was added, the relaxation of5groups were all enhanced, compared with every group of intestinal absorption solution, there was statistical significance in control group, P<0.01; the results indicated that the relaxation was also enhanced with the delayed absorption time of extracts of compound Guangtongxiao preparation and a time-effect relationship was observed.Conclusion The results show that the intestinal absorption solution of extracts of compound Guangtongxiao preparation may have significant relaxation activity with a manner of dose-effect and time-effect relationship.Part threeThe study in vitro on rat rectal absorption kinetics of three components in extracts of compound Guang tong xiao preparationObjective To investigate the absorption kinetics of tetrahydropalmatine (THP), berberine hydrochloride (Ber)and paeoniflorin (Pae) of Guangtongxiao extracts in rat rectal segment and to clarify the effective substances of intestinal absorption solutionMethods The non everted rectal sac model was adopted. Rectal sac fluid samples at different time points after administration of three concentrations of Guangtongxiao extracts were collected and detected by LC-MS/MS, and the absorption rate constants (Ka) to describe the character of absorption was calculated.Results The absorption of THP, Ber and Pae was linearity in different concentration, and the square of coefficient correlation exceed0.95. The Ka of THP kept at the same level when the extracts in different concentrations (P>0.05), which consistent with first order process. The Ka of Ber and Pae increased along with the raised dosage of Guangtongxiao extracts (P<0.01), which conformed to the zero order process.Conclusion The absorption of THP in rat’s rectal is a first-order process with the passive diffusion mechanism. The absorption of Ber and Pae in rat’s rectal is not the simple passive diffusion mechanism, and transporters carrier may involve in mediating. THP, Ber and Pae can be concentrated in rectal and absorbed into the blood to exert their pharmacological effects.Part fourEffect and mechanism of THP and Pae on the tension of isolated mice rectal ringObjective To observe the effect and mechanism of THP and Pae on the tension of isolated mice rectal ring.Methods Isometric tension was recorded in response to drugs in organ bath. Effects of THP(5.73x10-6M～1.72×10-3M) and Pae(3.3×10-6M～1×10-3M) on the rectal ring tone of high KCl(60mmol·L-1) and ACh (10-6mol·L-1) pre-constricted mice rectal ring were determined. To explore the mechanism, adrenergic receptor inhibitor phentolamine (10-6mol·L-1) and propranolol(10-5mol·L-1) were used. Evaluate the effect of THP and Pae on the ACh-induced contraction in an incubation period of Ca2+-free K solution. Following an incubation period of K+-rich and Ca2+-free K solution, control concentration-response curves (CRCs) of CaCl2were obtained. When the control CRCs of CaCl2were found superimposable (usually after two cycles), the tissue was pretreated with the THP and Pae to test the possible calcium channel blocking effect. The CRCs of CaCl2were reconstructed in the presence of different concentrations of the test material.Results THP and Pae caused concentration relaxation of rectal rings pre-constricted with high KCl and ACh. Pae and THP caused inhibition of high K+-induced contractions, with respective EC50values of8.67×10-5M (6.68×10-5-1.13×10-4),1.41×10-4M (1.24×10-4×-1.61×10-4); and in ACh group the EC50values were6.15×10-5M (4.47×10-5-8.45×10-5),1.31×10-4M (1.22×10-4-1.42×10-4). Pretreatment with phentolamine and propranolol could not abolish the relaxation response to Pae and THP. In Ca2+-free K solution, THP and Pae could relax ACh-constricted mice rectal ring. The calcium channel blockade (CCB) activity was further confirmed when pretreatment of the tissue with THP and Pae caused a rightward shift with maximal efficiency decreased in the Ca2+concentration-response curves (CRCs), similar to verapamil.Conclusion These data indicate that the anti-spasmolytic effects of the THP and Pae are mediated through the presence of CCB-like constituent(s) by blocking voltage-dependent calcium channel, intracellular calcium delivering and external calcium influx.
|
PDF Full Text Request