| Background Anal pain and urinary retention are common symptoms after procedure for prolapse and hemorrhoids(PPH)surgery in patients with damp-heat injection hemorrhoids,which cause certain problems for patients’ postoperative recovery and quality of life.Postoperative analgesia and improvement of urinary retention symptoms are problems that clinicians are committed to breaking through.The concept of treatment based on syndrome differentiation in traditional Chinese medicine believes that pathogenesis is the mechanism of disease development and outcome,which is not limited to a certain disease or location,implying that different diseases with the same pathogenesis can be treated in the same way or with the same prescriptions,namely “treating different disease with the same treatment”.Treating different diseases with the same treatment is not only a common method of traditional Chinese medicine,but also an important embodiment of dialectical treatment of traditional Chinese medicine,which guides us to diagnose and treat diseases.Chinese medicine suggests that the pathogenesis of anal pain and urinary retention after damp-heat type hemorrhoids after PPH is damp-heat combined with stasis,which has the same pathogenesis as a variety of damp-heat and stasis-type scorching diseases such as chronic prostatitis.The treatment with Qianliean Suppository to the latter disease presents significant curative effect.At present,it has confirmed by many studies that the mechanism of Qianlieanshuan treating such diseases is to reduce the irritation of inflammatory response to surrounding tissues by inhibiting the release of local inflammatory factors,which can reduce inflammation and edema of the rectum,prostate,bladder neck and urethral sphincter, and relieve muscle spasms.Purpose To prove that the application of Qianliean Suppository in the treatment with postoperative anal pain and urinary retention of damp-heat injection hemorrhoids PPH is clinically effective,based on the concept of "different diseases with the same treatment",and its mechanism analyzed from the molecular level.Methods Clincal research1.Normal information Thirty patients with damp-heat injection hemorrhoids(in line with the selective criteria)were all inpatients from the Anorectal Department of Gaozhou Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine.The patients were divided into two groups: a treatment group and a control group by a randomized single-blind control method.15 cases per group,the age,gender and course of disease were statistically processed.2.Treatment PPH was given to both the treatment group and the control group.The treatment group was applied one piece of Qianliean Suppository before the end of the operation,about 4cm in the rectum.In the morning and evening after the operation,Qianlieanshuan was replaced to the anus at the same position when was changed the dressing every morning and night for 7 days.The control group was given a blank control.3.Observation(1)Observation indicators: the degree of dysuria,lower abdomen condition,urination time,and pain degree are recorded by scoring.(2)Detection indicators: Enzyme-linked immunosorbent assay(ELISA)detects serum IL-6 and TNF-α expression levels.Basic experiment1.BAL/C mice were used to established a model of hemorrhoids and a model of hemorrhoids combined with urinary retention.The mice were randomly divided into blank group,hemorrhoids combined with urinary retention model group,hemorrhoids combined with urinary retention model + low-concentration medication intervention group,hemorrhoids combined with urinary retention model + high-concentration medication intervention group,6 mice per group.(1)ELISA was applied to detect the expression levels of serum inflammatory factors TNF-α,IL-2,hs-CRP.(2)Flow cytometry was given to detect the levels of CD4+Foxp3+Treg and CD4+IL-17A+T cells.(3)Western Blot was used to detect the levels of related proteins JAK2,p-STAT3,SOCS.(4)Immunohistochemical method was applied to detected the expression level of VEGF in mucosal layer.2.An animal model of hemorrhoids combined with urinary retention pain was established with BAL/C mice.The mice were randomly divided into blank group,hemorrhoids combined with urinary retention pain model group,hemorrhoids combined with urinary retention pain model + low-concentration medication intervention group,hemorrhoids combined with urinary retention pain model + high-concentration medication intervention group,6 mice per group.(1)Mouse writhing reaction experiment: the drug was stuffed into the mouse rectum,and a0.6% glacial acetic acid solution was injected into the mouse’s abdominal cavity after 20 minutes,and the number of "writhing" in each mouse within 15 minutes of chemical stimulation was observed and recorded.(2)Pressure analgesia experiment on the tail root of the mouse: first all the hair in tails of the mice was removed,then the mouse was put in a fixed tube with the tail exposed and a symbol marked at the spot 1 cm from the tail root.The pressure gradually increased in the experiment and took the tail retraction as the pain response,and the reading(g)on the scale indicator was written down.The pressure weight reflected the pain threshold,and the basic threshold was measured first,following by drug intervention,pain response was respectively measured 0.5h,1h,2h,3h,4h,a total of 5 times,after the administration of the mice to calculate the percentage increase in pain threshold to compare the analgesic effect.(3)Mouse plantar swelling experiment: a mark was made on the right hind plantar joint of each mouse,and 0.1 m L of 1% fresh egg white was injected into the plantar fascia of the foot to cause inflammation.The paw volume was measured 1,2,3,4,and 5 hours after inflammation,and the swelling degree and inflammation inhibition rate were calculated.(4)ELISA was used to detect the expression levels of cytokines IFN-γ,IL-17 A,IL-4 and IL-10 secreted by T cells.(5)Real-time quantitative polymerase chain reaction(RT-q PCR)was applied to detect the m RAN expression levels of T-cell specific transcription factors T-bet,RORγt,GATA3 and Foxp3.Results Clinical research1.Before treatment,there was no statistical difference in gender,age,and course of disease between the two groups of patients(P=1.000,0.343,1.000,P>0.05,respectively),and they were comparable.2.The effect of two groups of patients on urinary retention after treatment: for urination symptom invaluation,the treatment group was 7 cases with 0 point,8 cases with 1 point,the control group 7 cases with 0 points,4 cases with 1 point,3 cases with 2 points,and 1case 3 wiht points,suggesting that the treatment group was better than that of the control group(P<0.05);for the lower abdomen symptom invaluation,the treatment group had 11 cases with 0 points,4 cases with 1 point,and the control group had 8 cases with 0 points,3cases with 1 point,3 cases with 2 points,1 case with 3 points,implying that the effect of the treatment group was significantly better than that of the control group(P<0.01);for urination time evulation,the treatment group was 7 cases with 0 points,8 cases with 1 point,control group was 7 cases with 0 points,4 cases with 1 point,3 cases with 2 points and 1case with 3 points,presenting that the urination time in the treatment group was significantly shorter(P<0.01).3.The effect of the two groups on analgesia after treatment: the treatment group had 10 cases with VAS score of 0-3 point and 5 cases with 4-6 points;the control group had 9cases with 0-3 points,4 cases with 4-6 points,2 cases wiht 7-9 In,showing that the pain intensity of the treatment group was significantly lower than that of the control group,indicating that the analgesic effect was obvious(P<0.01).4.Laboratory indicators: There was no significant difference in serum IL-6 and TNF-αbetween the two groups of patients preoperation(P>0.05).The IL-6 and TNF-α in the treatment group were significantly reduced postoperation and the concentrations are both2 d postoperation<1d postoperation,the difference is statistically significant(P<0.01)Basic experiment1.The efffects of Qianliean Suppository on urodynamics:(1)The first contractile urination time of the bladder significantly prolongs in the model group(P<0.01),and significantly shortens in the low-dose group(P<0.01)and the high-dose administration group(P<0.01);(2)The basal intravescial pressure during the urinary retention period,the basal intravescial pressure after the first intrvesical contractile urination,and the basal intravescial pressure30 minutes after the first intrvesical contractile urination in the model group was significantly increased(P<0.01),but significantly reduced in low dose Group(P<0.01)and the high-dose administration group(P<0.01);(3)The micturition threshold pressure and micturition pressure of the first intrvesical contractile urination were significantly increased in the model group(P<0.01),but the difference between the drug group and model administration group was not statistically significant in high and low doses group(P>0.05);(4)The micturition threshold pressure and micturition pressure at 30 minutes after the first intrvesical contractile urination were significantly increased in the model group(P<0.01),significantly reduced in the high-dose administration group(P<0.01),but no statistical difference in the low-dose group(P>0.05);(5)Unstable contraction occurrence after urination,compared with the model group,the low-dose group had no statistically significant difference(P>0.05),the unstable contraction after urination in the high-dose administration group was significantly reduced(P<0.01).2.The effects of Qianliean Suppository on related inflammatory factors in serum:(1)TNF-α,the level of TNF-α in the model group was significantly increased(P<0.01),in the high-dose administration group significantly reduced(P<0.01)and no significant difference in the low-dose administration group(P>0.05);(2)IL-2,the level of the model group was significantly reduced(P<0.01),high-dose administration group increased(P<0.05)and the low-dose administration group was not significantly different from it(P>0.05);(3)The level of hs-CRP in the model group increased significantly(P<0.01),and both of the level of the low-dose administration group(P<0.01)and the high-dose administration group(P<0.01)decreased significantly.3.The effect of Qianliean Suppository on differentiation of helper T lymphocytes(Th1/Th17)and regulatory CD4+/CD8+ T lymphocytes:(1)Helper T lymphocytes(Th1/Th17),the ratio of the model group was significantly increased(P<0.01),both of low-dose group(P<0.01)and the high-dose group(P<0.01)was significantly lower than that;(2)Regulatory CD4+/CD8+ T cells,the model group’s regulatory ratio was significantly lower(P<0.01),compared with the model group,the ratio of low-dose administration group(P<0.01)and high-dose administration group(P<0.01)increased significantly.4.The effect of Qianliean Suppository on the expression of related proteins:(1)The relative expression of JAK2 protein in the model group was significantly up-regulated(P<0.01),the low-dose group was down-regulated(P<0.05),and the high-dose group was significantly down-regulated(P<0.01);(2)The relative expression of p-STAT3 protein in the model group was significantly up-regulated(P<0.01),and both in the low-dose group (P<0.01)and high-dose group(P<0.01)was significantly down-regulated,in a manner of the magnitude of the down-regulation positively correlated with the dose;(3)The SOCS3 protein expression in the model group was significant up-regulation(P<0.01),both in low-dose group(P<0.01)and high-dose group(P<0.01)were significantly down-regulated,in a manner of down-regulation positively positively correlated with dose.5.The effect of Qianliean Suppository on the expression level of VEGF in the mucosal layer: the expression of VEGF in the model group was significantly increased(P<0.01),the low-dose group(P<0.01)and the high-dose group(P<0.01)significantly decreased.6.The analgesic effect of Qianliean Suppository:(1)Qianliean Suppository reduces the writhing reaction in mice.Compared with the number of writhing reactions in the blank group,the model group significantly increased to P<0.01),the low-dose group(P<0.01)and the high-dose group(P<0.01)were significantly reduced;(2)Qianliean Suppository significantly increased the pain threshold of mice(P<0.01),the high-dose group significantly increased the pain threshold than the low-dose group(P<0.05);treated with the same dose,the longer the compression time is given,the higher the pain threshold represents(P<0.01);(3)Qianliean Suppository improves mouse plantar swelling degree.When intervening for 1h and 2h,compared with the blank group,the swelling degree of the model group significantly increases(P<0.01).Compared with the model group,it decreases in low-dose group(P<0.01)and high-dose group(P<0.01)significantly.3 h after intervention,compared with the blank group,the swelling degree of the model group significantly increases(P<0.01).Compared with the model group,the low-dose group reduced(P<0.05),and the high-dose group significantly reduced(P<0.01).At 4h and 5h after intervention,compared with the blank group,the swelling degree of the model group was significantly increased(P<0.01),compared with the model group,there was no statistical difference in the low-dose group(P>0.05),but the swelling degree of the high-dose group was significantly reduced(P<0.01).7.The effect of Qianliean Suppository on Th cell secretion of cytokines:(1)The level of IFN-γ in the model group was significantly up-regulated(P<0.01),and the level in the low-dose and high-dose groups was significantly down-regulated(both P<0.01);(2)The model group The level of IL-17 A was significantly increased(P<0.01),and the level of the low-dose group and the high-dose group was significantly decreased(all P<0.01);(3)The difference between the model group,low-dose and high-dose groups was not statistically significant(P>0.05);(4)IL-10 levels in the model group were significantly down-regulated,the low-dose group was significantly up-regulated(P<0.01),and the high-dose group was not as significantly up-regulated as the low-dose group(P<0.05).8.The effect of Qianliean Suppository on Th cell-specific m RNA expression:(1)T-bet m RNA expression in the model group was significantly increased(P<0.01),the expression in the low-dose group was decreased(P<0.05),and the high-dose group was decreased more significantly(P<0.01);(2)The RORγt m RNA expression in the model group was significantly increased(P<0.01),but there was no significant difference neither in the low-dose nor high-dose Qianlieanshuan group(all P>0.05);(3)The GATA3 m RNA expression in the model group decreased(P<0.01),the high-dose group increased significantly(P<0.01),and the low-dose group had no significant difference(P>0.05);(4)The expression of Foxp3 m RNA in the model group had no significant difference(P>0.05),the low-dose group significantly decreased(P<0.01),and the high-dose group increased(P<0.05).Conclusions1.Qianliean Suppository has a good clinical effect on urinary retention and anal pain after procedure for prolapse and hemorrhoids for damp-heat injection hemorrhoids.2.Through animal experiments on exploring the mechanism of Qianlieanshuan in the treatment of urinary retention and anal pain after procedure for prolapse and hemorrhoids for damp-heat injection hemorrhoids,it finds that Qianliean Suppository can inhibit TNF-α,hs-CRP and other pro-inflammatory factors and inflammation factor release,while promoting the release of IL-2 immune factors.3.Qianliean Suppository inhibits SOCS signal transduction,negative-feedback regulates JAK2/STAT3 signal pathway,and indirectly regulates Th17/Treg balance in urinary retention mice,and then regulates the expression of inflammatory factors,finally improves urinary retention.4.Qianliean Suppository down-regulates inflammatory factors to improve the symptoms of urinary retention and pain by down-regulating the Th1 specific transcription factor T-bet and up-regulating the important transcription factor GATA3 that induces Th2 differentiation,while regulating the expression of the transcription factor Foxp3,thereby regulating Th17/Treg The balance between to achieve. |
