Effects of different transdermal penetration enhancers applied to herbal cake-partitioned moxibustion on liver lipids,HSL and HMG-CoA reductase in hyperlipidemia rabbits

Objective:To observe the effects of laurocapram and borneol as transdermal penetration enhancers applied to herbal cake-partitioned moxibustion on liver lipids,hormone-sensitive lipase(HSL)and hydroxymethylglutaryl CoA(HMG-CoA)reductase in hyperlipidemia rabbits.Methods:Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method,with 8 rats in each group.Rabbits in the blank group were fed routinely with a normal diet;rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model.Rabbits in the blank and the model groups were not given any intervention.After the model was prepared successfully,rabbits in the non-transdermal penetration enhancer group received herbal cake-partitioned moxibustion without transdermal penetration enhancers;rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively.After 4 weeks of treatment,the serum was isolated and enzyme-linked immunosorbent assay(ELISA)was applied for the detection of HSL and HMG-CoA reductase.The liver tissues were isolated,and total cholesterol(TC)and triglycerides(TG)were measured by enzymatic methods.One-step method was applied for high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)detection,and transmission turbidimetry was for apolipoprotein A1(Apo-A1)and apolipoprotein B(Apo-B)detection.Results:The serum concentrations of the drugs in the laurocapram and the borneol groups were significantly higher than those in the non-transdermal penetration enhancer group(both P<0.05);all drug penetrations in the borneol group were significantly higher than those in the laurocapram group(both P<0.05),except for tanshinoneⅡA.Compared with the non-transdermal penetration enhancer group,the HSL was significantly increased while the HMG-CoA reductase was significantly decreased in the laurocapram and the borneol groups(both P<0.05);between groups,the HSL in the borneol group was significantly higher than that in the laurocapram group(P<0.05).Compared with the blank group,the levels of LDL-C,TG,TC and Apo-B in rabbit liver were significantly increased in the model group(P<0.05);compared with the model group,the levels of LDL-C,TG,TC and Apo-B in the non-transdermal penetration enhancer,the laurocapram,and the borneol groups were significantly decreased(all P<0.05);between groups,the TG and TC in the laurocapram group and the LDL-C,TG,TC and Apo-B in the borneol group were significantly lower than those in the non-transdermal penetration enhancer group(all P<0.05),and the TG,LDL-C and Apo-B in the borneol group were significantly lower than those in the laurocapram group(all P<0.05).Compared with the blank group,the HDL-C and Apo-A1 were significantly decreased in the model group(both P<0.05),while compared with the model group,the HDL-C and Apo-A1 were significantly increased in the non-transdermal penetration enhancer,the laurocapram,and the borneol groups(all P<0.05).Between groups,the Apo-A1 in the laurocapram group,the HDL-C and Apo-A1 in the borneol group were significantly higher than those in the non-transdermal penetration enhancer group(all P<0.05).Conclusion:The application of laurocapram and borneol,as transdermal penetration enhancers,in herbal cake-partitioned moxibustion can promote the penetration of the drugs in the herbal cake,increase the levels of HDL-C and Apo-A1,improve the metabolism of HSL and HMG-CoA reductase,and also simultaneously reduce the levels of TC,TG,LDL-C and Apo-B in the liver.The transdermal penetration enhancement effect of borneol is slightly better than or equivalent to that of laurocapram.

Influence of herbal cake-partitioned moxibustion on lumbar functions and inflammatory factors in patients with lumbar disc herniation due to kidney deficiency and blood stasis

Objective To observe the efficacy of herbal cake-partitioned moxibustion for lumbar disc herniation(LDH)due to kidney deficiency and blood stasis and observe the influence of this method on lumbar functions and inflammatory factors in patients with this condition.Methods A total of 120 LDH patients who met the inclusion criteria were randomly divided into three groups,including a herbal cake-partitioned moxibustion group,a flour cake-partitioned moxibustion group,and a Western medication group,with 40 patients in each group.The patients in the Western medication group were treated with diflunisal tablets,0.5 g per dose,2 doses a day.Those in the herbal cake-partitioned moxibustion group were treated with additional herbal cake-partitioned moxibustion group at Back-Shu Points and Jiaji(EX-B2)Points once a day.Those in the flour cake-partitioned moxibustion group were treated with the same methods as in the herbal cake-partitioned moxibustion group,except that the herbal cake was replaced by a flour cake.All the patients were treated for 10 d.After treatment,the scores of the visual analog scale(VAS)and Japanese Orthopaedic Association(JOA)and the changes of the interleukin(IL)-6,tumor necrosis factor(TNF)-α,and substance P(SP)levels were observed,and the efficacy was evaluated.Results After treatment,the VAS score and the levels of IL-6,TNF-α,and SP were lower than those before treatment,and the JOA score was higher than that before treatment in the three groups,indicating intra-group statistical significance(P<0.05).The VAS score and the levels of IL-6,TNF-α,and SP of the herbal cake-partitioned moxibustion group were lower than those of the flour cake-partitioned moxibustion group and the Western medication group,while the JOA score of the herbal cake-partitioned moxibustion group was higher than that of the other two groups,indicating inter-group statistical significance(P<0.05).The total effective rate of the herbal cake-partitioned moxibustion group was 92.5%,higher than that of the flour cake-partitioned moxibustion group(80.0%)and the Western medication group(72.5%),indicating inter-group statistical significance(P<0.05).Conclusion On the basis of Western oral medication,additional herbal cake-partitioned moxibustion can alleviate the pain and improve the lumbar functions in patients with LDH due to kidney deficiency and blood stasis.The efficacy of the integrated method is better than that of either flour cake-partitioned moxibustion or Western medication alone,which may be related to the reduction of serum inflammatory factors.

Therapeutic observation on herbal cake-partitioned moxibustion for dysmenorrhea due to deficiency cold

Objective:To observe the clinical efficacy of herbal cake-partitioned moxibustion for dysmenorrhea due to deficiency cold.Methods:A total of 70 patients with dysmenorrhea who met the inclusion criteria were randomized into a mild moxibustion group and a herbal cake-partitioned moxibustion group by the random number table,with 35 cases in each group.Shenque(CV 8),Zhongji(CV 3)and bilateral Zigong(EX-CA 1)were selected for both groups.The treatment continued for 3 menstrual cycles.The visual analog scale(VAS)and COX menstrual symptom scale(CMSS)were scored in both groups before treatment,after treatment and at the end of the 3rd menstrual cycle after treatment.The clinical efficacy was evaluated at the end of the 3rd menstrual cycle after treatment.Results:After treatment,the clinical efficacy of the herbal cake-partitioned moxibustion group had the tendency to be superior to that of the mild moxibustion group,while there was no statistically significant difference in the overall efficacy between the two groups(P>0.05).The VAS and CMSS scores after treatment and at the follow-up were significantly lower than those before treatment in both groups(all P<0.05).At the follow-up,the VAS scores in both groups had no significant intra-group differences from those after treatment(both P>0.05).The CMSS scores in both groups were significantly lower than those after treatment(both P<0.05).The VAS scores at the follow-up of both groups had no statistical differences from those after treatment(both P>0.05).After treatment,the CMSS score in the herbal cake-partitioned moxibustion group was significantly lower than that in the mild moxibustion group(P<0.05).At the follow-up,there were no statistical differences in the CMSS score between the two groups(P>0.05).Conclusion:The herbal cake-partitioned moxibustion has the same therapeutic efficacy for dysmenorrhea as the mild moxibustion;the two moxibustion methods can significantly improve the concomitant symptoms of dysmenorrhea,and the herbal cake-partitioned moxibustion is little better.

Effect of herbal cake-partitioned moxibustion on Leptin/JAK#STAT3 in lipid-lowering pathway of hyperlipidemia rabbits

Objective:To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits,and to explore the possible mechanism.Methods:Forty New-Zea I a nd rabbits were ran domly divided into 5 groups using the ran dom nu mber table method,with 8 rats in each group.Rabbits in the blank group were fed routinely with normal diet;rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model.Rabbits in the blank and the model groups were not treated.After the model was prepared,rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer;rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively.After 4 weeks of treatment,serum was collected for enzyme-linked immunosorbent assay(ELISA),and the liver tissues were isolated for imm uno histochemistry,qua ntitative polymerase chain reactio n(qPCR)and Western-blotting(WB)detecti on.Results:Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group(P<0.05);compared with the model group,lepti n was significa ntly in creased in the non-tran sdermal absorpti on enhanee。the laurocapram and the borneol groups(all P<0.05);compared with the non-transdermal absorption enhancer group,leptin was significantly increased in the laurocapram group and the borneol group(both P<0.05);there was no significant differenee in leptin between the laurocapram and the borneol groups(P>0.05).The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin,Janus kinase 2(JAK2)and signal transducer and activator of transcription 3(STOT3)in the model group were significantly lower than those in the blank group(all P<0.05);compared with the model group,the mRNA expressions of leptin,leptin receptor(LR),JAK2 and S1AT3 in the non-transdermal absorptio n enhan cer,the laurocapram and the born eol groups were significantly in creased(all P<0.05);compared with the non-transdermal absorption enhancer group,the mRNA expressions of leptin,LR,JAK2 and S77VT3 in the laurocapram and the bor neol groups were sign ificantly in creased(all P<0.05);compared with the laurocapram group,the mRNA expressi ons of lepti n,LR,JAK2 and SW3 in the bor neol group were significa ntly in creased(P<0.05).The trend of immun ohistochemistry and WB detecti on results was basically con siste nt with the qPCR assay results.The immuno histochemistry and WB detection results of phosphorylated JAK2(phospho-JAK2)and phosphorylated S7AT3(phospho-STAT3)were basically consistent with those of JAK2 and S7AT3.Conclusion:The molecular expression of Leptin/JAK"S7AT3 pathway in the hyperlipidemia model rabbits was decreased.The molecular expression of Leptin/JAK0STCT3 pathway was significantly increased after the herbal cake-partitioned moxibustion.The application of laurocapram and borneol,as transdermal absorption enhancers,in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK^SIAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.

Effects of herbal cake-partitioned moxibustion on the expression of thyroid autophagy-related factors LC3B and Beclin-1 in rats with autoimmune thyroiditis

Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere reduced in the herbal cake-partitioned moxibustion group(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group(P<0.05).The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion:Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb.This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.

Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206,AMPK and TSC2 in rats with Crohn disease

Objective:To explore the mechanism of herbal cake-partitioned moxibustion in Crohn disease(CD)treatment by observing the effect of herbal cake-partitioned moxibustion on protein expressions of colonic M2 macrophage marker CD206,AMP-activated protein kinase(AMPK)and tuberous sclerosis complex(TSC)2.Methods:Twenty-six specific pathogen free male rats were randomly divided into a normal group,a model group and a herbal cake-partitioned moxibustion group.The CD model was prepared by enema with the mixture of 5%(W/V)2,4,6-trinitrobenzene sulfonic acid(TNBS)and 50%ethanol at 2:1(volume ratio).After the model was successfully prepared,rats in the herbal cake-partitioned moxibustion group received herbal cake-partitioned moxibustion at Qihai(CV 6)and bilateral Tianshu(ST 25).Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat colon;immunohistochemical technique was used to detect the expression of colonic CD206 protein;Western blot,immuno fluorescence,and real-time fluorescence quantitative polymerase chain reaction(RTqPCR)technologies were used to detect the protein and mRNA expressions of colonic AMPK and TSC2.Results:Compared with the normal group,rats in the model group showed damaged colonic mucosa,missing of the epithelial layer;thickened submucosa,vascular proliferation,massive infiltration of monocytes and lymphocytes,and cracked ulcers that reached the muscle layer.Rats in the herbal cake-partitioned moxibustion group showed reduced intestinal inflammation and healing intestinal epithelium ulcers.Compared with the normal group,rat colonic CD206 protein expression,and the protein and mRNA expressions of colonic AMPK and TSC2 were decreased in the model group(all P<0.Ol);compared with the model group,rat colonic CD206 protei n expression was in creased(P<Q.Q1),as well as the protein and mRNA expressions of AMPK and TSC2 in the herbal cake-partitioned moxibustion(all P<0.05).Conclusion:Herbal cake-partitioned moxibustion can reduce intestinal inflammation in CD rats,increase colonic CD206 protein expression,and up-regulate the protein and mRNA expressions of colonic AMPK and TSC2.

Effect of moxibustion on expressions of HO-1 and MCP-3 protein in colon of rats with Crohn's disease

Objective： To observe the effect of moxibustion therapy on heme oxygenase-1（HO-1） and monocyte chemoattractant protein-3（MCP-3） protein expressions in the colonic mucosa of rats with Crohn＇s disease（CD）, and to explore the intestinal mucosal immune mechanism of moxibustion therapy in treating CD. Methods： The CD rat model was established using the internationally accepted Morris method. The rats were randomly divided into a model group, a herbal cake-partitioned moxibustion group, a mild moxibustion group, a cigarette moxibustion group and a hot compress group, which were compared with the normal group. Except the normal group and the model group, rats in the other groups accepted different moxibustion therapies on bilateral Tianshu（ST 25）. Hematoxylin-eosin（HE） staining was conducted and the pathological changes of the colon were observed under light microscope; the expressions of HO-1 and MCP-3 protein in rat＇s colonic mucosa were determined by immunohisto-chemistry. Results： Compared with the normal group, rats in the model group showed mucosal defect, villus destruction or loss, submucosal congestion and edema, glandular destruction or disappearance, reduced goblet cells, ulcer formation, significantly increased positive target area and positive target integral optical density of HO-1 and MCP-3 protein expression（all P〈0.01）. After treatment, compared with the model group, colonic mucosa was significantly improved in the herbal cake-partitioned moxibustion group and the mild moxibustion group, which mainly showed that the intestinal glands were arranged regularly, ulcer surfaces were covered by the neoformative epitheliums, or intestinal ulcers were replaced by the nascent granulation tissue, and submucosal edema was alleviated, with a small amount of inflammatory cell infiltration. The total areas and the integral optical densities of the positive targets for rat＇s colonic mucosa HO-1 and MCP-3 protein expressions were decreased（all P〈0.01）. Compared with the cigarette moxibustion group and the hot compress group, the total areas and the integral optical densities of the positive targets for rat＇s colonic mucosa HO-1 and MCP-3 protein expressions were significantly decreased（all P〈0.01） in the herbal cake-partitioned moxibustion group and the mild moxibustion group. Conclusion： Herbal cake-partitioned moxibustion and mild moxibustion can significantly improve the inflammatory response of colonic mucosa in CD rats. It can down-regulate the expressions of HO-1 and MCP-3 proteins in the colonic mucosa of CD rats, which may be one of the mechanism in intestinal mucosal immunity caused by moxibustion therapy.

Experimental observation of effects of acupoints,cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature

Objective:To observe the effects of acupoints,cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature,and to provide references for the clinical standardization of moxibustion amount.Methods:The 42 big-ear white rabbits were divided into 6 groups according to the random number table method,a 2-cone direct moxibustion group,a 2-cone direct moxibustion group,a 3-cone direct moxibustion group,a 1-cone herbal cake-partitioned moxibustion group,a 2-cone herbal cake-partitioned moxibustion group,and a 3-cone herbal cake-partitioned moxibustion group,with 7 rabbits in each group.Shenque(CV 8);Shenshu(BL 23)and Zusanli(ST 36)were used in each group,but the moxibustion methods,cone numbers and durations of moxibustion were different・Rabbits in each group received moxibustion once every other day for 5 times in total.During the intervention,a thermoelectricity coupled probe and a temperature recorder were used to record the real-time-acupoint skin temperature and the temperature at different time points,so as to observe,analyze and process the real-time changes in the temperature difference between the surface and inside of acu point skin.Results:For herbal cake-partitioned moxibustion,the best temperature for cone changing was(46.38±0.51)℃when the highest surface temperature was(49.20±0.52)℃;the multi-factor comparison of acupoint×cone number×time and acupoint×moxibustion method×time showed that time×acupoint,time×moxibustion method and cone number×acupoint had interactive effects(all P<0.05).Comparing skin temperature differences between different cone numbers at the same acupoint,Shenque(CV 8)on the 1st and the 5th days,Shenshu(BL 23)on the 3rd and the 7th days,Zusanli(ST 36)on the 1st and the 9th days of experiment showed statistically significant differences(all P<0.05).The skin temperature comparison of different moxibustion methods at the same acupoint all had statistical differences(all Pv0.05),except for Shenque(CV 8)before moxibustion,Shenshu(BL 23)before moxibustion and on the 5th day;Zusanli(ST 36)only showed statistical differences on the 5th and 7th days(both P<0.05).The skin temperature differences of different acupoints after moxibustion in the 1-cone,2-cone and 3-cone groups were statistically different(all P<0.05);direct moxibustion and herbal cake-partitioned moxibustion at different acupoints were all statistically different(all P<0.05).Conclusion:Cone changing temperature under the same specifications of herbal cake-partitioned moxibustion was confirmed.Temperature differenee between surface and inside of different acupoint skin at the same maximum temperature was significantly different due to the cone numbers and moxibustion methods,which showed the highest at Shenshu(BL 23),the sec ond at Shenque(CV 8),and the lowest at Zusanli(ST 36).The in fluence of acupoint factor should be considered to determine the quantitative indicators of moxibustion.

Effect of herb-partitioned moxibustion in improving tight junctions of intestinal epithelium in Crohn disease mediated by TNF-α-NF-κB-MLCK pathway

Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)-myosin-light-chain kinase(MLCK)pathway.Methods:Forty-eight male Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model control(MC)group,an HPM group and a mesalazine(MESA)group,with 12 rats in each group.Trinitrobenzene sulfonic acid(TNBS)was administered to establish CD models.When the model was confirmed a success,the HPM group rats were treated with HPM at Tianshu(ST 25)and Qihai(CV 6),while the MESA group rats were given MESA solution by lavage.When the intervention finished,the colonic epithelial tissues were separated,purified and cultured in each group to establish the intestinal epithelial barrier model in vitro,and TNF-αwas added(100 ng/mL)in the culture medium and maintained for 24 h to establish an increased epithelial permeability model.Transepithelial electrical resistance(TEER)was used to examine the permeability of the barrier;Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway;immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium.Results:After TNF-αinduction,compared with the MC+TNF-αgroup,the TEER value increased significantly in the HPM+TNF-αand MESA+TNF-αgroups(both P<0.001);the expressions of nuclear factor kappa B(NF-κB)p65,MLCK,myosin light chain(MLC),tumor necrosis factor receptor-associated factor 6(TRAF6)and receptor interaction protein-1(RIP1)decreased significantly(P<0.01 or P<0.05),and the expression of zinc finger protein A20(A20)increased significantly(P<0.01);the expressions of occludin,claudin-1,zonula occludens protein 1(ZO-1)and F-actin also increased significantly(all P<0.01).Compared with the MESA+TNF-αgroup,the expressions of MLC,occludin,claudin-1,ZO-1 and F-actin increased significantly in the HPM+TNF-αgroup(P<0.01 or P<0.05).Conclusion:HPM can protect or repair the damage of intestinal epithelial barrier in CD rats,which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.

Investigating the influence of moxibustion on colonic mucosal barrier in rats with dextran sulfate sodium-induced ulcerative colitis

Objective:To observe the effect of moxibustion on the colonic mucosal barrier of rats with ulcerative colitis(UC)induced by dextran sulfate sodium(DSS).Methods:Forty male Sprague-Dawley rats were randomly divided into a normal group and a modeling group,with 20 rats in each group.Rats in the modeling group were subjected to preparing experimental UC models by drinking 4%DSS for seven consecutive days.Two modeled rats and two normal rats were randomly selected for model identification.After the success of UC model was confirmed,the remaining 18 modeled rats were randomly divided into three groups,a model group,a model+herbal cake-partitioned moxibustion group,and a model+mild moxibustion group,with six rats in each group;the remaining normal rats were randomly divided into three groups,a normal group,a normal+herbal cake-partitioned moxibustion group,and a normal+mild moxibustion group,with six rats in each group.After 7 d of intervention with the herbal cake-partitioned moxibustion or the mild moxibustion,hematoxylin-eosin(HE)staining technique was used to observe the pathological changes of colon tissue under a light microscope;Western blotting and/or immunohistochemical techniques were used to detect the protein expression levels of Occludin,Claudin,junction adhesion molecular 1(JAM1),mucin 2(MUC2),and transforming growth factor beta1(TGF-β1)in rat colon tissue.Results:Compared with the normal group,the colon tissue was severely damaged,the pathological score was significantly increased,and the protein expression levels of Occludin,Claudin,JAM1,MUC2,and TGF-β1 were significantly decreased in the model group(P<0.01);while there were no significant differences in the colonic histopathological score,protein expression levels of Occludin,Claudin,JAM1,MUC2,and TGF-β1 in the normal+herbal cake-partitioned moxibustion group and the normal+mild moxibustion group(P>0.05).Compared with the model group,the model+herbal cake-partitioned moxibustion group and the model+mild moxibustion group showed repaired colon tissue,ulcer healing,significantly reduced pathological score,and significantly increased protein expression levels of JAM1,MUC2,and TGF-β1(P<0.05);the Occludin protein expression level in the colon tissue of the model+mild moxibustion group was increased(P<0.01).Conclusion:Neither herbal cake-partitioned moxibustion nor mild moxibustion influences the colonic histopathology and intestinal mucosal barrier-related protein expression in the normal rats;both herbal cake-partitioned moxibustion and mild moxibustion can up-regulate the protein expression levels of JAM1,MUC2,and TGF-β1 in the colon tissue of UC rats.Mild moxibustion can up-regulate Occludin protein expression.This may be a mechanism of moxibustion in reducing colonic mucosa inflammation in UC.

Effects of moxibustion on the P2X7R/STAT3/VEGF pathway in rats with colitis-associated colon cancer

Objective:To observe the effects of herb-partitioned moxibustion and ginger-partitioned moxibustion on the growth of colon tumors in rats with colitis-associated colon cancer(CACC),and explore the mechanism of moxibustion intervening CACC through the purinergic receptor P2X ligand-gated ion channel 7(P2X7R)/signal transducer and activator of transcription 3(STAT3)/vascular endothelial growth factor(VEGF)pathway.Methods:A total of 26 male Sprague-Dawley rats were selected.According to the random number table method,6 rats were selected as the normal group.The remaining 20 rats were injected intraperitoneally with azoxymethane(AOM)combined with oral dextran sodium sulfate(DSS)to prepare the CACC model.After the model was successfully established,2 rats were randomly selected for model identification.The remaining 18 rats which were successfully modeled were randomly divided into a model group,a herb-partitioned moxibustion group and a ginger-partitioned moxibustion group,with 6 rats in each group.Moxibustion intervention was performed in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group at Qihai(CV 6)and bilateral Tianshu(ST 25).Moxibustion was performed twice at each point each time,once a day,at a 1-day interval after 6 consecutive interventions,for a total of 30 interventions.After intervention,the colon tumor load,pathological change and histopathological score were observed.Immunohistochemistry was used to detect the expressions of VEGF,P2X7R,phospho-STAT3(p-STAT3),and nuclear factor-kappa B p65(NF-κB p65)proteins in rat colon tissue.Western blot was used to detect the levels of p-STAT3 and NF-κB p65 proteins in rat colon tissue.Results:Compared with the normal group,the colon tumor load and histopathological score in the model group were significantly increased(both P<0.001),and different grades of dysplasia were observed in colon tissue from the model group,reaching the degree of adenocarcinoma;the expression level of P2X7R protein in colon tissue was significantly decreased(P<0.001),and the expression levels of p-STAT3,NF-κB p65 and VEGF proteins were significantly increased(all P<0.001)in the model group.Compared with the model group,the colon tumor load,colon histopathological score and the levels of p-STAT3,NF-κB p65 and VEGF proteins in colon tissue were significantly decreased(all P<0.05)in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group while the expression levels of P2X7R protein in colon tissue were significantly increased(both P<0.05).Conclusion:Both herb-partitioned moxibustion and ginger-partitioned moxibustion can reduce the colon tumor load in CACC rats and delay the progression of colon adenomas.The mechanism may be mediated by the P2X7R/STAT3 pathway to inhibit STAT3 phosphorylation,thereby reducing VEGF protein expression.