Experimental Study On The Prevention And Treatment Of Delayed Diarrhea In Mice Treated By Sishen Pill

Objective:Delayed diarrhea is the main dose-limiting toxicity of irinotecan(Irinotecan;trade name:Camptorcin-11,CPT-11),which seriously affects the quality of life of patients and even leads to treatment interruption and affect the efficacy of patients.There is still a lack of effective prevention and treatment methods.In this study,we established CPT-11-induced delayed diarrhea in mice to study the preventive effect of Sishen pills on CPT-11-induced delayed diarrhea and to explore its possible mechanism.Methods:1)Establishment and grouping of delayed diarrhea model.Randomly fifty healthy male ICR mice were grouped,including diarrhea model group,Sishen pill low(0.6g·kg-1),medium(1.2g·kg-1)and high(2.4g·kg-1)dose group,normal control group,10 mice in each group.The mice except normal control group were injected CPT-11(35mg·kg-1·d-1)via caudal vein daily,for 4 continuous days.The normal control group was injected with the equal volume of NS.Suspension of Sishen Pill powder was administered via gavage in each treated group before Tail vein injection daily for 7 consecutive days.Normal control group was received equal volume of distilled water by gavage,the same as diarrhea model group.2)The severity of delayed diarrhea,diet and mental activity status were daily observed and recorded;3)Sample collection.After the last 24 hours of gavage,abdominal aorta bleed were collected under 10%chloral hydrate anesthesia;and then the colon tissue and cecal contents were sampled after the necks of mice were pulled off.4)Detection of samples.The serum levels of IL-1? and TNF-? in were measured by ELISA;The concentrations of P-glucuronidase(P-G)in cecal contents were detected by colorimetry;Escherichia coli of cecal contents were cultured,and the number of colonies was counted;Colon tissue was fixed,sliced and stained by HE,and the histopathology of colonic mucosa was surveyed under regular optical microscope.Results:1)The occurrence of delayed diarrhea.After the injection of CPT-11 in the tail vein of the mice,delayed diarrhea occurred after 24h,and the diarrhea was 100%in the diarrhea model group.Especially the incidence of severe diarrhea was 80%.But,the incidence of severe diarrhea was 40%in the middle and high dose groups,and compared with the diarrhea model group,the degree was significantly reduced(P =0.021,P = 0.019).2)Damage of colonic mucosa.The colonic mucosal injury were lower in the middle and high dose groups(P<0.05).3)The changes of serum IL-1? in mice.The levels of IL-1? in the medium and high dose group was respectively(72.61 ± 10.15)/g·L-1 and(71.86 ± 8.78)/g·L-1,significantly below those in the diarrhea model group(P<0.01).4)The changes of serum TNF-a in mice.The serum TNF-a levels in the medium and high dose group of Sishen pill were significantly decreased than those in the diarrhea model group(P<0.01),was separately(71.50± 3.39)/g·L-1,(69.43 ± 6.23)/g·L-1 and(109.50 ± 4.21)/g·L-1.5)The level of intestinal p-G in mice.The expression of ?-G in the medium and high dose group[(1.109 ± 0.055)U and(1.100 ±0.063)U]was significantly decreased than that in the diarrhea model group(1.438 ± 0.022)U(P<0.01).6)The colonies number of Escherichia coli in the cecum.The Escherichia coli colonization of the cecum in the normal control group,the diarrhea model group and the each dose group of Sishen pill was respectively(31.2 ± 2.9)× 1013/g,(31.4± 3.2)×1013/g,(29.5 ± 2.2)×1013/g,(30.4 ± 3.5)×1013/g and(31.4 ± 2.7)×1013/g,and There was no significant change in each group(P>0.05).Conclusion:Sishen pills have some preventive and mitigation effects on CPT-11-induced delayed diarrhea in mice,and its mechanism may be to decrease the levels of ?-G,and to reduce the levels of IL-1? and TNF-? in serum.