| BackgroundIn recent years,the incidence of urinary stones has been rising,which has occupied the first place in clinical department of urology disease incidence.There were nearly 200/10,0000 cases of new cases each year in China.Urinary system stone(also known for urinary tract stones or urolithiasis)includes kidney stones,ureteral calculus,calculus of urethra,bladder stones,causing inflammation,urethral obstruction,edema and hydronephrosis.Urinary system stone is one of the important diseases to human health hazards.Although Urinary system stone has been a hundred years of history,it is still difficult to clarify,and the occurrence and recurrence rates are high.The recurrence rates are 6%,14%,22.8%,30% in half year,1-year,2-year and 10-year,and the recurrence rate is still increased.Although diagnosis and treatment technology in the past decades have made considerable progress,the morbidity and recurrence rates remain stubbornly high and shows an upward trend.The reason is mainly due to the pathogenesis of kidney stones is not clear.Therefore,convenient and stable experimental animal oxalic acid calcium crystallization model is an important tool for the study of kidney stones and to explore the mechanism of emergence and prevention and cure of crystal of kidney.Traditional Chinese medicine in the treatment of urinary stones has a long history due to its curative effect,less adverse reactions.Christina loosestrife herb belongs to the family Fabaceae family members,including 349 species of plants.Some of them can be used as medicine.Christina loosestrife herb plant extracts in vitro and in vivo reveal the function of anti-stones,anti-inflammatory,cytotoxicity,anti-diabetic etc.At present in the clinic,christina loosestrife herb is the most commonly in treatment of urinary calculi.Recently,commercial medicinal materials are mainly divided into herba lysimachia,Sichuan lysimachia,lysimachia grass and Jiangxi lysimachia.In addition,the pharmacological function of herba lysimachia is most effective of the all.Part 1 The biological function of different active fraction of herba lysimachia in alleviate the kidney injury by crystallization of calcium oxalatObjective: To evaluate the effects of different active fraction of herba lysimachiae in mice kidney injury by crystallization of calcium oxalate in vivo.Methods: 60 adult male C57BL/6 mice were randomly divided into six groups,including control group(A group),purely crystalline group(B group),petroleum intervention group(C group),ethyl acetat intervention group(D group),n-butanoicacid intervention group(E group),aqueous solution intervention group(F group).Mice of group A were injected normal saline.Mice of group B were injected crystalline(100 mg/kg).Mice of group C were injected crystalline(100 mg/kg)+ petroleum(20 m L/kg).Mice of group D were injected crystalline(100 mg/kg)+ ethyl acetat(20 m L/kg).Mice of group E were injected crystalline(100mg/kg)+ n-butanoicacid(20 m L/kg).Mice of group F were injected crystalline(100mg/kg)+ aqueous solution(20 m L/kg).At 5d,mice were sacrificed and their blood and kidneys tissue were harvested to analyse the serum creatinine,urea nitrogen,calcium concentration levels.Furthermore,we observed the location of crystallization of calcium oxalate in vivo kidney tissue and evaluated the pathological changes by microscopy.Results: Level of serum creatinine,urea nitrogen,calcium concentration were significant lower in C,D,E group compared with B group(P<0.05).Meanwhile,the results of crystallization of calcium oxalate and pathological changes are much lower than those in B group(P<0.05).Conclusion: Three extract part of herba lysimachiae(petroleum,ethyl acetat,n-butanoicacid)reduce crystallization of calcium oxalate content in the kidney tubules,which would protect kidney.Part 2 The biological mechanism of different active fraction of herba lysimachia in alleviate the kidney injury by crystallization of calcium oxalateObjective: To evaluate the effects of different active fraction of herba lysimachiae in mice kidney injury by crystallization of calcium oxalate in vivo.Methods: 60 adult male C57BL/6 mice were randomly divided into six groups,including control group(A group), purely crystalline group(B group),petroleum intervention group(C group),ethyl acetat intervention group(D group),n-butanoicacid intervention group(E group),aqueous solution intervention group(F group).Mice of group A were injected normal saline.Mice of group B were injected crystalline(100 mg/kg).Mice of group C were injected crystalline(100 mg/kg)+ petroleum(20 ml/kg).Mice of group D were injected crystalline(100 mg/kg)+ ethyl acetat(20 ml/kg).Mice of group E were injected crystalline(100mg/kg)+ n-butanoicacid(20 ml/kg).Mice of group F were injected crystalline(100 mg/kg)+ aqueous solution(20 ml/kg).At 5d,mice were sacrificed and their blood and kidneys tissue were harvested to analyse.The expression levels of CAT,Cu Zn-SOD,GSH,MDA,TRPV5,Calbindin-D28,osteopontin,CD44,Ki67 and TUNEL were detected by real-time polymerase chain reaction(RT-PCR)and immunohistochemistry.Results: Levels of CAT,Cu Zn-SOD,GSH,osteopontin,CD44,TRPV5,Calbindin-D28 and Ki67 were significant higher in C,D,E group compared with B group(P<0.05).The contents of MDA and TUNEL were lower in C,D,E group compared with B group(P<0.05).Conclusion: Three extract part of herba lysimachiae(petroleum,ethyl acetat,n-butanoicacid)reduce crystallization of calcium oxalate content,promoted proliferation,and inhibit oxidative stress and apoptosis in the kidney tubules,which would protect kidney. |
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