| ObjectiveTo clear that rhizoma coptis chinensis compatibility can reduce cinnabarinduced kidney damage,to make a thorough inquiry the meterial base and mechanism of rhizoma coptis chinensis compatibility reduce cinnabar-induced kidney damage preliminarily,to provide a theoretical basis for explaining the cinnabar-containing formula are reasonable and scientific.Methods30 Healthy SD female rats(130~160 g)were selected,and randomly divided into 6 groups(5 per group),that was,control group,rhizoma coptis chinensis group,cinnabar group,rhizoma coptis chinensis and cinnabar group,berberine and cinnabar group,coptisine and cinnabar group.The rats were respectively given by gavage 0.5% sodium carboxymethyl cellulose,3.33g/kg rhizoma coptis chinensis,1.00 g/kg cinnabar,1.00 g/kg cinnabar & 3.33g/kg rhizoma coptis chinensis,1 g/kg cinnabar & 0.23 g/kg berberine,1.00 g/kg cinnabar & 0.08 g/kg coptisine,once a day for 12 weeks.During the experiment,the rats were weighed twice a week,and the dosage according to body weight adjustment.The situation in rats such as behavior,diet,appearance,excretion,etc,were observed everyday.After the end of 12 weeks of intragastric gavage,rats were put into metabolic cages alone and fasting couldn't help but water,collecting 24 h urine.The blood and kidney samples were collected when the rats were under anesthesia.The mercury content in the blood,the kidney andurine were determined by Zeeman atomic absorption mercury analyzer.The concentrations of Serum Creatinine(SCr)and Blood Uric Acid(BUA)were measured by Automatic Biochemical Detector.Kidney slice were made,and pathological changes were observed under light microscope by HE staining and Masson staining.Gene expression levels of TNF-??IL-1??IL-4 and IL-10 were tested by Real-time fluorescent quantitative(q-PCR).Protein expression levels of TNF-??IL-1??IL-4 and IL-10 were tested by enzyme-linked immuno sorbent assay(ELISA).The data were represented by x ąs,and analyzed by SPSS19.0statistical software.The comparison among the groups using single factor analysis of variance(One-Way ANOVA)or repeated measures of variance,and the difference between two groups was analyzed with S-N-K(equal variances assumed)or Dunnett's T3 analysis(equal variances not assumed).Drug interactions were analyzed with 2×2 Univariate.Test level is 0.05.Results1.In general,the weight of rats increased normally during the experimental period,there was no significant difference in each groups.The feces of rats in cinnabar group,rhizoma coptis chinensis & cinnabar group,berberine &cinnabar group,coptisine & cinnabar group were red;The feces in rhizoma coptis chinensis & cinnabar group,rhizoma coptis chinensis group were thin,and shape of feces in other groups was no obvious abnormality.2.After 12 weeks of continuous administration,blood mercury(BHg),kidney mercury(KHg),and urinary mercury(UHg)levels in the cinnabar group were significantly higher than those in the control group and the rhizoma coptis chinensis group(P<0.05).The content of BHg,KHg,and UHg in the rhizoma coptis chinensis group were not significantly different from those in the control group.Coptis & cinnabar the content of blood mercury,kidney mercury and urine mercury in rats have interaction effects(P<0.05).The content of BHg,KHg and UHg levels in the three compatibility groups,that was,rhizoma coptischinensis & cinnabar group,berberine & cinnabar group,and Coptisine &cinnabar group,were all lower than those in the cinnabar group.The difference was statistically significant.The content of BHg and KHg levels in the three compatibility groups were berberine & cinnabar group>coptisine & cinnabar group>rhizoma coptis chinensis & cinnabar group.The differences in each groups were statistically significant(P<0.05).There was no significant difference in UHg content between these three groups.3.After 12 weeks of continuous administration,the content of SCr in cinnabar were higher than Control group and rhizoma coptis chinensis group(P<0.05),there was no significant difference between the three compatibility groups.The content of BUA in cinnabar were higher than control group and rhizoma coptis chinensis group(P<0.05);The content of BUA in rhizoma coptis chinensis & Cinnabar group and coptisine & cinnabar group were lower than the cinnabar group,the difference was statistically significant(P<0.05),there was no significant difference between the berberine &cinnabar group and the cinnabar group;There was no significant difference between coptisine & cinnabar group and rhizoma coptis chinensis & Cinnabar group.The analysis of factorial variance showed that taking cinnabar had an effect on the levels of SCr and BUA in rats,taking rhizoma coptis chinensis had no effect on the levels of SCr and BUA in rats.Rhizoma coptis chinensis compatibility with cinnabar had interaction effects on SCr and BUA levels in rats.4.After 12 weeks of continuous administration,Under the light microscopy kidney pathology staining results showed that,In the cinnabar group,kidney pathology changes were characterized by extensive inflammatory cell infiltration,the part of kindey proliferated collagen fibers,part of the kidney dilated tubules and protein casts in dilated tubules,kidney medulla appeared mesh-like structure were observed;In rhizoma coptis chinensis group explicit showed epithelial cell vacuolation,protein casts in tubules occasionally were observed,kindey proliferated collagen fibers were not observed;In rhizoma coptis chinensis & Cinnabar group,berberine & cinnabar group,coptisine & cinnabar group,kidney pathology changes were characterized by inflammatory cell infiltration occasionally,part of kidney tubules were fade away,protein casts occasionally were observed,The extent of fibrous tissue proliferation was slighter than that were observed in cinnabar group.The results of fiber hyperplasia score showed that the cinnabar group was higher than the control group and rhizoma coptis chinensis group,and the difference was statistically significant(P<0.05).The fiber proliferative degree scores of the three compatibility groups were all lower than the cinnabar group,and the difference was statistically significant(P< 0.05);Compared with rhizoma coptis chinensis &cinnabar group,the score of fibrosis of berberine & cinnabar group and coptisine & cinnabar group increased,the difference was statistically significant(P<0.05).5.After 12 weeks of continuous administration,the gene expression levels of cytokines TNF-?,IL-1?,IL-4 and IL-10 in kidney tissue of rats in cinnabar group were higher than those in control group and rhizoma coptis chinensis group,and the difference was statistically significant(P<0.05).Compared with the cinnabar group,the gene expression of TNF-? and IL-1? in the three compatibility groups was down-regulated(P<0.05),and there was no significant difference among the three compatibility groups.Compared with Cinnabar group,IL-4 gene expression was increased in rhizoma coptis chinensis &Cinnabar group group and coptisine & cinnabar group(P<0.05),and there was no significant difference between berberine & cinnabar group and cinnabar group,IL-4 gene expression levels in coptisine cinnabar group.It was lower than rhizoma coptis chinensis & Cinnabar group(P<0.05).Compared with the cinnabar group,the gene expression of IL-10 in the kidney of cinnabar group was increased(P<0.05).There was no significant difference between theberberine & cinnabar group,coptisine & cinnabar group and cinnabar group.The analysis of factorial variance showed that taking rhizoma coptis chinensis and cinnabar had an effect on the gene expression levels of cytokines TNF-?,IL-1?,IL-4,and IL-10 in rat kidney tissue respectively;Rhizoma coptis chinensis used with cinnabar,there was an interaction between the gene expression levels of cytokines TNF-?,IL-1? and IL-4 in rat kidney tissue,and there was no interaction effect on the gene expression levels of cytokine IL-10 in rat kidney tissue.6.After 12 weeks of continuous administration,the protein expression levels of cytokines TNF-?,IL-1?,IL-4 and IL-10 in kidney tissue of rats in cinnabar group were higher than those in control group and rhizoma coptis chinensis group,and the difference was statistically significant(P<0.05).Compared with the cinnabar group,the protein expression levels of TNF-? and IL-1? in the three compatibility groups was down-regulated(P<0.05),and there was no significant difference among the three compatibility groups.Compared with Cinnabar group,cytokines IL-4 protein expression levels was increased in rhizoma coptis chinensis & Cinnabar group and coptisine & cinnabar group(P<0.05),and there was no significant difference between berberine &cinnabar group and cinnabar group.Compared with the cinnabar group,the protein expression levels of IL-10 in the kidney of cinnabar group was increased(P<0.05),and there was no significant difference between the berberine &cinnabar group,coptisine & cinnabar group and cinnabar group.The analysis of factorial variance showed that taking rhizoma coptis chinensis and cinnabar had an effect on the protein expression levels of cytokines TNF-?,IL-1?,IL-4,and IL-10 in rat kidney tissue respectively,and taking rhizoma coptis chinensis used with cinnabar,there was an interaction between the protein expression levels of cytokines TNF-?,IL-1?,IL-4 and IL-10 in rat kidney tissue.ConclusionsThe long-term oral administration of cinnabar in large quantities was likely to cause mercury accumulation in the kidneys of rats and kidney tissue damage.Rhizoma Coptis chinensis and cinnabar compatibility were possible reducing cinnabar induced the nephrotoxicity,and the main components of berberine and berberine are the material basis for the compatibility to reduce the poison.The mechanism of rhizoma coptidis in alleviating cinnabar-induceed nephrotoxicity may be: reducing blood mercury absorptive capacity in rats,thereby reducing the kidney mercury accumulation;Down-regulate the expression of pro-inflammatory cytokines IL-1?,TNF-? and up-regulate the expression of anti-inflammatory cytokines IL-4,IL-10. |
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