中国全科医学
中國全科醫學
중국전과의학
CHINESE GENERAL PRACTICE
2015年
21期
2542-2547
,共6页
王丽%王兰辉%孟翠巧%李立萍%宋广昊%任路平%李国风
王麗%王蘭輝%孟翠巧%李立萍%宋廣昊%任路平%李國風
왕려%왕란휘%맹취교%리립평%송엄호%임로평%리국풍
硫化氢%肝炎%炎性反应%大鼠
硫化氫%肝炎%炎性反應%大鼠
류화경%간염%염성반응%대서
Hydrogen sulfide%Hepatitis%Inflammatory reaction%Rats
目的：观察硫化氢（ H2 S）对非酒精性脂肪性肝炎（ NASH）大鼠炎性因子水平的影响，探讨H2 S防治NASH的可能机制。方法2013年1月—2014年7月选用SPF级雄性SD大鼠60只，按照随机数字表法分为对照组、NASH模型组、硫氢化钠（ NaHS）干预组，各20只。对照组给予普通饲料， NASH模型组给予高脂饲料， NaHS干预组给予高脂饲料的同时腹腔注射H2S供体NaHS （56 mg? kg-1? d-1），对照组和NASH模型组每日腹腔注射等量0.9％氯化钠溶液。8周后处死所有大鼠，留取静脉血及肝脏组织。检测血清及肝脏生化指标〔血清丙氨酸氨基转移酶（ALT）﹑天冬氨酸氨基转移酶（AST）、三酰甘油（TG）、总胆固醇（TC）及肝脏组织TG、 TC〕﹑血浆及肝脏组织H2 S水平、肝脏组织氧化应激指标〔丙二醛（ MDA）、超氧化物歧化酶（ SOD）〕； HE染色观察肝脏组织病理学变化；检测肝脏组织肿瘤坏死因子α（TNF-α）、白介素6（IL-6） mRNA表达水平及肝脏组织TNF-α、 IL-6、核因子κB抑制因子α（IκBα）、核因子κB p65（NF-κB p65）表达水平。结果 NASH模型组和NaHS干预组血清ALT、 AST、 TG、TC水平及肝脏组织TG、 TC水平均高于对照组，且NaHS干预组以上指标均低于NASH模型组（ P＜0.05）。 NASH模型组血浆H2 S水平和肝脏组织H2 S水平均低于对照组及NaHS干预组（ P＜0.05）。与对照组比较， NASH模型组和NaHS干预组肝脏组织MDA水平升高， SOD水平降低（ P＜0.05）；与NASH模型组比较， NaHS干预组MDA水平降低， SOD水平升高（P＜0.05）。与对照组比较， NASH模型组肝脏组织结构紊乱，呈弥漫性脂肪变性、肝细胞灶性坏死，并可见淋巴细胞和中性粒细胞浸润； NaHS干预组肝脏组织脂肪变性、坏死及炎性反应程度较NASH模型组明显减轻。NASH模型组和NaHS干预组肝脏组织TNF-α、 IL-6 mRNA表达水平和TNF-α、 IL-6表达水平均高于对照组，且NASH模型组以上指标均高于NaHS干预组（ P＜0.05）。与对照组比较， NASH模型组和NaHS干预组细胞质IκBα表达水平均降低，细胞核NF-κB p65表达水平均升高（P＜0.05）；与NASH模型组比较， NaHS干预组细胞质IκBα表达水平升高，细胞核NF-κB p65表达水平降低（P＜0.05）。结论 H2S能够有效改善NASH大鼠肝脏组织病理变化，并抑制炎性反应，这可能是H2 S发挥治疗作用的机制之一。
目的：觀察硫化氫（ H2 S）對非酒精性脂肪性肝炎（ NASH）大鼠炎性因子水平的影響，探討H2 S防治NASH的可能機製。方法2013年1月—2014年7月選用SPF級雄性SD大鼠60隻，按照隨機數字錶法分為對照組、NASH模型組、硫氫化鈉（ NaHS）榦預組，各20隻。對照組給予普通飼料， NASH模型組給予高脂飼料， NaHS榦預組給予高脂飼料的同時腹腔註射H2S供體NaHS （56 mg? kg-1? d-1），對照組和NASH模型組每日腹腔註射等量0.9％氯化鈉溶液。8週後處死所有大鼠，留取靜脈血及肝髒組織。檢測血清及肝髒生化指標〔血清丙氨痠氨基轉移酶（ALT）﹑天鼕氨痠氨基轉移酶（AST）、三酰甘油（TG）、總膽固醇（TC）及肝髒組織TG、 TC〕﹑血漿及肝髒組織H2 S水平、肝髒組織氧化應激指標〔丙二醛（ MDA）、超氧化物歧化酶（ SOD）〕； HE染色觀察肝髒組織病理學變化；檢測肝髒組織腫瘤壞死因子α（TNF-α）、白介素6（IL-6） mRNA錶達水平及肝髒組織TNF-α、 IL-6、覈因子κB抑製因子α（IκBα）、覈因子κB p65（NF-κB p65）錶達水平。結果 NASH模型組和NaHS榦預組血清ALT、 AST、 TG、TC水平及肝髒組織TG、 TC水平均高于對照組，且NaHS榦預組以上指標均低于NASH模型組（ P＜0.05）。 NASH模型組血漿H2 S水平和肝髒組織H2 S水平均低于對照組及NaHS榦預組（ P＜0.05）。與對照組比較， NASH模型組和NaHS榦預組肝髒組織MDA水平升高， SOD水平降低（ P＜0.05）；與NASH模型組比較， NaHS榦預組MDA水平降低， SOD水平升高（P＜0.05）。與對照組比較， NASH模型組肝髒組織結構紊亂，呈瀰漫性脂肪變性、肝細胞竈性壞死，併可見淋巴細胞和中性粒細胞浸潤； NaHS榦預組肝髒組織脂肪變性、壞死及炎性反應程度較NASH模型組明顯減輕。NASH模型組和NaHS榦預組肝髒組織TNF-α、 IL-6 mRNA錶達水平和TNF-α、 IL-6錶達水平均高于對照組，且NASH模型組以上指標均高于NaHS榦預組（ P＜0.05）。與對照組比較， NASH模型組和NaHS榦預組細胞質IκBα錶達水平均降低，細胞覈NF-κB p65錶達水平均升高（P＜0.05）；與NASH模型組比較， NaHS榦預組細胞質IκBα錶達水平升高，細胞覈NF-κB p65錶達水平降低（P＜0.05）。結論 H2S能夠有效改善NASH大鼠肝髒組織病理變化，併抑製炎性反應，這可能是H2 S髮揮治療作用的機製之一。
목적：관찰류화경（ H2 S）대비주정성지방성간염（ NASH）대서염성인자수평적영향，탐토H2 S방치NASH적가능궤제。방법2013년1월—2014년7월선용SPF급웅성SD대서60지，안조수궤수자표법분위대조조、NASH모형조、류경화납（ NaHS）간예조，각20지。대조조급여보통사료， NASH모형조급여고지사료， NaHS간예조급여고지사료적동시복강주사H2S공체NaHS （56 mg? kg-1? d-1），대조조화NASH모형조매일복강주사등량0.9％록화납용액。8주후처사소유대서，류취정맥혈급간장조직。검측혈청급간장생화지표〔혈청병안산안기전이매（ALT）﹑천동안산안기전이매（AST）、삼선감유（TG）、총담고순（TC）급간장조직TG、 TC〕﹑혈장급간장조직H2 S수평、간장조직양화응격지표〔병이철（ MDA）、초양화물기화매（ SOD）〕； HE염색관찰간장조직병이학변화；검측간장조직종류배사인자α（TNF-α）、백개소6（IL-6） mRNA표체수평급간장조직TNF-α、 IL-6、핵인자κB억제인자α（IκBα）、핵인자κB p65（NF-κB p65）표체수평。결과 NASH모형조화NaHS간예조혈청ALT、 AST、 TG、TC수평급간장조직TG、 TC수평균고우대조조，차NaHS간예조이상지표균저우NASH모형조（ P＜0.05）。 NASH모형조혈장H2 S수평화간장조직H2 S수평균저우대조조급NaHS간예조（ P＜0.05）。여대조조비교， NASH모형조화NaHS간예조간장조직MDA수평승고， SOD수평강저（ P＜0.05）；여NASH모형조비교， NaHS간예조MDA수평강저， SOD수평승고（P＜0.05）。여대조조비교， NASH모형조간장조직결구문란，정미만성지방변성、간세포조성배사，병가견림파세포화중성립세포침윤； NaHS간예조간장조직지방변성、배사급염성반응정도교NASH모형조명현감경。NASH모형조화NaHS간예조간장조직TNF-α、 IL-6 mRNA표체수평화TNF-α、 IL-6표체수평균고우대조조，차NASH모형조이상지표균고우NaHS간예조（ P＜0.05）。여대조조비교， NASH모형조화NaHS간예조세포질IκBα표체수평균강저，세포핵NF-κB p65표체수평균승고（P＜0.05）；여NASH모형조비교， NaHS간예조세포질IκBα표체수평승고，세포핵NF-κB p65표체수평강저（P＜0.05）。결론 H2S능구유효개선NASH대서간장조직병리변화，병억제염성반응，저가능시H2 S발휘치료작용적궤제지일。
Objective To observe the effect of hydrogen sulfide ( H2 S) on the levels of inflammatory cytokines in liver tissue of rats with nonalcoholic steatosishepatitis (NASH), and to explore the mechanisms of the anti-NASH effect of H2S. Methods 60 male SD rats at SPF level were randomly and equally divided into control group, NASH model group and NaHS intervention group.Rats in control group were fed with normal diet, rats in NASH model group were fed with high fat diet, rats in NaHS intervention group were fed with high fat diet and were injected intraperitoneally with NaHS (56 mg? kg-1? d-1 ) , rats in control group and NASH model group were injected intraperitoneally with 0.9% sodium chloride solution.At the end of the 8th week, rats in each group were killed to obtain venous blood and liver tissue samples.The serum level of alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST ) , triglyceride ( TG ) , cholesterol ( TC ) and H2 S, the levels of TG, TC, malondialdehyde ( MDA) , superoxide dismutase ( SOD ) and H2 S in liver tissues were detected by kit.The histopathological changes of the liver tissues were observed under microscope with HE staining.The mRNA and protein expression levels of TNF-αand IL-6 in liver tissues were detected.The protein expression levels of IκBαand NF-κB p65 in the liver tissues were detected.Results The serum levels of ALT, AST, TG, TC, and the levels of TG, TC in hepatic tissues in NASH model group and NaHS intervention group were significantly higher than those in control group respectively, the above indicators in NaHS intervention group were significantly lower than those in NASH model group respectively ( P<0.05) .The levels of H2 S in blood and hepatic tissues in NASH model group were significantly lower than those in NaHS intervention group and control group respectively ( P <0.05 ) .The level of MDA in hepatic tissues in NASH model group and NaHS intervention group was significantly higher than that in control group respectively, whereas the level of SOD in hepatic tissues in NASH model group and NaHS intervention group was significantly lower than that in control group respectively ( P<0.05 ) .The MDA level in NASH model group was significantly lower than that in NaHS intervention group, and the SOD level in NASH model group was significantly higher than that in NaHS intervention group ( P<0.05 ) .Compared with the liver tissues of rats in control group, the liver tissues of rats in NASH model group showed structure disturbance, macrovesicular steatosis, localized necrosis, neutrephil and lymphocytes infiltration.The levels of macrovesicular steatosis, necrosis, and inflammatory reaction in hepatic tissues in NaHS intervention group were significantly lower than those in NASH model group respectively.The mRNA and protein expression levels of TNF-αand IL-6 in liver tissues in NASH model group and NaHS intervention group were significantly higher than those in control group respectively, and the above indicators in NaHS intervention group were significantly lower than those in NASH model group respectively ( P<0.05) .The protein expression level of IκBαin cytoplasm in NaHS intervention group and NASH model group was significantly lower than that in control group respectively, the protein expression level of NF-κB p65 in cell nucleus in NaHS intervention group and NASH model group was significantly higher than that in control group respectively (P<0.05) .The protein expression level of IκBαin cytoplasm in NaHS intervention group was significantly higher than that in NASH model group, the protein expression level of NF-κB p65 in cell nucleus in NaHS intervention group was significantly lower than that in NASH model group ( P <0.05 ) .Conclusion H2 S is effective in improving the pathological changes in hepatic tissues, and it can inhibit inflammatory reaction, which might be one of the mechanisms of the anti-NASH effect of H2 S.
