Experimental Study, Heat-producing Cell Phenotype Changes In Barriers To The Dominant Spleen Deficiency Disease

Spleen yang deficiency is a representative clinic deficient one.For yang deficiency is related to the hypothermogenesis,spleen deficiency is related to the indigestion,So those could be found that the biomolecules of spleen yang deficiency and the regulative thermones bridging yang and spleen deficiencies.1 AimThe aim of this study is to explore the pathogenesis of spleen yang deficiency.A crosstalk between the cold due to yang deficiency and the indigestion due to spleen deficiency is to be explored by utilizing the two pathological processes of hypothermogenesis and indigestion.The dystransformation of thermogenic cell phenotype in rat with spleen yang deficiency would be understood well.2 MethodsAccording to the two pathological processes of hypothermogenesis and indigestion,the spleen yang deficiency model of rats in the same litter was replicated with partal thyroidectomy at dl- interscapular brown adipose tissueexcision-feding a fat diet from d₅₆,andits cold features were evaluated with the therapeutic effects of thermogenesis-promoting decoction.(1) Isoproterenol-induced thermogenesis:Isoproterenol optimic dose for the thermogenesis of rat at thermoneutrality(28℃) was conformed by using no-linear dose-effect relationship with Prism 4.(2)Etiological factors erosstalking in rats with spleen yang deficiency:After being born(d₁),rats were given the intraperitoneal injection with a series ratio of ¹³¹I at thermoneutrality(28℃)(developmental hypothermogenesis).,They were fed with higher lipid 24 h then fasted 24 h interval from d₅₆,senno sides were given intragastrically for 14 d to make them sensitive to digestive load from d₇₀(acquired hypothermogenesis).The core temperatures induced with isoproterenol in rats at thermoneutrality(28℃) were detected.The correlative analysis between the digestive load and thyroidectomy.Crosstalk between congenital and postnatal pathogen was confirmed by using no-linear dose-effect analyzing.(3)Scale evaluation of severity in rats with spleen yang deficiency:Rat model of spleen yang deficiency was replicated with partal thyroidectomy and three kinds of digestive loads.A scale for evaluating manefastations in the rat model was set up from clinic patients with the spleen yang deficient shydromes.The severities were evaluated with the scale at four endpoints.Corelations between manefastations in the model rats were analyzied with no-linear dose-effect,the coordination between experimental rats and clinic patients was calculated on the theory frequency. (4) Acquired etiology in rats with spleen yang deficiency:Five littermates in each litter were bred sequentially and randomized into five groups,lnterscapular brown adipose tissue was removed from the rats at d₄₂ after birth.Rats were fed cholesterol ester from d₅₆, then intragastrically administrated a decoction of medical herbs to promote or inhibit chronic the thermogenesis from d₇₀.plasma ghrelin was detected on radioimmunoassay,plasma metabolite levels with use of an automatic chemistry analyzer,and ghrelin level in the gastric mucous membrane on immunohistochemistry and morphometry.The effects of traditional Chinese medicine Fuzi-Lizhong Pill was confirmed by t-test.(5)Developmental etiology in rats with spleen yang deficiency:Nine littermates in each litter were bred sequentially and randomized into 9 groups.Thyroidectomy in hypothyroidism littermates was went with ¹³¹iodine to destroy partly thyroidal tissue at d₁ after birth,interscapular brown adipose tissue was removed at d₄₂,littermates were fed with more cholesterol ester from d₅₆,then administered intragastrically a decoction of medical herbs to promote or inhibit chronic thermogenesis from d₇₀.Plasma ghrelin or thyroid-related hormone did on radioimmunoassay,plasma metabolite levels with use of an automatic chemistry analyzer,and ghrelin level in the gastric mucous membrane on immunohisto chemistry and morphometry,he potency data were used in statistical analysis.The effects of traditional Chinese medicine Fuzi-Lizhong Pill was confirmed by t-test.(6)Both etiologies in rats with spleen yang deficiency:Nine littermates in each litter were bred sequentially and randomized into 9 groups.Thyroidectomy in hypothyroidism littermates was went with ¹³¹iodine to destroy partly thyroidal tissue at dl after birth, interscapular brown adipose tissue was removed at d₄₂,littermates were fed with more cholesterol ester from d₅₆,then administered intragastrically a decoction of medical herbs to promote or inhibit chronic thermogenesis from d₇₀.Thermogenesis,plasma ghrelin, thyroid-related hormone,fatty degeneration in adrenal gland,and B lymphocyte loss in spleen were observed.The hypo- thyroid potency data were derives from the primary data.The variation between various groups t-test was evaluated in t-test3 ResultsBoth of acquired and developmental etiologies involved in the pathogenic vicious circle of the spleen yang deficient syndrome in rats.(1)Isoprenaline-inducesed thermogenesis:Ones of isoproterenol ED₅₀ from core temperature as the peak and the area,were 0.28 mg·kg^-1 and 0.106 mg·kg^-1,respectively. Those from chest skin temperatures were 0.066 mg·kg^-1 and 0.070 mg·kg^-1,and from extrimity skin temperatures were 0.071 mg·kg^-1 and 0.072 mg·kg^-1.There was all positive correlation between them.(2)Etiological factors crosstalking in rats with spleen yang deficiency:There was negative correlation between rat temperature changes(peaks and areas) induced with isoproterenol before administrated senna and after,Hypothermogenesis in rats aggravates the ingestion caused by administrated senna(The ¹³¹I-ED₅₀ of thermogenic peak in rats were decreased from 21201 nCi·g^-1 to 4064nCi·g^-1.The ¹³¹I- ED₅₀ of thermogenic area in rats were decreased from 19209 nCi·g^-1 to 9027 nCi·g^-1).(3)Scale evaluation of severity in rats with spleen yang deficiency:The closed linear relationship was found between severity of the yang deficienct and spleen deficiency with the degree of hypothyrodism(P＜0.01).The correlations are significant between the weight loss and cold extremities,less activity and cold extremities,weight loss and less activity(P＜0.05); eating less and diarrhea,eating less and abdominal pain(P＜0.01);So did between eating less and less activity,ating less and weight loss,cold extremities and abdominal pain(P＜0.05).(4)Acquired etiology in rats with spleen yang deficiency:Removed interscapularbrown adipose tissue in rats showed significantly decreased thermogenic peak by3.18%and 3.38%(P＜0.01),increased plasma ghrelin level by 129.88%(P＜0.01),and decreased ghrelin level in the gastric mucous membrane by 34.30%.(P＜0.01),The plasma ghrelin level and the existing gastric ghrelin level were inversely correlated,increased fatty degeneration in adrenal gland(P＜0.05)and B lymphocyte loss in spleen lost(P＜0.05).These changes were supported by levels of plasma thyroid hormones or metabolites affected by the thermogenic decoctions from selected medical herbs..(5)Developmental etiology in rats with spleen yang deficiency:Removed interscapular brown adipose tissue in eu- or hypothyroidism rats showed significantly decreased thermogenesis.the hypothyroidism aggravated the decrease by 67.62%(P＜0.01).The removed interscapular brown adipose tissue in hypothyroidism rats demonstrated significantly increased plasma ghrelin level and decreased gastric ghrelin level(P＜0.01).The fat diet aggravated the increase by158.95%(P＜0.01 ).(6)Both etiologies in rats with spleen yang deficiency:Hypothyroid potency data are used here to simplify the changes in hypothyroid rats for digging out the hypothermogenic biochemistry and its clinic application.Removed interscapular brown adipose tissue in rats showed thermogenesis peak decreased(-0.70±0.36),Fuzi-Lizhong Pill increased it (-0.48±0.16,P＜0.01) and the Huanglian-Jiedu decoction aggravated it(-1.07±0.53,P＜0.01); and the Fuzi-Lizhong Pill increase it from the Huanglian-Jiedu decoction(P＜0.01).Removed interscapular brown adipose tissue in rats showed thermogenesis peak decreased (-0.45±0.14),Fuzi-Lizhong Pill increased it(-0.48±0.16,P＜0.05) and the Huanglian-Jiedu decoction aggravated it(-1.07±0.59,P＜0.01);and the Fuzi-Lizhong Pill increase it from the Huanglian-Jiedu decoction(P＜0.01);Plasma ghrelin lever showed the Fuzi-Lizhong Pill (-0.69±1.79) lower than the Huanglian-Jiedu decoction(-1.58±3.19,P＜0.05).The gastric ghrelin lever showed the Fuzi-Lizhong Pill(0.40±0.38) more than the Huanglian-Jiedu decoction(-0.51±0.22,P＜0.05).4 ConclusionsBoth of acquired and developmental etiologies involved in the pathogenic vicious circle of the spleen yang deficient syndrome in rats.The leading role of spleen yang deficiency etiology is thermogenic cell phenotype transforma_tion disorder.(1) The dose-effect relationship of the thermogenesis induced with isoproterenol in rats represents thermogenesis in rats.(2) The linear relationship was found between the degrees of hypothyrodism and hypothermogenesis;and digestive loads had been considered to aggravate hypothermogenesis in rats with hypothyroidism.(3) Scale evaluation of severity inrats of spleen yang deficiency:Above more than 32% manifestations were associated with both of rats and patients suffered from the spleen yang deficient syndrome.(4)Removed interscapular brown adipose tissue in rats showed significantly increased plasma ghrelin,A crosstalk between hypothermogenesis and indigestion.(5)The developmental etiology of the spleen yang deficiency aggravated hypothermogenesis and increased plasma ghrelin level in rats.(6)The etiology of the spleen yang deficiency was developmental one,ghrelin involved in it.Fuzi-Lizhong Pill relieved hypothermogenesis and ghrelin secretion,but the antithermogenesis aggravated them.The dystransformation of thermogenic cell phenotype plays a key role in the spleen yang deficiency,and ghrelin,as an endocrine hormone,crosstalking in hypothermogenesis and indigestion.