The Pharmacodynamics Research And Acute Toxicity Experiment Of Geng Kang Keli For Tonifying Kidney

Menopause syndrome is caused by the change of female sex hormones series of syndrome,and can lead to physical,emotional and social barriers,every woman have to undergo during this period,just have different symptom severity,but China’s large population base,the high incidence of a disease,menopause syndrome caused attention.Modern medical treatment is given priority to with hormone treatment,clinical effect is fast,but its use timing,treatment duration,dosage form,side effects,etc.are very controversial.People have paid more attention on good curative effect,multiple treatments and less side effects of Chinese medicine,recently,with the rise of Chinese medicine,Chinese medicine compound development research has become hot spot.Many scholars at home and abroad explore the effect and mechanism of traditional Chinese medicine compound,from clinical randomized controlled trials,animal experiment,sensors and actuators,gene research studies,and mainly through experimental research of traditional Chinese medicine compound on regulation of reproductive endocrine,oxidation ageing,enhancing sedative hypnotic effect,enhancing the immune function,improving learning and memory function,take the main animal model of natural aging model,castrated model,chemical drug induced model,the cell model,etc.Geng Kang Keli is the medicine that has been used clinical for almost 30 years by the Tutor Wang,which consists of prepared rehmannia root and cistanche deserticola,etc.that could nourish the kidney and essence,and balance the Yin and Yang(two opposing principles in nature,the former is feminine and negative while the latter is masculine and positive in Chinese philosophy medicine)in humans’ body.Geng Kang Keli is mainly designed for patients who are deficiency of the kidney with menopause syndrome.The TCM pathogenesis of menopause syndrome is based on deficiency of the kidney,Geng Kang Keli works from tonifying kidney and takes good care of other viscera.Purpose:1.Study the effects of Geng Kang Keli on ovariectomized rats’ serum sex hormones,the hypothalamus single amine neurotransmitter,3-endorphin,antioxidant function related materials,and the uterus.2.Observe and study the toxic effects conducted in certain period by three times of filling the stomach to give Geng Kang Keli for SD rats within 24 hours and get to know the acute toxic effect doses.Method:1.Pharmacodynamics study of Geng Kang Keli:Choose 80 SD female rats and randomly divided into 8 groups(each group have 10 rats):blank control group,sham operation group,the model group,the pentanoic acid estradiol group,the KunTai capsule group,the Geng Kang Keli low-dose group,Geng Kang Keli middle-dose group,Geng Kang Keli high-dose group.Except for the blank control group and sham operation group,all the other six groups’ rats did surgical excision of bilateral ovaries.Each group had four-week intervention(Blank control group,sham operation group and model group were given distilled water),adopted ELISA method to determine the rats’ serum levels of sex hormones,hypothalamic neurotransmitter contents(5-HT、5-HIAA、DA、NE),β-EP,antioxidant function related materials(SOD、MDA、GSH-Px),took out the uterus,calculated organ index and detected the thickness of endometrium.SPSS17.0 was used to do statistical analysis.2.Acute toxicology experiment of Geng Kang Keli:The maximum dosage method.According to the weight after stratification,40 female rats were randomly divided into two groups[the subject group(148.5g crude drug/kg)and the solvent control group(pure water)]of 20 rats for each.Each group was continuously filled the stomach to be given the medicine for three times within one day based on the standard 30ml/kg and there is a four-hour interval each time.In the continuous four hours after giving the medicine,observed the toxic symptoms and its start time,etc.Measured the weights on D0,D1,D7 and D14.Continuously observed 14 days after treatment,which observed the rats’ appearance,behavior,mental condition,response to stimulation,secretion,excreta,etc.for one time respectively in the morning and in the afternoon.All the rats were generally anatomized and the changes of their organ size,color and quality were observed by naked eyes.If there were the change of any organ size,color and quality,histopathological examination would be conducted.Results:1.Experimental study of Geng Kang Keli on ovariectomized rats(1)The effects of Geng Kang Keli on ovariectomized rats’ serum sex hormones:① Compared with the blank control group、sham operation group,model group rats serum E2 levels were significantly reduced(P<0.01),LH,GnRH,T content significantly increased(P<0.01),FSH levels increased significantly(P<0.05,)and blank group compared with sham operation group,there was no statistically significant difference content of FSH,LH,E2,GnRH and T levels(P>0.05).② Compared with the model group after treatment,the Geng Kang Keli high-dose group’ s rats have increased serum E2 levels(P<0.01),decreased FSH、LH、GnRH and T levels(P<0.05、P<0.01);the Geng Kang Keli middle-dose group’ s rats have increased E2 levels(P<0.05)and decreased FSH、LH、GnRH、and T levels(P<0.05);the Geng Kang Keli low-dose group’ s serum sex hormone levels difference has no statistical significance(P>0.05).③ The FSH、LH、GnRH and T levels of Geng Kang Keli middle and high-dose group’ s rats has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group,(P>0.05);the E2 levels of middle and high-dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05).④ The T、E2 levels of Geng Kang Keli low-dose group’ s rats has statistical significance with blank group and sham operation group(P<0.05);The FSH、LH、GnRH levels of Geng Kang Keli middle and high-dose group’ s rats has no statistical significance with blank group and sham operation group(P>0.05).⑤ The mean of high-dose group’ s rats was superior to the low and middle-dose group’ s rats in serum sex hormones level.(2)The effects of Geng Kang Keli on ovariectomized rats’ the hypothalamus single amine neurotransmitter:① Compared with the blank control group、sham operation group,the 5-HT、5-HIAA、DA and NE levels of model group rats significantly increased(P<0.01),and blank group compared with sham operation group,there was no statistically significant difference content of 5-HT、5-HIAA、DA and NE levels(P>0.05).② Compared with the model group after treatment,the Geng Kang Keli high-dose group’s rats have greatly decreased hypothalamus 5-HT、5-HIAA、DA and NE levels(P<0.01);the Geng Kang Keli middle-dose group’ s rats have decreased hypothalamus 5-HT、5-HIAA、DA and NE levels(P<0.05、P<0.01);the Geng Kang Keli low-dose group’ s rats has no statistical significance(P>0.05).③ the 5-HT、DA levels of low-dose group has statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P<0.05),there has better effect than low-dose group;the 5-HIAA、NE levels of low dose group and the 5-HT、5-HIAA、DA and NE levels of middle and high-dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05).④ The NE levels of low-dose group has statistical significance with blank control group and sham operation group(P<0.05),The 5-HT、5-HIAA and DA levels of low-dose group has no statistical significance with blank control group(P>0.05),the 5-HT、5-HIAA、DA and NE levels of middle and high-dose group has no statistical significance with blank group and sham operation group(P>0.05).⑤ The mean of high-dose group’ s rats was superior to the low and middle-dose group’ s rats in hypothalamus single amine neurotransmitter.(3)The effects of Geng Kang Keli on ovariectomized rats’ β-EP level:① Compared with the blank control group、sham operation group,model group rats β-EP levels were significantly reduced(R<0.01),and blank group compared with sham operation group,there was no statistically significant difference content of β-EP levels(P>0.05).② Compared with the model group after treatment,the Geng Kang Keli middle and high-dose group’ s rats has increased hypothalamus p-EP levels(P>0.05);the Geng Kang Keli low-dose group’ s rats have no statistical significance in β-EP levels(P>0.05).③ The β-EP level of middle and high-dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05).④ The β-EP level of low-dose group has statistical significance with blank group and sham operation group(P<0.05).The β-EP level of middle and high-dose group has no statistical significance with blank group and sham operation group(P>0.05).⑤ The mean of high-dose group’ s rats was superior to the low and middle-dose group’ s rats in β-EP level.(4)The effects of Geng Kang Keli on ovariectomized rats’ antioxidant function related materials:① Compared with the blank control group、sham operation group:Compared with the blank control group,model group rats serum SOD、GSH-Px levels were significantly reduced(P<0.01);Compared with the sham operation group,model group rats serum SOD、GSH-Px levels were significantly reduced(P<0.05、P<0.01);Compared with the blank control group、sham operation group,MDA content significantly increased(P<0.01),and blank group compared with sham operation group,there was no statistically significant difference content ofSOD、GSH-Pxand MDA levels(P>0.05).② Compared with the model group after treatment,the Geng Kang Keli high-dose group’s rats have increased serum SOD、GSH-Px levels(P<0.01)and greatly decreased MDA levels(P<0.01);the Geng Kang Keli middle-dose group’ s rats have significantly increased serum SOD、GSH-Px levels(P<0.01)and decreased MDA levels(P<0.05);the Geng Kang Keli low-dose group’ s rats have increased serum GSH-Px levels(P<0.05)and the other indicators’ levels have no statistically significant difference(P>0.05).③ The SOD、MDA levels of middle and high-dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05),theGSH-Px level of three dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05).④ The SOD、MDA levels of middle and high-dose group has no statistical significance with blank group andsham operation group(P>0.05),The GSH-Px levels of three dose group has no statistical significance with blank group andsham operation group(P>0.05).⑤ The mean of high-dose group’s rats was superior to the low and middle-dose group’ s rats in SOD、MDA and GSH-Px level.(5)The effects of Geng Kang Keli on ovariectomized rats’ uterus:① Compared with the blank control group、sham operation group,model group rats uterus coefficients were significantly reduced(P<0.01),endometrial thickness significantly reduced(P<0.01),and blank group compared with sham operation group,there was no statistically significant difference(P>0.05).② Compared with the model group after treatment,the Geng Kang Keli high-dose group’ s rats have increased uterus coefficients(P<0.05)and endometrial thickness(P<0.05);the Geng Kang Kelilow and middle-dose group’ s rats has no statistical significance in the uterus coefficients and endometrial thickness(P>0.05).③ The uterus coefficients and endometrial thickness of three dose group has no statistical significance with the pentanoic acid estradiol group and KunTai capsule group(P>0.05).④ Compared with blank group,The uterus coefficients and endometrial thickness of three dose group has statistical significance(P<0.05,P<0.01);compared with sham operation group,The uterus coefficients and endometrial thickness of low and middle-dose group has statistical significance(P<0.05,P<0.01),The endometrial thickness of high-dose group has no statistical significance(P>0.05).⑤ The mean of high-dose group’s rats was superior to the low and middle-dose group’ s rats in the uterus coefficients and endometrial thickness.2.Acute toxicology experiment of Geng Kang KeliIn this period,rats have good mental condition and there is no abnormal performance on their appearance.Rats have normal behaviors and there is no abnormal secretion and excreta.No rat died till the end of the experiment.Compare the weight of the subject group with the solvent control group on D0,D1,D,and D14,there is no statistical significance(P>0.05).Subjects group and solvent control group rats to put to death at the end of the observation period,Observation of the rats’ surface structure is complete,the rats’ hair were smooth,without injury and ulceration,There was no bleeding point and pigmentation in the skin,abdomen and anocelia has no effusion,the organ position is normal,the volume,color and texture of the organs were not found to have obvious pathological changes and abnormalities.Conclusion:1.Geng Kang Keli could regulate castrated model rats’ relevant sex hormones and improve reproductive endocrine,could enhance the uterus function through increasing the uterus coefficients and endometrial thickness;Geng Kang Keli could improve the plant nerve dysfunction through regulating the hypothalamus single amine neurotransmitter(5-HT、5-HIAA、DA、NE)and p-EP level;Geng Kang Keli could improve the antioxidant levels(SOD、GSH-Px)and reduce the hamful product(MDA),proving to have antioxidation function.2.Geng Kang Keli(148.5g crude drug/kg,179 times of the proposed clinical dose)showed no toxic effects on SD rats by three times of filling the stomach to give medicine within 24 hours.