| Objective:The effect of" Kang feng shi ling pian " on ovarian function in rats with CIA (collagen induced arthritis) was observed by animal experiment. To explore the toxic dose of the drug in rats, and try to explore the mechanism of its effect on ovarian function, and provide more theoretical reference for clinical application of the Kang feng shi ling pianMethods:10 healthy female Wistar rats were randomly divided into 65 groups as blank control group. For the remaining 55 rats were injected with bovine II collagen injection molding, and the model was successful and the normal CIA rats were randomly selected for the follow-up experiments in 50 rows. The 50 CIA rats were randomly divided into 5 groups:the low dose group, the positive control group and the model control group, and the 3 groups were randomly divided into two groups. Each drug group was administered orally, and the model control group and blank control group were given normal saline for 28 days. During administration, the daily vaginal shedding cells were examined and recorded, and the rest of the ovarian function was observed. The basic organs such as serum sex hormone, ovary and uterus were observed. Weight calculation of the organ index of ovary uterus. The morphological changes of ovarian tissue were observed under microscope and the total number of follicles and corpus corpus. Serum sex hormone levels were detected by enzyme-linked immunosorbent assay (ELLSA). The expression of ovarian BCL-2 and GnRHR was determined by immunohistochemistry using monoclonal antibody test kit. Statistical analysis and comparison of each index, determine the effect of each drug group on ovarian function.Results:l)compared with the control group, there were no significant differences in the indexes of ovarian function in the model control group. (P> 0.05)2) in the three dose levels of the Kang feng shi ling pian , only the high dose of the CIA rat’s ovary organ index was down regulated (P<0.05), and no significant difference (P<0.01). Compared with the model control group, the levels of the uterine organ index of CIA rats (P>0.05) were not affected by each administration group.3) antirheumatic spirit in three dose levels, high and low dose of CIA rat estrous cycle produced the effect (P<0.05), appear to extend and disorder, which extend the estrous cycle in the high dose group and disorder appear earlier, but with the model control group than in the high and low dose groups dynamic cycle changes were not significant difference (P<0.01).4) the high dose group of the three dose levels in the dose levels of the CIA rats (P <0.05), and no significant difference (P<0.01).5) antirheumatic spirit of the high dose group of CIA rat FSH and FSH/LH mean produced significantly up-regulated effect (P<0.01); with the model control group, Kang feng shi ling pian, low dose and Tripterygium wilfordii positive control group (50mg/kg/D) on serum FSH were the number of levels were of no significant influence (P> 0.05). Compared with the model control group, the serum levels of LH and E2 were not significantly affected by the administration group (P>0.05).6) compared with the model control group, the expression of Bcl-2 in the ovary was not significantly affected by the three dose groups and the 50mg/kg/d of the dose groups (P>0.05).7) and model control group, antirheumatic spirit in three dose groups were not on ovarian GnRHR expression have significant effect (P>0.05) and positive control group of Tripterygium wilfordii on GnRHR were the number of levels of cut did not reach statistical significance (P> 0.05).Conclusion:Compared with the control group, no significant differences were found in the ovarian function of the CIA model control group, which showed that the CIA model had no significant effect on ovarian function in rats.Antirheumatic spirit in three dose levels, only high doses (9 times clinical dosage) of ovarian tissue of CIA rats caused substantial damage (ovarian index decreased, endoscopic ovarian tissue ultrastructure see atrophy degeneration), and disrupt or prolonged estrous cycle of rats. At the same time with respect to the other dose level of the Kang feng shi ling pian, only the high dose group caused a reduction in the number of ovarian corpus. The dose group also raised the level of serum FSH level in rats (P< 0.01). Compared with the model control group, the levels of serum LH and E2 were not significantly affected (including the positive control group) (P> 0.5). From the FSH/LH ratio, compared with the model group, there was a significant difference in the ratio of high dose group (P< 0.01). The result shows that the Kang feng shi ling pian (9 times of clinical dose) can cause loss of ovarian function and decrease the ability of ovarian reserve. But the expression of BCL-2 and GnRHR in ovary was observed, and there was no significant difference between each group and model control group (P> 0.05). Show Kang feng shi ling pian tablet in a certain dose, on ovarian function influence the main mechanism of May and the expression of Bcl-2 in granulosa cell apoptosis pathway independent of ovarian granulosa cell GnRHR expression had no significant effect.In summary, Kang feng shi ling pian tablets clinical dose,3 times of dosage (low, middle dose group) of ovarian function in rats caused no obvious effect. Antirheumatic spirit tablet in clinical 9 times dose (high dose group) of ovarian function in rats caused by wear and tear, showed a lower ovarian reserve function (DOR). But relative Lei Gong Teng multi glycoside tablet positive control group (5.3 times clinical dosage), and Kang feng shi ling pian tablet high dose (9 times clinical dosage) on ovarian function loss degree is light. |
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